Silk was widely investigated as a promising scaffold material in ligament tissue engineering. Although a variety of silk scaffolds were developed for the regeneration of anterior cruciate ligament (ACL) in vitro and in vivo, more investigations should be performed in large animals to translate these findings into clinical applications. The aim of this study is to evaluate the feasibility of using silk-based ACL scaffolds to regenerate damaged ACLs in porcine model. The microstructural organization, tissue regeneration as well as ligament-bone interface of silk implants were evaluated with scanning electron microscopy, micro-computerized tomography, histological and immunohistochemical staining at three and six months postoperatively. The results demonstrated that silk fibers in the ACL scaffolds organized in parallel similar with collagen fibers in native ligaments, which facilitated and guided the penetration of newly regenerated tissue into the pores among silk fibers. Collagen production especially collagen I in silk implants significantly increased from three to six months, and was gradually close to the level of native ligaments. At implant-bone interface, indirect ligament-bone insertion was observed at three months and substantial Sharpey's fibers formed at six months. The results indicated that the silk-based ACL scaffold provides a promising tissue engineering approach for ACL regeneration.
Tissue Engineering of the Anterior Cruciate Ligament and Meniscus Using Acellularized Scaffolds
