Potentiation of cryopreserved rat adipose-derived multipotent mesenchymal stromal cells by BMP-12 in vitro for the treatment of tendinopathy

Home/2018, Vol. 6, No. 2/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

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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1. Boyer MI, Goldfarb CA, Gelberman RH. Recent progress in flexor tendon healing: the modulation of tendon healing with rehabilitation variables. Journal of Hand Therapy. 2005; 18(2):80-85.
https://doi.org/10.1197/j.jht.2005.01.009
PMid:15891963
2. Graham R. Tendinopathy – from basic science to treatment. Nature Clinical Practice Rheumatology. 2008; 4(2):82-89.
https://doi.org/10.1038/ncprheum0700
PMid:18235537
3. Maffulli N. Overuse tendon conditions: time to change a confusing terminology. Arthroscopy: The Journal of Arthroscopic & Related Surgery. 1998; 14(8):840-843.
https://doi.org/10.1016/S0749-8063(98)70021-0
4. Sharma P, Maffulli N. Tendon injury and tendinopathy: healing and repair. JBJS. 2005; 87(1):187-202.
https://doi.org/10.2106/JBJS.D.01850
https://doi.org/10.2106/00004623-200501000-00030
5. Dyment NA, Galloway JL. Regenerative biology of tendon: mechanisms for renewal and repair. Current molecular biology reports. 2015; 1(3):124-131.
https://doi.org/10.1007/s40610-015-0021-3
PMid:26389023 PMCid:PMC4570727
6. Tozer S, Duprez D. Tendon and ligament: development, repair and disease. Birth Defects Res C Embryo Today. 2005; 75(3):226-236.
https://doi.org/10.1002/bdrc.20049
PMid:16187327
7. Docheva D, Müller SA, Majewski M, Evans CH. Biologics for tendon repair. Advanced drug delivery reviews. 2015; 84:222-239.
https://doi.org/10.1016/j.addr.2014.11.015
PMid:25446135 PMCid:PMC4519231
8. Voleti PB, Buckley MR, Soslowsky LJ. Tendon healing: repair and regeneration. Annu Rev Biomed Eng. 2012; 14:47-71.
https://doi.org/10.1146/annurev-bioeng-071811-150122
PMid:22809137
9. Fang F, Lake S. Experimental evaluation of multiscale tendon mechanics. Journal of Orthopaedic Research. 2017; 35(7):1353-1365.
https://doi.org/10.1002/jor.23488
PMid:27878999
11. Hegyi B, Sagi1 B, Kovacs J. Identical, similar or different? Learning about immunomodulatory function of mesenchymal stem cells isolated from various mouse tissues: bone marrow, spleen, thymus and aorta wall. Int Immunology. 2010; 22(7):551-559.
https://doi.org/10.1093/intimm/dxq039
PMid:20497958
12. Volkova N, Yukhta M, Goltsev A. Cryopreserved Mesenchymal Stem Cells Stimulate Regeneration in an Intervertebral Disc. Biomedicines. 2015; 3(3):237-247.
https://doi.org/10.3390/biomedicines3030237
PMid:28536410 PMCid:PMC5344241
13. Rybachuk ОА, Кyryk VМ, Poberezhny PA, et al. Еffect of the bone marrow multipotent mesenchimal stromal cells to the neural tissue after ischemic injury in vitro. Cell and Organ Transplantology. 2014; 2(1):74-78.
https://doi.org/10.22494/COT.V2I1.38
14. Volkova NА, Yukhta MS, Yurchuk ТА, et al. Multipotent mesenchymal stromal cells of bone marrow in therapy of chronic inflammation of murine ovaries. Biotechnologia Acta. 2014; 7(5):35-42.
https://doi.org/10.15407/biotech7.05.035
15. Molloy T, Wang Y, Murrell GAC. The roles of growth factors in tendon and ligament healing. Sports Medicine. 2003; 33(5):381-394.
https://doi.org/10.2165/00007256-200333050-00004
PMid:12696985
16. Inada M, Katagiri Т, Akiyama S, et al. Bone morphogenetic protein-12 and -13 inhibit terminal differentiation of myoblasts, but do not induce their differentiation into osteoblasts. Biochem Biophys Res Commun. 1996; 222(2):317-322.
https://doi.org/10.1006/bbrc.1996.0742
PMid:8670203
17. Costa MA, Wu C, Pham BV, et al. Tissue engineering of flexor tendons: optimization of tenocyte proliferation using growth factor supplementation. Tissue engineering. 2006; 12(7):1937-1943.
https://doi.org/10.1089/ten.2006.12.1937
PMid:16889523
18. Folkman J, Klagsbrun M. Angiogenic factors. Science. 1987; 235(4787):442-447.
https://doi.org/10.1126/science.2432664
PMid:2432664
19. Dai L, Hu X, Zhang X, et al. Different tenogenic differentiation capacities of different mesenchymal stem cells in the presence of BMP-12. Journal of translational medicine. 2015; 13(1):200.
https://doi.org/10.1186/s12967-015-0560-7
PMid:26104414 PMCid:PMC4479325
20. Volkovа NA, Yukhta MS, Goltsev AN. Morphological and functional characteristics of cryopreserved multipotent mesenchymal stromal cells from bone marrow, adipose tissue and tendons. Cell and Organ Transplantology. 2016; 4(2):200-205.
https://doi.org/10.22494/cot.v4i2.64
22. Violini S, Ramelli P, Pisani LF, et al. Horse bone marrow mesenchymal stem cells express embryo stem cell markers and show the ability for tenogenic differentiation by in vitro exposure to BMP-12. BMC cell biology. 2009; 10(1):29.
https://doi.org/10.1186/1471-2121-10-29
PMid:19383177 PMCid:PMC2678092
23. Zarychta-Wiśniewska W, Burdzinska A, Kulesza A, et al. BMP-12 activates tenogenic pathway in human adipose stem cells and affects their immunomodulatory and secretory properties. BMC cell biology. 2017; 18(1):13.
https://doi.org/10.1186/s12860-017-0129-9
PMid:28214472 PMCid:PMC5316159
24. Liu J, Tao X, Chen L, et al. CTGF positively regulates BMP-12 induced tenogenic differentiation of tendon stem cells and signaling. Cellular Physiology and Biochemistry. 2015; 35(5):1831-1845.
https://doi.org/10.1159/000373994
PMid:25833297
25. Mossman T. Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol Methods. 1983; 65(1-2):55-63.
https://doi.org/10.1016/0022-1759(83)90303-4
26. Council of Europe [France]. European convention for the protection of vertebrate animals used for experimental and other scientific purposes. Strasbourg, 18.III.1986. Available from: http://conventions.coe.int/treaty/en/Treaties/Word/123.doc
29. Stewart A, Guan H, Yang K. BMP-3 promotes mesenchymal stem cell proliferation through the TGF-beta/activin signaling pathway. J Cell Physiol. 2010; 223(3):658-66.
PMid:20143330
30. Rodrigues M, Griffith LG, Wells. Growth factor regulation of proliferation and survival of multipotential stromal cells. Stem cell research & therapy. 2010; 1(4):32.
https://doi.org/10.1186/scrt32
PMid:20977782 PMCid:PMC2983445

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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