豬皮基質及聚胺基甲酸酯複合膜於人工皮膚之研究

Abstract

本研究為設計理想之創傷披覆材料以及人工皮膚以應用於燒燙傷時之需要。在創傷披覆材料的設計上，由於聚胺基甲酸酯應用於傷口覆蓋已有很長之時間，且為生物相容性佳之產品，所以用聚胺基甲酸酯（polyurethane）為此研究之薄膜以控制水氣穿透、體液排出。由於PU薄膜若製作非常薄時，本身容易沾黏，因此於PU薄膜上黏合不織布，以支撐PU薄膜，使得取用容易，簡稱PU/Nw複合膜。另外於PU薄膜內加入抗菌劑silver sulfadiazine，以抑制傷口之感染。本實驗之複合膜有三種，分別為PU200/Nw、PU300/Nw、PU400/Nw（200、300、400為製膜時模具的厚度，單位為μm）。PU200/Nw複合膜其平均水氣穿透率可達到3114 g/m2/day， PU300/Nw為2243 g/m2/day，PU400/Nw為1743 g/m2/day。這三種複合膜可控制不同的水氣穿透率以因應各種狀況。在機械性質方面，加入不織布提高了PU膜的支撐力，因此取用較方便。另外本實驗之複合膜在藥物釋放速率，含50μg/cm2之PU200/Nw複合膜在8小時與16小時有隨時間釋放，而在16小時至24小時漸趨平緩，為藥物釋放量最高的ㄧ組。而在抗菌性實驗中，由於和控制組不含抗菌劑的PU300/Nw複合膜比較，含抗菌劑的複合膜，除了含25μg/cm2 AgSD 之PU300/Nw和含50μg/cm2 AgSD之PU400/Nw複合膜兩種抗菌情形不佳以外，其它均有良好的抗菌性。 而在人工皮膚方面，本實驗欲研究出培養纖維母細胞、角質細胞於豬真皮基質之人工皮膚。實驗結果得知，在培養纖維母細胞方面，以真皮面的組織間隙較大，細胞可長入基質中，而基底膜面的組織間隙較小，纖維母細胞容易長成一層。纖維母細胞植入的數量與後來角質細胞的生長有密切的關係，若纖維母細胞長得太多，或長至基底膜面，都會影響角質細胞的生長，因此這一點仍待研究。而角質細胞則較適合生長於基底膜面，把兩種細胞用基底膜分開，可能為較好的一種培養方法。而另外以胃蛋白酵素處理豬真皮基質，希望降低豬真皮基質於植入別種動物體中的抗原性。但在此實驗中，以胃蛋白酵素處理豬真皮基質的時間並未與細胞生長有明顯的關係，但在培養纖維母細胞十四天後，測其細胞活性，處理後的細胞活性均較未處理來的高，這個原因仍待探討，而關於減低植入動物體之抗原性必須做動物實驗才能知道。但以酵素處理後冷凍乾燥，確實會增加豬真皮基質的組織間隙。由本研究得知，植入人類細胞於豬真皮基質作為人工皮膚是可行的，未來可繼續做探討。

In this research, some wound dressing materials and artificial skin have been studied for application on burns. To decrease water loss from the wound and bacterial invasion, a wound dressing consisted of polyurethane membrane and nonwoven has been prepared. Antimicrobial agent silver sulfadiazine have been impregnated into the polyurethane membrane to inhibit bacterial infection by releasing antimicrobial agent. The average vapor permeability of polyurethane/ nonwoven composite membranes (PU/Nw) can be controlled by the film thickness according to different degree of burns. The vapor permeability for PU200/Nw is 3114 g/m2/day, for PU300/Nw is 2243 g/m2/day and for PU400/Nw is 1743 g/m2/day. All the wound dressings have no cytotoxicity to the fibroblasts. The concentration 50μg/cm2 of silver sulfadiazine is enough to against Pseudomonas aeruginosa. In this research, a porcine dermal matrix with human fibroblasts and keratinocytes has been cultivated. The porcine dermal matrix is a good scaffold for cell growth. The dermal side of matrix is suitable for fibroblasts and the basement membrane side is suitable for keratinocytes. The number of fibroblasts grew on the matrix increase as increasing fibroblasts seeding density. Migration of fibroblasts in the matrix can be improved by treating with pepsin. This is because that the tissue space in the matrix became loose after pepsin treated. From above mention, a potential application on the artificial skin can be acquired by cultivating fibroblasts and keratinocytes into the porcine dermal matrix.