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RELIABILITY AND VALIDITY OF MEASURING IMPLANT STABILITY WITH RESONANCE FREQUENCY ANALYSIS Georgios Katsavrias An Abstract Presented to the Faculty of the Graduate School of Saint Louis University in Partial Fulfillment of the Requirement for the Degree of Master of Science in Dentistry 2009 Abstract Purpose: To evaluate the reliability and validity of resonance frequency as a method for measuring the stability of miniscrew implants. Methods: One hundred and twenty miniscrew implants were randomly allocated to synthetic bone blocks of different densities: 40 low [cortical part: 38.1pcf, cancelous part: 30.0pcf ] density, 40 medium [cortical part: 40.6pcf, cancelous part: 31.8pcf] density, and 40 high [cortical part: 50.1pcf, cancelous part: 31.8pcf] density. For each implant insertion torque was measured using Mecmesin Advanced Force and Torque Indicator©. Pullout strength was measured using the Instron© Machine, Model 1011 and applying a vertical force at 20mm/min until failure. Resonance frequency was measured three times parallel and three times perpendicular to the long axis of each bone block. Results: Intraclass correlations based on multiple measures of resonance frequency ranged from 0.953 to 0.992; single measure intraclass correlations ranged from 0.870 to 0.977. Analysis of variance showed significant effects of bone density on resonance frequency; post-hoc tests showed differences for high density only. Insertion torque and pullout strength demonstrated statistically significant differences between all three densities. The two orientations showed no significant differences for low and medium densities, the high density bone showed a 1 significant (p= .049) orientation effect. Independent of bone density, correlations between resonance frequency, insertion torque, and pullout strength were low and showed no consistent pattern. Conclusions: Resonance frequency is a reliable measure. It is also a valid measure that is related with bone density. As expected for homogeneous synthetic bone, resonance frequency shows little or no relationship with orientation, insertion torque, or pullout strength. 2 RELIABILITY AND VALIDITY OF MEASURING IMPLANT STABILITY WITH RESONANCE FREQUENCY ANALYSIS Georgios Katsavrias A Thesis Presented to the Faculty of the Graduate School of Saint Louis University in Partial Fulfillment of the Requirement for the Degree of Master of Science in Dentistry 2009 COMMITTEE IN CHARGE OF CANDIDACY: Adjunct Professor Peter Buschang, Advisor Professor Rolf G. Behrents Assistant Professor Ki Beom Kim Assistant Professor Donald R. Oliver i DEDICATION I would like to dedicate this study to my beloved family for their constant support and encouragement towards my goals. To my mother, for devoting a huge amount of her personal time to me at critical points in my carrier. To my father, for encouraging me to try harder throughout my education. To my brothers Stamatis and Iasonas, for showing me the need of persistence in life and teaching me about the need for acceptance of diversity. ii ACKNOWLEDGEMENTS Dr. Ana Claudia Melo for supporting me from the beginning and throughout the whole study despite of all the occurring difficulties. Without her support this study would not be possible. Dr. Buschang, for helping me substantially in the design, the statistical analysis and the interpretation of my results. Dr. Behrents, for being available whenever I needed his guidance and his support. For picking out for me all the small details which would make a significant difference in the end, encouraging me to work further. Dr. Kim, for his time, interest and help in the support of my thesis. Dr. Oliver, for his insightful comments on my work, keeping me on my toes throughout the duration of the study and being sure that I have paid attention to every single detail. Dr. Araujo’s family for their substantial support when this study was in jeopardy. Without their help this project would surely be dropped. Neodent for sponsoring me with their miniscrew implants. iii Table of Contents List of Tables.........................................vi List of Figures.......................................vii CHAPTER 1: INTRODUCTION.................................1 CHAPTER 2: LITERATURE REVIEW............................7 Miniscrew Implants in Orthodontics...........7 Uses and Importance of Miniscrew Implants..................................7 Failure of Miniscrews.....................9 Primary and Secondary Stability..........13 Techniques to Measure Stability.............18 Invasive Methods.........................19 Histologic and Histomorphometric Technique.............................19 Cutting Torque Resistance Analysis....22 Reverse/Removal Torque Value..........23 Insertion Torque Analysis.............24 Pullout Test..........................26 Non-Invasive.............................28 Radiographic Analysis.................28 Finite Element Analysis...............29 Percussion Test.......................32 Pulsed Oscillation Waveform...........33 Impact Hammer Method..................34 Resonance Frequency Analysis..........38 Summary.....................................49 Reference List..............................51 CHAPTER 3: JOURNAL ARTICLE.............................71 Abstract....................................71 Introduction................................72 Materials and Methods.......................76 Miniscrew Implants.......................76 Synthetic Bone...........................77 Insertion Torque.........................78 Resonance Frequency Analysis.............79 Pullout Strength.........................79 Statistical Analyses........................80 Results.....................................81 Reliability..............................81 Validity.................................82 Stabilization of Blocks..................84 Discussion..................................84 Conclusions.................................90 References..................................91 iv Appendix I.............................................96 Appendix II...........................................102 Vita Auctoris.........................................106 v List of Tables Table 1: Comparison of resonance frequency (RF) of three consecutive measurements [taken parallel (║) and perpendicular (⊥)to the long side of the rectangular synthetic bone block] for the high, medium and low density bone blocks............97 Table 2: Reliabilities of resonance frequency (RF) measures evaluated using intraclass correlations for single measurements and Cronbach’s Alpha for multiple measurements.........................98 Table 3: Comparison of resonance frequency (RF measurement averages [taken parallel(║) and perpendicular (⊥) to the long side of the rectangular synthetic bone block] insertion torque (IT) and the pullout strength (POS) for the high, medium and low density bone blocks..99 Table 4: Correlations between resonance frequency (RF) measurements [taken parallel(║) and perpendicular (⊥)to the long side of the rectangular synthetic bone block],insertion torque (IT) and pullout strength (POS) for the high, medium and low density synthetic bone blocks.......................................100 Table 5: Comparison of resonance frequency (RF) measurements [taken parallel (║) and perpendicular (⊥)to the long side of the rectangular synthetic bone block] with the medium density block clamped and not clamped......................................101 vi LIST OF FIGURES Figures 1: Miniscrew Implant and Custom PulloutBase...103 Figures 2: Synthetic bone.............................103 Figures 3: Miniscrew Implant Driver...................104 Figures 4: Resonance frequency analysis measurements..104 Figures 5: Comparisons of resonance frequencies of three consecutive measurements...................105 Figures 6: Scatterplots...............................105 vii Chapter 1: Introduction One of the most important changes in the way orthodontic treatment is executed occurred with the introduction of miniscrew implants, by providing better control of tooth movement. Their relative stability under the application of substantial force makes it possible for the orthodontist to diminish the negative consequences of the force system being applied. Their advantages plus their ease of use are the major reasons that miniscrew implants have been quickly and extensively accepted by the orthodontic community. Their generalized usage has revealed that one of the major problems connected with miniscrews is the degree to which they fail.1 Research has suggested multiple factors may be responsible for failures including mobility of the miniscrew, overheating of the bone during drilling, gingival inflammation, and placement in areas with nonkeratinized mucosa. Great efforts have been made to reduce the number of failures and increase the ability to predict the implants most likely to fail. Very important in this regard is the effort to increase our understanding of primary stability. With adequate primary stability, the adjacent bone can remodel and enhance secondary stability. Poor bone quality or random events like trauma can increase 1 the likelihood of necrosis, reducing effective stability of the screw through the formation of woven bone around the implant.2 Woven bone is poorly organised and its structure does not provide adequate strength for supporting loads,3 thus lowering the implants ability to resist forces. A technique that can estimate the physical properties of the peri-implant bone will allow us to time loading accordingly and avoid loading when the quality of the bone around the implant is not optimum. One of the most established measures of stability is insertion torque, which is the rotational force required to insert a screw into bone. Insertion force is transferred through the screw and becomes a compressive force onto the adjacent bone. A minimal level of insertion torque is required to achieve primary stability. Excessive torque during placement can cause damage to the adjacent bone and increases the likelyhood of screw failure. According to Motoyoshi et al., there is a range of acceptable insertion torques outside of which the implants might be expected to fail.4 Another well established method for evaluating stability is the pullout test. Pullout tests have been used extensively to measure biomechanical holding power and gauge the amount of stability that has been attained.5-7 2 A relationship between insertion torque and pullout strength has been observed and a connection with screw stability has been established.8 Although insertion torque has been found to correlate linearly with pullout strength, there are some limits. Studies have shown that excessive insertion torque can cause osseous damage and lead to a decrease in pullout strength.9,10 As insertion torque increases, pullout strength increases up to a point, after which pullout strength decreases and the likelyhood of failure increases. The decrease in pullout strength occurs due to excessive compression that causes trauma to the adjacent bone and increases the likelihood of failure.11 Finally, extreme insertion torque could lead to screw distortion or breakage and, ultimately, failure.11 Failure is determined by the amount of movement of the implant,12 establishing a way to measure implant movement will offer us a way to predict failure. Currently there is no technique available that allows assessment of a miniscrew’s stability after insertion and before removal. However, non-invasive methods for measuring dental implant stability have been available for almost a decade.13,14 The two methods most commonly used are resonance frequency analysis (Osstell Mentor, Göteborg, Sweden) and dampening capacity assessment (Periotest, Modautal, Germany). A number of studies have shown that resonance frequency analysis with 3 the Osstell device is the best method quantifying implant stability.15,16 The Periotest instrument shows a greater measurement error in clinical application (intraclass correlation 0.88), thus making the Osstell a more reliable option to be used clinically(intraclass correlation 0.99).17 The Osstell Mentor device measures implant stability based on the resonance frequency analysis technique. This technique utilizes a bending test of the implant-bone complex with a transducer applying an extremely small bending force. A fixed lateral force is applied to the implant and its displacement is measured, thus mimicking the clinical loading condition, albeit of a much reduced magnitude.18 The most recent version of the resonance frequency analysis is wireless, where a metal rod (peg) connected to the implant by a screw connection. The peg has a small magnet attached to its top, which is excited by magnetic pulses generated by a handheld computer. The magnetic pulses, of known frequency, force the peg to oscillate in the direction of the transducer.18 The peg vibrates in two directions, which are approximately perpendicular to each other. Vibration is measured in the directions that produces the highest and lowest resonance frequencies.18 Thus, two implant stability values are provided, one high and one low. In this way, 4 circumferential estimation of the implants stability can be made.18 This method has proven useful for dental implants in research and clinical applications. The dental implant literature has shown that Osstell measurements correlate with bone-to-implant contact,19 bone density measured from CT scans,20,21 and insertion torque.20,22 Osstell measurements have also helped in establishing individual healing curves of implants after load initiation.21,23-26 Sequential measurements have shown that the Osstell device can be used to quantify ongoing osseointegration.27 Osstell measurements are most influenced by changes in the stiffness of alveolar bone.28 Despite of the fact that the Osstell device has been used extensively with dental implants, it has not yet been used with miniscrew implants. Until recently, it was not possible to mount a peg on miniscrews. The purpose of this study is to investigate if the resonance frequency analysis can be used to evaluate the stability (primary and secondary) of orthodontic implants. To validate the Osstell Mentor device, the measurements will be correlated with insertion torque and pullout strength (from mini implants that are placed in synthetic bone with known and homogenous density). The primary objective of this study is to establish the reliability of the Osstell used with miniscrew implants, 5 by repeating the Osstell measurements multiple times, the reliability of the technique will be tested. This novel, non-invasive, measurement technique could prove useful in helping orthodontists better understand the healing procedures of bone around miniscrews. 6 Chapter 2:Literature Review Miniscrew Implants in Orthodontics Uses and Importance of Miniscrew Implants Maintaining anchorage has been a challenge in orthodontics. In fact, being able to achieve adequate anchorage during treatment is an important determinant of the treatment outcome.29 According to the third law of Newton, teeth connected by an active appliance are subjected to equal and opposite forces. It is, however, not desirable in most cases to perform equal and opposite tooth movements. Various methods have been introduced to maintain anchorage. These methods include the application of Tweed mechanics,30 segmented mechanics,31 bioprogressive technique,32 bi-dimensional technique,33 archwires with differential sizes and shapes, uprighting springs,34 intra-arch elastics and headgears with various vectors of action. The last two methods are the approaches most commonly used, but they depend heavily on patient compliance.35 In noncompliant patients, there may be unpredictable tooth movements, which could lead to unfavorable treatment results. Miniscrew implants are becoming increasingly popular in orthodontics,36,37 because they provide skeletal 7 anchorage for orthodontic tooth movements.38-42 The possibility of moving miniscrews is minimal due to the absence of a periodontal ligament. Animal and human studies have provided the basis for clinical use.43-45 Creekmore and Eklund46 first suggested that such screws could be used for orthodontic applications. The biomechanical principles of this approach are very similar to those applied in traditional anchorage, making their utilization in treatment easier. However, the possibility of using a stable anchorage point has extended the treatment possibilities for orthodontic patients. For example, miniscrews make treatment possible when a significant number of teeth are missing, when existing bone support is inadequate, and in adults or some adolescent patients, who refuse to wear headgears. In the past, when miniscrews were not available, if the patient had simultaneous prosthetic implant restorations dental implants would be utilized for orthodontic purposes. When dental implants were used in such a way a healing period was considered necessary.47-53 The time period for healing was based upon Branemark’s suggestion that endosseous dental implants should remain unloaded for three months in the mandible and for six months in the maxilla in order for osseointegration to occur.54 The same trend, of delayed loading, continued in the first studies published for miniscrew impants. Those 8 older clinical studies using miniscrews applied forces ranging from 30 gm-250 gm, with a delay in the application of forces ranging from 4 to 36 weeks. The authors believed that longer healing periods were necessary to ensure longterm success.48,55,56 Later studies showed that shorter healing periods did not compromise implant stability.38,53,57,58 Presently, most miniscrew implants are loaded immediately. According to Buchter et al., miniscrew implants can be loaded immediately without impairing their stability if moments of forces are maintained below 900 cN·mm .59 Failure of Miniscrews The number of miniscrew failures plays a significant role in the orthodontists’ decision to utilize them in everyday practice. The literature reports success rate for miniscrews ranging from 83.9% to 91.6%.4,60,61 Due to the absence of studies about the influence of systemic factors upon the failure rate of miniscrew implants some conclusions can be made from the dental implant literature. Research with dental implants have shown that failures are due to a number of host factors that can interfere with the healing process.62 These factors include: osteoporosis, uncontrolled diabetes, smoking, and parafunctional habits (this is more 9 problematic with dental implants that support crowns in bruxist patients). Another group of factors that can cause failures in miniscrew implants relate to the surgical procedures or techniques used at the time of the placement. For example, Kang et al. noted that the failure rate for miniscrews that contacted the roots was 79.2%, compared to an 8.3% failure rate in alveolar bone.63 In a recent survey by Buschang and coworkers it was found that practitioners who experienced significantly more failures in miniscrews did not measure insertion torque nor the forces applied to the implants.64 Problems may also arise due to improper selection of the implant site, based on the quality of bone. A dental implant at a site of very low bone quality loaded with orthodontic forces might be subject to movement during the first few days of loading, and then either settle into a fixed position after a few weeks or fail.49 In such cases, failure has been related to microfracture or microcracks of the peri-implant bone and bone remodeling on the tension and compression sides of the miniscrew.49 Placement of the implant in very dense bone may also reduce the chances of implant survival due to excessive pressure on the surrounding bone, as expressed by excessive insertion torque values.4 Also, premature loads have been shown histologically to produce a layer of fibrous tissue between the bone and implant.65,66 Increased 10 failure rates have also been related to the side into which miniscrews have been inserted (with the left side showing more failures than the right) and to the jaw (with the mandible showing more failures than the maxilla).67 Failure may also be the result of overheating at the time of the implant placement. Bone overheating during drilling may cause an inflammatory infiltration of interposed soft tissue that has been correlated with complete loosening or failure of the implant. It has been shown that temperatures above 47oC can cause bone damage.68,69 Heat generation may be due to excessive pressure of the drill on the bone surface, and to the use of worn drills.70 Finally, pilot hole diameter has been related to the survival of the implant. Pilot holes that are too small increase the pressure to the surrounding bone, leading to possible necrosis;71 pilot holes that are too large will increase initial micro-motion and inhibit the healing process, both of which could dramatically decrease the holding power of the screw/bone system.72,73 Management factors, pertaining to the doctor and the patient after the implant’s placement are also important. These include poor home care, poor oral hygiene which may lead to inflammation and infection, and excessive load during the treatment (900 cN·mm).59 General oral hygiene does not affect miniscrew success as much as hygiene around the implant.61 Inefficient hygiene ultimately leads 11 to peri-implantitis which has been shown to be an important risk factor in dental-implant failure.74 Local inflammation can be exaggerated not only by oral hygiene but also by non-keratinized soft tissue around the neck of the implant. A recent study suggested that nonkeratinized mucosa was a risk factor for miniscrews.60 Park et al. showed that local inflammation can damage the bone surrounding the neck of screw implants.61 With progressive damage of the cortical bone, screw stability can be compromised.74 Costa and coworkers also suggest that a force system that generates a force in the unscrewing direction causes implants to failure.38 Buchter et al. reported loosening of implants subjected to forces of 900 cN·mm (force 300 cN and 3mm lever arm).59 They related implant failure to the tipping movements. None of the implants that were tested moved through the bone, but three of them showed slight tipping in the direction of the applied force (an observation that agrees with the observations of Liou et al.75). It is also worth mentioning that their reference implants, which had no forces applied to them, showed significantly higher removal torque than loaded implants at the end of the study. Along the same lines, Isidor found that all forces may not be tolerated by the crestal bone.76 High strain values, above 6700 μstrain, resulted in peri-implant bone 12 resorption and a negative balance between bone apposition and resorption.76 The ability of an implant to withstand such negative effects is not the same throughout its utilization. Implants stability is related initially to mechanical retention and ultimately to the amount of healing that takes places around the implant. Primary and Secondary Stability The stability of an implant can be defined as the implant’s capacity to withstand loading. Since loading can be lateral, axial or rotational, stability can be divided, mechanically, into at least three types.77 Biologically, stability can be categorized as primary, mechanical, stability and secondary stability, associated with bone remodeling around the implant. The events that occur during a dental implant’s osseointegration have been described by Scwartz and Boyan.78 Right after implant placement, serum proteins adhere to its surface. During the first three days, mesenchymal cells attach to the implant and proliferate. By day six osteoid is produced. After two weeks, matrix calcification is complete and by three weeks remodeling is well underway. 13 Similar stages have been reported by Berglundh and coworkers, who used implants with a wound chamber, that were placed in Labrador dog mandibles.79 After two hours the wound chamber was occupied by a coagulum of erythrocytes, neutrophils and macrophages in a fibrin network. After four days, osteoclasts and mesenchymal cells were observed along the implants surface; vascular structures and densely packed connective tissue cells were found within the wound chamber. By day seven new woven bone started to appear on the implant surface, along with vascular units. The trabeculae were lined with osteoblasts and there was a provisional matrix, which had collagen fibrils and vasculature. At two weeks, newly formed bone appeared to be extending from the parent bone. At four weeks, marked formation of woven bone and lamellar bone was seen. After eight to twelve weeks there were marked signs of remodeling within the wound chamber. Fluorescent bone labels and microradiography have been used to demonstrate that woven bone forms close to the implant surface after approximately three days of healing.80 Due to the inferior physical properties of woven bone, in comparison to lamellar bone, loading of the implant at this phase of healing has a greater risk for the implant to fail. The primary stability of screws used to enhance orthodontic anchorage is largely determined by the 14 cortical plate; primary stability may be in the form of monocortical or bicortical anchorage.81 The density of the cortical bone greatly influences the distribution of stress from the screw to the bone. Dense bone restricts the stress in the cortical area, whereas less dense cortical bone distributes the stress along the entire length of the miniscrew.82,83 Relative motion between the implant body and the surrounding bone during the early phase of healing is considered to be an important risk factor for early implant loss due to failure of osseointegration.84,85 Primary stability is important because micromotion at the implant-to-bone interface can result in the formation of fibrous tissue, as demonstrated with blade-type implants.65,66 It is important to recognize that pluripotential bone cells can differentiate into fibrous tissue, cartilage and bone.86 Fibrous tissue cannot be converted to bone; once the fibrous tissue forms between the bone-to-implant interface the osseointegration process is inhibited and the implant is destined to fail.87 Following the placement of an endosseous implant (dental or miniscrew implant), primary mechanical stability (from the interlocking of the screw threads in the peri-implant bone) gradually decreases and secondary (biologic) stability gradually increases. The transition from primary mechanical stability to secondary stability, 15 provided by newly formed osseointegrated bone, takes place during early wound healing.88 To study healing Morinaga et al. used Ti-coated plastic implants placed in the tibiae of rats, which they followed for 28 days by means of light microscopy, transmission electron microscopy and micro computed tomography.88 The plastic implants were free of threads, but they were the size of commonly used orthodontic implants, with a diameter of 1.6 mm and a length of 7 mm. They noted that bone formation started a small distance away from the implant and after 28 days, covered the implant continuously.88 During the transition from primary (mechanical interlocking of screw threads in peri-implant bone) to secondary stability (from the newly formed bone around the implant), there was a period of time during which osteoclastic activity decreased stability, because the formation of new bone has not yet occurred.89 Luzi and coworkers noted a decrease in bone-to-implant contact between 1 week and 1 month for loaded miniscrew implants.89 Due to remodeling, there is gradual replacement of peri-implant bone, with new bone formation near the implant surface.90 According to Frost, if the load of the implant falls into the window of adaptation of the surrounding bone, then the adjustment of the bone’s architecture will lead into an increase of strength in that area.91 Jing et al. placed miniscrews in 16 rabbit tibiaes and found that placement removal torque and pullout force increase significantly after four weeks, histomorphometric analyses showed new bone around the miniscrew.7 Prager and coworkers showed no significant differences in bone-to-implant contact and mineral apposition rate between pre-drilled and self drilling miniscrew implants,90 which agrees with the findings of Luzi and coworkers.89 While the bone remodeling and loading occur simultaneously, the interaction of these two factors seems to be critical for the survival of the implant. 17 Techniques to Measure Stability Stability, an indirect indication of osseointegration, is a measure of implant’s resistance to movement.71 Quantification of implant stability at various time points provides significant information about individual healing times.92 The available methods for studying stability can be categorized as invasive, which interfere with the osseointegration process of the implant, and noninvasive, which do not. Invasive methods include histologic and histomorphometric evaluations, cutting torque resistance analysis, insertion torque and pullout tests. Unlike all other methods, histologic and histomorphometric evaluations evaluate osseointegration directly. Thus, they provide the best methods for establishing secondary stability. The reliability of these methods, however, has not been evaluated. The same applies for the other invasive methods. The non-invasive methods include radiographic evaluations of the implant, three dimensional finite element analysis, percussion tests, pulsed oscillation waveforms, impact hammer tests and resonance frequency analyses. With the exception of the last two, the reliability of the noninvasive methods has also not been tested. It has been suggested that 18 resonance frequency analysis is the most reliable method for dental implants.17 Invasive Methods Histologic and Histomorphometric Technique The implant’s stability (i.e. its anchorage potential) can be estimated indirectly by examining the bone-implant interface. With a microscope the boneimplant interface can be studied, cell proliferation and local bone morphology can be observed, and the implant’s capability to resist movement can be estimated. Histomorphometry is commonly used as a quantitative method for establishing the percentage of bone to implant contact from ground sections of implants.80,93,94 Typical parameters measured include percentage of bone contact and the bone area within the threads. In addition, the number of osteocytes can be counted. Histomorphometric studies of miniscrew implants have also been conducted. Vannet showed that the amount of osseointegration, estimated based on bone-to-implant contact, was independent of loading time and location.95 His findings support previous animal studies in evaluating miniscrew impants96-99 and dental implants.83-85 Melsen and Lang showed that there was a significant 19 increase in bone-to-implant contact and in bone density after six months.96 The bone in close contact to the miniscrew implant was characterized as woven. Melsen and Costa studied, histologically, immediately loaded implants placed into the infrazygomatic crest and the mandibular symphysis of four adult male Macaca Fascicularis monkeys.57 They observed no difference in the amount of bone-to-implant contact between screws loaded with 25 grams and those loaded with 50 grams. Bone-toimplant contact increased over time. After one month of loading, 21% of the screw was in contact with bone; after six months, the bone-to-implant contact increased to 60%. Bone density also increased over time, from 8% in the first month to 50% in the sixth month. The bone in close contact to the mini implant was characterized as woven and, was in most cases, distinguishable from the surrounding lamellar bone. Melsen and Lang,96 who evaluated the amount of bone turnover adjacent to miniscrew implants, showed that bone density was generally higher within 1 mm of the miniscrew implants surface than at a distance of more than 1 mm. The bone close to the screw was mostly woven. Similar observations have been made by Deguchi et al., who reported continuous turnover of bone in dental implants that were used as anchor units for orthodontic treatment.53 20 Ultrastructural studies performed on decalcified specimens that were sectioned for transmission electron microscopy indicated that bone closely approximated the surface of titanium implants (inserted in the rabbit tibia for 12 months) but that it was never in a direct contact with the implant.100 However, it is not possible from decalcified sections to make observations about the presence of mineralized tissue and its relation to the surface of the implant. While histologic and histomorphometric measurements provide the most objective estimates of osseointegration, they are of no use clinically. First, they require the extraction of the implant with the surrounding bone, which is unethical to perform clinically. Destruction of the implant site also prevents sequential or longitudinal measurements from being taken. Sequential measurements are necessary to fully understand the healing process associated with secondary stability. Finally, a microscopic image does not provide any information about the physical properties of bone, such as stiffness around the implant, which also can affect the implants anchorage potential.101 21 Cutting Torque Resistance Analysis Cutting torque resistance analysis was originally developed by Johansson and Strid102 and later improved by Friberg et al.103,104 It is based on the energy (J/mm3) required for an electric motor to cut off a unit volume of bone during implant surgery. This energy has been shown to be significantly correlated with bone density, which has been suggested as one of the factors that influences implant stability.105 Cutting torque resistance analysis can be used to identify areas of low bone density and to quantify bone hardness during the lowspeed insertion of implants. In orthodontics cutting torque has also been used to improve the clinician’s ability to detect root contact when placing mini implants.106 The major limitation of cutting torque resistance analysis is that it does not give any information on bone quality until the osteotomy site has been prepared. Furthermore, it does not allow longitudinal changes in bone quality to be assessed. Its primary use, therefore, lies in estimating primary stability indirectly, through the quantification of bone hardness, before placement. 22 Reverse/Removal Torque Value The reverse torque test was first proposed by Roberts et al.3 and developed further by Johansson and Albrektsson.93,107 It measures the critical torque threshold when the bone-implant contact is broken. Removal torque provides information on the degree of bone-to-implant contact in a given implant. Reverse torque values, in combination with histologic evaluations, have shown that greater bone-toimplant contact can be achieved with longer healing times.93 With surface treated (Hydroxyapatite coated) miniscrew implants, the longer the period of time after initial placement, the higher the torque.108 Okazaki et al. used removal torque to evaluate the stability of miniscrew implants placed in dog femurs. They inserted 1.2 mm diameter miniscrew implants using 1.0 mm and 1.2 mm pilot holes and showed that the removal torque values, compared to initial insertion torque measurements, increased for the implants placed in the 1.2 mm pilot holes and decreased in the 1.0 mm pilot holes. As a result the removal torque values 6, 9 and 12 weeks post insertion were almost equal for the two pilot holes.109 Removal torque has been criticized as being destructive. Branemark110 cautioned about the risk of irreversible plastic deformation within peri-implant bone 23 and of implant failure when unnecessary load is applied to an implant that is still undergoing osseointegration. A lower threshold (i.e., the value under which an implant is destined to fail) for removal torque has yet to be supported scientifically. Subjecting implants placed in bone of low quality, may result in a shearing of bone-toimplant contact and cause implants to irretrievably fail. Insertion Torque Analysis Insertion torque analysis quantifies the amount of force that is applied to the implant as it is inserted. Implant placement insertion torque is initially minimal, and increases rapidly until the cortical layer is fully engaged. As the implant is driven into the bone repeated measurements are taken and a graph is often produced. The maximum value is attained when the head of the screw makes contact with the cortical plate. Countersink friction between the head of the screw and the cortical plate explains why insertion torque increases dramatically at the end of implant placement.111 The analysis consists of finding the maximum insertion torque value when the screw head contacts the cortical plate. Further insertion of the screw beyond that point leads to fracture of the implant or stripping of the surrounding 24 bone; eventually, the screw spins freely in the hole with its holding strength severely limited.112 This test has been generally well accepted and has been used for evaluating various implant designs.113 Insertion torque has been found to correlate with bone density,114 which has in turn been shown to be correlated to implant stability.114 In other words, insertion torque measurements allow assumptions to be made about the quality of bone that supports the implant. Insertion torque has been shown to increase as the thickness of the cortical bone increases.115 In a study by Lim and coworkers, insertion torque significantly increased with increasing screw length and diameter.116 Values of insertion torque less than 15 Ncm have been related to failure of both machined and sandblasted miniscrew implants.117 Extreme insertion torque values, either too high or too low, have also been related to implant failure.4 However, insertion torque has been shown to be limited in certain applications. With self-drilling miniscrews, insertion torque may not reflect differences in the cortical bone thickness.115 Moreover, it is impossible to estimate the quality of the bone until you actually start implant insertion. As such, insertion torque measurements cannot be used for the selection of implant sites. Additionaly, insertion torque does not 25 offer the possibility of sequential measurements without damaging the bone-to-implant interface; as such it cannot be used to follow implant healing and osseointegration procedures. Pullout Test A pullout test is another indirect test of an implant’s anchorage potential. It usually measures the tensional force, applied vertically to the surface of bone into which an implant has been inserted, necessary to pull the implant out of bone. The force is applied parallel to the long-axis of the implant. Pullout tests have been extensively used in other medical fields, such as otolaryngology,118 orthopedics,119,120 as well as for the evaluation of dental implants.121 Pullout strength has typically been used to evaluate the design of implants and the mechanical interface between bone and implants.122 Chapman and colleagues showed that the pullout force of an implant is proportional to the length of the thread engaged in the bone, its major diameter and its thread depth and inversely proportional to the thread pitch.123 In orthodontics, Huja et al. showed that pullout measurements are significantly higher in the posterior part of the mandible of dogs specimens than in the 26 anterior part.124 They also found a weak correlation between pullout and cortical bone thickness. Salmoria et al. found that pullout decreases over time, which they related to the resorption of the cortical plate.125 Their findings have been confirmed by others.126 Because they could not establish a relationship between insertion torque and cortical bone thickness, Salmoria et al. concluded that pullout measurements are more efficient (easier to show difference) than insertion torque.125 Miniscrew pullout tests have also been used to evaluate different designs. Carano et al., who studied three different designs of miniscrews of the same dimensions, concluded that screws with asymmetric cut show higher pullout values.127 Leung et al.128 found that cylindrical 2.0 mm miniscrews connected with mini-plates produced significantly higher pullout forces than miniscrews of lesser diameters. Pullout tests suffer from the same limitations as insertion torque. Since the procedure is invasive, the implant site is destroyed after the test has been performed, making it impossible to use pullout tests to evaluate the implant-bone interface periodically. Because it cannot be used in normal clinical situations, this test is limited to laboratory experiments. 27 Non-Invasive Radiographic Analysis Radiographic analysis was one of the first methods applied to evaluate implants after they had been placed. The density, and therefore the physical properties of the surrounding bone, can be indirectly estimated through radiographs. This method is more commonly used and more efficient with dental implants, due to their position after placement. Dental implants are oriented with their long axis parallel to the long axis of the surrounding teeth, which makes them well suited for radiographic analysis. Bitewing radiographs are used to measure crestal bone level, because it has been suggested as an important indicator for implant success.129 However the resolution of bitewing radiographs is not to evaluate either primary or secondary stability. There is also the possibility of distortion, if the central x-ray tube is not positioned parallel to the structures of interest. Bone loss usually starts from the facial and then extends to the mesiodistal surfaces of the implant.130 Conventional periapical and panoramic views do not provide information about facial bone levels. 28 Regular radiographs cannot be used to quantify bone quality nor density. They can be used to assess changes in bone mineral only when there are decreases that exceed 40% of the initial mineralization.131 It may be that computer assisted measurements of bone level change or three dimensional estimations of radio-opacity will prove to be the best ways to use radiographic information.132 Finite Element Analysis Two and three dimensional finite element models provide a computer simulated, theoretical analysis, based on known material properties. Young’s Modulus, the Poisson ratio and bone density are typically properties used. By altering boundary conditions, such as the bone level, finite element modeling can theoretically be used to calculate the anticipated stresses and strains in various simulated peri implant bone levels.133,134 Finite element modeling has been used to study the stress and strain provided by miniscrew implants.82,83 A study done by Dalstra and Melsen used the finite element method for stress/strain analysis of complex bone-implant interactions.83 They used two three-dimensional models, one that was a geometrical representation of the actual bone-miniscrew interface (from micro-computed tomography) 29 and one that was used to study the influence of cortical bone and trabecular bone density on local strain distributions. It was shown that the miniscrew was displaced in a tipping mode, with the screw apex moving in the opposite direction of the screw head. This generates compressive stresses in the opposite direction of the applied force. In general, the stress levels were higher in the cortical bone than in the underlying trabecular bone. The opposite was the case for the strain values. Higher strains were observed in the trabecular bone. The same finite element study83 showed that the miniscrew implants dimensions and geometry play a significant role in the transfer of load from the implant to the bone. The diameter and the length of the implant were especially important. If the diameter is doubled, the stresses in the screw, loaded under bending, decreased by a factor of eight. On the other hand, the strains in the surrounding bone decreased only by a factor of two, and the effective surface area of bone-toimplant contact doubled. The overall deformation mode of the screw remained the same. As far as the length is concerned, the part of the miniscrew that was out of the bone was deformed in a tipping mode. The further it extended from bone, the higher the lever effect of the applied force on the miniscrew implant. On the other 30 hand, the less the miniscrew implant extended out of the bone, the lower the resistance of the surrounding trabecular bone against deformation, leading to higher bone strains. This could create a potential risk of loosening. Miyajima et al. performed another finite element analysis using two different miniscrew implant models, one was 1 mm in diameter and 5 mm long, and the other was 2 mm in diameter and 15 mm long.82 The miniscrews were placed in virtual alveolar bone model that was 10 mm wide, 10 mm deep, 30 mm tall, with the top 2 mm consisting of cortical cone. The finite element analysis revealed that the stress distributions and deflections were similar for the two miniscrew models, although the values were much larger for the smaller screw. The maximum stress in the alveolar bone was again found to be distributed in the cervical region, which approximates the first two revolutions of the miniscrew threads. The same study also showed that horizontal forces applied to the miniscrew produces greater stress in both the alveolar bone and the miniscrew,82 the vertical forces applied to the miniscrew were distributed over a larger area, and consequently produced less stress. Finally, Miyajima et al. noted that the further away from the surface of the bone that orthodontic forces are applied, 31 the higher the stress in both the alveolar bone and the miniscrew.82 A serious limitation of finite element modeling is that it is a theoretical approach, based on assumptions derived from average bone properties. It is essentially a static analysis that is difficult to apply in clinical situations. Percussion Test A percussion test is one of the simplest methods that can be used to estimate osseointegration. It is based upon vibrational acoustic science and impact response theory. A clinical judgement about osseointegration is made based on the sound heard upon percussion with a metallic instrument. A ringing sound indicates successful osseointegration, whereas a “dull” sound indicates no osseointegration. However, this method heavily relies on the clinician’s experience level and it is very subjective. It has not been used experimentally and is difficult to standardize clinically. 32 Pulsed Oscillation Waveform Kaneko et al. used a pulsed oscillation waveform to analyze the mechanical vibrational characteristics of the bone-to-implant interface using forced excitation of a steady-state wave.135 Pulsed oscillation waveform is based on the frequency and amplitude of the implant vibration induced by a small pulsed force.135-138 This system consists of an acoustoelectric driver, an acoustoelectric receiver, a pulse generator and an oscilloscope. Both the acoustoelectric driver and the acoustoelectric receiver consist of a piezoelectric element and a puncture needle. A multifrequency pulsed force of about 1 kHz is applied to the implant by lightly touching it with two fine needles connected to piezoelectric elements. Resonance and vibration generated from the bone-implant interface of an excited implant are picked up and displayed on an oscilloscope. An in vitro study showed that the sensitivity of the pulsed oscillation waveform test depended on load directions and positions; sensitivity was low for the assessment of implant rigidity.136 33 Impact Hammer Method The impact hammer method is an improved version of the percussion test. By enhancing the response detection using various devices, such as a microphone, an accelerometer, or a strain gauge, and by processing the detected response with fast fourier transform, it becomes possible to quantify and qualify the response wave in terms of dislocation, speed, acceleration, stress, distortion, sound and other physical properties. The Dental Mobility Checker (Osaka, Japan) was originally developed by Aoki and Hirakawa. It detected the level of tooth mobility by converting the rigidity of teeth and alveolar bone into acoustic signals. The dental mobility checker uses a small impact hammer as an excitation device and has been shown to provide quite stable measurement for osseointegrated implants.139 The two most common difficulties with the Dental Mobility Checker are double tapping and the ability to attain constant excitation. The Periotest has been reported to be a reliable method for determining implant stability.140-144 Periotest uses an electromagnetically driven and electronically controlled metallic tapping rod located in a handpiece. The implant’s response to striking is measured by a small accelerometer incorporated into the head of the device. 34 Contact time between the test object and tapping rod is measured and then converted to the Periotest value. The Periotest and Dental Mobility Checker are similar. They both use a transient impulse as an excitation force, and in both cases the analysis is conducted over time. Also, they were both developed for measuring natural tooth mobility.140 Reports using the Periotest for measuring dental implant stability have noted a lack of sensitivity.142,145 Periotest permits a very wide dynamic range [ -8 to +50 (Periotest Values-PTV)] of possible responses, which is necessary due to variation in natural tooth mobility.140,142 However, the dynamic range of implant mobility is very limited (-5 to +5), which reduces the sensitivity of these devices.140 Studies have indicated that Periotest values of clinically stable implants fall within an even narrower range(-4 to -2 or -4 to +2).146,147 Values measured with Periotest are significantly influenced by excitation conditions, such as position and direction; the device is also sensitive to technical errors. As noted in the Periotest user’s manual, “The Periotest measurement must be made in the midbuccal direction” and “During measurement the Periotest handpiece must always be held perpendicular to the tooth axes”.148 Considering the intraoral environment, and the pengrip-shaped handpiece of the Periotest, it can be used 35 quite easily for the anterior region. However, its use in the molar region is extremely difficult due to the presence of buccal mucosa. Derhami et al. used a fixing device to hold the handpiece at the correct angle.149 The fixing device was used for in vitro measurements of a cranial bone model. It has been shown that “in vitro” Periotest measurements are reliable15 and can be used to identify peri-implant bone loss in millimiter increments.16 In addition to the previous studies, longterm term data have shown that the Periotest provides objective clinical measurements of bone-implant anchorage.150,151 Aparicio used the Periotest to measure implant stability and found a direct correlation between Periotest values and the degree of initial osseointegration.152 Later studies have produced similar findings.150,151 However, other researchers have concluded that Periotest values cannot be used to identify an implant that may or may not be adequately osseointegrated.153 Despite these controversies, there has been interest in using the Periotest for evaluating miniscrew stability. For example, Orquin and colleagues used the Periotest to show that neither the length nor the diameter of miniscrew influence their primary stability.154 The reliability (intraclass correlation) for 36 the Periotest was found to be 0.88 in a clinical17 and 0.86 in a laboratory environment.15 Periotest measurements are limited because they are strongly dependent to the orientation of the excitation source and the striking point. Periotest measurements have been shown to increase 1.5 units for each millimeter from the marginal bone in abutment or reference point height.145 In vitro and in vivo experiments have demonstrated that the influence of the striking point on Periotest values is much greater than the effect of increased implant length, due to marginal bone resorption, or even other excitation conditions, such as the angle of the handpiece or repercussion of a rod.145,149 Unfortunately, controlling these factors is extremely difficult. Despite some positive claims for the Periotest,142,152 the prognostic accuracy of the Periotest for implant stability has been criticized for a lack of resolution, poor sensitivity and susceptibility to operator variables.155 37 Resonance Frequency Analysis In resonance frequency analysis the implant, through various ways (such as physical contact or excitation with magnetic forces), is forced to oscillate. The frequency that causes the implant to oscillate at maximum amplitude is registered as the implant’s resonance frequency. A physical system can have as many resonance frequencies as it has degrees of freedom; each degree of freedom can vibrate as a harmonic oscillator and have its own mode of vibration. A mode of vibration is a characteristic pattern or shape in which a mechanical system will vibrate. Most systems have many modes of vibration, and it is the task of the modal analysis to determine these shapes. The actual vibration of a structure is always a combination of all the vibration modes. But they need not all be excited to the same degree. While resonance frequency has shown to be of limited clinical value if single readings are used to evaluate osseointegration, they are useful if sequential measurements are taken.27 There are two commercially available devices used to evaluate the resonance frequency of implants placed into bone. Their main difference is in the way they excite the implant. The Implomates device (Taipei, Taiwan) has been studied extensively by Huang et al.24,156-159 Huang et al. 38 conducted their first study in a laboratory environment.158 They used an impulse hammer to excite the implant, which was secured in a clamp stand, and recorded the results with an acoustic sensor.158 The results showed that the resonance frequency values correlated with the height of the clamping level and the magnitude of the clamping torque.158 In another study by Huang et al., the resonance frequency was evaluated in vitro and also theoretically, through the use of a finite model analysis.24 The study showed that the resonance frequency of the implant was affected by its marginal bone characteristics, including type, density and level.24 It was also shown that finite element analysis provides reliable results for the analysis of the vibrational characteristics of a dental implant.24 Their results were repeated in another “in vitro” study by Huang et al.160 Chang et al.159 used the same type of device for detecting resonance frequency of dental implants. Their results showed that the device can be used for evaluating bone union during osseointegration.159 They maintained the orientation of the device when evaluating resonance frequency in the buccolingual direction.159 The frequencies increased as clamping torque and exposed height of the implants increased.159 Their results also correlated with the Osstell device.159 Results substantiate the notion that higher initial resonance 39 frequency values are related to higher final resonance frequency values taken 12 weeks after implant placement.156 A major disadvantage of the Implomates technique is that when a structure is excited transiently by tapping, with a hammer for an example, it oscillates in all its modes of vibration, with the strengths of the different modes being dependent on the nature of the impulse.13 The structural response is a function of natural frequencies of all the modes of the system.13 It is very important to restrict the oscillation in the first mode of resonance, which is the mode that is a function of the length of the beam and its stiffness(i.e., the two factors of greatest clinical importance).13 The Osstell Mentor device also uses resonance frequency for evaluating the stability of dental implants.161,162 It excites the implant with a steady-state sinusoidal method, which is the traditional method of measuring the natural frequencies and the corresponding damping factors of a structure.13 The Osstell device has been found to be better than the Periotest device as a mean of measuring dental implant stability in the clinical (intraclass correlation coefficients for the implant stability quotient based on Mentor was found 0.99 and 0.88 for the Periotest)17 and in a laboratory environments (intraclass correlation coefficients for the 40 implant stability quotient was found to be 0.99, compared to 0.86 for the Periotest).15 Current resonance frequency analysis systems are battery driven and use third generation transducers that are precalibrated by the manufacturer.18 The results are presented as the implant stability quotient rather than in hertz, as measured by the systems. The implant stability quotient is based on the underlying resonance frequency and ranges from 1 (lowest stability) to 100 (highest stability). This index is only valid for a transducer calibrated for a specific type of implant.163 The first and second generation transducers had to be calibrated in order for their measurements to be comparable. Each transducer had its own fundamental resonance frequency.18 Precalibrated transducers are available for different implant systems, making resonance frequency analysis measurements comparable, irrespective of the type of implant measured. Without precalibration, the Osstell transducer would not be suited for quantitative comparison of the implant stability.164,165 If the transducer is not precalibrated for a particular type of implant, then the resonance frequency value should be used instead.163 The third generation transducer is wireless, with a metal rod (a peg) that is screwed into the implant. The peg has a small magnet attached to its top which is 41 excited by magnetic pulses generated by a handheld computer. The peg vibrates in two directions, which are approximately perpendicular to each other. The vibration is measured in the direction that gives the highest resonance frequency, as well as in the direction that gives the lowest resonance frequency. Thus, two implant stability quotient values are provided. It has been shown that changes in dental implant stability measured with the newer magnetic, third generation, device correlates well with those of the electronic, second generation device, even though the values cannot be directly compared.166 It should be noted that the transducer’s orientation influences the measurement. As such it is important to standardize the orientation when taking sequential measurements.18,163,167 Implant resonance frequency analysis stability readings vary depending on which orientation the measurements are made using the transducer. If the orientation of the transducer is maintained, the reliability has been shown to be excellent in the clinical environment17 as well as in laboratory situations15 (both studies found intraclass correlation coefficients of 0.99). Recently, Brouwers and coworkers, who studied the reliability for the second generation transducer, reported intraclass correlations of 0.46 and 42 0.77 for intra-observer and inter-observer reliability with this particular device.168 In an in vitro study Ito et al.169 used sets of three screws to stabilize an implant at four different levels. Resonance frequency decreased when the most coronal screws were unscrewed. This suggests that the marginal region is the most important for the implant stability. These findings have been confirmed by Rodrigo et al., who noted higher resonance frequency measurements in specimens where the cortical bone had been maintained than when it had been eliminated.170 Finite element analysis by Deng et al. also showed that coronal osseointegration models produce slightly higher resonance frequencies than models that had other patterns of osseointegration.171 There are a substantial number of studies that have shown correlations between resonance frequency analysis and various other measures of dental implant stability. In a clinical study by Lopez et al., resonance frequency analysis was found to be significantly correlated with insertion torque (r=.284) implant diameter and negatively related to implant length.172 Guncu et al. found negative relations between the radiographic bone level and implant stability for both immediately and conventionally loaded dental implants.173 For conventionally loaded implants, the relationship reached significant levels at 6 43 months.173 O’Sullivan et al. reported a statistically significant difference in insertion torque and resonance frequency for implants that were placed in type IV bone, when compared with dental implants that were placed in type II and III.174 Tozum et al. used the third generation transducer to study the implant stability quotient, insertion torque175 and vertical bone defects.176 They found significant correlation between the first two ( r = .76) in laboratory conditions175 and a significant negative correlation between the implant stability quotient and vertical bone defects in clinical environments (r = -.464).176 Turkyilmaz et al. showed moderate correlations between resonance frequency analysis and bone density (r= .557), as measured in Hounsfield units from a CT scan and insertion torque(highest was r=.853 and lowest r=.78).114,177,178, 179,180 Degidi reported significant positive correlations between resonance frequency values and implant diameter, implant length and the diameter of bur used to create the pilot hole.181 Stenport et al. found a significant difference in resonance frequency measurement in rabbits that were hGH treated when compared to rabbits that were not.182 Finally, Wook and coworkers found a correlation between resonance frequency and the density of the surrounding bone(r= .829).183 44 The use of resonance frequency analysis as a means of evaluating osseointegration has been theoretically confirmed by finite element analyses, which show that resonance frequency increases as the interfacial bone stiffness increases.171,184 Wang and colleagues used finite element analysis to show that bi-cortical anchorage increases axial bending resonance frequency values.185 Pattijn et al. also conducted a finite element analysis study, followed by some in vitro and in vivo measurements with the second generation Osstell device. According to their results the adapted transducer does not always measure the first bending mode of the system.163 Their finite element analysis suggested that several resonance frequencies lie within the measurement range of the transducer. Relationships between resonance frequency and histomorphometric measurements have also been established. Using resonance frequency analysis, Zhou et al. were able to distinguish between sand blasted/acid etched and machined implants.19 They also showed that increased bone-to-implant contact during healing was correlated with the implant stability quotient.19 Correlation between histomorphometric data and the implant stability quotient was also observed by Scarano et al. in a sample of 7 implants.186 Kim et al., who compared different types of dental implants and surgical 45 placement techniques, found that the implant stability quotient and the bone-to-implant contact increased significantly between 0 to 8 weeks for all groups tested.187 A long-term study by Gualini et al. found very small changes in implant stability, as measured with resonance frequency, over a five year observation period.188 A number of researchers have been able to quantify a loss of dental implant stability after placement.79 Lopez et al., who used resonance frequency to evaluate the healing process with dental implants,189 noted a marked decrease in stability during the fourth week, which was followed by increases of resonance frequency through the tenth week. A similar decrease in resonance frequency was noted around the third week by Stadlinger et al. for dental implants with various surface coatings.190 They also noted increases thereafter. Also based on resonance frequency, Strand et al. observed a decrease between the 1st and 9th week for machined implants,191 while Gleuser et al. observed a decrease between the 1st and 8th week for implants that had their surface processed.192 Becker and colleagues noted that implants may have a high or low value of resonance frequency initially.193 Those that start with high values, later showed lower values; those that started with low values showed an increase.193 46 A number of studies have focused on the potential of using resonance frequency for making clinical decisions about implants. Scarano and coworkers showed that implant stability quotient values below 36 could be used to identify irretrievably failed implants.194 West et al. also found that resonance frequency analysis was useful when deciding about the timing of loading after the implant has been left unloaded for a period of time.195 They were able to identify reductions in stability initially and delay implant loading until greater stability has been attained.195 Seong et al., who measured the resonance frequency of dental implants in different anatomical regions of fresh human cadavers, found that mandibular implants had significantly higher stability than maxillary implants, with implants placed in the posterior maxilla being the least stable.196 There have also been a number of studies that were not able to estalish correlations between resonance frequency and implant stability. Veltri and colleagues observed a relationship between the implant stability quotient and marginal bone levels, but the correlation was not statistically significant.167 Cunha et al. evaluated two dental implant systems using resonance frequency analysis and cutting torque.197 They were not able to identify a correlation between the two measurements. Friberg and coworkers also failed to show 47 any correlation between cutting torque and implant stability quotient.104 They did, however, note a significant correlation between cutting torque and the implant stability quotient in the upper/crestal third of the implants (similar to the findings of Ito et al.28). Rasmusson et al. were not able to establish relationship between bone to implant contact and resonance frequency.198 Balleri et al. examined 45 implants after one year of loading and did not find any significant correlations between implant length, marginal bone level and resonance frequency. 199 An older study by Zix and coworkers showed no significant differences in the resonance frequency between immediately loaded and delay loaded implants.200 There also was no statistical difference in the implant stability quotient between the anterior and posterior segments of the maxilla. Ostman et al.201 also found no differences between immediately loaded implants and delayed loaded implants. Ramakrisha and Nayar reached the same conclusion based on a sample of eight implants.202 Schliepake et al., who evaluated resonance frequency, histomorphometric data, bone density and insertion torque, did not find any correlations between the aforementioned measurements.203 Ito et al. were also not able to identify a correlation between resonance frequency and histomorphometric data.28 Nawas et al. concluded that neither insertion torque nor resonance 48 frequency were predictive of implant loss.204 Finally, Huwiler and coworkers, who took weekly measurements of resonance frequency, were not able to identify significant changes over time.205 They did note that measurements increased during the first week, then decreased during the second and third week and finally increased again during the fourth. Summary With their increased popularity, miniscrew implants will soon be implemented extensively in everyday clinical practice. Their possible failure will heavily influence the outcome and efficiency of the treatment. As such, an in vivo method to evaluate miniscrew implant stability would hold great clinical implications. By quantifying stability, it would be possible to follow the changes that occur during the transition from primary to secondary stability. Changes in stability due to inflammation of the peri-implant tissues, overloading etc. could also be evaluated. Resonance frequency has been used to quantify implant stability in dental implants for the last ten years. There is substantial support for the method in the literature, especially when compared to other available methods. Resonance frequency analysis also holds great potential for quantifying the 49 stability of miniscrew implants. This project will test the validity of this approach with miniscrew implants by comparing resonance frequency with insertion torque and pullout test values. 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Glauser R, Lundgren A, Gottlow J, Sennerby L, Portmann M, Ruhstaller P et al. Immediate occlusal loading of Branemark TiUnite™ implants placed predominantly in soft bone: 1-year results of a prospective clinical study. Clin Implant Dent Relat Res 2003;5:47-56. 193. Becker W, Sennerby L, Bedrossian E, Becker B, Lucchini J. Implant stability measurements for implants placed at the time of extraction: a cohort, prospective clinical trial. J Periodontol 2005;76:391-397. 194. Scarano A, Carinci F, Quaranta A, Iezzi G, Piattelli M, Piattelli A. Correlation between implant stability quotient (ISQ) with clinical and histological aspects of dental implants removed for mobility. Int J Immunopathol Pharmacol 2007;20:33-36. 68 195. West J, Oates T. Identification of stability changes for immediately placed dental implants. Int J Oral Maxillofac Implants 2007;22:623-630. 196. Seong W, Holte J, Holtan J, Olin P, Hodges J, Ko C. Initial stability measurement of dental implants placed in different anatomical regions of fresh human cadaver jawbone. J Prosthet Dent 2008;99:425-434. 197. Cunha H, Francischone C, Filho H, de Oliveira R. A Comparison Between Cutting Torque and Resonance Frequency in the Assessment of Primary Stability and Final Torque Capacity of Standard and TiUnite Single-Tooth Implants Under Immediate Loading. Int J Oral Maxillofac Implants 2004;19:578-585. 198. Rasmusson L, Kahnberg K, Tan A. Effects of Implant Design and Surface on Bone Regeneration and Implant Stability: An Experimental Study in the Dog Mandible. Clin Implant Dent Relat Res 2001;3:2-8. 199. Balleri P, Cozzolino A, Ghelli L, Momicchioli G, Varriale A. Stability Measurements of Osseointegrated Implants Using Osstell in Partially Edentulous Jaws after 1 Year of Loading: A Pilot Study. Clin Implant Dent Relat Res 2002;4:128-132. 200. Zix J, Kessler-Liechti G, Mericske-Stern R. Stability measurements of 1-stage implants in the maxilla by means of resonance frequency analysis: a pilot study. Int J Oral Maxillofac Implants 2005;20:747-752. 201. Ostman P, Hellman M, Sennerby L. Direct Implant Loading in the Edentulous Maxilla Using a Bone DensityAdapted Surgical Protocol and Primary Implant Stability Criteria for Inclusion. Clin Implant Dent Relat Res 2005;7:60-69. 202. R Ramakrishna, Nayar S. Clinical assessment of primary stability of endosseous implants placed in the incisor region, using resonance frequency analysis methodology: An in vivo study. Indian J Dent Res 2007;18:168-172. 203. Schliephake H, Sewing A, Aref A. Resonance frequency measurements of implant stability in the dog mandible: experimental comparison with histomorphometric data. Int J Oral Maxillofac Surg 2006;35:941-946. 69 204. Al-Nawas B, Wagner W, KA G. Insertion torque and resonance frequency analysis of dental implant systems in an animal model with loaded implants. Int J Oral Maxillofac Implants 2006;21:726-732. 205. Huwiler M, Pjetursson B, Bosshardt D, Salvi G, Lang N. Resonance frequency analysis in relation to jawbone characteristics and during early healing of implant installation. Clinical Oral Implants Research 2007;18:275-280. 70 Chapter 3: Journal Article Abstract Purpose: To evaluate the reliability and validity of resonance frequency as a method for measuring the stability of miniscrew implants. Methods: One hundred and twenty miniscrew implants were randomly allocated to synthetic bone blocks of different densities: 40 low [cortical part: 38.1pcf, cancelous part: 30.0pcf ] density, 40 medium [cortical part: 40.6pcf, cancelous part: 31.8pcf] density, and 40 high [cortical part: 50.1pcf, cancelous part: 31.8pcf] density. For each implant insertion torque was measured using Mecmesin Advanced Force and Torque Indicator©. Pullout strength was measured using the Instron© Machine, Model 1011 and applying a vertical force at 20mm/min until failure. Resonance frequency was measured three times parallel and three times perpendicular to the long axis of each bone block. Results: Intraclass correlations based on multiple measures of resonance frequency ranged from 0.953 to 0.992; single measure intraclass correlations ranged from 0.870 to 0.977. Analysis of variance showed significant effects of bone density on resonance frequency; post-hoc tests showed differences for high density only. Insertion torque and pullout strength demonstrated statistically 71 significant differences between all three densities. The two orientations showed no significant differences for low and medium densities, the high density bone showed a significant (p= .049) orientation effect. Independent of bone density, correlations between resonance frequency, insertion torque, and pullout strength were low and showed no consistent pattern. Conclusions: Resonance frequency is a reliable measure. It is also a valid measure that is related with bone density. As expected for homogeneous synthetic bone, resonance frequency shows little or no relationship with orientation, insertion torque, or pullout strength. Introduction Because miniscrew implants provide skeletal anchorage for tooth movements,1,2 they are becoming increasingly popular in orthodontics.3,4 They can be easily implemented for orthodontic tooth movement using established biomechanic principles. Miniscrews hold the potential for making treatment more efficient and expanding the treatment possibilities.4 The literature reports success rates for miniscrews ranging from 83.9% to 91.6%.5-7 Failures may be caused by general factors, local factors or erroneous decisions at the time of placement or during utilization.8 Stability is 72 ultimately related to the healing process that takes place around the implant.9 Primary stability is also important because micromotion at the implant-to-bone interface can result in the formation of fibrous tissue, which leads to implant failure.10,11 Relative motion between the implant body and the surrounding bone during the early phase of healing is considered to be an important risk factor for early implant loss due to failure of osseointegration.12,13 Various techniques and devices have been suggested for evaluating of the stability of miniscrew implants.14 Some of the most commonly used methods used to evaluate stability are insertion torque, removal torque and pullout strength.6,15-17 Wu and coworkers, who evaluated miniscrew implant stability from 0 to 8 weeks, noted that removal torque and pullout strength increased over time, with statistically significant increases in removal torque after 4 weeks. Due to their invasive nature, neither of these methods can be used to evaluate stability longitudinally while the bone is healing. In order to evaluate healing of miniscrews of individual patients, a non-invasive technique that can be applied clinically is needed. The dental implant literature has shown that resonance frequency analysis holds great potential for evaluating stability in vivo.18-20 Resonance frequency 73 analysis has been found to be a reliable, with repeated measures of dental implants showing intraclass correlations of 0.99 in controlled laboratory settings21 and 0.99 in clinical situations.22 In living bone, resonance frequencies have been shown to be correlated with insertion torque (at least r= .78, p<.05)23 and bone density (at least r=.79, p<.05).23 Resonance frequency analysis is routinely used to make clinical decisions about dental implants. For example, Scarano and coworkers have provided a threshold value stability, below which dental implants might be expected to fail.24 West and coworkers used resonance frequencies to identify initial reductions in stability and subsequent increases in stability in order to determine the best time to load dental implants of individual patients.25 One of the most commonly used devices to evaluate resonance frequency is the Osstell Mentor. It uses a peg that is screwed into the head of the implant to measure resonance frequency. The peg holds a small magnet that is excited by magnetic pulses. The signal is captured with a transducer, processed with fast fourier transforms, measured in hertz, and shown on the instrument as the implant stability quotients.26 The implant stability quotient is pre-calibrated for specific combinations of implants and pegs. Because implants are often not been pre-calibrated, it has been recommended that hertz rather 74 than the implant stability quotient should be used when reporting resonance frequences.27 It has been shown that resonance frequencies are influenced by the design of the peg, the stiffness of the implant fixture and its interface, and the effective length of the implant above the marginal bone level.26 While resonance frequency analysis holds great potential, its application with smaller implants remains limited. Gedrange and coworkers used resonance frequency analysis to show that there was no difference in stability between 4 mm and 6 mm Straumann palatal implants.28 Recently, Jackson and coworkers, who also used resonance frequency analysis with palatal implants (3.3mm diameter, 4-6mm length), showed that immediately loaded implants were significantly less stable than implants that remained unloaded for eight weeks.29 Resonance frequency analysis has also been used to show that there are no differences in stability between cylindrical and conical miniscrew implants.30 Most recently, Veltri et al used resonance frequency to show that there were no significant differences between three commercially available miniscrew implants.31 The reliability of resonance frequencies of miniscrew implants has not been investigated. The purpose of this project was to determine the reliability and the validity of using resonance frequency 75 analysis with miniscrew implants. The following working hypotheses were tested: 1. Multiple resonance frequency measures taken on the same specimen are reliable and show no systematic errors. 2. There are differences in the resonance frequency between synthetic bone of low, medium and high density. 3. There are no correlations between resonance frequency analysis, insertion torque and pullout test. 4. There are no differences in resonance frequencies associated with the orientation of the device. Materials and Methods Mini Screw Implants The miniscrews that were used for this study were the Ancoragem Ortodontica made by Neodent© (Curitiba, Brazil). They were 11 mm long, with an external diameter of 1.6 mm, an internal diameter of 0.9 mm, and a 0.7 mm pitch (Figure 1a, Appendix II). The implant has a conical shape and was self drilling. The head had been modified with an inner thread of maximum diameter of 1.1 mm 76 (Figure 1b, all figures in appendix II), for insertion of the Osstell Mentor Smartpeg Type A3 (Figure 1c). Synthetic Bone The miniscrews were placed in synthetic bone blocks made by Sawbones© (Vashon, Washington). The synthetic bone blocks included three combinations of cortical and cancellous bone of differing densities, including low density [cortical part: 38.1pcf, cancellous part: 30.0pcf], medium density [cortical part: 40.6pcf, cancellous part: 31.8pcf], and high density [cortical part: 50.1pcf, cancellous part: 31.8pcf]. The cortical bone was 2 mm thick and fixed to the cancellous bone with 40 pcf rigid polyurethane foam. The densities of the synthetic bone selected for this study were chosen to represent the human mandible, which has been reported to be 41.2pcf (0.66 g/cc).32 Previous studies have shown that synthetic bone is a good substitute for real bone.33 The homogenous nature of synthetic bone is essential for controling the variability of bone properties found in natural bone. Each block of synthetic bone (170 mm x 120 mm x 42 mm) was marked so that the surface delimited 20 squares (30 mm x 30 mm). A miniscrew was inserted into the center of each square (Figure 2a). There were two blocks per 77 density (high, medium, low), with a total of six blocks and 120 miniscrews. The synthetic bone blocks were embedded in plaster stone to stabilize the blocks with two vices. The compression forces of the vices were applied to the plaster stone so as not to interfere with the resonance frequency measurement (Figure 2b). Insertion Torque A guide for the miniscrew driver was placed over the bone block to standardize miniscrew insertion. It ensured that each screw was inserted in the same direction; it also prevented wobble of the driver during insertion (Figure 3a). Each miniscrew was inserted manually with the hand driver until only one thread remained exposed above the surface of the cortical bone. The hand drive was then replaced by the Mecmesin Advanced Force and Torque Indicator© (Mecmesin, Ltd, West Sussex, UK) (Figure 3b) and the miniscrew was inserted until the point right before the head touched the synthetic bone. This was done to prevent countersink friction. The maximum value of insertion torque was measured in N/cm. 78 Resonance Frequency Analysis After the miniscrews had been inserted into the synthetic bone, the Osstell Smartpeg was fitted onto the head of each miniscrew with finger pressure, according to the manufacturer’s instructions. It has been previously shown that tightening the transducer 10 Ncm or more has no significant effect on resonance frequency of implants.18 Once the Smartpeg was fitted, sequential measurements of the resonance frequency were taken using the Osstell Mentor© transducer (Göteborg, Sweden). Three measurements were taken parallel to the long side of the synthetic bone block and three more were taken parallel to the short side of the synthetic bone block (Figure 4. These two sets of measurements were perpendicular to each other. All resonance frequency measures were reported in hertz. Pullout Strength To measure pullout strength, each bone block was sectioned into 20 smaller blocks (12mm x 12mm x 12mm), with one implant in the center of each block. Each of the smaller blocks was placed in a base specially designed to secure the block and fit the Instron© Machine, Model 1011 (Instron Corp, Canton, MA). The base had a lid that secured each block of bone during pullout testing (Figure 79 1d). The miniscrews was attached to the Instron using a custom made grappling device designed to fit the head of the miniscrew. A vertical force of 20mm/min was oriented parallel to the long axis of the miniscrew and applied until failure. Peak load at failure was obtained from the readout and recorded in kilograms. Statistical Analyses Skewness and kurtosis statistics showed that the variables were normally distributed. Repeated measures analyses of variance were used to evaluate systematic differences between the three measures of resonance frequency. Cronbach’s Alpha was used to estimate the reliability of the three measures taken on each miniscrew; intraclass correlations were used to evaluate the reliability of single measures of resonance frequency. Validity of the resonance frequency measures was evaluated two ways. Assuming that valid measures are able to identify differences that should exist (discriminant validity), analyses of variance were used to evaluate differences in resonance frequency associated with bone density. Based on the fact that the synthetic bone is homogeneous in its material properties, valid measures of resonance frequency should not be related to other 80 measures of stability and should be independent of the orientation from which the measure were taken (convergent validity). To this end, paired t-tests were used to compare differences in orientation and correlations were used to evaluate associations between resonance frequency, insertion torque and pullout force. Results Reliability Analysis of variance showed no significant differences between the three measurements taken parallel and perpendicular to the long axis of the bone block regardless of density (Table 1; Figure 5, Appendix I and II). Multiple measure intraclass correlations ranged from 0.953 to 0.992 and single measure reliabilities range from 0.870 to 0.977(Table 2, all tables are to be found in Appendix I). All of the intraclass correlations were statistically highly significant (p< .001). The lowest reliability (0.870) was found for single measurements in the medium density synthetic bone block; the highest reliability (0.992) was found for multiple measurements in the low density synthetic bone block. The multiple 81 measurements were generally more reliable than the single measurements in all instances. Validity Based on the mean values of the three resonance frequency measurements, paired t-tests showed that there was a statistically significant difference between the parallel and perpendicular orientation for the medium density bone (p=0.049). There was no statistically significant (p>.05) difference due to the orientation of the device for low and high density bone. Analyses of variance demonstrated significant difference in resonanance frequency between the three bone densities. Post hoc tests showed differences for high density only. There was no significant difference in resonance frequency between the low and the medium density bone (Table 3). Analyses of variance also showed significant effects of bone density on insertion torque and pullout strength, with significant differences between all three densities (Table 3). With the three bone densities combined (ie. not controlling for density) there was a moderately high correlation (r= 0.830) between insertion torque and pullout strength (Table 4; Figure 6). Insertion torque was also significantly (p<.01) related with the parallel 82 resonance frequency measures (r= 0.471), as well as the perpendicular resonance frequency measures (r= 0.337). Pullout strength showed significant correlations with both the parallel (r= 0.490) and perpendicular (r= 0.327) resonance frequency measures. Independent of bone density (i.e. with the three densities evaluated separately), correlations between resonance frequency, insertion torque and pullout strength were limited and showed no consistent pattern (Table 4). Only six of the fifteen possible correlations were statistically significant (p<.05). The medium density bone blocks showed low correlations between resonance frequency and insertion torque (r=0.336 and r=0.338 for the parallel and perpendicular orientations, respectively). Resonance frequency was also related to pullout strength in the medium density bone; the high density bone showed significant correlations between resonance frequency and pullout strength for the parallel orientation only (r=.401). Insertion torque was significantly related to pullout strength (r=.412) only in the low density bone blocks. 83 Stabilization of Blocks Regardless of orientation there were no statistically significant differences in resonance frequency between the miniscrew implants that had been stabilized and those that had not been stabilized (Table 5). Discussion The results showed high level of reliability when using the Osstell Mentor with miniscrew implants. The lowest intraclass correlation was 0.953 for multiple measurements and 0.870 for single measurements. As multiple measurements were more reliable than single measures, it may be preferable to measure resonance frequency multiple times and average the measures in order to obtain the most reliable estimates. While reliability has not previously been established for miniscrew implants, studies have shown similar or lower levels of reliability for dental implants. In a clinical study, Zix and coworkers reported an intraclass correlation of 0.99 for the Mentor.22 Lachmann and coworkers, in a laboratory study, also found an intraclass correlation coefficient of 0.99.21 On the other hand, Brouwers and coworkers, reported an intraclass correlation of 0.46.34 However, Brouwers and coworkers 84 studied dental implants that had been placed in dry edentulous mandibles that had not been stabilized, which holds important implications for the clinical application of resonance frequency measures. Based on the present findings, the Osstell Mentor can reliably be used to quantify the stability of miniscrew implants. The resonance frequency recorded for miniscrew implants in this study was substantially lower than previously reported for miniscrews. Veltri and coworkers reported resonance frequencies of 6000 hertz (compared with the 2000 hertz found in this study) for three types of miniscrew implants placed in rabbit femoral condyles.35 However, their miniscrews had an adapter for the Osstell transducer soldered onto the miniscrew head, which could have affected the resonance frequencies. They also used a previous generation of the device, the frequencies of which have been shown not to correlate with the newer generation devices.36 Moreover, their miniscrews were fully inserted while those in the present study were not, which also could have produced different frequency values.26 As expected, insertion torque, pullout strength and resonance frequency were significantly related to bone density. While these relationships have not been previously demonstrated for miniscrews, it has been shown that resonance frequencies of dental are correlated with 85 insertion torque,23,37,38 bone density.23,37,39 The relationship between resonance frequency and pullout strength has not been previously investigated. Pullout strength has been found to correlate with bone density40 and insertion torque.41 The existence of relationships between the measurements taken and bone density is important because they make it possible to indirectly estimate the process of mineralization around the implant. Resonance frequencies were not significantly different between the low and medium density bone. This may be due to the fact that there was relatively little difference in bone properties between low and medium density bone. Low density was made from a cortical part of 38.1 pcf and a cancellous part of 30.0 pcf, while medium density was made from a cortical part of 40.6 pcf and a cancellous part of 31.8 pcf. There was a total 4.3 pcf difference between the low and medium density bone, while the high and the medium difference was 10.5 pcf. It is also possible that the device may not be as sensitive in the lower end of the frequency spectrum. The Osstell Mentor measures resonance frequencies between 1000 and 10,000 hz, and the measurements taken were clearly at the lower end of that spectrum. Associations between resonance frequency, insertion torque, and pullout strength and bone density have been previously reported for dental implants. However, most 86 studies reporting correlations did not control for variation in bone properties, and correlations that do not control for extraneous sources are by definition spurious. In a cadaver study, Turkyilmaz and coworkers found that resonance frequency was correlated both with insertion torque (r= 0.78) and bone density (r= .853).23 When bone density was not controlled, the present study also showed significant, albeit slightly lower, correlations between resonance frequency and insertion torque. Similarly, Song and coworkers found correlations ranging from 0.72 to 0.81 between insertion torque and pullout strength in Sawbones, but they included bone of different cortical thicknesses.41 The present study showed a correlation of .83 between insertion torque and pullout strength when all three densities were included in the calculations (i.e. did not control for density). The present findings emphasize the importance of controlling extraneous variation when evaluating associations between stability measures. When variation in density was controlled, there was little or no association between resonance frequency, insertion torque and pullout strength. Only six of the fifteen possible correlations were statistically significant (Table 4). Importantly, correlations should not be expected in homogeneous bone. If the technical aspects could be totally controlled without error, all of 87 the resonance frequencies should be identical. The weak correlations identified simply indicate that the procedures were not controlled and that there was error. Most important, there was no pattern of association evident and the associations identified were weak. Since there was no distinct pattern of across the three density groups, the correllations could have be due to lack of technical control, especially in the medium density group that should the greatest number of association. Moreover, the lack of association between insertion torque and pullout strength suggests that the correlations were not due to variation in stability. Because the lack of association in homogeneous bone is expected, the present finding serve to validate resonance frequency measurements with miniscrew implants. Resonance frequencies have been previously shown to differ in real bone, depending on the orientation of the transducer at the time of the measurement.26,36,42-44 With homogeneous bone, difference should not be expected. The effect of orientation of the measurement has not been previously evaluated with sawbone. While there were no differences for low and medium density bone, the high density bone showed a significant difference (p=.049). However, after a Bonferroni adjustment for multiple measurements, this difference was not statistically significant. Because no differences were found and no 88 differences were expected, this further serves to further validate the use of resonance frequency with miniscrew implants. While no significant effects were found in the present study, it is important to stabilize specimens when measuring resonance frequency. Lack of stability interferes with the measurement and can lead to erroneous results. Pattjin et al showed that the nearer the clamping vice is to the implant the higher the measurement that is measured.42 The present study stabilized the bone blocks indirectly using plaster stone, the weight of which was evidently sufficient to stabilize the miniscrews. 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Med Eng Phys 2007;29:182-190. 43. Veltri M, Balleri P, Ferrari M. Influence of Transducer Orientation on OsstellTM Stability Measurements of Osseointegrated Implants. Clin Implant Dent Relat Res 2007;9:60-64. 44. Fischer K, Stenberg T, Hedin M, Sennerby L. Five-year results from a randomized, controlled trial on early and delayed loading of implants supporting full-arch prosthesis in the edentulous maxilla. Clin Oral Implants Res 2008;19:433-441. 95 Appendix I: Tables 96 Table 1. Comparison of resonance frequency (RF) of three consecutive measurements [taken parallel (║) and perpendicular (⊥)to the long side of the rectangular synthetic bone block] for the high, medium and low density bone blocks. 97 RF ║ (Hz) RF ⊥ (Hz) Low Medium High Low Medium High 1st Measurement Mean SD CV (%) 2nd Measurement Mean SD CV (%) 3rd Measurement Mean SD CV (%) 1888.4 1896.6 2015.0 1898.2 1906.1 1996.7 1878.0 1897.9 2028.6 1896.4 1894.6 1988.5 1886.0 1902.3 2024.6 1901.5 1896.1 1994.6 131.7 103.0 96.9 126.7 102.7 81.9 6.9 5.4 4.8 6.7 5.4 4.1 139.9 95.4 90.4 125.5 111.8 90.0 7.4 5.0 4.4 6.6 5.9 4.5 129.6 94.2 90.8 129.0 110.8 85.6 6.9 5.0 4.5 6.8 5.8 4.3 ANOVA P Value .529 .840 .068 .399 .131 .278 Table 2. Reliabilities of resonance frequency (RF) measures evaluated using intraclass correlations for single measurements and Cronbach’s Alpha for multiple measurements. RF ║ RF ⊥ 98 Single Multiple Single Multiple Low Density Estimated P Value .928 <.001 .975 <.001 .977 <.001 .992 <.001 Medium Density Estimated P Value .870 <.001 .953 <.001 .947 <.001 .982 <.001 High Density Estimated .912 .969 .886 .959 P Value <.001 <.001 <.001 <.001 Table 3. Comparison of resonance frequency (RF) measurement averages [taken parallel (║) and perpendicular (⊥) to the long side of the rectangular synthetic bone block] insertion torque (IT) and the pullout strength (POS) for the high, medium and low density bone blocks. 99 Mean RF ║ Average(Hz) 1884.5 RF ⊥ Average(Hz) 1898.7 IT (Ncm) 18.2 POS (Kg) 45.5 Low Density SD CV (%) 130.5 6.9 126.1 6.6 1.2 6.6 2.1 4.6 Medium Density Mean SD 1898.9 97.5 1898.9 108.4 21.0 1.4 54.5 1.8 CV (%) 5.1 5.7 6.5 3.4 Mean 2022.7 1993.3 24.5 61.2 High Density SD 92.7 85.8 2.2 3.4 CV (%) 4.6 4.3 8.9 5.5 Difference P Value <.001 <.001 <.001 <.001 Table 4. Correlations between resonance frequency (RF) measurements [taken parallel (║) and perpendicular (⊥)to the long side of the rectangular synthetic bone block],insertion torque (IT) and pullout strength (POS) for the high, medium and low density synthetic bone blocks separately and combined. 100 RF ║ w/ IT Low Density r (prob) .006 (.972) Medium Density r (prob) .336 (.034)* High Density r (prob) .194 (.231) All Densities r (prob) .471 (<.001)* RF ⊥ w/ IT -.082 (.614) .338 (.033)* .057 (.726) .337 (<.001)* RF ║ w/ POS .027 (.867) .388 (.013)* .401 (.010)* .490 (<.001)* RF ⊥ w/ POS -.138 (.395) .335 (.035)* .134 (.409) .327 (<.001)* IT w/ POS .412 (.008)* .096 (.557) .251 (.118) .830 (<.001)* Table 5. Comparison of resonance frequency (RF) measurements [taken parallel (║) and perpendicular (⊥)to the long side of the rectangular synthetic bone block] with the medium density block clamped and not clamped. st 101 RF║ 1 RF║ 2nd RF║ 3rd RF║ Average RF ⊥ 1st RF ⊥ 2nd RF ⊥ 3rd RF ⊥ Average Mean 1875.8 1893.8 1893.8 1887.8 1916.4 1915.1 1907.0 1912.8 Not Clamped SD CV (%) 119.4 6.4 119.6 6.3 119.6 6.3 119.5 6.3 151.1 7.9 126.2 6.6 142.6 7.5 140.0 7.3 Mean 1945.9 1942.2 1932.7 1950.3 1936.4 1909.0 1927.9 1924.4 Clamped SD 134.5 118.9 110.4 121.3 127.4 124.0 141.6 131.0 Difference CV (%) P Value 6.9 .083 6.1 .108 5.7 .254 6.2 .101 6.6 .464 6.5 .802 7.3 .466 6.8 .627 Appendix II: Figures 102 a b c d Figures 1: Miniscrew Implant and Custom Pullout Base a) Neodent miniscrew implant, b) Head modified for the attachment of the Osstell peg, c) Miniscrew implant with the Osstell peg, d) Custom base to hold implants during pullout test. a b Figures 2:Synthetic Bone a)Synthetic bone marked for implant placement, b)Synthetic bone block stabilized with two vices on laboratory bench. 103 b a Figure 3: Miniscrew Implant Driver a) Hand driver with customized placement device, b)Insertion torque measuring device. a b Figure 4: Resonance frequency analysis measurements a) parralel and b) b) perpendicular to the long side of the synthetic bone block. 104 a b c 3rd 2100 2050 2000 1950 1900 1850 1800 1750 1700 2100 2050 2000 1950 1900 1850 1800 1750 1700 d 2nd 3rd 1st Hz 1st Hz e 1st 2nd 3rd 1st 2nd 2nd 3rd 2100 2050 2000 1950 1900 1850 1800 1750 1700 f 3rd Hz 2100 2050 2000 1950 1900 1850 1800 1750 1700 2nd Hz Hz Hz 1st 2100 2050 2000 1950 1900 1850 1800 1750 1700 1st 2nd 3rd 2100 2050 2000 1950 1900 1850 1800 1750 1700 Figure 5. Comparisons of resonance frequencies of three consecutive measurements [taken parallel (║ ) to the long side of the synthetic bone block for the a) low, b) medium and c) high density; comparison of resonance frequencies of three consecutive measurements [taken perpendicular (⊥) to the long side of the synthetic bone block for the d) low, e) medium and f) high density a Low Medium b High Resonance Frequency Pullout Strength 70 65 60 55 50 45 Low Medium High 2200 2000 1800 1600 1400 40 150 170 190 210 230 250 270 150 290 170 190 210 230 250 270 290 Insertion Torque Insertion Torque Resonance Frequency c Low Medium High 2200 2000 1800 1600 1400 40 45 50 55 60 65 70 Pullout Strength Figure 6. Scatterplots of a) pullout strength with insertion torque, b) resonance frequency with insertion torque, and c) resonance frequency with pullout strength. 105 Vita Auctoris Georgios Katsavrias was born in Marousi, Athens on the 3rd of June 1979. He was raised in Chalandri, Athens. He attended primary and secondary school in Chalandri, Athens and high school in Penteli, Athens. He entered the Dental School of the University of Athens in September, 1999 and graduated in 2005. In June, 2006 he came to Saint Louis, USA to attend the postgraduate orthodontic program of Saint Louis University from which he graduated in June, 2009. 106