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可降解镁植入物对单培养和三培养成骨细胞的影响。

Impact of degradable magnesium implants on osteocytes in single and triple cultures.

机构信息

Centre for Translational Bone, Joint- and Soft Tissue Research, Technische Universität Dresden, Medical Faculty and University Hospital, 01307 Dresden, Germany.

Helmholtz-Zentrum Hereon, Institute of Metallic Biomaterials, 21502 Geesthacht, Germany.

出版信息

Biomater Adv. 2022 Mar;134:112692. doi: 10.1016/j.msec.2022.112692. Epub 2022 Feb 4.

Abstract

In vitro triple cultures of human primary osteoblasts, osteocytes and osteoclasts can potentially help to analyze the effect of drugs and degradation products of biomaterials as a model for native bone tissue. In the present study, degradation products of Magnesium (Mg), which has been successfully applied in the biomedical field, were studied with respect to their impact on bone cell morphology and differentiation both in osteocyte single cultures and in the triple culture model. Fluorescence microscopic and gene expression analysis, analysis of osteoclast- and osteoblast-specific enzyme activities as well as osteocalcin protein expression were performed separately for the three cell types after cultivation in triple culture in the presence of extracts, containing 5 and 10 mM Mg. All three cell species were viable in the presence of the extracts and did not show morphological changes compared to the Mg-free control. Osteoblasts and osteoclasts did not show significant changes in gene expression of ALPL, BSPII, osteocalcin, TRAP, CTSK and CA2. Likewise on protein level, no significant changes in ALP-, TRAP-, CTSK- and CAII activities were detected. Osteocytes showed a significant downregulation of MEPE, which codes for a protein playing an important role in regulation of phosphate homeostasis by osteocytes. This study is the first to analyze the effects of Mg degradation products on primary osteocytes in vitro, both in single and triple culture. Even if promoting effects on the three examined bone cell species were not found in the applied triple culture setup, it was shown, that Mg degradation products do not interfere with the activity of osteoblasts, osteoclasts and osteocytes in vitro.

摘要

在体外将人原代成骨细胞、骨细胞和破骨细胞进行三重培养,可作为天然骨组织模型,用于分析药物和生物材料降解产物的作用。在本研究中,研究了镁(Mg)降解产物对成骨细胞单培养和三重培养模型中骨细胞形态和分化的影响,镁已成功应用于生物医学领域。在三重培养中培养后,分别对三种细胞类型进行荧光显微镜和基因表达分析、破骨细胞和成骨细胞特异性酶活性以及骨钙素蛋白表达分析,以研究其对骨细胞形态和分化的影响。在提取物存在的情况下,三种细胞类型均具有活力,与无镁对照相比,形态没有变化。与无镁对照相比,成骨细胞和破骨细胞的基因表达 ALPL、BSPII、骨钙素、TRAP、CTSK 和 CA2 没有显著变化。同样在蛋白水平上,ALP、TRAP、CTSK 和 CAII 活性也没有显著变化。骨细胞中 MEPE 的表达明显下调,MEPE 编码的蛋白在骨细胞调节磷酸盐稳态中起重要作用。这项研究首次分析了 Mg 降解产物对原代骨细胞的体外影响,包括单培养和三重培养。即使在应用的三重培养体系中没有发现对三种被检测的骨细胞有促进作用,但表明 Mg 降解产物不会干扰体外成骨细胞、破骨细胞和骨细胞的活性。

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