Vijayalekha Ashwathi, Anandasadagopan Suresh Kumar, Gopal Thiyagarajan, Durai Saravanan, Anumaiya Vandhana, Pandurangan Ashok Kumar
School of Life Sciences, B.S. Abdur Rahman Crescent Institute of Science and Technology, Chennai, Tamil Nadu, India.
Biochemistry & Biotechnology Laboratory, CSIR-Central Leather Research Institute, Chennai, India.
J Biomed Mater Res A. 2025 Aug;113(8):e37977. doi: 10.1002/jbm.a.37977.
Osteochondral defects (OCDs) present significant clinical challenges, necessitating scaffolds that effectively regenerate both cartilage and subchondral bone. We developed a bilayer scaffold using fish collagen extracted from Catla catla skin to overcome the limitations of conventional biomaterials, such as mammalian collagen and synthetic polymers, which often suffer from immunogenic risks, poor bioactivity, or inadequate structural integration. The scaffold is comprised of collagen/fibrin (CC/FIB) for the articular cartilage layer and collagen/sodium citrate/hydroxyapatite (CC/NAC/HAP) for the subchondral bone layer, which is cross-linked with citric acid. Physicochemical characterization confirmed scaffold integration, enhanced thermal stability, and a porous architecture. The scaffold demonstrated optimal porosity (63.12%), degradation (62.08% over 28 days), superior swelling potential, and enhanced bio-mineralization in simulated body fluid. In vitro studies using MG-63 osteoblast-like cells and MC3T3-E1 cells showed high biocompatibility, increased alkaline phosphatase activity, and enhanced calcium deposition (33.73 ± 0.53 μg/mg of protein at 21 days). Gene expression analysis revealed upregulation of osteogenic (COL I ~23-fold, RUNX-2 ~15-fold, OCN ~8-fold) and chondrogenic (COL II ~12-fold, SOX-9 ~10-fold, ACAN ~6-fold) markers, confirming osteochondral regeneration potential. In vivo studies involving the implantation of 3 mm femoral trochlear OCDs in albino Wistar rats (n = 3 per group) resulted in substantial bone and cartilage regeneration, with complete defect closure by 12 weeks. Radiographic and histological assessments at 4, 8, and 12 weeks confirmed well-organized osteochondral repair, demonstrating superior regenerative capability compared to control groups. This study establishes the novelty of the fish collagen-based bilayer scaffold as a promising candidate for osteochondral tissue engineering, supporting effective cartilage and subchondral bone regeneration in OCD treatment.
骨软骨缺损（OCDs）带来了重大的临床挑战，因此需要能够有效再生软骨和软骨下骨的支架。我们利用从印度鲮鱼皮肤中提取的鱼胶原蛋白开发了一种双层支架，以克服传统生物材料（如哺乳动物胶原蛋白和合成聚合物）的局限性，这些传统材料常常存在免疫原性风险、生物活性差或结构整合不足的问题。该支架由用于关节软骨层的胶原蛋白/纤维蛋白（CC/FIB）和用于软骨下骨层的胶原蛋白/柠檬酸钠/羟基磷灰石（CC/NAC/HAP）组成，后者通过柠檬酸交联。物理化学表征证实了支架的整合性、增强的热稳定性和多孔结构。该支架表现出最佳孔隙率（63.12%）、降解率（28天内为62.08%）、优异的溶胀潜力以及在模拟体液中增强的生物矿化作用。使用MG-63成骨样细胞和MC3T3-E1细胞进行的体外研究表明，其具有高生物相容性、碱性磷酸酶活性增加以及钙沉积增强（21天时为33.73±0.53μg/mg蛋白质）。基因表达分析显示成骨标志物（COL I约23倍、RUNX-2约15倍、OCN约8倍）和软骨生成标志物（COL II约12倍、SOX-9约10倍、ACAN约6倍）上调，证实了骨软骨再生潜力。在白化Wistar大鼠（每组n = 3）中植入3毫米股骨滑车OCDs的体内研究导致了大量的骨和软骨再生，到12周时缺损完全闭合。在4周、8周和12周时进行的影像学和组织学评估证实了组织良好的骨软骨修复，与对照组相比显示出卓越的再生能力。本研究确立了基于鱼胶原蛋白的双层支架的新颖性，作为骨软骨组织工程中有前景的候选材料，为OCD治疗中有效的软骨和软骨下骨再生提供了支持。
