去細胞化脂肪組織/蠶絲蛋白基質及微球體於脂肪組織工程上的應用

Abstract

先天畸形、外傷及腫瘤移除等各種原因常會造成軟組織缺陷，進而影響病患的心理及人際關係，故整形重建外科一直都受到高度重視；而由於醫學的進步，人們對醫療品質的要求及形體外觀的重視也日益提高，故醫學美容的需求也隨之增加。然而，脂肪組織工程現今面臨的問題為無法持久維持移植後的脂肪體積，因此發展同時具有功能性及美學感的軟組織填充物極為重要。 在本篇論文中，我們利用去細胞化脂肪基質與蠶絲蛋白混合作為支架複合材料，並透過EDC交聯的方式減緩脂肪基質成分的流失速率。將脂肪基質中大部分的細胞及脂質去除後，約有80 %的collagen及30 %的GAGs能被保留，且殘留的活性成分也已被證實能誘導細胞分化，而經水解後的蠶絲蛋白亦有促使細胞增生的能力；另外，這兩種材料都具有高生物相容性，且混合後可增加整體的機械性質。 在in vitro實驗中，我們可以觀察到3T3-L1和脂肪幹細胞都順利地貼附於混合支架上，且於培養幾天後GAGs分泌量皆可增加30 %~ 45 %，與純蠶絲所製備的支架間存有顯著差異。而在in vivo實驗中，我們發現相較於純蠶絲製備的微球體，脂肪基質/蠶絲蛋白微球體更能有效地促使脂肪細胞增生及分化，顯示用脂肪基質和蠶絲蛋白做成的複合支架應用於皮膚填充及軟組織再生具有其發展潛力存在。

The soft tissue defects caused by congenital malformation, trauma, tumor removal and other various reasons affect the patient's psychology and interpersonal relationship, so it has been gaining popularity for Plastic and Reconstructive Surgery. Due to advances in medicine, the emphasis on the requirements of the quality of medical care and physical appearance is increasing, and it also increases the demand for medical cosmetic. However, current challenge of adipose tissue engineering failed to maintain volume of adipose after transplantation, so it is important to find the soft fillers that have both functionality and aesthetics. In the study, we used decellularized porcine adipose matrix and silk fibroin as composite materials for scaffolds. The loss of the ECM content mixed in the scaffolds was slowed down by cross-linking. Eighty percentage collagen and thirty percentage GAG contents were retained after removing most cells and lipids from porcine adipose tissue. These bioactive contents have been proved to induce cell differentiation, and hydrolyzed silk fibroin also has the ability to promote cell proliferation. In addition, both of these materials have high biocompatibility, and they can increase the overall mechanical properties after blending. In vitro experiments, we observed that 3T3-L1 and adipose stem cells attached to the composite scaffolds successfully, and their GAG content increased 30 % to 45 % after culture several days, and there was remarkable difference between the composite scaffolds and the silk fibroin scaffolds. In vivo experiments, we observed that adipose matrix-silk fibroin composite microspheres were more effective to promote adipose stem cells proliferation and differentiation than silk fibroin microspheres. There are great potentials for the application of these hybrid materials in dermal fillers and soft tissue regeneration.