S.V. Nadezhdin - Ph.D. (Biol.), Associate Professor, Department of Biology, Belgorod National Research University
Yu.Ye. Burda - Ph.D. (Med.), Associate Professor, Department of Pharmacology, Belgorod National Research University
E.V. Zubareva - Ph.D. (Biol.), Associate Professor, Department of Biology, Belgorod National Research University
E-mail address: email@example.com
V.V. Botvin - Head of Laboratory of Polymers and Composite Materials, National Research Tomsk State University
L.A. Pokrovskaya - Head of Medical and Pharmaceutical Projects, Engineering Chemical Technological Сenter, National Research Tomsk State University
A.D. Latypov - Junior Research Scientist, Laboratory of Polymers and Composite Materials, National Research Tomsk State University
N.N. Volkovnyak - Junior Research Scientist, Laboratory of Biomaterials, Scientific-Educational and Innovative Centre «Nanostructured Materials and Nanotechnologies, Belgorod National Research University
E.A. Grebtsova - Engineer, Laboratory of Biomaterials, Scientific-Educational and Innovative Centre «Nanostructured Ma-terials and Nanotechnologies», Belgorod National Research University
V.S. Beliaeva - Student, Belgorod National Research University
Osteoplastic materials on the basis of lactide and glycolide copolymers and calcium phosphates are characterized by the properties of lactide and glycolide copolymers to dissolve gradually in physiological liquids of the organism during the for-mation of new tissue and to remove from the organism while calcium phosphates are characterized by the osteoconductive and osteoinductive properties.
The purpose of the research was the estimation of biocompatibility, biodegradation and osteogenic properties of composite material on the basis of lactide and glycolide copolymers and nanohydroxyapatite (nHAP).
During the investigation the composite material on the basis of lactide and glycolide copolymers and nanohydroxyapatite had been produced.
In vitro experiments were carried out on the mesenchymal stem cells (MSC) of the rats red bone marrow. Differentiation of the MSC in osteogenic direction was estimated by the calcium accumulation in the cells, with this purpose the cells were fixed with 10% formalin solution and dyed with 10% alizarin red solution after two weeks of cultivation.
In vivo experiments performed in accordance with requirements of ISO 10993-2:2006, ISO 10993-6:2007. The research was carried out on 8 male rats of an inbred Wistar strain, each weighing less than 350 g. Plates from composite material were inoculated under the skin at the back of animals. Length of the experiment was 21 day. Animals were sacrificed by an overdose of ether vapors. Histological specimens were prepared from the composite material covered with connective-tissue capsule.
During investigation of dried plates of composite material using SEM it was revealed that nHAP particles in copolymer ma-trix represented as conglomerates with the size of 5-20 µm, there were air sinuses with the size of 50-100 µm in the struc-ture of composite material.
In the in vitro experiments it was determined that material could be exposed to biodegradation, characterized as biocom-patible because mesenchymal stem cells actively proliferated and moved all over the plate crawling on and covering the fragments of the material. As a result of cells staining with alizarin red it was determined that calcium phosphate accumu-lated in the cells. Loci of ossification were discovered and it was the evidence of MSC differentiation in an osteogenic direc-tion.
In the in vivo experiments during macroscopic study of the place of subcutaneous inoculation of composite material in ex-perimental group external signs of inflammatory reaction and destructive changes of cutaneous covering at the defect zone were not discovered.
During the investigation it was determined that composite material on the basis of lactide and glycolide copolymer and na-nosized hydroxyapatite was biocompatible, had osteoconductive properties, could be exposed to biodegradation in the in vitro and in vivo experiments. Presence of pores and air sinuses in the matrix of the material promoted its biodegradation and stepwise invasion of connective-tissue structures and cellular elements deep into the material. Decomposition products of composite material matrix did not give negative influence on cyto/histotypic differentiation of cells and tissues. During biodegradation of the copolymer particles of hydroxyapatite become bioavailable and this potentiate differentiation of mesenchymal stem cells in osteogenic direction.
Composite material based on lactide and glycolide copolymers and nanohydroxyapatite can be used for development of porous three-dimensional matrixes which can serve as carrier of cellular material during bone defect replacement in osteo-regenerative therapy.
- Dimitriou R., Mataliotakis G.I., Calori G.M., Giannoudis P.V. The role of barrier membranes for guided bone regeneration and restoration of large bone defects: current experimental and clinical evidence // BMC Med. 2012. V. 10. P. 81. http://www.biomedcentral.com/1741-7015/ 10/81/prepub (data obrascheniya: 05.08.2017).
- Uskoković V., Hoover C., Vukomanović M., Uskoković D.P., Desai T.A. Osteogenic and antimicrobial nanoparticulate calcium phosphate and poly-(D,L-lactide-co-glycolide) powders for the treatment of osteomyelitis // Mater. Sci. Eng. C. Mater. Biol. Appl. 2013. V. 33. № 6. P. 33623373.
- Mandal B.B., Grinberg A., Gil E.S., Panilaitis B., Kaplan D.L. High-strength silk protein scaffolds for bone repair // Proc. Natl. Acad. Sci. USA. 2012. V. 109. № 20. P. 76997704. www.pnas. org/cgi/doi/10.1073/pnas.1119474109 (data obrascheniya: 05.08.2017).
- Gentile P., Chiono V., Carmagnola I., Hatton P.V. An overview of poly(lactic-co-glycolic) acid (PLGA)-based biomaterials for bone tissue engi¬neering // Int. J. Mol. Sci. 2014. V. 15. № 3. P. 36403659. doi:10.3390/ijms15033640 (data obrascheniya: 05.08.2017).
- Wang P., Zhao L., Liu J., Weir M.D., Zhou X., Xu H.H. Bone tissue engineering via nanostructured calcium phosphate biomaterials and stem cells // Bone Res. 2014. V. 2. P. 14017. doi:10.1038/ bo-neres.2014.17 (data obrascheniya: 05.08.2017).
- Ignjatovic N., Ajdukovic Z., Uskokovic D. New biocomposite [biphasic calcium phosphate/poly-DL-lactide-co-glycolide/biostimulative agent] filler for reconstruction of bone tissue changed by osteoporosis // J. Mater. Sci. Mater. Med. 2005. V. 16. № 7. P. 621626.
- Электронный ресурс: http://www.sigmaald¬rich. com/materials-science/biomaterials.html. (data obrascheniya: 05.08.2017).
- Gunatillake P.A., Adhikari R. Biodegradable synthetic polymers for tissue engineering // Eur. Cell. Mater. 2003. V. 5. P. 116.
- Ali S.A., Doherty P.J., Williams D.F. Molecular biointeractions of biomedical polymers with extracellular exudate and inflammatory cells and their effects on the biocompatibility, in vivo // Biomaterials. 1994. V. 15. № 10. P. 779785.
- Shih Y.R.V., Hwang Y.S., Phadke A., Kang H., Hwang N.S. Caro E.J., Nguyen S., Siu M., Theodorakis E.A., Gianneschi N.C., Vecchio K.S., Chien S., Lee O.K., Varghese S. Calcium phosphate-bearing matrices induce osteogenic differentiation of stem cells through adenosine signaling // Proc. Natl. Acad. Sci. USA. 2014. V. 111. № 3. P. 990995.
- Wójtowicz J., Leszczyńska J., Chróścicka A., Slósarczyk A., Paszkiewicz Z., Zima A., Rożniatowski K., Jeleń P., Lewandowska-Szumieł M. Comparative in vitro study of calcium phosphate ceramics for their potency as scaffolds for tissue engineering // Biomed. Mater. Eng. 2014. V. 24. № 3. P. 16091623.
- Kurzina I.A., Pukhova I.V., Botvin V.V., Davydova D.V., Filimoshkin A.G., Savkin K.P., Oskomov K.V., less Oks E.M. New materials based on polylactide modified with silver and carbon ions // AIP Conference Proceedings. 2015. V. 1688. Is. 1. P. 030033-1–030033-7.
- Patent № 2342319 (RF). Sposob polucheniya nanorazmernogo gidroksiapatita / Yu.R. Kolobov, N.N. Volkovnyak, M.B. Ivanov.
- Pinaev G.P., Bogdanova M.S. Metodyi kultivirovaniya kletok. SPb.: Izd-vo Politehn. un-ta. 2008. 278 s.
- Nadezhdin S.V., Zubareva E.V., Burda Yu.E., Kolobov Yu.R., Ivanov M.B., Hramov G.V., Afanasev A.Yu. Vliyanie svoystv poverhnosti implantatov na formirovanie kostnoy tkani v teste ektopicheskogo osteogeneza // Byulleten eksperimentalnoy biologii i meditsinyi. 2016. T. 162. № 12. S. 786
- Birmingham E., Niebur G.L., McHugh P.E., Shaw G., Barry F.P., McNamara L.M. Osteogenic differentiation of mesenchymal stem cells is regulated by osteocyte and osteoblast cells in a simplified bone niche // Eur. Cell. Mater. 2012. V. 23. P. 1327.
- Nadezhdin S.V., Kovaleva M.G., Kolpakov A.Ya., Zubareva E.V., Horolskaya E.N. Otsenka biosovmestimosti i biorezistentnosti zagotovok implantatov iz nikelida titana s modifitsirovannyimi nanorazmernyimi poverhnostnyimi sloyami v opyitah in vivo // Biomeditsina. 2016. № 1. S. 95.
- Avtandilov G.G. Meditsinskaya morfometriya. M.: Meditsina. 1990. 384 s.