FIBROUS TISSUE FORMATION OF INTRACUTANEOUS INJECTION OF TGF-β AND L-ASCORBIC ACID TREATED SKIN FIBROBLASTS

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We investigated effects of intracutaneous injection of transforming growth factor-beta 1 (TGF-β1) and L-ascorbic acid treated-skin fibroblasts in order to develop the treatment of skin wrinkles. Human skin fibroblasts were treated with 0-50 ng/ml of TGF-β1 and/or 0-400μM L-ascorbic acid. To determine optimal concentration of TGF-β1 and L-ascorbic acid, collagen synthesis and proliferation of fibroblasts were analyzed by employing XTT assay and western blot. Maximum collagen synthesis and proliferation of fibroblasts were observed at the dose of 50μM of L-ascorbic acid. TGF-β1 increased collagen synthesis and proliferation of fibroblasts to the same extent at the range of 5-50 ng/ml. When fibroblasts were treated with both 5 ng/ml TGF-β1 and 50μM L-ascorbic acid, collagen synthesis and proliferation of fibroblasts were significantly increased, suggesting that these concentrations were suitable for the future in vivo study. Fibroblasts treated with 5 ng/ml TGF-β1 and 50μM L-ascorbic acid were injected into the subcutis of nude mice. Five days after injection small fibrous tissues were found in the subcutis of the mice. The size of fibrous tissue was larger than that of controls with none treated-fibroblasts. In the next study organ culture system of human skin was employed. The cultured human skins were injected with the fibroblasts treated with both 5 ng/ml TGF-β1 and 50μM L-ascorbic acid and were cultured. In the skins injected with the treated fibroblasts, the size of fibrous tissue was 10-fold larger than that of none treated-fibroblasts on day 4. Similar fibrous tissue formation was observed in the injected skins on day 7. Newly synthesized collagen was found in the skins injected with the treated fibroblasts. Cutaneous or subcutaneous injection of cultured fibroblasts stimulated by TGF-β1 and L-ascorbic acid may be useful for the treatment of skin wrinkles.
名古屋市立大学の論文
2003-02-28

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