Department of Orthopaedics, The First Affiliated Hospital of Soochow University, Suzhou, 215006, China; Orthopaedic Institute, Medical College, Soochow University, Suzhou, 215007, China; The Experimental Center, Cancer Hospital of the University of Chinese Academy of Sciences, Hangzhou, 310022, China.
Department of Orthopaedics, The First Affiliated Hospital of Soochow University, Suzhou, 215006, China; Orthopaedic Institute, Medical College, Soochow University, Suzhou, 215007, China.
Mater Sci Eng C Mater Biol Appl. 2020 Jan;106:110286. doi: 10.1016/j.msec.2019.110286. Epub 2019 Oct 7.
Autologous chondrocyte implantation (ACI) is a promising approach to repair cartilage defects; however, the cartilage trauma-induced inflammatory environment compromises its clinical outcomes. Cell-derived decellularized extracellular matrix (DECM) has been used as a culture substrate for mesenchymal stem cells (MSCs) to improve the cell proliferation and lineage-specific differentiation. In this study, DECM deposited by synovium-derived MSCs was used as an in vitro expansion system for rabbit articular chondrocytes and the response of DECM-expanded chondrocytes to pro-inflammatory cytokines such as interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF-α) was evaluated. Compared with those grown on tissue culture polystyrene (TCPS), the proliferation rate was significantly improved in DECM-expanded chondrocytes. TCPS- and DECM-expanded chondrocytes were isolated and induced to redifferentiation in a high-density pellet culture. DECM-expanded chondrocytes exerted a stronger resistance to 1 ng/mL of IL-1β than TCPS-expanded cells, but the production of cartilage matrix in both groups was inhibited by 5 ng/mL of IL-1β. When exposed to 1 or 5 ng/mL of TNF-α, DECM-expanded chondrocytes showed higher levels of cartilage matrix synthesis than TCPS-expanded cells. In addition, the gene expression of IL-1β- or TNF-α-induced matrix degrading enzymes (MMP3, MMP9, MMP13, and ADAMTS5) was significantly lower in DECM-expanded chondrocytes than TCPS-expanded cells. Furthermore, we found that SIRT1 inhibition by nicotinamide completely counteracted the protective effect of DECM on chondrocytes in the presence of IL-1β or TNF-α, indicating that the SIRT1 signaling pathway was involved in the DECM-mediated enhancement of anti-inflammatory properties of chondrocytes. Taken together, this work suggests that stem cell-derived DECM is a superior culture substrate for in vitro chondrocyte expansion by improving proliferation and enhancing the anti-inflammatory properties of chondrocytes. DECM-expanded chondrocytes with enhanced anti-inflammatory properties hold great potential in clinically ACI-based cartilage repair.
自体软骨细胞移植（ACI）是修复软骨缺损的一种很有前途的方法；然而，软骨创伤诱导的炎症环境会影响其临床效果。细胞来源的去细胞细胞外基质（DECM）已被用作间充质干细胞（MSCs）的培养底物，以提高细胞增殖和谱系特异性分化。在这项研究中，滑膜来源的 MSC 沉积的 DECM 被用作兔关节软骨细胞的体外扩增系统，并评估了 DECM 扩增的软骨细胞对促炎细胞因子（如白细胞介素-1β（IL-1β）和肿瘤坏死因子-α（TNF-α））的反应。与在组织培养聚苯乙烯（TCPS）上生长的软骨细胞相比，DECM 扩增的软骨细胞的增殖率显著提高。将 TCPS 和 DECM 扩增的软骨细胞分离出来，并在高密度微球培养中诱导其再分化。与 TCPS 扩增的细胞相比，DECM 扩增的软骨细胞对 1ng/mL 的 IL-1β表现出更强的抵抗力，但两组细胞的软骨基质生成均被 5ng/mL 的 IL-1β抑制。当暴露于 1 或 5ng/mL 的 TNF-α时，DECM 扩增的软骨细胞的软骨基质合成水平高于 TCPS 扩增的细胞。此外，DECM 扩增的软骨细胞中，IL-1β或 TNF-α诱导的基质降解酶（MMP3、MMP9、MMP13 和 ADAMTS5）的基因表达明显低于 TCPS 扩增的细胞。此外，我们发现烟酰胺抑制 SIRT1 完全抵消了 DECM 在存在 IL-1β或 TNF-α时对软骨细胞的保护作用，表明 SIRT1 信号通路参与了 DECM 介导的增强软骨细胞抗炎特性的过程。综上所述，这项工作表明，干细胞来源的 DECM 通过提高增殖和增强软骨细胞的抗炎特性，是一种更好的体外软骨细胞扩增培养底物。具有增强抗炎特性的 DECM 扩增软骨细胞在基于 ACI 的临床软骨修复中具有很大的潜力。
