Effect of Hydrostatic Pressure on Bone Regeneration Using Human Mesenchymal Stem Cells
Effect of Hydrostatic Pressure on Bone Regeneration Using Human Mesenchymal Stem Cells
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DOI：
10.1089/ten.tea.2012.0064
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发表时间：
2012-10-01
影响因子：
4.1
通讯作者：
Ogawa, Rei
中科院分区：
文献类型：
--
作者：
Huang, Chenyu;Ogawa, Rei
背景资料：力学是骨组织工程中继细胞、支架、生长因子之后的第四要素。生物处理器的发展使得模拟生成三维（3D）骨结构所需的体内力学成为可能。然而，虽然静水压力（HP）是一个占主导地位的和恒定的机械应变在体内的骨细胞，很少有人知道的HP通过灌注生物处理器应用于体外人骨髓间充质干细胞（hMSC）behaviors.Methods的效果：hMSC进行原代培养三个通道，然后被接种到羟基磷灰石（HA）支架。将支架在自动化生物处理器中在循环HP下孵育3周。暴露于大气压力（AP）的支架用作比较物。成骨分化培养基用于HP和AP组。在孵育前和孵育后1、2、3周，分别取支架进行组织学、免疫组化和基因表达分析。结果：AP组支架表面仅发现细胞，而HP组则均匀分布于整个支架。免疫组化分析显示，HP组表达较高水平的骨钙素（OC），骨桥蛋白（OP），骨连接蛋白（ON），胶原1型（Col 1）比AP组在3周的过程。基因表达分析显示，HP组在1周、2周和3周时间点的ON、Col 1、碱性磷酸酶和整合素β 5表达水平高于AP组。HP组在2周和3周时间点也表达了较高水平的核心结合因子α-1（Cbfa 1），在1周时间点表达了较高水平的OP和OC。他们的增殖细胞核抗原水平较低，在1-和2-week timepoints.Conclusions：HP增强细胞活力，提高成骨分化和成熟，虽然有点牺牲的增殖和自我更新的间充质干细胞。未观察到生物处理器诱导的HP对骨再生的可能负面影响。此外，在HP刺激后，机械传导分子整合素β 5以高水平表达，并且可能在体外HP驱动的骨生成期间增强迁移、促进分化并抑制破骨细胞成熟。
Background: Mechanics is increasingly being recognized as the fourth essential factor in bone tissue engineering next to cell, scaffold, and growth factors. The development of bioprocessors has made it possible to simulate the in vivo mechanics that are needed to generate three-dimensional (3D) bone constructs. However, although hydrostatic pressure (HP) is a dominant and constant mechanical strain on bone cells in vivo, little is known about the effect of HP applied via perfusion bioprocessors on in vitro human bone marrow derived mesenchymal stem cell (hMSC) behavior.Methods: hMSCs underwent primary culture for three passages before being seeded into hydroxyapatite (HA) scaffolds. The scaffolds were incubated for 3 weeks in an automated bioprocessor under cyclic HP. Scaffolds exposed to atmospheric pressure (AP) served as the comparator. Osteogenic differentiation medium was employed for both the HP and AP groups. Immediately before and 1, 2, and 3 weeks after incubation, the scaffolds were harvested for histological, immunohistochemical, and gene expression analyses.Results: Cells were only found in the AP scaffold surfaces, whereas in the HP group, they were distributed evenly throughout the scaffolds. Immunohistochemical analysis revealed that the HP group expressed higher levels of osteocalcin (OC), osteopontin (OP), osteonectin (ON), and collagen type 1 (Col1) than the AP group during the 3-week process. Gene expression analysis revealed that the HP group expressed higher levels of ON, Col1, alkaline phosphatase, and integrin beta 5 than the AP group at the 1-, 2-, and 3-week timepoints. The HP group also expressed higher levels of core-binding factor alpha-1 (Cbfa1) at the 2- and 3-week timepoints and higher levels of OP and OC at the 1-week timepoint. Their proliferating cell nuclear antigen levels were lower at the 1- and 2-week timepoints.Conclusions: HP enhances cellular viability and improves osteogenic differentiation and maturation, although somewhat at the expense of proliferation and self-renewal of MSCs. Possible negative effects of the bioprocessor-induced HP on bone regeneration were not observed. Further, the mechanotransductive molecule integrin beta 5 was expressed at high levels after HP stimulation and may enhance migration, promote differentiation, and inhibit osteoclast maturation during HP-driven osteogenesis in vitro.