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首页> 外文期刊>Materials science & engineering >An axial distribution of seeding, proliferation, and osteogenic differentiation of MC3T3-E1 cells and rat bone marrow-derived mesenchymal stem cells across a 3D Thai silk fibroin/gelatin/hydroxyapatite scaffold in a perfusion bioreactor
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An axial distribution of seeding, proliferation, and osteogenic differentiation of MC3T3-E1 cells and rat bone marrow-derived mesenchymal stem cells across a 3D Thai silk fibroin/gelatin/hydroxyapatite scaffold in a perfusion bioreactor

机译：灌注生物反应器中3D Thai丝素蛋白/明胶/羟基磷灰石支架上MC3T3-E1细胞和大鼠骨髓间充质干细胞的播种，增殖和成骨分化的轴向分布

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摘要

In cell culture, a perfusion bioreactor provides effective transportation of nutrients, oxygen, and waste removal to and from the core of the scaffold. In addition, it provides mechanical stimuli for enhancing osteogenic differentiation. In this study, we used an axial distribution of cell numbers, alkaline phosphatase (ALP) enzyme activity, and calcium content across 4 cross-sections of 10 mm thick scaffold, made of Thai silk fibroin (SF)/gelatin (G)/hydroxyapatite (HA), as a tool to evaluate the suitable perfusion flow rate. These evaluations cover all cellular developmental phases starting from seeding, to proliferation, and later osteogenic differentiation. Mouse pre-osteoblastic MC3T3-E1 cell lines were used as a cell model during seeding and proliferation. The bioreactor seeded scaffold provided more uniform cell distribution across the scaffold compared to centrifugal and agitation seeding, while the overall number of adhered cells from bioreactor seeding was slightly lower than agitation seeding. The dynamic culture using 1 ml/min perfusion flow rate (initial shear stress of 0.1 dyn/cm~2) enabled statistically higher MC3T3-E1 proliferation, ALP activity, and calcium deposition than those observed in the static-culturing condition. However, the perfusion flow rate of 1 ml/min seemed not to be enough for enhancing ALP expression across all sections of the scaffold. Rat bone marrow derived stromal cells (rMSC) were used in the detachment test and osteogenic differentiation. It was found that perfusion flow rate of 5 ml/min caused statistically higher cell detachment than that of 1 and 3 ml/min. The perfusion flow rate of 3 ml/min gave the highest rMSC osteogenic differentiation on a SF/G/HA scaffold than other flow rates, as observed from the significantly highest number of ALP enzyme activity and the calcium content without any significant cell growth. In addition, all of these parameters were evenly distributed across all scaffold sections.
机译：在细胞培养中，灌注生物反应器可将营养物，氧气和废物清除有效地运入和运出支架的核心。此外，它提供了机械刺激以增强成骨分化。在这项研究中，我们使用了由泰国丝素蛋白（SF）/明胶（G）/羟基磷灰石制成的10 mm厚支架的4个横截面的细胞数量，碱性磷酸酶（ALP）酶活性和钙含量的轴向分布（HA），作为评估合适的灌注流速的工具。这些评估涵盖了从播种到增殖再到成骨分化的所有细胞发育阶段。小鼠成骨前MC3T3-E1细胞系在接种和增殖过程中用作细胞模型。与离心和搅拌接种相比，生物反应器接种的支架在整个支架上提供了更均匀的细胞分布，而来自生物反应器接种的粘附细胞的总数略低于搅拌接种。动态培养使用1 ml / min的灌注流量（初始剪切应力为0.1 dyn / cm〜2），与静态培养条件下相比，MC3T3-E1的增殖，ALP活性和钙沉积在统计学上更高。然而，1 ml / min的灌注流速似乎不足以增强支架所有部分的ALP表达。大鼠骨髓来源的基质细胞（rMSC）用于分离测试和成骨分化。发现5 ml / min的灌注流速与1 ml和3 ml / min相比引起统计学上更高的细胞脱离。 3 ml / min的灌注流速比其他流速在SF / G / HA支架上产生了最高的rMSC成骨分化，这是从ALP酶活性的显着最高值和钙含量无明显细胞生长观察到的。此外，所有这些参数均均匀地分布在所有脚手架部分。

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