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

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 Ra = 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.

Zn-Doped CaP-Based Coatings on Ti–6Al–4V and Ti–6Al–7Nb Alloys Prepared by Magnetron Sputtering: Controllable Biodegradation, Bacteriostatic, and Osteogenic Activities

“…Informed consent for the procedure was obtained from the volunteer prior to his participation in the study, according to [26]. Isolation of stromal cells and confirmation of their multilineage differentiation into osteoblasts, chondrocytes, and adipocytes were performed as previously described [27,28].…”

Calcium Chelidonate: Semi-Synthesis, Crystallography, and Osteoinductive Activity In Vitro and In Vivo

“…1 on 28 February 2019) were obtained. A stromal vascular fraction (SVF) and processed lipoaspirate (PLA) were isolated according to Zuk et al (2001) [ 47 ] as described previously [ 48 ]. To define the multipotent origin of viable fibroblast-like adherent PLA cells, the minimal criteria of the International Society for Cellular Therapy (ISCT) and the International Federation for Adipose Therapeutics and Science (IFATS) [ 49 , 50 ] were used.…”

“…Cell immunophenotype (CD105, CD73, and CD90 positive markers (>90%) and lack (<2%) expression of CD45, CD34, CD20, or CD14) were detected. Multilineage differentiation of cells into osteoblasts, adipocytes and chondrocytes with reagent using a StemPro ® Differentiation Kit (Thermo Fisher Scientific, Waltham, MA, USA) as well differential staining by alizarin red S (Sigma-Aldrich, St. Louis, MO, USA), alcian blue (Sigma-Aldrich), and oil red (Sigma-Aldrich) was confirmed after 21 days of culture as described elsewhere [ 32 , 48 ]. Thus, the isolated cells constitute a pool of human adipose-derived multipotent mesenchymal stromal/stem cells (hAMMSCs).…”

“…There were a few reports stating that CaP scaffolds without cells had not new bone formation after subcutaneous implantation [ 48 , 54 ]. Hence, well–known variant of ectopic test with syngeneic bone marrow showed reproducible (75–100%) bone/marrow remodeling on micro-arc CaP coating implanted subcutaneously [ 42 , 55 ].…”

“…For implantation, 10 titanium plates ((10 × 10 × 1) mm 3 ) were prepared from pure titanium VT1-0. Titanium plates were coated with calcium phosphate (CaP) by microarc oxidation (MAO) using the Microarc 3.0 system (ISPMS SB RAS, Tomsk, Russia) as described earlier [ 48 ]. Before the experiment, the plates were sterilized by dry heat in the Binder FD53 oven (Binder GmbH, Tuttlingen, Germany) at 453 K for 1 h.…”