褪黑凝膠於傷口修復及美白之應用

Abstract

本研究欲發展一種整合各種能在黑色素生成及沈澱的不同階段進行抑制的試劑及傷口修復試劑的褪黑修復凝膠(APH gel)，並從傷口照護的早期至後期階段來預防及治療傷口發炎反應所引起的發炎後過分黑色素沈澱（PIH）。首先，我們利用B16F10黑色素瘤細胞與人類黑色素細胞-角質細胞共培養系統測試了一些鎖定的美白成分，包含幾丁聚醣的衍生物、凱索尼酸(CA)、N-乙醯基葡萄糖胺(NAG)、熊果素、火龍果莖(DFS)萃取物和菸鹼酸(NCA)。在一定的濃度範圍內，它們皆能有效的抑制細胞黑色素生成或是抑制黑色素小體的傳遞。接著，我們進一步測試各成分的最佳濃度之共同效應。結果顯示，N-乙醯基葡萄糖胺、熊果素、火龍果莖萃取物三者相加的抑制黑色素程度最強烈，且並不會降低細胞存活率。另一方面，凱索尼酸與火龍果莖萃取物相加可能會產生分子間作用力而導致兩者抑制黑色素能力皆降低。然而，在共培養的實驗中，N-乙醯基葡萄糖胺、熊果素、菸鹼酸不加火龍果莖萃取物的抑制黑色素沈澱效果反而最佳，因此建議在四種成分之共同效應中，火龍果莖萃取物的濃度可能還要再降低。此外，我們也測試了L-抗壞血酸-2-磷酸(AA2P)於誘導人類皮膚纖維母細胞生成膠原蛋白的能力，以達到幫助傷口修復之目的。最後，我們提出了兩種最佳的組合成分於明膠膠中並應用於傷口修復。儘管我們建立的四種由傷口引發的發炎後過分黑色素沈澱之天竺鼠動物模型尚未完美，但因為我們在此研究中對各傷口種類及性質有了更深的了解，未來要找到一個可行的動物模型來驗證我們的APH gel之發明是可行的。

This study aimed to preclude and treat injury-induced post inflammatory hyperpigmentation (PIH) from the early to the late stage of wound care by incorporating depigmenting agents responsible for each inhibition states and wound healing agent in our anti-pigmentation healing gel (APH-gel). First, we examined some targeted lightening agents, including chitosan derivatives, chitosonic acid (CA), N-acetyl-D-glucosamine (NAG), arbutin, dragon fruit stem (DFS) extract, and nicotinamide (NCA) by B16F10 melanoma cells and human melanocyte-keratinocyte co-culture system. They all inhibited melanogenesis or melanosome transfer of cells under certain concentrations. Next, we tested their combined effects under their own best dosages. The results showed that the combination of NAG, arbutin, and DFS exhibited the strongest inhibitory effect on melanogenesis without diminishing cell viabilities. On the other hand, the combination of CA and DFS may undergo intermolecular interactions that hinder both of their efficacies. In co-culture experiments, however, the combination of NAG, arbutin, NCA was better without DFS for anti-pigmentation, thus a lower dosage of DFS was suggested. We also determined the best concentration of ascorbic acid-2-phosphate (AA2P) on collagen induction of human skin fibroblasts for healing purpose. Finally, we hypothesized two best gelatin gel formulas for wound healing applications. Although the four wound-induced PIH guinea pig models we established were still not perfect, finding a feasible animal model for future in vivo testing of our invention is possible with our new insights to different wound types and properties.