M C McCormack1, K C Nowak, R J Koch. 1. Division of Otolaryngology-Head and Neck Surgery, Stanford University Medical Center, Stanford, Calif 94305-5328, USA.
Abstract
OBJECTIVE: To evaluate the effect of copper tripeptide and tretinoin on normal and keloid-producing dermal fibroblasts in a serum-free in vitro model. The cellular response was described in terms of viability and secretion of basic fibroblast growth factor (bFGF) and transforming growth factor-beta1 (TGF-beta1). METHODS: Primary cell lines were established from patient facial skin samples obtained during surgery and plated in serum-free media. At 0 hour, copper tripeptide (1 x 10 (-9) mol/L), tretinoin (1 x 10 (-5) mol/L), or appropriate control vehicle was added. Cell counts and viability were established at 24, 72, and 120 hours. Supernatants were collected at the same intervals and were assessed for bFGF and TGF-beta1 concentrations using the enzyme-linked immunosorbent assay technique. RESULTS: Cell lines showed viability between 86% and 96% (mean, 92%) throughout the experiment. Tretinoin-treated normal fibroblasts secreted more bFGF than did controls at 24 hours (P<.05). Tretinoin-treated keloid-producing fibroblasts secreted more TGF-beta1 than did controls at 120 hours (P<.05). Keloid-producing fibroblasts treated with copper tripeptide secreted less TGF-beta1 than did controls at 24 hours (P<.05); a similar trend was observed in normal fibroblasts. CONCLUSIONS: Normal fibroblasts treated with tretinoin produced more bFGF than did controls, and this might partially explain the clinically observed tightening effects of tretinoin. Normal and keloid-producing dermal fibroblasts treated with copper tripeptide secreted less TGF-beta1 than did controls, suggesting a possible clinical use for decreasing excessive scar formation.
OBJECTIVE: To evaluate the effect of copper tripeptide and tretinoin on normal and keloid-producing dermal fibroblasts in a serum-free in vitro model. The cellular response was described in terms of viability and secretion of basic fibroblast growth factor (bFGF) and transforming growth factor-beta1 (TGF-beta1). METHODS: Primary cell lines were established from patient facial skin samples obtained during surgery and plated in serum-free media. At 0 hour, copper tripeptide (1 x 10 (-9) mol/L), tretinoin (1 x 10 (-5) mol/L), or appropriate control vehicle was added. Cell counts and viability were established at 24, 72, and 120 hours. Supernatants were collected at the same intervals and were assessed for bFGF and TGF-beta1 concentrations using the enzyme-linked immunosorbent assay technique. RESULTS: Cell lines showed viability between 86% and 96% (mean, 92%) throughout the experiment. Tretinoin-treated normal fibroblasts secreted more bFGF than did controls at 24 hours (P<.05). Tretinoin-treated keloid-producing fibroblasts secreted more TGF-beta1 than did controls at 120 hours (P<.05). Keloid-producing fibroblasts treated with copper tripeptide secreted less TGF-beta1 than did controls at 24 hours (P<.05); a similar trend was observed in normal fibroblasts. CONCLUSIONS: Normal fibroblasts treated with tretinoin produced more bFGF than did controls, and this might partially explain the clinically observed tightening effects of tretinoin. Normal and keloid-producing dermal fibroblasts treated with copper tripeptide secreted less TGF-beta1 than did controls, suggesting a possible clinical use for decreasing excessive scar formation.