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

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

doi:10.3390/ma13194398

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

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

Authors

Igor A Khlusov^{1

2

3}, Larisa S Litvinova¹, Valeria V Shupletsova¹, Olga G Khaziakhmatova¹, Vladimir V Malashchenko¹, Kristina A Yurova¹, Egor O Shunkin¹, Vasilii V Krivosheev¹, Ekaterina D Porokhova³, Anastasiia E Sizikova³, Linara A Safiullina³, Elena V Legostaeva⁴, Ekaterina G Komarova⁴, Yurii P Sharkeev^{4

5}

Affiliations

¹ Center for Immunology and Cell Biotechnology, Immanuel Kant Baltic Federal University, 236029 Kaliningrad, Russia.
² Research School of Chemistry and Applied Biomedical Sciences, National Research Tomsk Polytechnic University, 634050 Tomsk, Russia.
³ Department of Morphology and General Pathology, Siberian State Medical University, 634050 Tomsk, Russia.
⁴ Laboratory of Physics of Nanostructured Biocomposites, Institute of Strength Physics and Materials Science SB RAS (ISPMS SB RAS), 634055 Tomsk, Russia.
⁵ Research School of High-Energy Physics, National Research Tomsk Polytechnic University, 634050 Tomsk, Russia.

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 R_a = 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.

Keywords: cytokine/chemokine secretion; human cells; immunophenotype; in vitro modeling; mice; microarc calcium phosphate coating; motility; osteogenic differentiation; subcutaneous behavior of implants; surface morphology.

Grants and funding

16-15-10031/Russian Science Foundation