Harrison Caroline A, Gossiel Fatma, Layton Christopher M, Bullock Anthony J, Johnson Timothy, Blumsohn Aubrey, MacNeil Sheila
Department of Tissue Engineering, Kroto Institute, University of Sheffield, UK.
Tissue Eng. 2006 Nov;12(11):3119-33. doi: 10.1089/ten.2006.12.3119.
Skin graft contraction leading to loss of joint mobility and cosmetic deformity remains a major clinical problem. In this study we used a tissue-engineered model of human skin, based on sterilized human adult dermis seeded with keratinocytes and fibroblasts, which contracts by up to 60% over 28 days in vitro, as a model to investigate the mechanism of skin contraction. Pharmacologic agents modifying collagen synthesis, degradation, and cross-linking were examined for their effect on contraction. Collagen synthesis and degradation were determined using immunoassay techniques. The results show that skin contraction was not dependent on inhibition of collagen synthesis or stimulation of collagen degradation, but was related to collagen remodelling. Thus, reducing dermal pliability with glutaraldehyde inhibited the ability of cells to contract the dermis. So did inhibition of matrix metalloproteinases and inhibition of lysyl oxidase-mediated collagen cross-linking, but not transglutaminase-mediated cross-linking. In summary, this in vitro model of human skin has allowed us to identify specific cross-linking pathways as possible pharmacologic targets for prevention of graft contracture in vivo.
皮肤移植收缩导致关节活动度丧失和美容畸形仍然是一个主要的临床问题。在本研究中，我们使用了一种基于接种角质形成细胞和成纤维细胞的无菌成人人类真皮的组织工程化人类皮肤模型，该模型在体外28天内收缩高达60%，作为研究皮肤收缩机制的模型。研究了改变胶原蛋白合成、降解和交联的药物制剂对收缩的影响。使用免疫测定技术测定胶原蛋白的合成和降解。结果表明，皮肤收缩不依赖于胶原蛋白合成的抑制或胶原蛋白降解的刺激，而是与胶原蛋白重塑有关。因此，用戊二醛降低真皮柔韧性会抑制细胞收缩真皮的能力。抑制基质金属蛋白酶和抑制赖氨酰氧化酶介导的胶原蛋白交联也有同样的效果，但转谷氨酰胺酶介导的交联则没有。总之，这种人类皮肤体外模型使我们能够确定特定的交联途径，作为预防体内移植挛缩的可能药物靶点。
