Department of Biomedical Sciences, Graduate School of Ajou University, 206 World Cup-Ro, Yeongtong-Gu, Suwon, Republic of Korea.
Wake Forest Institute for Regenerative Medicine, Wake Forest School of Medicine, Winston-Salem, NC, 27157, USA.
Tissue Eng Regen Med. 2024 Feb;21(2):341-351. doi: 10.1007/s13770-023-00593-2. Epub 2023 Oct 19.
Current tendon and ligament reconstruction surgeries rely on scar tissue healing which differs from native bone-to-tendon interface (BTI) tissue. We aimed to engineer Synovium-derived mesenchymal stem cells (Sy-MSCs) based scaffold-free fibrocartilage constructs and investigate in vivo bone-tendon interface (BTI) healing efficacy in a rat anterior cruciate ligament (ACL) reconstruction model.
Sy-MSCs were isolated from knee joint of rats. Scaffold-free sy-MSC constructs were fabricated and cultured in differentiation media including TGF-β-only, CTGF-only, and TGF-β + CTGF. Collagenase treatment on tendon grafts was optimized to improve cell-to-graft integration. The effects of fibrocartilage differentiation and collagenase treatment on BTI integration was assessed by conducting histological staining, cell adhesion assay, and tensile testing. Finally, histological and biomechanical analyses were used to evaluate in vivo efficacy of fibrocartilage construct in a rat ACL reconstruction model.
Fibrocartilage-like features were observed with in the scaffold-free sy-MSC constructs when applying TGF-β and CTGF concurrently. Fifteen minutes collagenase treatment increased cellular attachment 1.9-fold compared to the Control group without affecting tensile strength. The failure stress was highest in the Col + D + group (22.494 ± 13.74 Kpa) compared to other groups at integration analysis in vitro. The ACL Recon + FC group exhibited a significant 88% increase in estimated stiffness (p = 0.0102) compared to the ACL Recon group at the 4-week postoperative period.
Scaffold-free, fibrocartilage engineering together with tendon collagenase treatment enhanced fibrocartilaginous BTI healing in ACL reconstruction.
目前的肌腱和韧带重建手术依赖于瘢痕组织愈合，这与天然的骨-肌腱界面（BTI）组织不同。我们旨在基于滑膜衍生间充质干细胞（Sy-MSCs）构建无支架纤维软骨构建体，并在大鼠前交叉韧带（ACL）重建模型中研究体内骨-肌腱界面（BTI）愈合效果。
从大鼠膝关节中分离出 Sy-MSCs。构建无支架 Sy-MSC 构建体，并在分化培养基中进行培养，包括仅 TGF-β、仅 CTGF 和 TGF-β+CTGF。优化肌腱移植物上的胶原酶处理以改善细胞与移植物的整合。通过进行组织学染色、细胞黏附试验和拉伸试验来评估纤维软骨分化和胶原酶处理对 BTI 整合的影响。最后，使用组织学和生物力学分析来评估纤维软骨构建体在大鼠 ACL 重建模型中的体内效果。
当同时应用 TGF-β和 CTGF 时，无支架 Sy-MSC 构建体中观察到纤维软骨样特征。与对照组相比，15 分钟的胶原酶处理将细胞黏附增加了 1.9 倍，而不会影响拉伸强度。在体外整合分析中，Col+D+组的失效应力最高（22.494±13.74 Kpa）。与 ACL Recon 组相比，ACL Recon+FC 组在术后 4 周时估计刚度增加了 88%（p=0.0102）。
无支架、纤维软骨工程以及肌腱胶原酶处理增强了 ACL 重建中的纤维软骨 BTI 愈合。
