Karnataka. Annexure II proforma for registration of subjects for dissertation name of the candidate and address



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RAJIV GANDHI UNIVERSITY OF HEALTH SCIENCES

BANGALORE, KARNATAKA.
ANNEXURE II
PROFORMA FOR REGISTRATION OF SUBJECTS FOR

DISSERTATION

1. NAME OF THE CANDIDATE AND ADDRESS:

DR. GEORGY JAMES.

P.M.N.M.DENTAL COLLEGE & HOSPITAL

BAGALKOT-587101

KARNATAKA.


2. NAME OF THE INSTITUTION:

P.M.N.M.DENTAL COLLEGE & HOSPITAL


BAGALKOT-587101
KARNATAKA.


3. COURSE OF STUDY AND SUBJECT:

M.D.S (Master of Dental Surgery)

CONSERVATIVE DENTISTRY AND ENDODONTICS


4. DATE OF ADMISSION TO COURSE:

31st MAY 2010.




5. TITLE OF TOPIC:
Comparative evaluation of maintenance of cell viability of two experimental transport mediums “oral rehydration solution” & “coconut water” with Hank’s balanced salt solution and milk for transportation of an avulsed tooth.
6. BRIEF RESUME OF THE INTENTED WORK

6.1 NEED FOR THE STUDY:

Avulsion is a complex injury and accounts for approximately 16 % of all traumatic injuries in permanent dentition. After avulsion, the number of viable cells on the root surface decreases with increased drying time and may fail to demonstrate cell viability. Therefore the tooth should be stored in a medium that maintains periodontal ligament cell viability until definitive dental treatment can be accomplished. Several interim transport media for avulsed teeth have been investigated with respect to their capacity to preserve the vitality of PDL components & dental pulp tissue. The ideal storage medium should be able to preserve cell vitality, readily available/easily accessible & economical too. The biologically pure & sterile tender coconut water is rich in electrolytes. Moreover this natural isotonic fluid has become increasingly more available. The osmolality & composition of oral rehydration solution are almost similar to coconut water. Hence the purpose of this study is to evaluate the efficacy of oral rehydration solution & coconut water, in comparison with other traditional storage media like HBSS and milk




6.2 REVIEW OF LITERATURE:

In a study the viability of human PDL cells was determined when they were exposed for 1 hour to culture medium, milk, Hanks balanced salt solution, soft wear Opti free, and Solo care contact lens solutions, Gatorade and tap water at room temperature and on ice. Results indicated that a significantly higher number of cells survived and proliferated when exposed at 00 C. Culture medium and HBSS preserved significantly more cells than the other experimental solutions3.

In an in vitro study PDL cells were cultured from extracted human teeth and incubated over timed intervals of 15 to 210 minutes in oral rehydration fluid Gatorade and milks of varying fat content and saliva. Results using Gatorade indicated it to be not suitable as a temporary storage media for avulsed teeth. Whereas milk was indicated as an appropriate media for storage of avulsed teeth2

In an in vitro study PDL cells were obtained from extracted third molars and premolars. These cells were placed in culture plates containing milk, save –a- tooth, save a tooth supplemented with platelet derived growth factor (PDGF) and Gatorade. Results indicated that milk and save a tooth with PDGF are suitable for transport medium of avulsed teeth1.

In an in vitro study 70 freshly extracted human teeth were divided into 4 experimental groups to investigate the potential of a new storage medium , coconut water, in comparison with Propolis, Milk, Hanks balanced salt solution(HBSS), in maintaining viable periodontal ligament cells. Analysis showed that coconut water kept significantly more cells viable compared with Propolis, HBSS or milk4

In an in vitro study 55 freshly extracted human teeth were divided into 3 experimental groups to investigate the potential of a new storage medium, coconut water, in comparison with milk, Hanks balanced salt solution(HBSS), in maintaining viable periodontal ligament cells using collagenase assay. Analysis showed that coconut water may be better alternative to HBSS or milk in terms of maintaining PDL cell viability after avulsion and storage5.


6.3. OBJECTIVE OF THE STUDY:

The aim of this in vitro study is to evaluate the efficacy of oral rehydration solution & coconut water, in comparison with other traditional storage media like HBSS and milk in maintaining the viability of PDL cells


7. MATERIALS AND METHODS:


7.1 SOURCE OF DATA:

Human teeth with closed apices that are extracted for orthodontic purpose will be collected from the Department of Oral and Maxillofacial Surgery, P.M.N.M. Dental College, Bagalkot.


MATERIALS USED IN THE STUDY:


  • Collagenase II

  • Dispase II

  • Fetal bovine serum

  • Coconut water

  • Hanks balanced salt solution.

  • Milk

  • Oral rehydration solution

  • Trypan blue

ARMAMENTARIUM:

  • 15ml falcon tubes

  • Centrifugation machine

  • Micropipettes

  • Neubauer chamber

  • Light microscope with haemocytometer


7.2 METHODS OF COLLECTION OF DATA
SAMPLE SIZE:

Fifty teeth that are extracted for orthodontic purposes.



INCLUSION CRITERIA:

Human permanent teeth with closed apices will be collected. The age of patients should be within 35 years.


EXCLUSION CRITERIA:

Teeth extracted from patients with moderate to severe periodontitis or extensive caries will be excluded.




METHOD OF STORAGE OF TEETH:

Extractions will be performed as atraumatically as possible. Following extractions the coronal 3mm of the PDL will be scraped with a curette to remove the cells that may have been damaged. The teeth in the experimental group will be air dried for 30 minutes followed by a 45 minute immersion in one of the following five groups.




Group I

:

Coconut water

10 freshly extracted teeth will be stored in 10 ml of coconut water.

Group II

:

Milk

10 freshly extracted teeth will be stored in 10 ml of milk

Group III

:

Hanks balanced salt solution

10 freshly extracted teeth will be stored in 10 ml of HBSS

Group IV

:

Oral rehydrating solution

10 freshly extracted teeth will be stored in 10ml of oral rehydrating solution













Group V

:

Positive control

5 freshly extracted teeth will be immediately treated with Collagenase and Dispase

Group VI

:

Negative control

5 freshly extracted teeth will be bench dried for 8 hours with no follow up storage time and then placed in Collagenase and Dispase.

METHOD OF STUDY:

Each experimental tooth after drying and soaking will be incubated for 30 min in falcon tubes with 2.5ml solution of 0.2 mg/ml of collagenase II and 2.4 mg/ml solution of dispase grade II in phosphate buffered saline. After incubation, 50 μl of fetal bovine serum will be added to each tube. All the tubes will be centrifuged for 4 minutes. The supernatant will be removed with sterile micropipettes and the cells will be labeled with dye for determination of viability of periodontal ligament cells. The viable periodontal ligament cells will be counted under light microscope with 20X magnification.


7.3 DOES THE STUDY REQUIRE ANY INVESTIGATIONS OR INTERVENTIONS TO BE CONDUCTED ON PATIENTS OR OTHER HUMANS OR ANIMALS? IF SO PLEASE DESCRIBE BRIEFLY.

No.

7.4 HAS ETHICAL CLEARANCE BEEN OBTAINED FROM YOUR INSTITUTION IN CASE OF 7.3?

Nil.

8. LIST OF REFERENCES:


  1. Brian D. Olson, Jason M. Mailhot. Comparison of various transport media on human periodontal ligament cell viability. J Endod 1997; 23: 676-679.

  2. Orset M. Hakacz Sr, David L. Carnes Jr. Determination of periodontal ligament cell viability in the oral rehydration fluid Gatorade and milks of varying fat content. J Endod 1997; 23: 687-690.

  3. Sigalas E, Regan J.D. survival of human periodontal ligament cells in media proposed for transport of avulsed teeth. Dent Traumatol 2004; 20: 21-28.

  4. Velayutham Gopikrishna, Parvinder Singh Baweja. Comparison of coconut water, propolis, HBSS, Milk on PDL cell survival. J Endod 2008; 34(5): 587-589.

  5. Velayutham Gopikrishna, Toby Thomas. A quantitative analysis of coconut water: a new storage media for avulsed teeth. Oral Surg Oral Med Oral Pathol Oral Radiol Endod 2008;105:e61-e65





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