Department of Bioengineering, Northeastern University, Boston, MA, USA.
Department of Chemical Engineering, Northeastern University, Boston, MA, USA.
Cartilage. 2021 Dec;13(2_suppl):1214S-1228S. doi: 10.1177/1947603520988768. Epub 2021 Jan 20.
Advanced glycation end-product (AGE) accumulation is implicated in osteoarthritis (OA) pathogenesis in aging and diabetic populations. Here, we develop a representative nonenzymatic glycation-induced OA cartilage explant culture model and investigate the effectiveness of resveratrol, curcumin, and eugenol in inhibiting AGEs and the structural and biological hallmarks of cartilage degeneration.
Bovine cartilage explants were treated with AGE-bovine serum albumin, threose, and ribose to determine the optimal conditions that induce physiological levels of AGEs while maintaining chondrocyte viability. AGE crosslinks, tissue stiffness, cell viability, metabolism and senescence, nitrite release and loss of glycosaminoglycans were assessed. Explants were cotreated with resveratrol, curcumin, or eugenol to evaluate their anti-AGE properties. Blind docking analysis was conducted to estimate binding energies of drugs with collagen II.
Treatment with 100 mM ribose significantly increased AGE crosslink formation and tissue stiffness, resulting in reduced chondrocyte metabolism and enhanced senescence. Blind docking analysis revealed stronger binding energies of both resveratrol and curcumin than ribose, with glycation sites along a human collagen II fragment, indicating their increased likelihood of competitively inhibiting ribose activity. Resveratrol and curcumin, but not eugenol, successfully inhibited AGE crosslink formation and its associated downstream biological response.
We establish a cartilage explant model of OA that recapitulates several aspects of aged human cartilage. We find that resveratrol and curcumin are effective anti-AGE therapeutics with the potential to decelerate age-related and diabetes-induced OA. This nonenzymatic glycation-induced model provides a tool for screening OA drugs, to simultaneously evaluate AGE-induced biological and mechanical changes.
糖基化终产物（AGE）的积累与衰老和糖尿病人群中骨关节炎（OA）的发病机制有关。在这里，我们开发了一种具有代表性的非酶糖基化诱导 OA 软骨外植体培养模型，并研究了白藜芦醇、姜黄素和丁香酚抑制 AGE 以及软骨退变的结构和生物学特征的效果。
用 AGE-牛血清白蛋白、苏糖醇和核糖处理牛软骨外植体，以确定在保持软骨细胞活力的同时诱导生理水平 AGE 的最佳条件。评估 AGE 交联、组织硬度、细胞活力、代谢和衰老、亚硝酸盐释放和糖胺聚糖丢失。用白藜芦醇、姜黄素或丁香酚共同处理外植体，以评估它们的抗 AGE 特性。进行盲目对接分析以估计药物与胶原蛋白 II 的结合能。
用 100mM 核糖处理可显著增加 AGE 交联形成和组织硬度，导致软骨细胞代谢降低和衰老增强。盲目对接分析显示，白藜芦醇和姜黄素的结合能都比核糖强，并且在一个人胶原蛋白 II 片段上存在糖基化位点，表明它们更有可能竞争性抑制核糖的活性。白藜芦醇和姜黄素，但不是丁香酚，成功地抑制了 AGE 交联形成及其相关的下游生物学反应。
我们建立了一个 OA 软骨外植体模型，该模型再现了人类老年软骨的多个方面。我们发现，白藜芦醇和姜黄素是有效的抗 AGE 治疗药物，有可能减缓与年龄相关和糖尿病相关的 OA。这种非酶糖基化诱导的模型为筛选 OA 药物提供了一种工具，可同时评估 AGE 诱导的生物学和机械变化。
