Cell and Organ Transplantology. 2018; 6(2):131-136.
DOI: 10.22494/cot.v6i2.86
Potentiation of cryopreserved rat adipose-derived multipotent mesenchymal stromal cells by BMP-12 in vitro for the treatment of tendinopathy
Volkova N. O., Yukhta M.S., Goltsev A. M.
Institute of Problems of Cryobiology and Cryomedicine of the National Academy of Sciences of Ukraine, Kharkiv, Ukraine
Abstract
Bone morphogenetic proteins (BMP), which are part of TGF-b superfamily, have a stimulating effect on bone formation, as well as the proliferation of tenocytes and fibroblasts, which form the tendon tissue.
The purpose of the study was to determine the effect of BMP-12 on the morphofunctional characteristics of cryopreserved rat adipose tissuederived multipotent mesenchymal stromal cells (AT-MMSCs) and to investigate the effects of the use of AT-MMSCs pre-cultured with BMP-12 on the tendinopathy model.
Materials and methods. MMSCs were obtained from the adipose tissue of rats. Cryopreservation was carried out under the protection of 10 % DMSO with the addition of 20 % fetal bovine serum at a cooling rate of 1 °C/min to -80 °C and subsequent transfer to liquid nitrogen. When culturing AT-MMSCs, the BMP-12 was added at the concentration of 50 ng/mL. The ability of cells to proliferation (by MTT-test) and the synthesis of collagen I and III types were evaluated. Rats with moderate degenerative-dystrophic damage to the Achilles tendons were locally administered with 0.25·106 AT-MMSCs or AT-MMSCs+BMP-12 in the defect zone. The animals injected with saline provided control group. On the 21st day after the treatment, a histological, immunofluorescence and biomechanical studies were performed.
Results. The application of BMP-12 results in the decrease in proliferation of AT-MMSCs along with an increase in the relative number of cells that synthesize collagen I and III types relative to AT-MMSCs cultivated under standard conditions. The injection of AT-MMSCs promotes the activation of regenerative processes in damaged tendons. The use of AT-MMSCs+BMP-12 accelerates the histological structure, strength, and the content of collagen I and III types in the Achilles tendons of animals with degenerative-dystrophic damage compared to the cells without BMP-12.
Conclusions BMP-12 can be used as a stimulating agent for the tenogenic differentiation of AT-MMSCs before transplantation into the damaged tendon tissue.
Key words: adipose-derived multipotent mesenchymal stromal cells; BMP-12; collagen; tendinopathy; cell therapy
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Volkova N, Yukhta M, Goltsev A. Potentiation of cryopreserved rat adipose-derived multipotent mesenchymal stromal cells by BMP-12 in vitro for the treatment of tendinopathy. Cell and Organ Transplantology. 2018; 6(2):131-136. doi:10.22494/cot.v6i2.86
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