D.Y. Aleynik, I.N. Charykova, T. I. Sidorova

The paper presents the investigation of cytotoxicity of the new bone replacement material code-named «Combas». The base was bone non-demineralized collagen, crumbled into 500-700 mcm pieces «VladMiva», Belgorod), saturated with vessel endothelium growth factor under a developed methodology. (ZAO (JSVC) «Proteinsynthes», Moscow). To estimate the bone replacement material the authors chose a complex approach which allows to make conclusions on the effect of the drug on cell viability in the culture as well as on cell functional state. Cultures of 4-6 passage diploid fibroblasts functioned as a test culture. As the use of the drug suggests a long-time direct contact with the blood and tissue, the direct contact method was chosen for the investigation. The test process included estimation of culture state in the monolayer and cell morphology, the number of viable cells and their functional activity in the culture on change of proliferation and fibronectin synthesis function. Collagen-containing Collatamp EG, manufacturer «Syntcall GmbH, Germany, was used as a positive controller. The morphological estimation of the state of the mololayer on the whole and of separate cell structures was performed on the basis of visualization with the help of the microscope «Leica DM IL», equipped with a camera and software for archiving and culture growth control. (Leica IM 1000). For quantitative measuring of fibronectin, «Biokhimmak» enzyme sets were used. Optical density (OD) was registered by the enzyme analyzer «Sunrise» (Austria), using Magellan software, which allows to automatically build a calibration curve and determine concentration. A PC and Stadia 6.0 software were used for statistical analysis. The investigation found faults in the material at the stage of its development which allowed to take timely measures to enhance the quality of the new innovative drug. The complex approach proved that with consideration of the parameters under analysis bone replacement material «Combas» (Series 2) is not cytotoxic and can be admitted for in vivo studies.

 

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