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以溶菌酶敏感细菌纤维素为支架的体外软骨生成

In vitro chondrogenesis with lysozyme susceptible bacterial cellulose as a scaffold.

作者信息

Yadav Vikas, Sun Lin, Panilaitis Bruce, Kaplan David L

机构信息

Department of Biomedical Engineering, Tufts University, Medford, MA, USA.

Department of Chemical and Biological Engineering, Tufts University, Medford, MA, USA.

出版信息

J Tissue Eng Regen Med. 2015 Dec;9(12):E276-88. doi: 10.1002/term.1644. Epub 2013 Jan 11.

Abstract

A current focus of tissue engineering is the use of adult human mesenchymal stem cells (hMSCs) as an alternative to autologous chondrocytes for cartilage repair. Several natural and synthetic polymers (including cellulose) have been explored as a biomaterial scaffold for cartilage tissue engineering. While bacterial cellulose (BC) has been used in tissue engineering, its lack of degradability in vivo and high crystallinity restricts widespread applications in the field. Recently we reported the formation of a novel bacterial cellulose that is lysozyme-susceptible and -degradable in vivo from metabolically engineered Gluconacetobacter xylinus. Here we report the use of this modified bacterial cellulose (MBC) for cartilage tissue engineering using hMSCs. MBC's glucosaminoglycan-like chemistry, combined with in vivo degradability, suggested opportunities to exploit this novel polymer in cartilage tissue engineering. We have observed that, like BC, MBC scaffolds support cell attachment and proliferation. Chondrogenesis of hMSCs in the MBC scaffolds was demonstrated by real-time RT-PCR analysis for cartilage-specific extracellular matrix (ECM) markers (collagen type II, aggrecan and SOX9) as well as histological and immunohistochemical evaluations of cartilage-specific ECM markers. Further, the attachment, proliferation, and differentiation of hMSCs in MBC showed unique characteristics. For example, after 4 weeks of cultivation, the spatial cell arrangement and collagen type-II and ACAN distribution resembled those in native articular cartilage tissue, suggesting promise for these novel in vivo degradable scaffolds for chondrogenesis.

摘要

组织工程学当前的一个重点是利用成人人类间充质干细胞(hMSCs)作为自体软骨细胞的替代品用于软骨修复。几种天然和合成聚合物(包括纤维素)已被探索用作软骨组织工程的生物材料支架。虽然细菌纤维素(BC)已用于组织工程,但它在体内缺乏可降解性且结晶度高,限制了其在该领域的广泛应用。最近我们报道了通过代谢工程改造的木醋杆菌合成了一种新型的对溶菌酶敏感且在体内可降解的细菌纤维素。在此我们报道使用这种改性细菌纤维素(MBC)结合hMSCs用于软骨组织工程。MBC类似葡糖胺聚糖的化学性质,加上其在体内的可降解性,表明了在软骨组织工程中利用这种新型聚合物的可能性。我们观察到,与BC一样,MBC支架支持细胞附着和增殖。通过对软骨特异性细胞外基质(ECM)标志物(II型胶原蛋白、聚集蛋白聚糖和SOX9)的实时RT-PCR分析以及对软骨特异性ECM标志物的组织学和免疫组织化学评估,证实了hMSCs在MBC支架中的软骨形成。此外,hMSCs在MBC中的附着、增殖和分化表现出独特的特征。例如,培养4周后,细胞的空间排列以及II型胶原蛋白和聚集蛋白聚糖的分布类似于天然关节软骨组织中的情况,这表明这些新型的体内可降解支架在软骨形成方面具有应用前景。

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