Dynamics of indicators of markers of bone metabolism in bone defect replacement fabric equivalents of bone tissue on the basis of mmsc-at

  • A. V. Bambuliak Bukovinian State Medical University
  • N. B. Kuznyak Bukovinian State Medical University
  • R. R. Dmitrenko Bukovinian State Medical University
  • V. A. Goncharenko Bukovinian State Medical University
Keywords: multipotent mesenchymal stromal cells of adipose tissue, alkaline phosphatase, acid phosphatase

Abstract

Summary. Multipotent mesenchial stromal cells of adipose tissue (MMSC-АT) are capable of differentiation in the adipogenic, osteogenic, chondrogenic, endothelial, myogenic, hepatogenic, epithelial and neurogenic regions. Since bone tissue healing is done by replacing the defect with the connective tissue, our task was transplantation of multipotent stem cells, which in future will be differentiated into proper bone tissue. The questions of osteogenesis and processes of mineralization of jaw bone in dental interventions are relevant. To enzymes that are involved in the regulation of phosphorous-calcium metabolism and have a direct effect on bone resorption and regeneration processes (both physiological and reparative) that occur in the bone, include acid and alkaline phosphatase.

The aim of the study – to determine changes in bone remodeling rates when bone defects are filled with tissue equivalents of bone tissue based on multipotent mesenchymal stromal cells of adipose tissue (MMSC-AT).

Materials and Methods. The experiment was conducted on the Wistar line rats, weighing 200–250 grams, which were divided into VI groups. A bone defect model was formed in the parietal section of the skull of rats. The formed defect implanted the harvested material. The activity of alkaline phosphatase in blood of rats was investigated by a unified method using the kit "Alkaline Phosphatase-02-Vital" (fm "Vital Diagnostics, Spb", St. Petersburg). The total phosphorus acid in animal blood was investigated by photometric optimized kinetic method using the kit "Acid Phosphatase-02-Vital" (fm "Vital Diagnostics, Spb", St. Petersburg). Blood from the caudal vein of the animals was collected in 1, 2, 3 months of the experiment. The obtained results are processed statistically.

Results and Discussion. At the 90th day of observation in the blood of experimental animals, an increase in activity of alkaline phosphatase was investigated. At the same time, in the animals of groups IV and VI, the values of the studied index were maximum (14.49 ± 0.08) mmol/s • l, p1 - p2˂0.01 and 14.74 ± 0.09 mmol/s • l, p1 - p2˂0.01, p3˃0.05, p4˂0.01, respectively). In animals, the rest of the study groups, after 3 months of observation, the value of the analyzed parameters was lower than the data in the group I animals: 20.37 % in group II, p ˂ 0.01, 11.08 % – in group II, p – p1˂0.1, and by 18.91 % in group V, p˂0.01, p2-p3˂0.01, p2˃0.05. This tendency emphasizes that during this observation period, the activity of LF reaches the maximum values, contributing to the extracellular matrix and mucopolysaccharides synthesis, fibrillar protein synthesis and deposition of mineral deposits.

Conclusions. Thus, the studies conducted by us have shown the ability of multipotent mesenchymal fatty tissue cells (MMSC-AT) to stimulate osteogenesis processes, mainly affecting the mineralizing function, as evidenced by an increase in alkaline phosphatas / phosphorus acid.

References

Pittenger, M.F., Mackay, A.M., & Beck, S.C. (2015). Multilineage potential of adult human mesenchymal stem cells. Science, 284, 143-147.

Gimble, J., & Guilak, F. (2013). Adipose-derived adult stem cells: isolation, characterization, and differentiation potential. Cytotherapy, 5, 362-369.

Bongso, A., Fong, C.Y., & Ng, S.C. (2014). Isolation and culture of inner cell mass cells from human blastocysts. Human Reproduction, 9, 2110-2117.

Thomson, J.A., Itskovitz-Eldor, J., & Shapiro, S.S. (2014). Embryonic stem cell lines derived from human blastocysts. Science, 282, 1145-1147.

Yamanaka, S. (2017). Pluripotency and nuclear reprogramming. Philosophical Transactions of the Royal Society of London. Series B: Biological Sciences, 363, 2079-2087

Hillmann J.Z.(2015). Klin. Chem. Biochem., 9, 273.

Stepan, J.J., Silinkova-Malkova, E., & Havrenek, T. (2013). Relationship of plasma tartrate-resistant acid phosphatase to the bone isoenzyme of serum alkaline phosphatase in hyperparathyroidism. Clin. Chim. Acta., 133 (2), 189-200.

Goryachkovskiy, A.M. (2015). Klinicheskaya biokhimiya v laboratornoy diagnostike [The clinical biochemistry in laboratorial diagnostics]. Odessa: Ekologiya [in Russian].

Levitsky, A.P., Makarenko, O.A., & Denga, O.V. (2011). Eksperimentalnye metody issledovaniya stimulyatorov osteogeneza: metodicheskie rekomendatsii [The experimental methods of the study of osteogenesis stimulators]. Kyiv: GFTs MZU [in Russian].

Levytskyi, A.P., Makarenko, O.A., & Khodakov, I.V. (2013). Fermentatyvnyi metod otsinky stanu kistkovoi tkanyny [The enzymatic method of the estimation of the state of osseous tissue]. Оdеskyi medychnyi zhurnal – Odesa Medical Journal, 3, 17-21 [in Ukrainian].

Winter, A., Breit, S., & Parsch, D. (2014). Cartilage-like gene expression in differentiated human stem cell spheroids: a comparison of bone marrow-derived and adipose tissue-derived stromal cells. Arthritis and Rheumatism, 48, 418-429.

Mesimäki, K., Lindroos, B., & Törnwall, J. (2009). Novel maxillary reconstruction with ectopic bone formation by GMP adipose stem cells. International Journal of Oral and Maxillofacial Surgery, 38, 201-209.

Pereira, R.F., O’Hara, M.D., & Laptev, A.V. (2011). Marrow stromal cells as a source of progenitor cells for nonhematopoietic tissues in transgenic mice with a phenotype of osteogenesis imperfecta. Proceedings of the National Academy of Sciences of the United States of America, 95, 1142-1147.

Horwitz, E.M., Gordon, P.L., & Koo, W.K. (2016). Isolated allogeneic bone marrow-derived mesenchymal cells engraft and stimulate growth in children with osteogenesis imperfecta: implications for cell therapy of bone. Proceedings of the National Academy of Sciences of the United States of America, 99, 8932-8937.

Keating, A., Berkahn, L., & Filshie, R. (2015). A Phase I study of the transplantation of genetically marked autologous bone marrow stromal cells. Human Gene Therapy, 9, 591-600.

Published
2019-11-08
How to Cite
Bambuliak, A. V., Kuznyak, N. B., Dmitrenko, R. R., & Goncharenko, V. A. (2019). Dynamics of indicators of markers of bone metabolism in bone defect replacement fabric equivalents of bone tissue on the basis of mmsc-at. Clinical Dentistry, (3), 68-75. https://doi.org/10.11603/2311-9624.2019.3.10568
Section
Experimental researches