Polymeric biomaterials originally designed for regenerative medicine are increasingly used in pharmacy eg. in preclinical trials or in drug delivery systems. In the case of preclinical studies, the use of polymeric scaffolds may reduce the need for animal testing and the risk of early treatment failure in vivo. These improvements are the results of restoration of naturally occurring cell interactions. Previously used 2D cultures allowed the cell attachment to polystyrene surfaces but didn’t allow the formation of spatial interactions between cells. Scaffolds, made of polymeric biomaterials, ensure proper both cell-cell and cell-matrix interactions that mimic in vivo conditions. To ensure the optimal growth of cells, scaffolds are usually placed in suitable bioreactors which provide the proper flow of oxygen, nutrients and waste materials. These 3D cultures could be useful in drug discovery and enable the evaluation of the drug efficacy and cytotoxicity which among others corresponds with proliferative activity of cells. The reliable assessment of these properties is necessary in order to detect unsafe drugs and choose the best therapeutic candidate for clinical trials. The majority of manufactured scaffolds are characterized by structural heterogeneity. This property limits the ability to assess the proliferative status of cells using standard assays based on eg. dye detection. Due to these limitations, novel cell proliferation marker for molecular pharmacology is needed. The aim of the study was to evaluate the usefulness of replication-dependent subtype of histone H3 (H3.b) as a novel molecular marker of proliferation. The expression of replication-dependent subtypes of histone H3 is tightly correlated with cell cycle status. The experiment was carried out using normal human articular chondrocytes from knee – NHAC-kn. The results of the expression of histone H3.b (real time RT-PCR) were compared to results of the XTT and resazurin reduction. The obtained results were promising and indicated that H3.b may be a potential molecular marker of cell proliferation.

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