Download RAJIV GANDHI UNIVERSITY OF HEALTH SCIENCES

RAJIV GANDHI UNIVERSITY OF HEALTH SCIENCES
BANGALORE, KARNATAKA
PROFORMA FOR REGISTRATION OF SUBJECTS FOR DISSERTATION
1.
NAME OF THE CANDIDATE Dr. SOMA KRISHNA RAJA RAO BOGGARAPU
AND ADDRESS (in block letters)
POST GRADUATE STUDENT, DEPARTMENT
OF PEDODONTICS AND PREVENTIVE
DENTISTRY,
DR. SYAMALA REDDY DENTAL COLLEGE,
HOSPITAL & RESEARCH CENTRE,
#111/1, SGR COLLEGE MAIN ROAD,
MUNNEKOLALA EXTENSION,
MARATHAHALLI POST,
BANGALORE – 560037.
KARNATAKA
2.
NAME OF THE INSTITUTION
DR. SYAMALA REDDY DENTAL COLLEGE,
HOSPITAL & RESEARCH CENTRE,
MARATHAHALLI, BANGALORE- 560037
3.
COURSE OF
SUBJECT
STUDY
AND
MASTER OF DENTAL SURGERY IN
PEDODONTICS AND PREVENTIVE
DENTISTRY, (M.D.S)
4.
DATE OF ADMISSION TO THE 18th JULY 2013
COURSE
5.
TITLE OF THE TOPIC
COMAPARITIVE EVALUATION OF
VARIOUS STORAGE MEDIA VIZ. SOY
MILK, COCONUT WATER, EGG WHITE
HANK’S BALANCED SALT SOLUTION IN
MAINTENANCE OF VIABILITY OF
PERIODONTAL LIGAMENT
FIBROBLASTS.
BRIEF RESUME OF INTENDED WORK:
6.1 Need for the study:
Tooth avulsion is one of the most severe form of dental trauma. The incidence of tooth avulsion
ranges from 1% - 16% of all the traumatic injuries occurring in permanent dentition6.It occurs
most frequently during 8-12 years of age. Timely re-implantation of the avulsion is the most
appropriate management of avulsion of permanent teeth. Therefore, it is essential to maintain
the vitality of periodontal ligament cells (PDL) attached to the root portion of the tooth. From
the site and time of trauma leading to avulsion of tooth and time required to reach the dentists
office dictates the success of the re-implantation. Various storage media like Soy milk, Coconut
water, Egg white, Hank’s balanced salt solution (HBSS) are commercially available media used
to maintain the vitality of the pdl cells. Amongst the commercially available storage media, it is
found that HBSS is most suitable media. However, there is paucity of information on the
effectiveness of the storage media viz. Soy milk, Coconut Water, Egg white and HBSS with
respect to the viability of fibroblast cells.
The aim of the study is to come out with a suitable storage media which is effective in
maintaining the periodontal ligament fibroblasts viable with an appropriate time period and
which is easily available to the public.
6.2 Review of literature.
Thomas T et al (2008)1 conducted a study on Comparative evaluation of maintenance of cell
viability of an experimental transport media “coconut water” with Hank's balanced salt solution
and milk. They divided the storage media into three different groups. The three groups were
then divided into five subgroups, each denoting the storage time periods. They sub cultured the
cell line and standardized the number of cells by making a cell suspension in minimal essential
media in 5 different culture plates. They counted the cells using neubauer chamber and
statistically analyzed.
By this study, they came to an opinion that the coconut water can used as a better alternative to
HBSS.
Khademi AA et al (2008)2 conducted a study on the different experimental media namely: egg
white, milk, Hanks’ Balanced Salt Solution (HBSS) as the positive control, and tap water as the
negative control. The storage times were 1, 2, 4, 8, and 12 hours. Extracted premolar teeth of
healthy individuals were rinsed in the media. After trypsinization and subsequent treatment in
collagenase, cell viability was determined using trypan blue staining. The result of their study
was there was no difference in the cell viability between egg white and HBSS media, but there
was a statistically significant difference between the viability of PDL cells in egg white medium
in comparison with milk and water.
Ali SAH et al (2011)3 conducted a study on the effect of coconut water concentration on benchdried periodontal ligament cells.100 % mature coconut water (MCW) was better than 50%
dilutions obtained from mature or young coconuts.
MOURA CCG et al (2012)4 conducted a study on long shelf-life ultra-high temperature
whole cow milk (WM); long shelf-life ultra-high temperature skim cow milk (SKM);powdered
cow milk (PM); and soy milk (SM).
The results indicate that SM can be used as a more adequate storage medium for avulsed teeth.
Eskandarian T et al (2013)5 comparatively evaluated oral rehydration salt solutions (ORS) at
different concentrations as storage media for avulsed tooth. They results given by them elicits
that ORS with concentrations of 25% and 50% are more effective than ORS with 100%
concentration.
Sanghavi T et al (2013)6 conducted a study on the comparative efficacy of three storage media
coconut water, propolis and oral rehydration salt in maintaining the viability of periodontal
ligament cells. They selected 40 teeth with intact crown which are advised for orthodontic
extraction. They divided the teeth randomly into three experimental solution groups. They
divided the remaining 10 into positive and negative control groups.
The results of their prevailing study indicated that coconut water group demonstrated a
significantly higher number of viable PDL cells than propolis 50% and ORS. There was no
significant difference between coconut water and propolis 50% groups.
Bazmi BA et al (2013)7 published a review article on various storage media that can be used for
avulsed tooth.
They mentioned that Hank’s balanced salt solution was the most effective medium for
preserving viability, mitogenicity and clonogenic capacities of periodontal ligament cells for
upto 24 hours at 4◦C.
They also mentioned that when milk and egg white are compared as solutions for storing
avulsed teeth, and the results have shown that teeth stored in egg white for 6 to 10 hours had a
better incidence of repair than those stored in milk for the same amount of time.
Objective of the study
1. To compare and evaluate various storage media, soy milk, coconut water, egg white,
Hank’s balanced salt solution in maintaining the periodontal ligament fibroblasts
viability.
2. To compare the efficacy of the storage media in maintenance of pdl fibroblasts vitality at
different time intervals.
MATERIALS AND METHODS
7.1 Source of Data:
Teeth which indicated for orthodontic extraction purposes are are freshly extracted collected from the
Department of Oral and Maxillofacial Surgery, Dr.Syamala Reddy Dental College Hospital and
Research Center.
MATERIALS:
SOY MILK
COCONUT WATER
EGG WHITE
HANK’S BALANCED SALT SOLUTION
TEST TUBES(100mm×13mm)
SCALPEL BLADE NO:15
DULBECCO’S MODIFIED EAGLE’S MEDIUM
0.25% wt/vol TRYPHAN BLUE
7.2 Method of collection of data(including sampling procedure, if any):
Sample size: one hundred and sixty premolar teeth that are freshly extracted for orthodontic
purposes.
Criteria for selection of samples:
Inclusion Criteria:
The permanent either maxillary or mandibular first or second premolar teeth that are freshly
extracted for orthodontic purposes are collected.
Exclusion criteria:
Teeth with dental caries,
Teeth with Fractured crown or root,
Teeth with Periapical pathology,
Teeth with Periodontal pathology.
Procedure:
The sample to be collected immediately after extraction. The teeth are rinsed with normal saline
3 times and collected in a test tube containing Dulbecco’s modified eagles medium (DMEM).
They are transferred to laboratory within 30 minutes to 2 hours. It is transported to laboratory in
test tubes containing Dulbecco’s modified eagles medium (DMEM) placed in a flask made of
thermocol containing crushed ice.
The middle one third of the root surface is scrapped with a sterile scalpel blade no. 15 and
transferred to culture plates.
The cell suspension is incubated in culture plates using Dulbecco’s modified eagles medium
(DMEM).
The experimental groups are divided into four groups:
Group 1: Soy milk – 40 samples of extracted teeth
Group 2: Coconut water – 40 samples of extracted teeth
Group 3: Egg white – 40 samples of extracted teeth
Group 4: Hank’s balanced salt solution – 40 samples of extracted teeth
After incubation, the DMEM solution is replaced with the storage media. Each experimental
group is allotted with a time period of 15min, 30 min, 45 min, 60 min, and 120 min.
After the time period, 1ml of 0.25% wt/vol trypan blue solution is added to check the viability
of cells6.
.Results: The cells that are non-viable will take up the trypan blue as their cell wall becomes
permeable and turn up into blue color. The cells that are viable will not take up the trypan blue.
The results are thus tabulated for analysis.
Statistical Analysis :
One way ANNOVA test is employed for the statistical analysis.
7.3 Does the study require any investigation or intervention to be conducted on patients or other
Humans or Animals? If so, please describe briefly..
- NOT REQUIRED-
8.
List of references:
1. Thomas T, Gopi Krishna V, kandaswamy D. Comparative evaluation of maintenance of
cell viability of an experimental transport media “coconut water” with Hank's balanced salt
solution and milk, for transportation of an avulsed tooth: An in vitro cell culture study.
J Conserv Dent. 2008 Jan-Mar; 11(1): 22–29.
2. Khademi AA, Saei S, Mohajeri MR, Mirkheshti N, Ghassami F, Torabi nia N, Alavi SA.
A New Storage Medium for an Avulsed Tooth. J Contemp Dent Pract 2008 September;
(9)6:025-032.
3. Ali SAH, Jundi SA, Mhaidat N, Awadeh L, Naffa R. Effect of coconut water concentration
on survival of bench-dried periodontal ligament cells. International Journal of Clinical
Pediatric Dentistry, January-April 2011;4(1):9-13
4. Moura CCG, Soares PBF, Reis MVDP, Neto AJF, Soares CJ. Soy Milk as a Storage
Medium to Preserve Human Fibroblast Cell Viability: An In Vitro Study. Braz Dent J (2012)
23(5): 559-563.
5. Eskandarian T, Badakhsh S, Esmaeilpour T. The effectiveness of oral rehydration solution
at various concentrations as a storage media for avulsed teeth.
Iran Endod J.2013 Winter: 8(1): 22-24.
6. Sanghavi T, Shah N, Parekh V, singbal K. Evaluation and comparison of efficacy of three
different storage media, coconut water, propolis, and oral rehydration solution, in maintaining
the viability of periodontal ligament cells.
J Conserv Dent. 2013 Jan-Feb; 16(1): 71–74.
7. Bazmi BA, Singh AK, Kar S, Mubtasum H. Storage media for avulsed tooth. Indian
Journal of Multidisciplinary dentistry Vol.3 Issue.3 May-July 2013.
of
the DR.SOMA KRISHNA RAJARAO BOGGARAPU
9.
Signature
candidate:
10.
Remarks of guide:
11.
Name and designation of:
(in block letters)
11.1 Guide:
DR. KIRAN ARADHYA.
PROF. AND HEAD OF THE DEPARTMENT
DEPARTMENT OF PEDODONTICS
PREVENTIVE DENTISTRY,
AND
DR SYAMALA REDDY DENTAL COLLEGE,
HOSPITAL
AND
RESEARCH
CENTRE,
BANGALORE-37
11.2 Signature:
11.3 Co- guide (if any):
11.4 Signature:
11.5 Head of department
DR. KIRAN ARADHYA
PROFESSOR AND HEAD , DEPARTMENT OF
PEDODONTICS
AND
PREVENTIVE
DENTISRTY,
DR SYAMALA REDDY DENTAL COLLEGE,
HOSPITAL
AND
RESEARCH
CENTRE,
BANGALORE-37
11.6 Signature
12.
12.1 Remarks of the
Chairman and Principal
12.2 Signature: