Department of Bioengineering, İzmir Institute of Technology, İzmir, Turkey.
Faculty of Medicine, Department of Biochemistry, Dokuz Eylül University, İzmir, Turkey.
J Biomater Appl. 2021 Apr;35(9):1192-1207. doi: 10.1177/0885328220981189. Epub 2021 Jan 14.
Tissue engineering approaches which include a combination of cells and scaffold materials provide an alternative treatment for meniscus regeneration. Decellularization and recellularization techniques are potential treatment options for transplantation. Maintenance of the ultrastructure composition of the extracellular matrix and repopulation with cells are important factors in constructing a biological scaffold and eliminating immunological reactions.The aim of the study is to develop a method to obtain biological functional meniscus scaffolds for meniscus regeneration. For this purpose, meniscus tissue was decellularized by our modified method, a combination of physical, chemical, and enzymatic methods and then recellularized with a meniscal cell population composed of fibroblasts, chondrocytes and fibrochondrocytes that obtained from mesenchymal stem cells. Decellularized and recellularized meniscus scaffolds were analysed biochemically, biomechanically and histologically. Our results revealed that cellular components of the meniscus were successfully removed by preserving collagen and GAG structures without any significant loss in biomechanical properties. Recellularization results showed that the meniscal cells were localized in the empty lacuna on the decellularized meniscus, and also well distributed and proliferated consistently during the cell culture period (p < 0.05). Furthermore, a high amount of DNA, collagen, and GAG contents (p < 0.05) were obtained with the meniscal cell population in recellularized meniscus tissue.The study demonstrates that our decellularization and recellularization methods were effective to develop a biological functional meniscus scaffold and can mimic the meniscus tissue with structural and biochemical features. We predict that the obtained biological meniscus scaffolds may provide avoidance of adverse immune reactions and an appropriate microenvironment for allogeneic or xenogeneic recipients in the transplantation process. Therefore, as a promising candidate, the obtained biological meniscus scaffolds might be verified with a transplantation experiment.
组织工程方法包括细胞和支架材料的结合，为半月板再生提供了一种替代治疗方法。去细胞化和再细胞化技术是移植的潜在治疗选择。维持细胞外基质的超微结构组成并重新填充细胞是构建生物支架和消除免疫反应的重要因素。本研究旨在开发一种方法来获得用于半月板再生的生物功能半月板支架。为此，我们通过改良的方法（物理、化学和酶法的组合）对半月板组织进行去细胞化，然后用由纤维母细胞、软骨细胞和成纤维软骨细胞组成的半月板细胞群进行再细胞化，这些细胞群是从间充质干细胞中获得的。对去细胞化和再细胞化的半月板支架进行了生化、生物力学和组织学分析。我们的结果表明，通过保留胶原和 GAG 结构，成功去除了半月板的细胞成分，而生物力学性能没有明显损失。再细胞化结果表明，半月板细胞定位于去细胞化半月板的空腔中，并且在细胞培养期间均匀分布并持续增殖（p<0.05）。此外，在再细胞化的半月板组织中，半月板细胞群获得了大量的 DNA、胶原和 GAG 含量（p<0.05）。本研究表明，我们的去细胞化和再细胞化方法有效地开发了具有生物功能的半月板支架，可以模拟具有结构和生化特征的半月板组织。我们预测，获得的生物半月板支架可能在移植过程中为同种异体或异种受体提供避免不良免疫反应和适当的微环境。因此，作为一种有前途的候选物，获得的生物半月板支架可能需要通过移植实验进行验证。
