Mirsadraee Saeed, Wilcox Helen E, Watterson Kevin G, Kearney John N, Hunt John, Fisher John, Ingham Eileen
The Institute of Medical and Biological Engineering, University of Leeds, Leeds, United Kingdom.
J Surg Res. 2007 Dec;143(2):407-14. doi: 10.1016/j.jss.2007.01.026. Epub 2007 Jun 14.
Previous studies have shown successful decellularization of human pericardium without affecting the major structural components and strength of the matrix. The aim of this study was to assess the biocompatibility and reseeding potential of the acellular human pericardial scaffold.
Pericardia were treated sequentially with hypotonic buffer, sodium dodecyl sulfate, and a nuclease solution. The presence of cellular attachment factors after decellularization was evaluated using immunohistochemistry. The scaffold was seeded with dermal fibroblasts and cellular attachment to and numbers of cells penetrating were assessed over time. Biocompatibility was also evaluated following subcutaneous implantation into a mouse model for three months.
After decellularization, the scaffold stained positively for fibronectin, but collagen IV and laminin staining was reduced. Seeded fibroblasts attached to the mesothelial surface and were visualized in the tissue within a week of seeding. The majority of fibroblasts in the tissue were viable and there was evidence of remodeling of the matrix. Analysis of the explanted tissues from mice showed that fresh/frozen and glutaraldehyde-fixed pericardia were encapsulated with a thick layer of inflammatory cells and fibrous tissue. In contrast, the decellularized scaffold was infiltrated with myofibroblasts, CD34+ cells and macrophages, indicating a healthy repair process. Compared with the glutaraldehyde-fixed tissue, the calcium content of the fresh/frozen and decellularized pericardia was negligible.
The pericardial scaffold was biocompatible in vitro and in the mouse model in vivo.
先前的研究表明，人心包成功去细胞化后，其主要结构成分和基质强度未受影响。本研究的目的是评估脱细胞人心包支架的生物相容性和再接种潜力。
心包依次用低渗缓冲液、十二烷基硫酸钠和核酸酶溶液处理。使用免疫组织化学评估去细胞化后细胞附着因子的存在情况。将真皮成纤维细胞接种到支架上，并随时间评估细胞的附着情况和穿透细胞的数量。还通过皮下植入小鼠模型三个月来评估生物相容性。
去细胞化后，支架纤维连接蛋白染色呈阳性，但IV型胶原和层粘连蛋白染色减少。接种的成纤维细胞附着在间皮表面，接种一周内在组织中可见。组织中的大多数成纤维细胞存活，并且有基质重塑的证据。对小鼠取出组织的分析表明，新鲜/冷冻和戊二醛固定的心包被一层厚厚的炎性细胞和纤维组织包裹。相比之下，脱细胞支架中有肌成纤维细胞、CD34+细胞和巨噬细胞浸润，表明是一个健康的修复过程。与戊二醛固定的组织相比，新鲜/冷冻和脱细胞心包的钙含量可忽略不计。
心包支架在体外和体内小鼠模型中具有生物相容性。
