Berna YILMAZ, Biray Çığır AVCI, CUMHUR GÜNDÜZ

Effect of heparin on human skin fibroblast proliferation in vitro

Standart heparinin, kültüre edilmiş insan deri fibroblastlarının çoğalması üzerindeki doza bağlı etkisi, insan deri biyopsi materyalinin kullanımı amacıyla oluşturulan hücre kültürlerinde araştırılmıştır. Standart heparinin yüksek konsantrasyonlarının [1/2 - 1/64, (2500-78 IU/ml)] sitotoksik olduğu belirlenmiştir. Düşük konsantrasyonlarda [1/128 - 1/1024, (39-5 IU/ml)] ise heparin, hücrelerin çoğalma yolunda bir etki göstermiştir. Heparin konsantrasyonu ve kültüre edilmiş insan deri fibroblastlari arasındaki ilişki %90 olarak bulunmuştur ve bu sonuç istatistiksel olarak oldukça önemlidir( p

Anahtar Kelimeler:

Fibroblastlar, Hücre kültür teknikleri, Deri, Heparin

İn vitroda heparinin insan deri fibroblastlarının çoğalması üzerindeki etkisi

The dose-related effects of standard heparin on the proliferation of cultured human skin fibroblasts were investigated in the cell cultures which were established using human skin biopsy material. It was determined that the high concentrations [1/2 -1/64, (2500-78 IU/ml)] of standard heparin was cytotoxic. In low concentrations [1/128-1/1024, (39-5 IU/ml)], heparin showed an effect in proliferation way. The correlation between heparin concentration and the proliferation of cultured human skin fibroblasts was found to be 90 % and this result is statistically significant (p<0.001, r=0.9498). These results are discussed with the concerned literatures.

Keywords:

Fibroblasts, Cell Culture Techniques, Skin, Heparin,

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1. Denk PO, Knorr M. Effect of heparin on human corneal fibroblast proliferation in vitro with and without growth factor stimulation. Graefe's Arch Clin Exp Ophthalmol, 1999; 237: 342-347.
2. Garg HG, Thompson BT, Hales AC. Structural determinants of antiproliferative activity of heparin on pulmonary artery smooth muscle cells. Am J Physiol Lung Cell Mol Physiol, 2000;279: 779-789.
3. Del Vecchio PJ, Bizios R, Holleran LA, Judge TK, Pinto GL. Inhibition of human scleral fibroblast proliferation with heparin. Inverst Ophthalmol Vis Sci, 1988; 29:1272-1276.
4. Imaizumi-T, Jean- Louis-F, Dubertret-ML, Dubertret-L. Heparin induces fibroblast proliferation, cell matrix interaction and epidermal growth inhibition. Exp Dermatol,1996; 5: 89-95.
5. Johnson PR, Armour CL, Carey D, Black JL. Heparin and PGE2 inhibit DNA synthesis in human airway smooth muscle cells in culture. Am J Physiol, 1995; 269:514-519.
6. Tazawa S, Hayakawa Y, Ishikawa T, Niiya K, Sakuragawa N. Heparin stimulates the proliferation of bovine aortic endothelial cells probably through activation of endogenous basic fibroblast growth factor. Thromb Res, 1993; 72: 431-9.
7. Wong J, Wang N, Miller JW, Schumn JS. Modulation of human fibroblast activity by selected angiogenesis inhibitors. Exp Eye Res, 1994;58:439-51.
8. http://www.fibroblast.com/
9. Wright TC, Castellot JJ, Diamond JR, Karnovsky MJ. Regulation of cellular proliferation by heparin and heparan sulfate. Lane DA, Lindhal U ed. Heparin. Arnold, London, 1989: 295-316.
10. Yamashita Y, Nakagomi K, Takeda T, Hasegawa S, Mitsui Y. Effect of heparin on pulmanary fibrobiasts and vascular cells. Thorax, 1992; 47: 634-9.
11. Castellot JJ, Cochran D, Karnovsky M. Effects of heparin on vascular smooth muscle cells. I.Cell metabolisms. J Cell Physiol,1985; 124:21-28.
12.Letourneur.D, Caleb BL, Castellot JJ. Heparin Binding, Internalization and metabolism in vascular smooth muscle cells: I.Upregulation of heparin binding correlaties with antiproliferative activity. J Cell Physiol, 1995; 163: 61-79.
13. Heroult M, Bernard-Pierrot I, Delbe J, Hamma-Kourbaii Y, Katsoris P, Barritault D, Papadimitriou E, Plouet J, Courty J.Heparin affin regulatory peptide binds to vascular endothelial growth factor (VEGF) and inhibits VEGF-induced angiogenesis. Oncogene, 2004;23:1745-53.
14. Garg HG, Cindhuchao N, Quinn DA, Hales CA, Thanawiroon C, Capila I, Linhardt RJ.Heparin oligosaccharide sequence and size essential for inhibition of pulmonary artery smooth muscle cell proliferation.Carbohydr Res, 2002;337:2359-64.
15. Yua L, Quinna DA, Garga HG.Hales CA.Gene expression of cyclin-dependent kinase inhibitors and effect of heparin on their expression in mice with hypoxia-induced pulmonary hypertension. Biochem Biophys Res Commun, 2006;34:1565-72
16. Gosh TK, Eis PS, Mullaney JM, Ebert CL, Gill DL. Competitive, reversible and potent antagonism of Inositol 1,4,5-triphosphate-activated calcium release by heparin. J.Biol Chem, 1988; 263:11075-11079
17. Smith CD, Wen D, Mooberry SL, Chang KJ. Inhibiton of phosphatidyl-inositol-4-phosphate by heparin. Biochem J. ,1992; 109: 441-448.
18. Pukac LA, Castellot JJ, Wright TC, Caleb BL, Karnovsky MJ. Heparin inhibits c-fos and c-myc mRNA expression in vascular smooth muscle cells. J Biol Chem, 1992; 267: 3707-3711.
19. Reilly CF, Kindy MS, Brown KE, Rosenberg RD. Heparin prevents vascular smooth muscle progression through the G1 phase of the cell cycle. J Biol Chem, 1989; 264: 6990-6995.
20. Wright TC, Pukac LA, Castellot JJ, Diamond JR, Karnovsky MJ, Levine RA, Kim-Park HY, Campisi J. Heparin supresses the induction of c-fos and c-myc mRNA in murine fibrobiasts by selective inhibition of a protein kinase c-dependent pathway, 1994; 232: 355-360.