Secretion of Hematopoietic Niche Signal Molecules under Conditions of Osteogenic Differentiation of Multipotent Mesenchymal Stromal Cells Induced By Relief Calcium Phosphate Coating

2019 ◽  
Vol 13 (4) ◽  
pp. 341-348 ◽  
Author(s):  
L. S. Litvinova ◽  
V. V. Shupletsova ◽  
K. A. Yurova ◽  
O. G. Khaziakhmatova ◽  
N. M. Todosenko ◽  
...  
Keyword(s):  
Calcium Phosphate ◽  
Osteogenic Differentiation ◽  
Mesenchymal Stromal Cells ◽  
Stromal Cells ◽  
Multipotent Mesenchymal Stromal Cells ◽  
Calcium Phosphate Coating ◽  
Signal Molecules ◽  
Phosphate Coating ◽  
Hematopoietic Niche

scholarly journals Secretion of niche signal molecules in conditions of osteogenic differentiation of multipotent mesenchymal stromal cells induced by textured calcium phosphate coating

2019 ◽  
Vol 65 (4) ◽  
pp. 339-346
Author(s):  
L.S. Litvinova ◽  
V.V. Shupletsova ◽  
K.A. Yurova ◽  
O.G. Khaziakhmatova ◽  
N.M. Todosenko ◽  
...  
Keyword(s):  
Calcium Phosphate ◽  
Osteogenic Differentiation ◽  
Mesenchymal Stromal Cells ◽  
Stromal Cells ◽  
Multipotent Mesenchymal Stromal Cells ◽  
Calcium Phosphate Coating ◽  
Signal Molecules ◽  
Humoral Factors ◽  
Alizarin Red

Secretion of 21 cytokines, chemokines and growth factors (LIF, SCF, SDF-1a, SCGF-b, M-CSF, MCP-3, MIF, MIG, TRAIL, GRO-a; IL-1a, IL-2ra, IL-3, IL-12(p40), IL-16, IL-18, HGF, TNF-b, b-NGF, IFN-a2, CTACK) has been studied in vitro in the culture of human adipose-derived multipotent mesenchymal stromal cells (hAMMSCs) in conditions of its osteogenic differentiation caused by 14-day contact with calcium phosphate (CP) surface with different roughness. Bilateral X-ray amorphous CP coatings were prepared on the samples of commercially pure titanium in the anodal regime using a micro-arc method. An aqueous solution prepared from 20 wt% phosphoric acid, 6 wt% dissolved hydrohyapatite nanopowder (particle diameter 10-30 nm with single agglomerates up to 100 nm), and 9 wt% dissolved calcium carbonate was used to obtain CP coating. hAMMSCs isolated from lipoaspirate were co-cultured after 4 passages with the CP-coated samples at final concentration of 1.5´105 viable karyocytes per 1.5 mL of standard nutrition medium (without osteogenic stimulators) for 14 days (a determination of [CD45,34,14,20], CD73, CD90 и CD105 cell immunophenotype; an analysis of secretory activity) and 21 days (alizarin red S staining of culture) with medium replacement every 3-4 days. Under conditions of in vitro contact with rough CP coating hAMMSCs differentiated into osteoblasts synthesizing the mineralized bone matrix; this was accompanied by 2-3-fold increasing ratio of [CD45,34,14,20]+ hemopoietic cells. The following humoral factors of hemopoietic niches acted as the signal molecules escalating in vitro the hemopoietic base in 14 days of differentiating three-dimensional culture of hAMMSCs: either leukemia inhibitory factor (LIF) and stem cell factor (SCF) cytokines under mean index of CP roughness Ra=2.4-2.6 mm or stromal derived factor-1 (SDF-1a, CXCL12 chemokine) under Ra=3.1-4.4 mm.

scholarly journals Biologic Characteristics of Bone Substituting Tissue Engineering Construction Based on Calcium Phosphate Ceramics, Autologous Mesenchymal Stromal Cells and Fibrin Hydrogel

2015 ◽  
pp. 52-59
Author(s):  
V. E. Mamonov ◽  
A. G. Chemis ◽  
V. S. Komlev ◽  
A. L. Berkovskiy ◽  
E. M. Golubev ◽  
...  
Keyword(s):  
Tissue Engineering ◽  
Calcium Phosphate ◽  
Osteogenic Differentiation ◽  
Mesenchymal Stromal Cells ◽  
Stromal Cells ◽  
Multipotent Mesenchymal Stromal Cells ◽  
Orthotopic Implantation ◽  
Calcium Phosphate Ceramic ◽  
Engineering Construction

Biological characteristics of bone substituting tissue engineering construction (TEC) that contained porous calcium phosphate ceramic granulate (CPC) of phase structure ((tricalcium phosphate (TCP)), fibrin hydrogel and autologous multipotent mesenchymal stromal cells (auto-MMSC) induced and non-induced to osteogenic differentiation were studied in vivo. The following characteristics of TEC were determined: ability to transfer within its structure the viable auto-MMSC with preservation of their regeneration potential; ability to osteogenesis only under conditions of orthotopic implantation; ability of induced to osteogenic differentiation auto-MMSC to participate in the reparative processes for not more than within 6 weeks after implantation; negative affect of fibrin hydrogel on the osteoinductive properties of CPC within TCP structure. It was shown that to provide osteogenesis in the implanted TEC not only the viable auto-MMSC but simultaneous presence of osteoinductive and osteoconductive factors was required. No bone formation in a critical bone defect and in ectopic implantation takes place without observance of these conditions.

scholarly journals Biologic Characteristics of Bone Substituting Tissue Engineering Construction Based on Calcium Phosphate Ceramics, Autologous Mesenchymal Stromal Cells and Fibrin Hydrogel

2015 ◽  
Vol 22 (4) ◽  
pp. 52-59
Author(s):  
V. E Mamonov ◽  
A. G Chemis ◽  
V. S Komlev ◽  
A. L Berkovskiy ◽  
E. M Golubev ◽  
...  
Keyword(s):  
Tissue Engineering ◽  
Calcium Phosphate ◽  
Osteogenic Differentiation ◽  
Mesenchymal Stromal Cells ◽  
Stromal Cells ◽  
Multipotent Mesenchymal Stromal Cells ◽  
Orthotopic Implantation ◽  
Calcium Phosphate Ceramic ◽  
Engineering Construction

Biological characteristics of bone substituting tissue engineering construction (TEC) that contained porous calcium phosphate ceramic granulate (CPC) of phase structure ((tricalcium phosphate (TCP)), fibrin hydrogel and autologous multipotent mesenchymal stromal cells (auto-MMSC) induced and non-induced to osteogenic differentiation were studied in vivo. The following characteristics of TEC were determined: ability to transfer within its structure the viable auto-MMSC with preservation of their regeneration potential; ability to osteogenesis only under conditions of orthotopic implantation; ability of induced to osteogenic differentiation auto-MMSC to participate in the reparative processes for not more than within 6 weeks after implantation; negative affect of fibrin hydrogel on the osteoinductive properties of CPC within TCP structure. It was shown that to provide osteogenesis in the implanted TEC not only the viable auto-MMSC but simultaneous presence of osteoinductive and osteoconductive factors was required. No bone formation in a critical bone defect and in ectopic implantation takes place without observance of these conditions.

Induction of Osteogenic Differentiation of Multipotent Mesenchymal Stromal Cells from Human Adipose Tissue

2013 ◽  
Vol 155 (1) ◽  
pp. 145-150 ◽  
Author(s):  
L. V. Logovskaya ◽  
T. B. Bukharova ◽  
A. V. Volkov ◽  
E. B. Vikhrova ◽  
O. V. Makhnach ◽  
...  
Keyword(s):  
Adipose Tissue ◽  
Osteogenic Differentiation ◽  
Mesenchymal Stromal Cells ◽  
Stromal Cells ◽  
Human Adipose Tissue ◽  
Multipotent Mesenchymal Stromal Cells

scholarly journals Microscopic Investigation of Compatibility of Samples Containing Multipotent Mesenchimal Stromal Cells of Additive Tissue in Experimental Conditions

2020 ◽  
Vol 5 (6) ◽  
pp. 59-65
Author(s):  
A.V. Bambuliak ◽  
◽  
N.B. Kuzniak ◽  
R.R. Dmitrenko ◽  
S.V. Tkachik ◽  
...  
Keyword(s):  
Adipose Tissue ◽  
Osteogenic Differentiation ◽  
Mesenchymal Stromal Cells ◽  
Stromal Cells ◽  
Multipotent Mesenchymal Stromal Cells ◽  
Tooth Surface ◽  
Growth Surface ◽  
Experimental Conditions ◽  
Plasma Clot ◽  
Experimental Animals

The purpose of the study was to investigate the biocompatibility of samples containing multipotent mesenchymal stromal cells of adipose tissue to replace bone defects. Material and methods. The study was conducted at Bukovina State Medical University, Chernivtsi, Ukraine. Adipose tissue samples were obtained from the neck of 60 experimental animals (white Wistar rats). We selected 4 samples for the toxicological experiment, which allowed to establish the direct influence of factors in the contact of implantation material at the cellular level. Sample № 1 - Multipotent mesenchymal stromal cells of adipose tissue, which underwent osteogenic differentiation; № 2 - Multipotent mesenchymal stromal cells of adipose tissue with osteogenic differentiation with the addition of platelet-enriched blood plasma; № 3 - “Kolapan” with applied tissue culture of Multipotent mesenchymal stromal cells of adipose tissue cells, which underwent osteogenic differentiation; № 4 - "Kolapan" + Multipotent mesenchymal stromal cells of adipose tissue + platelet-enriched plasma. Multipotent mesenchymal stromal cells of adipose tissue were obtained by grinding adipose tissue of rats in 0.1% collagenase 1A [14]. The study of biocompatibility by cell culture in vitro was performed in accordance with the Working Instruction № 04/2013-VL. The cultures were investigated by the explantation method in a plasma clot in Karelian vials. In order to standardize the nature of growth, their zones were classified into compact, reticular and migrating cells of growing fibroblastic tissues. To assess the probability of the obtained results of the study we used a variation-statistical method of analysis using Microsoft Excel. Statistical calculation of the results of clinical and laboratory studies was carried out according to conventional methods. Results and discussion. Microscopic examination of the surface of samples with culture of fibroblasts showed their satisfactory adhesion on the tooth surface after 5 days of cultivation. In the study of sample № 1 (Multipotent mesenchymal stromal cells of adipose tissue, with osteogenic differentiation), it was noted that the structure of the cells acquired a rounded and oval shape, which indicated their destruction and damage. On the 5th day of observation, cells with numerical intussusception and processes were observed during visual examination of sample № 3 (“Kolapan”, with applied culture of Multipotent mesenchymal stromal cells of adipose tissue). In the study of samples № 4 (Multipotent mesenchymal stromal cells of adipose tissue + platelet-enriched plasma + "Kolapan") on the 5th day of research, signs of growth were manifested by migration of fibroblastic elements that had a spindle-shaped and polygonal shape, with the formation of the primary zone due to strands. On the 7th day of cultivation in experimental samples № 2, № 3, № 4 there was the formation of three growth zones: compact - from cells of polygonal and spindle-shaped form; reticulate - from strands and bundles of cells that were located reticulate and areas of single migrating elements of spindle-shaped. External characteristics and cell growth surface did not differ from control samples. On the 10th day of cultivation in the experimental samples, as well as in the control, the areas of compact and reticular growth zone and the zone of migrating fibroblasts were increased. At the same time, tissue-like growth of cells was observed. Visualization of compact and stack-like zones of the studied experimental samples revealed signs of the beginning of degenerative changes, which was characterized in the form of rounding of the shape and vacuolation of cells. This trend was most pronounced in samples № 2 and № 4. Conclusion. Thus, tissue equivalents of bone tissue based on Multipotent mesenchymal stromal cells of adipose tissue can be candidates for use in regenerative medicine, and studies of their application in experimental animals will provide an opportunity to expand the understanding of the characteristics of Multipotent mesenchymal stromal cells of adipose tissue in order to optimize their further clinical application and implement new approaches in different areas of dentistry

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