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首页> 外文期刊>Acta biomaterialia >Substrate stiffness and contractile behaviour modulate the functional maturation of osteoblasts on a collagen-GAG scaffold.
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Substrate stiffness and contractile behaviour modulate the functional maturation of osteoblasts on a collagen-GAG scaffold.

机译：基质刚度和收缩行为调节胶原GAG支架上成骨细胞的功能成熟。

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Anchorage-dependent cells respond to the mechanical and physical properties of biomaterials. One such cue is the mechanical stiffness of a material. We compared the osteogenic potential of collagen-glycosaminoglycan (CG) scaffolds with varying stiffness for up to 6 weeks in culture. The mechanical stiffness of CG scaffolds were varied by cross-linking by physical (dehydrothermal (DHT)) and chemical (1-ethyl-3-(3-dimethyl aminopropyl) carbodiimide (EDAC) and glutaraldehyde (GLUT)) methods. The results showed that all CG substrates allowed cellular attachment, infiltration and osteogenic differentiation. CG scaffolds treated with EDAC and GLUT were mechanically stiffer, retained their original scaffold structure and resisted cellular contraction. Consequently, they facilitated a 2-fold greater cell number, probably due to the pore architecture being maintained, allowing improved diffusion of nutrients. On the other hand, the less stiff substrates cross-linked with DHT allowed increased cell-mediated scaffold contraction, contracting by 70% following 6 weeks (P < 0.01) of culture. This reduction in scaffold area resulted in cells reaching the centre of the scaffold quicker up to 4 weeks; however, at 6 weeks all scaffolds showed similar levels of cellular infiltration, with higher cell numbers found on the stiffer EDAC- and GLUT-treated scaffolds. Analysis of osteogenesis showed that scaffolds cross-linked with DHT expressed higher levels of the late stage bone formation markers osteopontin and osteocalcin (P < 0.01) and increased levels of mineralisation. In conclusion, the more compliant CG scaffolds allowed cell-mediated contraction and supported a greater level of osteogenic maturation of MC3T3 cells, while the stiffer, non-contractible scaffolds resulted in lower levels of cell maturation, but higher cell numbers on the scaffold. Therefore, we found scaffold stiffness had different effects on differentiation and cell number whereby the increased cell-mediated contraction facilitated by the less stiff scaffolds positively modulated osteoblast differentiation while reducing cell numbers.
机译：锚定依赖性细胞对生物材料的机械和物理特性作出反应。一种提示就是材料的机械刚度。我们比较了不同硬度的胶原-糖胺聚糖（CG）支架在培养中长达6周的成骨潜力。 CG支架的机械刚度通过物理方法（脱水（DHT）和化学方法（1-乙基-3-（3-二甲基氨基丙基）碳二亚胺（EDAC）和戊二醛（GLUT））交联而变化。结果表明，所有CG底物均允许细胞附着，浸润和成骨分化。用EDAC和GLUT处理的CG支架在机械上更坚硬，保留了其原始支架结构并抵抗了细胞收缩。因此，它们促进了细胞数量增加2倍，这可能是由于维持了孔结构，从而改善了营养物质的扩散。另一方面，与DHT交联的较硬的底物使细胞介导的支架收缩增加，在培养6周后收缩70％（P <0.01）。支架面积的减少导致细胞在4周内更快到达支架中心。然而，在第6周，所有支架均表现出相似的细胞浸润水平，在较硬的EDAC和GLUT处理的支架上发现了更高的细胞数量。成骨分析表明，与DHT交联的支架表达了更高水平的晚期骨形成标志物骨桥蛋白和骨钙素（P <0.01），并增加了矿化水平。总之，更顺应的CG支架允许细胞介导的收缩并支持MC3T3细胞更高水平的成骨成熟，而较硬的，不可收缩的支架导致更低的细胞成熟水平，但支架上的细胞数量更高。因此，我们发现支架的刚度对分化和细胞数量具有不同的影响，由此，由刚度较小的支架促进的细胞介导的收缩增加可正向调节成骨细胞的分化，同时减少细胞数量。

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