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用于软骨组织工程的猪半月板脱细胞细胞外基质支架的研制与表征

Development and Characterization of Acellular Extracellular Matrix Scaffolds from Porcine Menisci for Use in Cartilage Tissue Engineering.

作者信息

Chen Ying-Chen, Chen Ray-Neng, Jhan Hua-Jing, Liu Der-Zen, Ho Hsiu-O, Mao Yong, Kohn Joachim, Sheu Ming-Thau

机构信息

1 School of Pharmacy, College of Pharmacy, Taipei Medical University , Taipei, Taiwan .

2 Department of Cosmetics Applications and Management, Mackay Junior College of Medicine , Nursing, and Management, Taipei, Taiwan .

出版信息

Tissue Eng Part C Methods. 2015 Sep;21(9):971-86. doi: 10.1089/ten.TEC.2015.0036. Epub 2015 Jun 10.

Abstract

Given the growing number of arthritis patients and the limitations of current treatments, there is great urgency to explore cartilage substitutes by tissue engineering. In this study, we developed a novel decellularization method for menisci to prepare acellular extracellular matrix (ECM) scaffolds with minimal adverse effects on the ECM. Among all the acid treatments, formic acid treatment removed most of the cellular contents and preserved the highest ECM contents in the decellularized porcine menisci. Compared with fresh porcine menisci, the content of DNA decreased to 4.10%±0.03%, and there was no significant damage to glycosaminoglycan (GAG) or collagen. Histological staining also confirmed the presence of ECM and the absence of cellularity. In addition, a highly hydrophilic scaffold with three-dimensional interconnected porous structure was fabricated from decellularized menisci tissue. Human chondrocytes showed enhanced cell proliferation and synthesis of chondrocyte ECM including type II collagen and GAG when cultured in this acellular scaffold. Moreover, the scaffold effectively supported chondrogenesis of human bone marrow-derived mesenchymal stem cells. Finally, in vivo implantation was conducted in rats to assess the biocompatibility of the scaffolds. No significant inflammatory response was observed. The acellular ECM scaffold provided a native environment for cells with diverse physiological functions to promote cell proliferation and new tissue formation. This study reported a novel way to prepare decellularized meniscus tissue and demonstrated the potential as scaffolds to support cartilage repair.

摘要

鉴于关节炎患者数量不断增加以及当前治疗方法的局限性,通过组织工程探索软骨替代物的需求极为迫切。在本研究中,我们开发了一种用于半月板的新型脱细胞方法,以制备对细胞外基质(ECM)影响最小的脱细胞细胞外基质支架。在所有酸处理中,甲酸处理去除了大部分细胞成分,并在脱细胞猪半月板中保留了最高的ECM含量。与新鲜猪半月板相比,DNA含量降至4.10%±0.03%,糖胺聚糖(GAG)或胶原蛋白没有明显损伤。组织学染色也证实了ECM的存在和细胞的缺失。此外,由脱细胞半月板组织制成了具有三维相互连接多孔结构的高亲水性支架。当在这种脱细胞支架中培养时,人软骨细胞显示出增强的细胞增殖以及软骨细胞ECM(包括II型胶原蛋白和GAG)的合成。此外,该支架有效地支持了人骨髓间充质干细胞的软骨形成。最后,在大鼠体内进行植入以评估支架的生物相容性。未观察到明显的炎症反应。脱细胞ECM支架为具有多种生理功能的细胞提供了天然环境,以促进细胞增殖和新组织形成。本研究报道了一种制备脱细胞半月板组织的新方法,并证明了其作为支持软骨修复支架的潜力。

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本文引用的文献

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The clinical status of cartilage tissue regeneration in humans.人类软骨组织再生的临床状况。
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