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Exp Clin Endocrinol Diabetes 2002; 110(4): 176-181
DOI: 10.1055/s-2002-32149
Articles

© Johann Ambrosius Barth

Effects of basic fibroblast growth factor (FGF-2) on proliferation of human skin fibroblasts in type II diabetes mellitus[*]

A. T. Grazul-Bilska 1, 2 , G. Luthra 3 , L. P. Reynolds 1, 2 , J. J. Bilski 2 , M. L. Johnson 1 , S. A. Adbullah 4 , D. A. Redmer 1 , K. M. Abdullah 3, 4
  • 1 Department of Animal and Range Sciences, North Dakota State University, Fargo
  • 2 Cell Biology Center, North Dakota State University, Fargo
  • 3 Department of Surgery, School of Medicine, University of North Dakota, Grand Forks
  • 4 Plastic Surgery Institute, Fargo, ND, USA
Further Information

Publication History

received 30 August 2001 first decision 21 November 2001

accepted 04 March 2002

Publication Date:
10 June 2002 (online)

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Summary

Skin fibroblasts from patients with diabetes mellitus display abnormalities in cell proliferation. The use of exogenous growth factors on diabetic wounds has been found to stimulate fibroblast proliferation and facilitate wound healing. However, the results of application of FGF-2 alone to diabetic wounds in clinical trials have been disappointing. The objective of this experiment was to study the effects of FGF-2 and media supplements on in vitro proliferation of skin fibroblasts from patients with type II diabetes and nondiabetic controls, and to evaluate the association between fibroblast proliferation and cAMP production. Fibroblast cell lines (n = 5 from diabetic and n = 5 from control individuals) were cultured in DMEM + 20% FBS for 7 days. Cells were then counted, plated into 24-well plates at a concentration of 2 × 104 cells/well and incubated for 24 h in DMEM with serum. The next day, medium was changed to serum-free DMEM alone or DMEM with supplements (albumin, transferrin, insulin and hydrocortisone). Cells were cultured in the presence or absence of varying doses of FGF-2 (0, 0.3, 1, 3, 10 and 30 ng/ml) for 72 hrs then counted and medium was collected for cAMP radioimmunoassay. The doubling time for cell number tended to be greater (p < 0.2) for diabetic fibroblasts than for control fibroblasts. The addition of supplements to the medium reduced (p < 0.05) the doubling time for both fibroblast types. FGF-2 stimulated (p < 0.05) proliferation of diabetic fibroblasts only in medium containing supplements. In contrast, FGF-2 stimulated proliferation of control fibroblasts in medium with or without supplements. The maximal effects of FGF-2 on fibroblast proliferation were greater (p < 0.02) in medium with supplements than in medium without supplements. The KD of FGF-2 for fibroblast proliferation was greater (p < 0.06) for diabetic than for control fibroblasts, and lower (p < 0.02) for medium with supplements than for medium without supplements. Fibroblasts from patients with diabetes mellitus produced more (p < 0.05) cAMP than control fibroblasts. These results demonstrate that FGF-2 requires the presence of supplements to enhance proliferation of fibroblasts from patients with type II diabetes mellitus. In addition, fibroblasts from diabetic patients showed a greater KD for FGF-2 in terms of cell proliferation. These data suggest a defective FGF receptor or down-regulation of the FGF receptor-mediated cascade that leads to cell proliferation. Identifying methods of reducing the KD of FGF-2 in stimulating the proliferation of diabetic fibroblasts may improve the clinical response of diabetic wounds to FGF-2.

Key words:

Diabetes mellitus - Fibroblasts - FGF-2 - cAMP - Wound healing

1 Supported by grants No. 1813-0496-2090 and No. 6371-8183-2661 from University of North Dakota, Grand Forks, ND to K.M.A. and A.T.G.-B.

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1 Supported by grants No. 1813-0496-2090 and No. 6371-8183-2661 from University of North Dakota, Grand Forks, ND to K.M.A. and A.T.G.-B.

A. T. Grazul-Bilska

Department of Animal and Range Sciences

Cell Biology Center

North Dakota State University

Fargo, ND, USA

Phone: + 701-231-7992

Fax: + 701-231-5090

Email: Anna.Grazul-Bilska@ndsu.nodak.edu