Rajiv Gandhi University of Health Sciences s310
RAJIV GANDHI UNIVERSITY OF HEALTH SCIENCES,
BANGALORE, KARNATAKA
ANNEXURE – II
PROFORMA FOR REGISTRATION OF SUBJECT FOR
DISSERTATION
1. /NAME OF THE CANDIDATE
AND ADDRESS(in block letters) / Dr. V.VENKATA RAMYA
DEPARTMENT OF ORAL AND MAXILLOFACIAL PATHOLOGY,
COLLEGE OF DENTAL SCIENCES,
DAVANGERE-577004
2. /
NAME OF THE INSTITUTION
/ COLLEGE OF DENTAL SCIENCES,DAVANGERE – 577004.
3. / COURSE OF THE STUDY AND SUBJECT / MASTER OF DENTAL SURGERY
ORAL AND MAXILLOFACIAL PATHOLOGY AND MICROBIOLOGY
4. / DATE OF ADMISSION TO COURSE / 07-06-2013
5. /
TITLE OF THE TOPIC
/ “STUDY OF COLLAGEN FIBERS IN LAMINA PROPRIA OF ORAL LICHEN PLANUS, CARCINOMA-IN-SITU AND EARLY INVASIVE SQUAMOUS CELL CARCINOMA USING PICRO SIRIUS RED STAIN.”6
7. /
BRIEF RESUME OF THE INTENDED WORK:
6.1 Need for study:
Lichen planus is a chronic mucocutaneous disease of autoimmune etiology commonly affecting the oral cavity besides the skin, genital mucosa, scalp and nails. It often presents as bilaterally symmetrical lesion most commonly involving the buccal mucosa followed by tongue, lips, gingiva and floor of the mouth.
Histopathologically lichen planus is diagnosed by liquefactive degeneration of basal cell layer and sub-epithelial band of inflammatory infiltrate predominantly comprised of lymphocytes.Although World Health Organization has classified oral lichen planus (OLP) as a Potentially Malignant Disorder there are still controversies regarding its malignant potential.1
It has been proved that the biologic behavior of many oral diseases like odontogenic cysts and tumors is dependent not only on the epithelium but also on the stroma supporting the epithelium of which collagen forms the major component.2
Numerous studies on the changes in collagen type IV & type VII at the basement membrane zone in lichen planus have been reported in the literature.3,4 However studies on the collagen fibers in the lamina propria of oral lichen planus are scanty.
Thus, the present study aims to evaluate the nature of collagen fibers in lamina propria of oral lichen planus using picrosirius red stain (PSR) and polarized microscope in comparison with buccal mucosa without any pathology related to exposure to tobacco and other oral carcinogens, carcinoma-in-situ (Ca in-situ) and early invasive squamous cell carcinoma (EISCC).
6.2 Review of literature:
Extracellular matrix is composed of collagenous proteins, non-collagenous proteins and proteoglycans. These proteins and molecules of extra cellular matrix influence the tumor behavior. Thus the stromal reaction also plays a key role in malignant transformation of potentially malignant disorders. 5
Collagen can be studied under various stains like Mallory, Masson Trichrome and Van Gieson but these stains fail to reveal thin collagen fibers. Collagen has a natural birefringence property which is enhanced by picrosirius red stain thus enabling differentiation of thin and thick fibers.6
The use of polarization microscopy to study collagen in PSR stained sections enhances the visibility of thin collagen fibers. Usually polarization colors of thin normal collagen fibers are green to greenish-yellow, where as thick fibers range from yellowish to red birefringence.7
Various studies on collagen using PSR stain to evaluate biologic behavior of lesions like odontogenic cysts and tumors, salivary gland tumors, various grades of epithelial dysplasia and squamous cell carcinoma have been reported in the literature. 2,8-10
Study of collagen fibers in various grades of squamous cell carcinoma showed the change from reddish orange to greenish yellow fibers from well differentiated to poorly differentiated squamous cell carcinoma, thus indicating that as the tumor progressed there is a change from the mature form of collagen to an immature form.10
6.3 Objectives of the study:
1. To determine the polarization colors of the collagen fibers in lamina propria of oral lichen planus, buccal mucosa without any pathology related to exposure to tobacco and other oral carcinogens, carcinoma-in-situ and early invasive squamous cell carcinoma using picrosirius red stain.
2. To compare the polarization colors of the collagen fibers in oral lichen planus, buccal mucosa without any pathology related to exposure to tobacco and other oral carcinogens, carcinoma-in-situ and early invasive squamous cell carcinoma.
MATERIALS AND METHODS
7.1 Source of data:
Paraffin embedded tissue blocks of OLP, Ca-in-situ and EISCC will be retrieved from the archives of Department of Oral Pathology and Microbiology, College of Dental Sciences, Davangere. For comparison, buccal mucosa without any pathology related to exposure to tobacco and other oral carcinogens will be included.7.2 Study methodology:
The study groups will be as follows
Study groups / Number of Cases
Group 1: Control Group / 20
Group 2: Oral lichen planus / 20
Group 3: Carcinoma-in-Situ / 20
Group 4: Early invasive squamous cell carcinoma / 20
Total number of samples / 80
Method of collection of data:
A minimum of 20 samples in each study group will be included in the study.
2 sections of 4-6µ thickness will be obtained from the archival paraffin embedded tissue blocks of each case. One section will be stained by Haematoxylin & Eosin for confirming the diagnosis. Other section will be stained by picrosirius red stain and observed under polarizing microscope for evaluating polarization colors of collagen.
The PSR stained sections will be observed under the polarizing microscope. The images will be captured using 3 chip CCD camera attached to Trinocular Research Microscope and all the images will be stored onto a computer. The thickness of fibers will be determined using proplus image analysis software and all measurements will be exported to microsoft excel sheet for further interpretation and statistical analysis.
Statistical analysis:
Descriptive summary statistics will be given for all the variables. Difference in
proportions will be tested using Chi-Square test and Kruskal- Wallis H test followed by
Mann-Whitney U test for intergroup comparison. Significance level will be set at
p< 0.05.
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, study will be conducted using paraffin embedded tissue blocks.7.4 Has ethical clearance been obtained from your institution in case of 7.3?
Submitted for institutional ethical clearance
LIST OF REFERENCES
1. Krutchkoff DJ, Cutler L, Laskowski S. Oral lichen planus: The evidence regarding potential malignant transformation. J Oral Pathol 1978;7(1):1-7.2. Vaidehi NN, Mandana D, Praveen SB, Selvamani M. Comparative Evaluation of Collagen fibers in Odontogenic Keratocyst and Unicystic Ameloblastoma. WJD 2011;2(2):125-7.
3. Jungell P. Immunoelectron microscopic study of the basement membrane in
oral lichen planus. J Cutan Pathol 1990;17(2):72-6.
4. Haapalainen T, Oksala O, Kallioinen M, Oikarinen A, Larjava H, Salo T.
Destruction of the epithelial anchoring system in lichen planus. J Invest
Dermatol 1995;105(1):100-3.
5. Shinohara M, Nakamur S, Harada T, Shimada M, Oka M. Mode of tumor invasion in oral squamous cell carcinoma: Improved grading based on immunohistochemical examination of extracellular matrices. Head Neck J Sci Spec 1996;18(2):153-9.
6. Parveen S, Ahmed SA, Tanveer SA. A Study of Orientation of Collagen
Fibers in Oral Submucous Fibrosis.IJSRP (Internet). 2013 Mar;3(3).
Available from:http://www.ijsrp.org/e-journal.html.
7. Rabau MY Dayan. Polarization microscopy of picrosirius red stained sections:
A useful method for qualitative evaluation of intestinal wall collagen. Histol
Histopath 1994;9:525-8.
8. Allon I, Vered M, Buchner A, Dayan D. Stromal differences in salivary gland
tumors of a common histopathogenesis but with different biological behavior:
a study with picrosirius red and polarizing microscopy. Acta Histochem
2006;108:259-64.
9. Yokoyama Megumi. Alterations in stromal reaction during tumor progression
in oral mucosa. J Hard Tissue Biology 2011;20(1):23-30.
10. Aparna V, Charu S. Evaluation of collagen in different grades of oral squamous cell carcinoma by using the picrosirius red stain- A histochemical study. J Clinc Diagn Res 2010;4:3444-49.
9. / SIGNATURE OF CANDIDATE
10. / REMARKS OF THE GUIDE / .
Study will be useful
11. / NAME AND DESIGNATION OF
(in block letters)
11.1 GUIDE
11.2 SIGNATURE
11.3 CO-GUIDE (if any)
11.4 SIGNATURE
11.5 HEAD OF THE
DEPARTMENT
11.6 SIGNATURE / Dr. NANDINI DB
PROFESSOR
DEPARTMENT OF ORAL AND MAXILLOFACIAL PATHOLOGY
AND MICROBIOLOGY,
COLLEGE OF DENTAL SCIENCES,
DAVANGERE - 577 004, KARNATAKA
Dr. MADHUSHANKARI G S
PROFESSOR AND HEAD,
DEPARTMENT OF ORAL AND MAXILLOFACIAL PATHOLOGY AND MICROBIOLOGY,
COLLEGE OF DENTAL SCIENCES,
DAVANGERE - 577 004, KARNATAKA
12. / 12.1 REMARKS OF THE
CHAIRMAN AND
PRINCIPAL
12.2 SIGNATURE