Artikel
VEGF increases human tenocyte growth and enhances tenocyte specific gene expression
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Veröffentlicht: | 3. September 2014 |
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Gliederung
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Purpose: It is not clear how exogenous vascular endothelial growth factor (VEGF) affects tendon regeneration. As VEGF is known to stimulate angiogenesis and to be a key factor of healing processes, its effects on in vitro tenocyte growth and alteration in gene expression was evalutated.
Methods: Tenocytes were obtained from cultures of human tendons. Cells were cultured for 7 days with varying VEGF-concentrations (0, 5ng/ml, 10ng/ml and 20ng/ml) with n=5. Cell growth was analysed under light microscopy using Neubauer counting chamber. Tenocyte gene expression was evaluated by realtime-‘polymerase chain reaction’ (rtPCR) for Collagen-1, Collagen-3, Tenascin C, Elastin and Scleraxis.
Results: Cell counting revealed 1,2x105/ml, 3,7x105/ml, 2x105/ml and 3 x105/ml for 0, 5ng/ml, 10ng/ml and 20ng/ml VEGF respectively (p=0.03 for VEGF 5ng/ml). Gene expression of Collagen-1 was 70x upregulated under 10ng/ml VEGF, there was no upregulation for Collagen-3 in either group. Gene expression of Tenascin-C was not upregulated in any of the experimental groups. Scleraxis was significantly upregulated in all experimental groups (40x under 0,5ng/ml, 50x under 10ng/ml and 16x under 20ng/ml, p=0,02; p=0,01; p=0,03 respectively). Elastin was upregulated 23x under 10ng/ml and 94x 20ng/ml (p=0,01; p=0,01 respectively).
Conclusions: VEGF could be beneficial for in vitro cultivation of human tenocytes as it enhances cell growth and increases tenocyte gene expression. Effects were most pronounced for 5ng/ml and 10ng/ml. Further experiments aim at confirming these data on a protein level.
Key words: human tendon healing, vascular endothelial growth factor, gene expression, rtPCR