Organically modified hydroxyapatite (ormoHAP) nanospheres stimulate the differentiation of osteoblast and osteoclast precursors: a co-culture study
Organically modified hydroxyapatite (ormoHAP) nanospheres stimulate the differentiation of osteoblast and osteoclast precursors: a co-culture study
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DOI：
10.1088/1748-605x/ab0fad
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发表时间：
2019-05-01
影响因子：
4
通讯作者：
Hanke, Thomas
中科院分区：
文献类型：
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作者：
Heinemann, Christiane;Heinemann, Sascha;Hanke, Thomas
将由电场辅助离子双迁移法制备的有机改性羟基磷灰石（ormoHAP）纳米球包埋在泡沫明胶中形成复合支架。已证明降解速率与交联度（40%，80%）以及支架的矿物质含量（0%，20%，40%）相关。分别来源于骨髓基质细胞和单核细胞的成骨细胞和破骨细胞的人共培养模型，在没有外部添加因子RANKL和M-CSF的情况下，运行长达42小时，以表征ormoHAP-明胶支架对共培养物的作用。通过定量生化方法（DNA、LDH、ALP、TRAP 5 b）、基因表达分析（ALP、BSP II、RANKL、IL-6、VTNR、CTSK、TRAP、OSCAR、CALCR）和共聚焦激光扫描显微镜（细胞核、肌动蛋白、CD 68、TRAP）进行检查。结果证实包埋在明胶基质中的ormoHAP增强了TRAP 5 b活性。作为反馈，成骨细胞ALP活性和BSP II基因表达增加。最后，提出了一系列细胞串扰行为，这可以解释所形成的重要共培养物的行为以及ormoHAP的存在和浓度的影响。
Isolated nanospheres consisting of organically modified hydroxyapatite (ormoHAP), prepared by an electric field-assisted ion double migration process, were embedded in foamed gelatin to form a composite scaffold. Degradation rates have been demonstrated to correlate with the crosslinking degree (40%, 80%) as well as with the mineral content of the scaffolds (0%, 20%, 40%). A human coculture model of osteoblasts and osteoclasts, derived from bone marrow stromal cells and monocytes, respectively, without external addition of the factors RANKL and M-CSF, was run for up to 42 din order to characterize the action of the ormoHAP-gelatin scaffolds on the co-culture. Examination was performed by quantitative biochemical methods (DNA, LDH, ALP, TRAP5b), gene expression analysis (ALP, BSP II, RANKL, IL-6, VTNR, CTSK, TRAP, OSCAR, CALCR) and confocal laser scanning microscopy (cell nuclei, actin, CD68, TRAP). Results confirm that ormoHAP embedded in the gelatin matrix enhanced TRAP 5b activity. As a feedback, ALP activity and gene expression of BSP II of osteoblasts increased. Finally, a sequence of cell cross-talk actions is suggested, which can explain the behavior of the formed vital co-culture and moreover the influence of the presence and concentration of ormoHAP.