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粗磷酸钙涂层与血液单核细胞对脂肪来源间充质干细胞的共刺激作用:体外作为体内组织修复模型的研究

Costimulatory Effect of Rough Calcium Phosphate Coating and Blood Mononuclear Cells on Adipose-Derived Mesenchymal Stem Cells In Vitro as a Model of In Vivo Tissue Repair.

作者信息

Khlusov Igor A, Litvinova Larisa S, Shupletsova Valeria V, Khaziakhmatova Olga G, Malashchenko Vladimir V, Yurova Kristina A, Shunkin Egor O, Krivosheev Vasilii V, Porokhova Ekaterina D, Sizikova Anastasiia E, Safiullina Linara A, Legostaeva Elena V, Komarova Ekaterina G, Sharkeev Yurii P

机构信息

Center for Immunology and Cell Biotechnology, Immanuel Kant Baltic Federal University, 236029 Kaliningrad, Russia.

Research School of Chemistry and Applied Biomedical Sciences, National Research Tomsk Polytechnic University, 634050 Tomsk, Russia.

出版信息

Materials (Basel). 2020 Oct 2;13(19):4398. doi: 10.3390/ma13194398.

Abstract

Calcium phosphate (CaP) materials do not always induce ectopic vascularization and bone formation; the reasons remain unclear, and there are active discussions of potential roles for post-implantation hematoma, circulating immune and stem cells, and pericytes, but studies on adipose-derived stem cells (AMSCs) in this context are lacking. The rough (average surface roughness = 2-5 µm) scaffold-like CaP coating deposited on pure titanium plates by the microarc oxidation method was used to investigate its subcutaneous vascularization in CBA/CaLac mice and in vitro effect on cellular and molecular crosstalk between human blood mononuclear cells (hBMNCs) and AMSCs (hAMSCs). Postoperative hematoma development on the CaP surface lasting 1-3 weeks may play a key role in the microvessel elongation and invasion into the CaP relief at the end of the 3rd week of injury and BMNC migration required for enhanced wound healing in mice. Satisfactory osteogenic and chondrogenic differentiation but poor adipogenic differentiation of hAMSCs on the rough CaP surface were detected in vitro by differential cell staining. The fractions of CD73 (62%), CD90 (0.24%), and CD105 (0.41%) BMNCs may be a source of autologous circulating stem/progenitor cells for the subcutis reparation, but allogenic hBMNC participation is mainly related to the effects of CD4 T cells co-stimulated with CaP coating on the in vitro recruitment of hAMSCs, their secretion of angiogenic and osteomodulatory molecules, and the increase in osteogenic features within the period of in vivo vascularization. Cellular and molecular crosstalk between BMNCs and AMSCs is a model of effective subcutis repair. Rough CaP surface enhanced angio- and osteogenic signaling between cells. We believe that preconditioning and/or co-transplantation of hAMSCs with hBMNCs may broaden their potential in applications related to post-implantation tissue repair and bone bioengineering caused by microarc CaP coating.

摘要

磷酸钙(CaP)材料并不总是能诱导异位血管生成和骨形成;其原因尚不清楚,目前对于植入后血肿、循环免疫细胞和干细胞以及周细胞的潜在作用存在积极的讨论,但在此背景下关于脂肪来源干细胞(AMSCs)的研究却很缺乏。采用微弧氧化法在纯钛板上沉积的粗糙(平均表面粗糙度 = 2 - 5 µm)支架状CaP涂层,用于研究其在CBA/CaLac小鼠皮下的血管生成情况以及对人血单核细胞(hBMNCs)和脂肪来源干细胞(hAMSCs)之间细胞和分子相互作用的体外影响。术后CaP表面持续1 - 3周的血肿形成可能在损伤后第3周结束时微血管伸长并侵入CaP凹陷以及小鼠伤口愈合增强所需的BMNC迁移中起关键作用。通过细胞差异染色在体外检测到hAMSCs在粗糙CaP表面具有令人满意的成骨和成软骨分化,但脂肪生成分化较差。CD73(62%)、CD90(0.24%)和CD105(0.41%)的BMNCs部分可能是皮下修复的自体循环干/祖细胞来源,但异体hBMNC的参与主要与CaP涂层共刺激的CD4 T细胞对hAMSCs的体外募集作用、它们分泌血管生成和骨调节分子以及体内血管生成期间成骨特征的增加有关。BMNCs和AMSCs之间的细胞和分子相互作用是有效的皮下修复模型。粗糙的CaP表面增强了细胞间的血管生成和骨生成信号。我们认为,hAMSCs与hBMNCs的预处理和/或共移植可能会拓宽它们在微弧CaP涂层引起的植入后组织修复和骨生物工程相关应用中的潜力。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3a37/7579197/cb737ea2d0c9/materials-13-04398-g001.jpg

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