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RAJIV GANDHI UNIVERSITY OF HEALTH SCIENCES,
BANGALORE, KARNATAKA
ANNEXURE II
PROFORMA FOR REGISTRATION OF SUBJECT FOR
DISSERTATION
1. NAME OF THE CANDIDATE
AND ADDRESS:
DR.CHAITANYA. R. UPPIN
DEPARTMENT OF PEDIATRIC AND
PREVENTIVE DENTISTRY,
P.M.N.M. DENTAL COLLEGE AND
HOSPITAL, BAGALKOT-587101
KARNATAKA.
2.NAME OF THE INSTITUTION:
P.M.N.M. DENTAL COLLEGE AND
HOSPITAL, BAGALKOT-587101
KARNATAKA
3.COURSE OF STUDY AND
SUBJECT:
4. DATE OF ADMISSION:
MASTER OF DENTAL SURGERY
(M.D.S.) IN PAEDODONTIA
31stMAY 2011
5.TITLE OF THE TOPIC:
“DETERMINATION OF ABO BLOOD GROUPS FROM DENTAL PULP AND
DENTIN IN DECIDUOUS TEETH”
6. BRIEF RESUME OF THE INTENDED WORK:
6.1 NEED FOR THE STUDY
Teeth are invaluable source of personal identification in the field of forensic
medicine as they are resistant to environmental assaults. Teeth can survive for long time
even after soft and skeletal tissues have been destroyed. Blood grouping from teeth could
be a source of personal identification. The use of blood group substance in medico-legal
examination is based on the fact that once a group is established in an individual it
remains unchanged throughout his/her life1. It was thought to be of interest to apply the
ABO blood grouping method to a calcified tissue such as dentin of teeth because it has
higher ratio of cell substances than bone2. Pulp tissue is protected from all sides from
dental hard tissue and also it has rich vascular supply, blood group antigens are bound to
be present in tooth pulp and therefore could be readily available for examination. Most of
the studies have used permanent tooth pulp for blood group determination1,7 but only few
studies have been undertaken to determine efficacy and reliability of blood grouping
using pulp of deciduous dentition. Hence the present study is taken up to determine blood
group antigens from pulp and dentin of deciduous tooth, which can be a valuable source
for personal identification, when permanent teeth are not available.
6.2 REVIEW OF LITERATURE:
A study was done to determine ABO blood grouping on the pulp, dentin and
enamel on 35 teeth using absorption elution technique. Twenty teeth were examined
within six weeks of extraction and 15 teeth after 6-10 months. It was found that blood
grouping on pulp gives fairly good results, whereas the possibilities for correct blood
grouping seem to be limited for dentin and debatable for enamel.3
In another study ABO blood group determination was done from dental pulp and
saliva for its use in Forensic Odontology. Hundred cases were studied using modified
Absorption Elution technique, 93 cases were positive for dental pulp and 79 were positive
for saliva.4
Thermo stability of ABO blood group antigens in human dental pulp was
described in a study conducted in the year 1978. The ability of the enamel and dentin to
protect the ABO antigens from thermal damage was estimated. The calculations shows
that dentin and enamel are poor insulators and give inadequate thermal protection to the
pulp when external temperature rises to 2000 c and more.5
A similar study was conducted at Mangalore for ABO blood group determination
from dental pulp and dentin from sixty teeth taken from dead bodies after 180 days, by
using Absorption Elution technique they were able to successfully determine the ABO
blood groups only from dental pulp but failed to determine the same from dentin.6
A comparative study was done to correlate the blood group determination by
dental pulp and dentin with that of socket blood collected from the same person. The
ABO blood group determined by dental pulp correlated well whereas the blood groups
from dentin failed to correlate.7
Another study conducted on 183 permanent and 77 deciduous extracted teeth
examined after 1-2 months by Elution technique proved that minute amount (6-10 mg) of
pulverized tooth material would be sufficient to determine the ABO blood groups.8
6.3 OBJECTIVES OF THE STUDY:
1. To determine ABO blood group from dental pulp and dentin of deciduous teeth
after storing the teeth for 6 months.
2. To determine ABO blood group from the socket blood.
3. Correlation of ABO blood group determined by dental pulp, dentin and socket
blood.
7. MATERIALS AND METHODS:
7.1 SOURCE OF THE DATA
60 children between age group 3-12 yrs coming to the Department of Pedodontics and
Preventive Dentistry, P. M. N. M. Dental College and Hospital, Bagalkot for extraction
of over retained teeth or extraction of deciduous teeth for orthodontic treatment are
randomly chosen for this study.
INCLUSION CRITERIA:
1. Over retained teeth.
2. Deciduous Teeth indicated for orthodontic extraction.
EXCLUSION CRITERIA:
1. Carious teeth.7
7.2 METHODS OF COLLECTING DATA
60 children are selected randomly coming to the Department of Pedodontics and
Preventive Dentistry, P. M. N. M. Dental College and Hospital, Bagalkot for extraction
of over retained teeth or extraction of deciduous teeth for orthodontic treatment.
MATERIALS:
1. Mouth mirror & Dental probe.
2. Anticoagulant solution EDTA
3. Fresh normal saline.
4. Antisera A and B.
5.
Gauze pieces, numbered bottles.
6. Lathe, carborundum disc, spoon excavator.
7. Straight fissure bur.
8. Centrifuge, Hot water bath, Incubator.
9. A and B Red Blood cell suspension.
10. Microscope
METHOD:
The study group shall comprise of 60 patients requiring extraction for overretained teeth or deciduous teeth for orthodontic reasons. A brief case history with
relevant medical history will be recorded and a detailed clinical examination under
artificial illumination will be done. Extraction procedure is carried under local anesthesia
following an aseptic precaution. After extraction, the socket is compressed with a sterile
gauze piece and socket blood will be collected for ABO blood group determination which
serves as a control group for the study. Anticoagulant is added to the socket blood and
one drop of this blood is placed on the slide and a drop of Antiserum-A is added, if
agglutination occurs on addition of Antiserum-A, blood group shall be recorded as A and
vice versa. The extracted teeth will be washed under running water and debris shall be
removed with the probe and wiped with gauze and kept in bottles which are numbered for
identification these teeth are stored for a span of 6 months7. The blood grouping is
performed by Absorption Elution procedure using dental pulp and dentin and later
compared with control sample7. The tooth is completely trimmed to remove the enamel
and cementum with lathe. The tooth is further split vertically with carborundum disc and
the dental pulp is scooped out with spoon excavator, the remaining tooth consisting of
dentin, shall be pulverized with straight fissure bur.
The pulverized tooth powder is put in two test tubes, to each of these test tubes
three drops of antiserum A and B shall be added and ensuring that the test samples are
being sufficiently soaked with antiserum for two and half hours and left standing at room
temperature. After removing antiserum, each sample shall be washed three times with
cold saline solution and centrifuged and the supernatant is sucked with pipette. Then two
drops of fresh saline is added to the sample and the test tube is heated in a water bath at
50-55°c for 10 minutes to elute the antibodies.
A drop of 0.5% A or B group RBC suspension is immediately put into each
respective test tube of known blood and the samples shall be put in humified recipient
and shall be incubated at 37°c for 30min to enhance agglutination and after this
procedure, it shall be centrifuged at 1,500- 2000rpm for 1 min. By gentle shaking of the
test tube the presence or absence of RBC agglutination shall be ascertained with
microscope at magnification of 100X. Actual blood groups determined from the socket
blood, dental pulp and dentin shall be correlated using appropriate statistical methods.
7.3 DOES THE STUDY REQUIRE ANY INVESTIGATION OR INTERVENTION
TO BE CONDUCTED ON PATIENTS OR OTHER HUMANS OR ANIMALS? IF
SO PLEASE BRIEFLY SPECIFY.
Yes.
7.4 HAS ETHICAL CLEARANCE BEEN OBTAINED FROM YOUR
INSTITUTION?
Yes, a copy is attached.
8. LIST OF REFERANCES:
1. Neiders ME. Blood group determinations in forensic dentistry. Dent Clin North
Am.1977; 21(1):99-111
2. Kramer Ivor RH. An examination of Dentin for A and B Bloodgroup antigens by
mixed agglutination technique. Proceedingsof Royal society of medicine. 1957;
50:677-78
3. Smeets B. ABO blood grouping on tooth material. Forensic Sci Int. 1991;
50(2):277-84.
4. Parekh, K Sansare, AG Malwankar, PG Gore. ABO blood group determination
from dental pulp and saliva for its use in forensic Odontology. Journal of Indian
Academy of Oral Medicine and Radiology 1994; 2:17-20.
5.
Anna-Krystyna Karszun. Thermo stability of ABH (O) blood antigens in human
teeth. Journal Forensic Science 1978; 11: 231-39.
6.
Dr. Mahabaleshshetty, Dr. Premlatha K. ABO blood grouping from tooth
material. Journal of Indian forensic medicine 2010; 32(4):336-338.
7. SushminiBallal, Maria Priscilla David. Determination of ABO blood grouping
from dentin and pulp. Pakistan Oral and dental journal 2011; 31( 1):3-6.
8.
Hideo Takata. Blood group identification from hard dental tissues by elution test.
International Journal of Forensic dentistry1974; 2(4):43-47.