THE INVESTIGATION OF DIRECT ACTION OF ENAMEL MATRIX PROTEINS ON OSTEOGENOUS PROGENITOR HUMAN BONE-MARROW CELLS IN VITRO

Braun I. E.

DENTISTRY

Scentific article

Generalized periodontitis (GP) became to be widely spread disease among other types of periodontal diseases, staying until present times as unsolved clinical problem because of completely untreated. The longterm chronic course of the GP became to be one of the main detail that accompanying by progressive loss of periodontal tissues. During this process such significant structures as: alveolar bone, periodontal ligament and root cement are injured by pathological process, whose are characterized by very slow reparation, need very longtime terms for regeneration that depends on condition of neighboring tissues of periodontal wound. The complicity of periodontal tissues structure and their anatomical connections during functioning determine the main problems in reaching aimed regeneration using modern methods. The tooth loss or loss of group of teeth still considered as frequent consequence of not forehanded treatment and as a result of GP further progression. However, according to modern investigations dedicated to periodontal tissues regeneration, it was stated that the periodontal regeneration can be reached successfully. Due to creation of appropriate micro surrounding conditions for cells, that can mimic the course of periodontal ligament formation process and cementogenesis, it became to be possible to reach regeneration of periodontal tissues. This became to be possible under action of enamel matrix proteins (EMD) in form of Emdogain. Plural scientific works of different authors showed that different types of cells including stromal progenitor cells of bone marrow can take part in regeneration of periodontal tissues. EMD are characterized by influencing on different cell types and also due to precipitation on root surface, they combine appropriate micro surrounding for cementoblasts, osteoblasts and their progenitors. The aim of current study was to investigate the direct action of “Amdogain” on osteogenous progenitor cells - colony-forming fibroblast units (CFFU) of human bone-marrow ex vivo and to evaluate presence of its osteoinductive properties. Cloning of CFFU of human bone-marrow was provided according to methodic of Fridenshtein O. Y. (1973) in modification of Astachova V. S. (1982). According to stated methodic, cancellous bone was taken for investigation. Under requested conditions, the investigation of colony growth of GFFU was provided. According to presented study, the investigation of osteogenous potential of GFFU was provided under action of “Amdogain” and “Pref-Gel” that both are main components recommended for usage in regenerative periodontal procedures. The 4 experimental series of CFFU cloning of human bone-marrow were provided: 1st group – with adding of etching gel “Pref-Gel” into feeding solution in cultural flacon; 2nd group – with adding of “Pref-Gel” with “Amdogain” into feeding solution in cultural flacon; 3rd group – with adding of “Amdogain” only into feeding solution in cultural flacon; 4th – without adding of any preparations into feeding solution in cultural flacon (control). The action was evaluated according to cloning effectiveness of CFFU of human bone-marrow among 105 nucleus-containing cells. 9 experimental and 6 control colonies were cultivated. The obtained results showed in 1st and 2nd experimental groups the growth of stromal fibroblasts wasn’t detected. The effectiveness of CFFU cloning = 0. In 3rd group in the mean 145 colonies of CFFU grew up with cloning effectiveness of 15,10 ± 0,95. In control group in the mean 120 colonies grew up. The cloning effectiveness was 12,48 ± 1,24 among 105 nucleus-containing cells. Ad oculus the colonies from control and investigated groups weren’t differ from each other. The presented investigated revealed that “Pref-Gel” and combination of “Pref-gel” with “Amdogain” completely depresses proliferation and differentiation of CFFU in human bone-marrow ex vivo. These data can show the direct chemical action on vital stromal bone cells due to action of “Pref-Gel” which is 24% EDTA. This chemical component may change pH in feeding solution in cultural flacon, leading to direct chemical stress in vital bone cells and their death. The obtained chemical changes were not enhanced under additional adding of “Amdogain” after into cultural flacon. However, separate adding of “Amdogain” into cultural flacon enhanced on 20,8% in comparison with control group, amount of CFFU colonies in bone-marrow, increasing specific gravity of multilayer colonies, giving evidence about presence of its osteoinductive properties.

generalized periodontits, regeneration, enamel matrix proteins, Emdogain, osteogenous progenitor human bone marrow cells, osteoinduction

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«Bulletin of problems biology and medicine» Issue 1 part 1 (126), 2016 year, 337-342 pages, index UDK 616.314.13-085:611-081.41:612.08