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 Effect on Osteogenic Differentiation of Genetically Modified IL4 or PDGF-BB Over-Expressing and IL4-PDGF-BB Co-Over-Expressing Bone Marrow-Derived Mesenchymal Stromal Cells In Vitro

2021 ◽  
Vol 8 (11) ◽  
pp. 165
Author(s):  
Masanori Tsubosaka ◽  
Masahiro Maruyama ◽  
Elijah Ejun Huang ◽  
Ning Zhang ◽  
Takeshi Utsunomiya ◽  
...  
Keyword(s):  
Cell Proliferation ◽  
Osteogenic Differentiation ◽  
Mesenchymal Stromal Cells ◽  
Stromal Cells ◽  
Early Stage ◽  
Genetically Modified ◽  
Interleukin 4 ◽  
Alizarin Red ◽  
Cell Based Therapy

The use of genetically modified (GM) mesenchymal stromal cells (MSCs) and preconditioned MSCs (pMSCs) may provide further opportunities to improve the outcome of core decompression (CD) for the treatment of early-stage osteonecrosis of the femoral head (ONFH). GM interleukin-4 (IL4) over-expressing MSCs (IL4-MSCs), platelet-derived growth factor (PDGF)-BB over-expressing MSCs (PDGF-BB-MSCs), and IL4-PDGF-BB co-over-expressing MSCs (IL4-PDGF-BB-MSCs) and their respective pMSCs were used in this in vitro study and compared with respect to cell proliferation and osteogenic differentiation. IL4-MSCs, PDGF-BB-MSCs, IL4-PDGF-BB-MSCs, and each pMSC treatment significantly increased cell proliferation compared to the MSC group alone. The percentage of Alizarin red-stained area in the IL4-MSC and IL4-pMSC groups was significantly lower than in the MSC group. However, the percentage of Alizarin red-stained area in the PDGF-BB-MSC group was significantly higher than in the MSC and PDGF-BB-pMSC groups. The percentage of Alizarin red-stained area in the IL4-PDGF-BB-pMSC was significantly higher than in the IL4-PDGF-BB-MSC group. There were no significant differences in the percentage of Alizarin red-stained area between the MSC and IL4-PDGF-BB-pMSC groups. The use of PDGF-BB-MSCs or IL4-PDGF-BB-pMSCs increased cell proliferation. Furthermore, PDGF-BB-MSCs promoted osteogenic differentiation. The addition of GM MSCs may provide a useful supplementary cell-based therapy to CD for treatment of ONFH.

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.

scholarly journals 3D-printed nerve guidance conduits multi-functionalized with canine multipotent mesenchymal stromal cells promote neuroregeneration after sciatic nerve injury in rats

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Diego Noé Rodríguez-Sánchez ◽  
Giovana Boff Araujo Pinto ◽  
Luciana Politti Cartarozzi ◽  
Alexandre Leite Rodrigues de Oliveira ◽  
Ana Livia Carvalho Bovolato ◽  
...  
Keyword(s):  
Growth Factor ◽  
Sciatic Nerve ◽  
Nerve Injury ◽  
Mesenchymal Stromal Cells ◽  
Stromal Cells ◽  
Multipotent Mesenchymal Stromal Cells ◽  
Sciatic Nerve Injury ◽  
Motor Deficits ◽  
3D Printed

Abstract Background Nerve injuries are debilitating, leading to long-term motor deficits. Remyelination and axonal growth are supported and enhanced by growth factor and cytokines. Combination of nerve guidance conduits (NGCs) with adipose-tissue-derived multipotent mesenchymal stromal cells (AdMSCs) has been performing promising strategy for nerve regeneration. Methods 3D-printed polycaprolactone (PCL)-NGCs were fabricated. Wistar rats subjected to critical sciatic nerve damage (12-mm gap) were divided into sham, autograft, PCL (empty NGC), and PCL + MSCs (NGC multi-functionalized with 106 canine AdMSCs embedded in heterologous fibrin biopolymer) groups. In vitro, the cells were characterized and directly stimulated with interferon-gamma to evaluate their neuroregeneration potential. In vivo, the sciatic and tibial functional indices were evaluated for 12 weeks. Gait analysis and nerve conduction velocity were analyzed after 8 and 12 weeks. Morphometric analysis was performed after 8 and 12 weeks following lesion development. Real-time PCR was performed to evaluate the neurotrophic factors BDNF, GDNF, and HGF, and the cytokine and IL-10. Immunohistochemical analysis for the p75NTR neurotrophic receptor, S100, and neurofilament was performed with the sciatic nerve. Results The inflammatory environment in vitro have increased the expression of neurotrophins BDNF, GDNF, HGF, and IL-10 in canine AdMSCs. Nerve guidance conduits multi-functionalized with canine AdMSCs embedded in HFB improved functional motor and electrophysiological recovery compared with PCL group after 12 weeks. However, the results were not significantly different than those obtained using autografts. These findings were associated with a shift in the regeneration process towards the formation of myelinated fibers. Increased immunostaining of BDNF, GDNF, and growth factor receptor p75NTR was associated with the upregulation of BDNF, GDNF, and HGF in the spinal cord of the PCL + MSCs group. A trend demonstrating higher reactivity of Schwann cells and axonal branching in the sciatic nerve was observed, and canine AdMSCs were engrafted at 30 days following repair. Conclusions 3D-printed NGCs multi-functionalized with canine AdMSCs embedded in heterologous fibrin biopolymer as cell scaffold exerted neuroregenerative effects. Our multimodal approach supports the trophic microenvironment, resulting in a pro-regenerative state after critical sciatic nerve injury in rats.

scholarly journals Optimizing Osteogenic Differentiation of Ovine Adipose-Derived Stem Cells by Osteogenic Induction Medium and FGFb, BMP2, or NELL1 In Vitro

2018 ◽  
Vol 2018 ◽  
pp. 1-9 ◽  
Author(s):  
Emil Østergaard Nielsen ◽  
Li Chen ◽  
Jonas Overgaard Hansen ◽  
Matilda Degn ◽  
Søren Overgaard ◽  
...  
Keyword(s):  
Osteogenic Differentiation ◽  
Stromal Cells ◽  
Basic Medium ◽  
Induction Medium ◽  
Control Analysis ◽  
Alizarin Red ◽  
Osteogenic Induction Medium ◽  
Osteogenic Induction ◽  
Fibroblast Growth Factor Basic

Although adipose-derived stromal cells (ADSCs) have been a major focus as an alternative to autologous bone graft in orthopedic surgery, bone formation potential of ADSCs is not well known and cytokines as osteogenic inducers on ADSCs are being investigated. This study aimed at isolating ADSCs from ovine adipose tissue (AT) and optimizing osteogenic differentiation of ovine ADSCs (oADSC) by culture medium and growth factors. Four AT samples were harvested from two female ovine (Texel/Gotland breed), and oADSCs were isolated and analyzed by flow cytometry for surface markers CD29, CD44, CD31, and CD45. Osteogenic differentiation was made in vitro by seeding oADSCs in osteogenic induction medium (OIM) containing fibroblast growth factor basic (FGFb), bone morphogenetic protein 2 (BMP2), or NEL-like molecule 1 (NELL1) in 4 different dosages (1, 10, 50, and 100 ng/ml, respectively). Basic medium (DMEM) was used as control. Analysis was made after 14 days by Alizarin red staining (ARS) and quantification. This study successfully harvested AT from ovine and verified isolated cells for minimal criteria for adipose stromal cells which suggests a feasible method for isolation of oADSCs. OIM showed significantly higher ARS to basic medium, and FGFb 10 ng/ml revealed significantly higher ARS to OIM alone after 14 days.

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