Sports Medicine Research Laboratory, Department of Orthopedic Surgery, Children's Hospital Boston, Harvard Medical School, 300 Longwood Avenue, Enders 1016, Boston, Massachusetts 02115, USA.
J Orthop Res. 2009 Dec;27(12):1612-8. doi: 10.1002/jor.20932.
Ruptures of the anterior cruciate ligament (ACL) are still associated with high rates of long-term complications, even in patients undergoing modern, state-of-the-art replacement. Tissue-engineering approaches have been shown to be of value in improving treatment of ACL ruptures. However, the success of tissue-engineering procedures depends on the choice of an appropriate biomaterial. Decellularized ACL tissue potentially combines the structural composition of the targeted tissue with a reduced risk of graft rejection or disease transmission. In this study, we tested the effectiveness of currently available decellularization methods based on TRITON-X, sodium dodecyl sulfate (SDS), and trypsin. After identifying the most effective decellularization method, the capacity for reseeding with ACL fibroblasts was studied. All decellularization protocols reduced DNA content, with TRITON-X treatment having the greatest effect. Concurrently, decellularization did not affect tissue collagen or total protein content, but did decrease glycosaminoglycan content. TRITON-X also resulted the least glycosaminoglycan depletion. Porcine ACL tissue after decellularization with TRITON-X could be successfully reseeded with human ACL fibroblasts as demonstrated by steady DNA content and increasing pro-collagen expression.
前交叉韧带 (ACL) 的断裂仍然与长期并发症的高发生率相关，即使在接受现代、最先进的 ACL 重建的患者中也是如此。组织工程方法已被证明对改善 ACL 断裂的治疗有价值。然而，组织工程程序的成功取决于选择合适的生物材料。脱细胞 ACL 组织可能将目标组织的结构组成与移植物排斥或疾病传播的风险降低相结合。在这项研究中，我们根据 Triton-X、十二烷基硫酸钠 (SDS) 和胰蛋白酶测试了当前可用的脱细胞方法的有效性。在确定了最有效的脱细胞方法后，研究了用 ACL 成纤维细胞再接种的能力。所有脱细胞方案均降低了 DNA 含量，其中 Triton-X 处理的效果最大。同时，脱细胞处理不会影响组织胶原或总蛋白含量，但会降低糖胺聚糖含量。Triton-X 还导致糖胺聚糖的耗竭最少。用 Triton-X 脱细胞后的猪 ACL 组织可以成功地用人 ACL 成纤维细胞再接种，这表现为稳定的 DNA 含量和增加的前胶原蛋白表达。
