Seo Young-Kwon, Yoon Hee-Hoon, Song Kye-Yong, Kwon Soon-Yong, Lee Hwa-Sung, Park Yong-Soon, Park Jung-Keug
Department of Chemical and Biochemical Engineering, Dongguk University, Phil-Dong, Seoul, Korea.
J Orthop Res. 2009 Apr;27(4):495-503. doi: 10.1002/jor.20752.
The purpose of this study was to evaluate the biocompatibility of silk and collagen-hyaluronan (HA) in vitro by assessing anterior cruciate ligament (ACL) cell and T-lymphocyte cultures on scaffolds. The use of composite scaffolds as artificial ligaments in ACL reconstruction and their effects on angiogenesis were evaluated in vivo. The silk scaffold was knitted by hand and dry coated with collagen-HA, whereas the composite silk scaffold was made by covering a silk scaffold with a lyophilized collagen-HA substrate. The initial attachment and proliferation of human ACL cells on the composite silk scaffold was superior to the attachment and proliferation observed on the silk scaffold. The immune response was higher in both scaffolds after 72 h (p < 0.05) compared with the control culture condition without scaffolding, as assessed by T-lymphocyte cultures in vitro. There was no significant difference in the immune response in vitro between the silk and composite silk scaffolds. Silk and composite silk scaffolds were implanted as artificial ligaments in ACLs removed from the knees of dogs, and they were harvested 6 weeks after implantation. On gross examination, the onset of an inflammatory tissue reaction, such as synovitis, was seen in both the silk scaffold and the composite silk scaffold groups. An histological evaluation of the artificial ligament implants revealed the presence of monocytes in the silk composite scaffold and the absence of giant cells in all cases. MT staining in the composite silk scaffold-grafted group showed granulation tissue consisting of fibroblasts, lymphocytes, monocytes, and collagen fibers. In addition, CD31 staining revealed the formation of new blood vessels. On the other hand, no reparative tissues, such as blood vessels, collagen, and cells, were observed in the silk scaffold-grafted group. These results suggest that the lyophilized collagen-HA substrate is biocompatible in vitro and enhances new blood vessel and cell migration in vivo.
本研究的目的是通过评估前交叉韧带（ACL）细胞和T淋巴细胞在支架上的培养情况，在体外评估丝绸与胶原-透明质酸（HA）的生物相容性。在体内评估复合支架作为人工韧带用于ACL重建的效果及其对血管生成的影响。丝绸支架采用手工编织并用胶原-HA进行干涂，而复合丝绸支架是通过用冻干的胶原-HA底物覆盖丝绸支架制成。人ACL细胞在复合丝绸支架上的初始附着和增殖优于在丝绸支架上观察到的附着和增殖。通过体外T淋巴细胞培养评估，与无支架的对照培养条件相比，72小时后两种支架中的免疫反应均较高（p<0.05）。丝绸支架和复合丝绸支架在体外的免疫反应没有显著差异。将丝绸和复合丝绸支架作为人工韧带植入从狗膝关节取出的ACL中，并在植入6周后收获。大体检查发现，丝绸支架组和复合丝绸支架组均出现了炎症组织反应，如滑膜炎。对人工韧带植入物的组织学评估显示，丝绸复合支架中有单核细胞存在，且所有病例均无巨细胞。复合丝绸支架移植组的MT染色显示有由成纤维细胞、淋巴细胞、单核细胞和胶原纤维组成的肉芽组织。此外，CD31染色显示有新血管形成。另一方面，在丝绸支架移植组中未观察到血管、胶原和细胞等修复组织。这些结果表明，冻干的胶原-HA底物在体外具有生物相容性，并能促进体内新血管形成和细胞迁移。
