Preparation and preliminary evaluation of bio-nanocomposites based on hydroxyapatites with antibacterial properties against anaerobic bacteria.

Hexagonal nanocrystalline powders of the non-doped Ca10(PO4)6(OH)2 as well as activated with Ag+ and Eu3+ ions were synthesized by using different wet chemistry methods. Moreover, the obtained hydroxyapatite was loaded with Ag0, as well as nitroimidazole antimicrobials: metronidazole and tinidazole. The structural properties of the products were analyzed by X-ray diffraction (XRD), scanning (SEM) and transmission (TEM) electron microscopy as well as infrared (IR) and Raman spectroscopy. The photoluminescence properties of the Eu3+ and Ag+ co-doped Ca10(PO4)6(OH)2 were characterized via the PL emission, excitation spectra and the luminescence decay curve. The antimicrobial activity of the obtained materials against Prevotella bivia and Parabacteroides distasonis was studied. The cytotoxicity assessment was carried out on the human osteosarcoma cell line (U2OS) as well as human red blood cells (RBC). The choice of the in vitro model was based on the fact that U2OS is a cancer cell line derived from bone tissue which is rich in apatites that play a pivotal role in the extracellular matrix formation. RBCs are the most abundant blood cells and they are used as a cell model in the study of biocompatibility of new prepared biocompounds with potential medical applications. The obtained multifunctional materials do not exhibit the haemolytic activity, therefore, they could be used as a promising antimicrobial agent and for anaerobic bacteria.

New approach to modification of poly (l-lactic acid) with nano-hydroxyapatite improving functionality of human adipose-derived stromal cells (hASCs) through increased viability and enhanced mitochondrial activity.

The aim of this study was to determine the cytocompatibility of poly (l-lactide) (PLLA) scaffolds fabricated using co-rotating twin screw extrusion technique and functionalized with different concentrations of nano-hydroxyapatite (nHAp). The efforts were aimed on the designing bioactive scaffolds improving the viability and metabolic activity of human adipose-derived multipotent stromal cells (hASCs). The in vitro study was designed to determine the optimal nHAp concentration, based on analysis of hASCs morphology, adhesion rate, as well as metabolic and proliferative potential. Initially, the PLLA filled with three different concentrations of the nHAp were tested i.e. 5%, 10% and 15 wt%. The obtained results indicated that the 10 wt% nHAp in the PLLA (10% nHAp/PLLA) matrices improved the adhesion and proliferation of the hASCs, what was in good agreement with the results of tensile properties of the composites. Further, we performed profound studies regarding the cytotoxicity of 10% nHAp/PLLA. The analysis included the evaluation of the biomaterial influence on viability, apoptosis-related markers expression profile and mitochondrial function. The cytocompatibility of 10% nHAp/PLLA scaffolds toward the hASCs was confirmed. The hASCs propagated on 10% nHAp/PLLA were more viable then those propagated on the plain PLLA. The level of pro-apoptotic markers, i.e. caspase-3 and Bax in cultures on 10% nHAp/PLLA was significantly decreased. Obtained results correlated with higher mitochondrial membrane potential of hASCs in those cultures. The obtained composites may improve therapeutic potential of hASCs via directing their adhesion, enhancing proliferation and viability as well as increasing mitochondrial potential, thus may find potential application in tissue engineering.