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J Bagh College Dentistry Vol. 21(1), 2009 Distribution and localization of… K S S BC BM Figure 5: Inflammatory gingival enlargement showing positive PAS reaction in keratinized epithelia (K), in granular layer (G) and in some layer of spinosum(S). Basement membrane (BM) shows positive stain too. PAS stain X 10 Figure 6: High magnifying view of inflammatory gingival enlargement showing negative PAS reaction in basal cell (BC) and Spinosum Cell (SC) while basement membrane and Lamina propria show positive reaction (arrow) PAS stain X 20 BM LP Figure 7: Inflammatory gingiva showing positive PAS stain in Basement membrane (BM) and in Lamina propria (LP). PAS stain X 20 Specimens from gingival of Phenytoin induced drug showed localization of positive PAS reaction in spinosum layer while negative stain showed in basement membrane and in connective tissue. (Figures 2-4). Histopathological slides for specimens of inflammatory gingival stained with PAS showed deep reddish purple stain in keratinized layer, granulosum layer and includes some layers of spinosum, basement membrane shows positive stain (figures 5,6). Figure 7 showed band of reddish purple stain in connective tissue underneath the epithelium. DISCUSSION The gingival tissue consist of epithelial cells, collagenous fibers, connective tissue cells, intercellular substance (ground substance) which includes glycogen, glycoprotein, proteoglycan and mucin. Capillaries, arterioles and venules as well as lymph vessels and nerves are also present. The macromolecular carbohydrate components in epithelium and connective tissue of gingival are secreted by epithelial cell and (13) fibroblast cells (respectively). The carbohydrate group includes glycogen, glycoprotein and proteoglycans detected by periodic acid Schiff (PAS) method. Changes in epithelial glycogen showed during inflammation and repair and even variation in keratinization may reflect the glycogen content of tissue (4) . Thus the present results showed an intense reaction of PAS stain in area of keratin and granular layer of inflammatory gingival which related to an increase of production of glycogen by epithelial cells basement membrane exhibit high PAS reaction which may relate to increase in fibronectin and lamanin (glycoprotein). As basement membrane separates between the epithelium and connective tissue, fibronectin secretes by fibroblast in connective tissue while lamanin secreted by epithelium cells. Both proteoglycans and glycoprotein in connective tissue undergo alteration in various pathological state, therefore during inflammation there is an increase in both glycoprotein and proteoglycan level correlated with pathological Oral Diagnosis 51
J Bagh College Dentistry Vol. 21(1), 2009 Distribution and localization of… behavior, induce fibroblast cell to secrete glycoprotein like fibronectin and proteoglycan, so the result illustrated the PAS reaction in basement membrane and in the connective tissue. (14) Results for induced Phenytoin gingival enlargement showed reddish purple stain of spinosum cell surface. This is contribute to the ability of histochemical dye (PAS) stain to bind to epithelium surface which constituents mainly glycoprotein and glycosamine glycan. In connective tissues PAS stain showed no reaction indicated of absences of increment in the ground substance in which fibroblast is responsible for synthesis it. Many studies use PAS method to investigate clinical signs related to injury and disease. Qualitative observations were corroborated by quantitative histochemistry of sections stained with PAS. In soft tissue injury showing high glycogen investigated with increased PAS stain when compared with healthy control. (15) Huang et al 1993 (16) demonstrated glycogen in tumor cell by PAS stain. Sexton and white in 1996 (17) used PAS stain for identification of glycoprotein in primary Ewings sarcoma. While Lee et al (18) 2002 used PAS stain to investigate progression of oral malignant tumor. REFERENCES 1. Trackman P, Kantarci A. Connective tissue metabolisum and gingival overgrowth. Crit Rev Oral Bio Med 2004; 15(3): 165-75. 2. Wright HJ, Chapple LC, Ptair C. Cervicular fluid level of TGFB1 in drug induced overgrowth. Arch Oral Biol 2004; 49: 421-5. 3. Pitiphate W, Kerbs A. Gingivitis may increase protein level in gingival. Periodontol 2006; 86: 1450-8. 4. Nurmennem PK, Pernu HE, Knuuttla LE. Mitotic activity of keratinocytes in nifedipine and immunosuppressive medication induced gingival overgrowth. J Periodontol 2001; 72: 167-73. 5. Lzakovie J, buchner Sa, Lavioe MC. Phenytoin induce proliferation n human gingival fibroblast. J Periodontol 2003; 74(11): 1625-33. 6. Hasegawa A, gotah S, Miyatak S. Mechanisum of human gingival overgrowth by phenytoin. J Periodontol 2004; 14:10-1. 7. Weinberg jk, Torone WD. Oral tissue chemical component. Arch Oral Biol 2007; 3(5): 12. 8. Laii RQ. Histopathological study of oral tissue. Chung Hu Ping li-Hsueh-Tsa-Chih 1993; 22(2): 92- 4. 9. Das SJ, Newman H, Olsen L. Up regulation of keratinocyte growth factor in gingival overgrowth. J Periodontol 2003;74:506-11. 10. Carranza FA, Newman mg, Takei HH. Clinical periodontology. 9 th edition WP Saunders Company 2002 281-95. 11. Seymour RA, Ellis JS, Thomasson JM. The effect of phenytoin on periodontal health of adult patients. J Clin Periodontol 1985; 12: 413-9. 12. Zugibe FT. Diagnostic histochemistry. 3 rd edition C.V.Mosby Company.1979. 277-8. 13. Scroggos M, roggli V, Fraire A. Eosinophilic intracytoplasmic globules in pulmonary adenocarcinoma; a histochemical, immunohistochemical study. Hum Pathol 1989; 20(9):845-9. 14. Morrison C, Prokoryn P, Piquero C. Oral tissue changes associated with squamous intra epithelial lesion. 15. Quiroz Rothe E, Novales M. Polysaccharide storage with back pain. Equine Vet J 2002; 34(2): 171-6. 16. Huang SH, Ko WS, Jen Sc. Intrathoracic extraskeletal Ewings sarcoma.Chung Hua Hsueh tsa Chih 1993;51(6):474-8. 17. Sexton C, White W. Primary Ewings sarcoma. Am J Dermatopathol 1996; 18(6): 601-5. 18. Lee Y, Nagai N, Siar C. Angioarchitecture of primary oral malignant.J Histochem Cytochem 2002; 50(11): 1555-62. Oral Diagnosis 52
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