The Laboratory of Orthopaedic Tissue Regeneration & Orthobiologics, Department of Bioengineering, Clemson University, Clemson, South Carolina, USA.
The Orthopaedic Bioengineering Laboratory, Department of Bioengineering, Clemson University, Charleston, South Carolina, USA.
J Biomed Mater Res A. 2024 Mar;112(3):421-435. doi: 10.1002/jbm.a.37639. Epub 2023 Nov 14.
Focal chondral defects (FCDs) significantly impede quality of life for patients and impose severe economic costs on society. One of the most promising treatment options-autologous matrix-induced chondrogenesis (AMIC)-could benefit from a scaffold that contains both of the primary cartilage matrix components-sulfated glycosaminoglycans (sGAGs) and collagen type II. Here, 17 different protocols were evaluated to determine the most optimum strategy for decellularizing (decelling) the bovine nucleus pulposus (bNP) to yield a natural biomaterial with a cartilaginous constituency. The resulting scaffold was then characterized with respect to its biochemistry, biomechanics and cytocompatibility. Results indicated that the optimal decell protocol involved pre-crosslinking the tissue prior to undergoing decell with trypsin and Triton X-100. The residual DNA content of the scaffold was found to be 32.64 ± 9.26 ng/mg dry wt. of tissue with sGAG and hydroxyproline (HYP) contents of 72.53 ± 16.43. and 78.38 ± 8.46 μg/mg dry wt. respectively. The dynamic viscoelastic properties were found to be preserved (complex modulus: 17.92-16.62 kPa across a range of frequencies) while the equilibrium properties were found to have significantly decreased (aggregate modulus: 11.51 ± 9.19 kPa) compared to the non-decelled fresh bNP tissue. Furthermore, the construct was also found to be cytocompatible with bone marrow stem cells (BMSCs). While it was not permissive of cellular infiltration, the BMSCs were still found to have lined the laser drilled channels in the scaffold. Taken together, the biomaterial developed herein could be a valuable addition to the AMIC family of scaffolds or serve as an off-the-shelf standalone option for cartilage repair.
局灶性软骨缺损 (FCD) 显著降低了患者的生活质量，并给社会带来了严重的经济负担。最有前途的治疗选择之一——自体基质诱导软骨形成术 (AMIC)——可能受益于一种包含两种主要软骨基质成分的支架，即硫酸化糖胺聚糖 (sGAG) 和 II 型胶原。在这里，评估了 17 种不同的方案，以确定最优化的去细胞化 (去细胞) 牛髓核 (bNP) 的策略，以获得具有软骨成分的天然生物材料。然后，根据其生物化学、生物力学和细胞相容性对所得支架进行了表征。结果表明，最优化的去细胞方案涉及在用胰蛋白酶和 Triton X-100 进行去细胞之前，预先交联组织。支架的残留 DNA 含量为 32.64 ± 9.26 ng/mg 组织干重，sGAG 和羟脯氨酸 (HYP) 含量分别为 72.53 ± 16.43 和 78.38 ± 8.46 μg/mg 组织干重。发现动态粘弹性特性得以保留（在一定频率范围内，复合模量为 17.92-16.62 kPa），而平衡特性显著降低（聚集模量为 11.51 ± 9.19 kPa）与非去细胞新鲜 bNP 组织相比。此外，该构建体还被发现与骨髓基质干细胞 (BMSCs) 具有细胞相容性。虽然它不允许细胞渗透，但仍发现 BMSCs 排列在支架中的激光钻通道中。总之，本文开发的生物材料可为 AMIC 支架家族增添有价值的选择，或作为软骨修复的即用型独立选择。
