Lü Wei-Dong, Zhang Ming, Wu Zhong-Shi, Hu Tie-Hui
Department of Thoracic and Cardiovascular Surgery, Second Xiangya Hospital of Central South University, Hunan Province, People's Republic of China.
Interact Cardiovasc Thorac Surg. 2009 Mar;8(3):301-5. doi: 10.1510/icvts.2008.194076. Epub 2008 Dec 12.
Decellularization means and altering crosslinking approaches were two promising alternatives for glutaraldehyde fixation to biological tissues. Bovine jugular veins (BJVs) were decellularized by a multi-step detergent-enzymatic extraction method, then photooxidatively crosslinked. Gross and histological integrity of which was retained. Ultrastructures showed integrity of collagen fibrils and elastic fibers, and a basement membrane free luminal surface. Mechanical strength test and tissue protein extraction assay demonstrated their tissue stability. After being pre-coated with gelatin, collagen IV and fibronectin, cultured human umbilical vein endothelial cells were planted in the luminal surface of decellularized plus photooxidized BJV patches for seven days. Endothelial cells were denser in pre-coated patches than in uncoated controls. A rat subcutaneous implantation model revealed more resistance against in vivo degradation for further crosslinked BJV patches than decellularized patches at 12-week retrieval. Host cells were all layer repopulated for both. Histological examination and content assay demonstrated collagen and glycosaminoglycan components synthesis for decellularized plus photooxidized BJV patches. Decellularized and photooxidatively crosslinked BJV patches possess tissue integrity, excellent in vitro and in vivo tissue stability and repopulation patterns. Thus, they have potentials as tissue engineering scaffolds in future cardiovascular surgery.
去细胞化方法和改变交联方法是戊二醛固定生物组织的两种有前景的替代方法。采用多步去污剂 - 酶提取法对牛颈静脉(BJVs)进行去细胞化处理,然后进行光氧化交联。其大体和组织学完整性得以保留。超微结构显示胶原纤维和弹性纤维完整,管腔表面无基底膜。力学强度测试和组织蛋白提取分析证明了它们的组织稳定性。在预先包被明胶、IV型胶原和纤连蛋白后,将培养的人脐静脉内皮细胞种植在去细胞化加光氧化的BJV补片管腔表面7天。预先包被的补片中内皮细胞比未包被的对照更密集。大鼠皮下植入模型显示,在12周取出时,进一步交联的BJV补片比去细胞化补片对体内降解具有更强的抵抗力。两种补片的宿主细胞均完全重新填充。组织学检查和含量分析证明去细胞化加光氧化的BJV补片有胶原和糖胺聚糖成分的合成。去细胞化并经光氧化交联的BJV补片具有组织完整性、优异的体外和体内组织稳定性以及重新填充模式。因此,它们在未来心血管手术中作为组织工程支架具有潜力。