Tissue Engineering and Biomaterials Laboratory, Department of Orthopedic Surgery, Mayo Clinic , Rochester, Minnesota.
Tissue Eng Part A. 2018 Feb;24(3-4):245-253. doi: 10.1089/ten.TEA.2016.0531. Epub 2017 Jun 29.
Intra-articular ligamentous injuries are typically unrepairable and have limited outcomes after graft reconstruction. A combination of porous polycaprolactone fumarate (PCLF) scaffolds with polyethylene terephthalate (PET) sutures was developed with the goal of regenerating intra-articular ligaments. Scaffolds were fabricated by injecting PCLF over three-dimensional-printed molds containing two strands of PET suture down its central pore followed by cross-linking. Scaffolds were seeded with human mesenchymal stem cells (MSCs) from adipose tissue. To demonstrate cell attachment and proliferation in culture, we performed live/dead staining and cell proliferation assays. These experiments showed that MSCs remain viable and continue to proliferate on the scaffolds in culture for at least 2 weeks. Bare scaffolds were then used to reconstruct the rabbit anterior-cruciate ligament (ACL), while control rabbits underwent semitendinosus autograft reconstruction. The specimens underwent micro-computed tomography (CT) imaging, histological examination, and biomechanical testing at 8 weeks. The ultimate pull-out strength of the PCLF-PET scaffolds and tendon autografts was initially 72 ± 30 N and to 45 ± 10 N, respectively (p < 0.06). On inspection after 8 weeks in vivo, the intra-articular portion of the PCLF-PET scaffolds was fragmented while the tendon autografts remained intact. Cross-sectional areas of bone tunnels in the PCLF-PET scaffolds (11.3 ± 1 mm) were enlarged compared to tendon autografts (3.8 ± 0.5 mm) (p < 0.004) as measured by micro-CT. These studies show that PET-reinforced PCLF scaffolds are capable of initial ACL reconstruction and supports stem cell growth. The intra-articular portion of the scaffold may need to be re-engineered to support their use in ligament regeneration.
关节内韧带损伤通常无法修复,并且在移植物重建后其结果有限。聚己内酯富马酸酯(PCLF)多孔支架与聚对苯二甲酸乙二醇酯(PET)缝线的结合是为了再生关节内韧带而开发的。支架是通过将 PCLF 注入包含两根 PET 缝线的三维打印模具中制成的,缝线沿着中央孔的中心放置,然后进行交联。支架用脂肪来源的间充质干细胞(MSCs)接种。为了在培养中证明细胞黏附和增殖,我们进行了活/死染色和细胞增殖测定。这些实验表明,MSCs 在培养中的支架上保持存活并继续增殖,至少持续 2 周。然后,将裸支架用于重建兔前交叉韧带(ACL),而对照兔则接受半腱肌腱自体移植重建。在 8 周时,对标本进行了微计算机断层扫描(CT)成像、组织学检查和生物力学测试。PCLF-PET 支架和肌腱自体移植物的最终拔出强度最初分别为 72±30N 和 45±10N(p<0.06)。在体内 8 周后检查时,PCLF-PET 支架的关节内部分碎裂,而肌腱自体移植物保持完整。PCLF-PET 支架中的骨隧道的横截面积(11.3±1mm)比肌腱自体移植物(3.8±0.5mm)(p<0.004)增大,这是通过微 CT 测量的。这些研究表明,PET 增强的 PCLF 支架能够进行初始 ACL 重建并支持干细胞生长。支架的关节内部分可能需要重新设计,以支持它们在韧带再生中的应用。