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软骨细胞外基质纳米颗粒存在下的软骨生成反应。

Chondrogenic response in presence of cartilage extracellular matrix nanoparticles.

机构信息

Department of Cellular Biotechnology, Cell Science Research Center, Royan Institute for Biotechnology, ACECR, Isfahan, Iran.

Department of Tissue Engineering, Najafabad Branch, Islamic Azad University, Najafabad, Iran.

出版信息

J Biomed Mater Res A. 2018 Sep;106(9):2463-2471. doi: 10.1002/jbm.a.36440.

Abstract

Current studies based on regenerative medicine suggest, decellularized extracellular matrix (DC-ECM) components can regulate cell phenotype. In this regard, it is believed, presence of cartilage extracellular matrix particles in culture condition could produce physical and biochemical supportive cues for chondrogenesis. In this study, DC-ECM nanoparticles with average size of 61.5± 22.4 nm were produced by decellularization and mechanical processing. Homogenous distribution and nanoscale size of yield particles were observed by scanning electron microscopy (SEM) and transmission electron microscopy (TEM) microscopy imaging. Chemical structure preservation of cartilage ECM after decellularization was also confirmed by typical Fourier transform infrared (FTIR) spectrum mapping. The influence these nanoparticles on chondrogenic response of chondrocyte cells was investigated by direct and indirect addition of nanoparticles to culture medium. A clinical devitalized cartilage powder (DV-ECM) was also used as a positive control. Totally, MTS results showed that direct and indirect presence of both DC-ECM and DV-ECM particles in culture medium enhanced cellular metabolic activity except on day one of culture. Furthermore, on day 21, SOX9 and COL2 expression of cultured chondrocytes in the medium containing DC-ECM nanoparticles were up-regulated in comparison to negative control, which was further confirmed by presence more frequent number of larger size lacunae in micromass spheroids. Our findings support the use of ECM nanoparticles as condition supplement in culture medium and injectable biomaterials, especially for cell-based therapies for cartilage regeneration. © 2018 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 106A:2463-2471, 2018.

摘要

目前基于再生医学的研究表明,去细胞细胞外基质(DC-ECM)成分可以调节细胞表型。在这方面,人们认为,在培养条件下存在软骨细胞外基质颗粒可以为软骨生成产生物理和生化支持线索。在这项研究中,通过去细胞化和机械处理生产出平均粒径为 61.5±22.4nm 的 DC-ECM 纳米颗粒。通过扫描电子显微镜(SEM)和透射电子显微镜(TEM)显微镜成像观察到均匀分布和纳米级尺寸的产率颗粒。通过典型的傅里叶变换红外(FTIR)图谱也证实了软骨 ECM 在去细胞化后的化学结构保存。通过直接和间接将纳米颗粒添加到培养基中研究了这些纳米颗粒对软骨细胞的软骨生成反应的影响。还使用了一种临床去细胞化软骨粉末(DV-ECM)作为阳性对照。总的来说,MTS 结果表明,直接和间接存在于培养基中的 DC-ECM 和 DV-ECM 颗粒除培养的第一天外均增强了细胞代谢活性。此外,在第 21 天,与阴性对照相比,含有 DC-ECM 纳米颗粒的培养基中培养的软骨细胞的 SOX9 和 COL2 表达上调,这进一步通过存在更大尺寸陷窝的更频繁数量得到证实微团球体。我们的研究结果支持将 ECM 纳米颗粒用作培养基和可注射生物材料中的条件补充物,特别是用于基于细胞的软骨再生治疗。 © 2018 Wiley Periodicals, Inc. J Biomed Mater Res Part A:106A:2463-2471,2018。

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