Extracellular matrix powder from cultured cartilage-like tissue as cell carrier for cartilage repair
Extracellular matrix powder from cultured cartilage-like tissue as cell carrier for cartilage repair
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来自培养的软骨样组织的细胞外基质粉末作为软骨修复的细胞载体
DOI：
10.1039/c7tb00640c
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发表时间：
2017
影响因子：
7
通讯作者：
Zhang Xingdong
中科院分区：
文献类型：
--
作者：
Teng Yingying;Li Xiupeng;Chen Yafang;Cai Hanxu;Cao Wanxu;Chen Xuening;Sun Yong;Liang Jie;Fan Yujiang;Zhang Xingdong
软骨细胞外基质（ECM）因其生物活性而成为一种有前途的软骨修复材料。然而，ECM的动物来源不可避免地增加了病原体感染的风险和产品质量的变异性。在本研究中，我们利用新颖的3D培养方法制备新型人工脱细胞基质粉末（DEMP），用于开发用于软骨组织工程的可注射、生物活性、可生物降解的细胞载体。该培养方法将悬滴培养与悬浮培养方法相结合，非常有效地产生软骨样组织（CLT）。通过这种方法，初始的2.3×106软骨细胞在两周内产生了高达58.22 mg湿重的CLT，通过染色技术和生化分析证明其含有丰富的糖胺聚糖（GAG）、II型胶原以及BMP-2和TGF-β1生长因子。随后，将两周龄的CLT脱细胞以制备人工DEMP。体外研究发现，即使没有外源TGF-β1，在DEMP上培养的间充质干细胞在三周后也能很好地分化为软骨细胞，并分泌丰富的GAG和II型胶原。体内研究表明，DEMP 不仅促进透明软骨的再生（1 个月时兔软骨下缺损中 GAG 和 II 型胶原的强烈染色表明这一点），而且有利于软骨下骨的再生（1 个月时骨向内生长：48.22%），微 CT 数据显示。总的来说，这些结果表明，通过这种培养方法制备的人工 DEMP 作为一种用于软骨修复的新型 ECM 材料具有巨大的潜力。
Cartilage extracellular matrix (ECM) is a promising material for cartilage repair because of its bioactivity. However, the animal source of ECM unavoidably increases the risk of pathogen infection and the variability of product quality. In this study, we utilized a novel 3D culture method to prepare a new type of artificial decellularized matrix powder (DEMP) for the development of injectable, bioactive, biodegradable cell carriers for cartilage tissue engineering. This culture method combined hanging drop culture with suspension culture method, and was very efficient to produce cartilage-like tissue (CLT). By this method, an initial 2.3 × 106 chondrocyte generated as much as 58.22 mg wet weight CLT at two weeks, which proved to contain abundant glycoaminoglycans (GAGs), type II collagen, and BMP-2 and TGF-β1 growth factors by staining techniques and biochemical analysis. Subsequently, the two-week-old CLT was decellularized to prepare the artificial DEMP. In an in vitro study, it was found that MSCs cultured on DEMP differentiated to chondrocytes very well and secreted rich GAGs and type II collagen at three weeks even without exogenous TGF-β1. The in vivo study demonstrated that the DEMP not only facilitated regeneration of hyaline cartilage, which was implied by the intense staining of GAGs and type II collagen in rabbit subchondral defects at 1 month, but also benefited the regeneration of subchondral bone (bone ingrowth at 1 month: 48.22%) as shown in micro-CT data. Collectively, these results suggest that the artificial DEMP prepared by this culture method holds great potential as a novel ECM material for cartilage repair.