Fast fabrication of stable cartilage-like tissue using collagen hydrogel microsphere culture
Fast fabrication of stable cartilage-like tissue using collagen hydrogel microsphere culture
复制标题
利用胶原水凝胶微球培养快速制造稳定的软骨样组织
DOI：
10.1039/c7tb02535a
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发表时间：
2017-12-14
影响因子：
7
通讯作者：
Zhang, Xingdong
中科院分区：
文献类型：
--
作者：
Liu, Jun;Yu, Cheng;Zhang, Xingdong
间充质干细胞(MSCs)越来越被认为是软骨修复的有效细胞来源。然而，由于MSC来源的软骨细胞表型的不稳定性和合成的软骨基质的骨化，MSCs再生稳定的软骨组织仍然具有挑战性。骨髓间充质干细胞向软骨分化的命运受其局部微环境的调控，微环境对细胞行为、软骨形成表型和基质合成至关重要。在本研究中，我们在三种不同的微环境中，包括细胞微球、胶原水凝胶微球(CHB)和胶原水凝胶微球(CHMS)，通过MSC的软骨形成来构建类软骨组织。培养15天后，CHM的细胞数增加到472.6%，而CHB减少了58.6%，颗粒减少了46.6%；导致CHM的大小增加了230%，而CHB减少了36.8%，颗粒增加了20.1%。组织学染色显示，颗粒中的糖胺多聚糖(GAG)比中成药中的糖胺聚糖(GAG)更密集，但不那么均匀。从第5天到第15天，CHB外区的GAG染色强于内区，但弱于颗粒和中成药。聚合酶链式反应结果显示，成软骨基因(AGG、COL2A1、SOX9)的表达明显高于颗粒细胞和CHB，而肥大基因(COL10A1)的表达明显低于颗粒细胞和CHB，提示CHM具有更好的软骨分化潜能和更稳定的表型。总之，本研究通过更好地模拟自然的MSC增殖过程和增强体外质量交换，突出了CHM微环境相对于CHB和颗粒的优势。在基于MSC的软骨组织再生中，CHM培养显示了构建稳定的软骨样组织的潜力。
Mesenchymal stem cells (MSCs) had been increasingly regarded as a potent cell source for cartilage repair. However, due to the instability of MSC-derived chondrocyte phenotype and ossification of the synthesised cartilage matrix, regenerating a stable cartilage tissue by MSCs is still challenging. The fate of chondrogenesis from MSCs is regulated by their local microenvironment, which is of vital importance to the cell behaviours, chondrogenic phenotype and matrix synthesis. In this study, we fabricated cartilage-like tissues by the chondrogenesis of MSC in three different microenvironments, including cell pellets, collagen hydrogel bulk (CHB) and collagen hydrogel microspheres (CHMs) in vitro. After 15 days in culture, the cell number was increased to 472.6% in CHMs, compared to a 58.6% decrease in CHB and a 46.6% decrease in pellets; resulting in a 230% increase in CHM size, but a 36.8% decrease in CHB and only a 20.1% increase in pellets. Histological staining demonstrated a more intensive but less homogeneous glycosaminoglycan (GAG) pattern in pellets than in CHMs. The outer area of CHB showed a stronger GAG staining than its inner area from day 5 to day 15, but the staining was weaker than that in both pellets and CHMs. The PCR results showed that CHMs achieved a significantly higher chondrogenic gene (AGG, COL2A1, SOX9) expression and a lower hypertrophic gene (COL10A1) expression than pellets and CHB, suggesting a better chondrogenic differentiation potential with a more stable phenotype in CHMs. In summary, this study highlights the advantages of CHM microenvironments over those of CHB and pellets by a better mimicking of the natural MSC proliferation process and enhancing mass exchange in vitro. The CHM culture demonstrated potential to fabricate stable cartilage-like tissue in MSC based cartilage tissue regeneration.