Construction and characterization of the ex-situ modified macroporous bacterial cellulose scaffold as a potential epidermal graft

Background: Skin is a 3-dimensional (3-D) tissue that mainly consists 2 layers, comprising the epidermis and dermis. Skin tissue engineering scaffolds are used commonly as 3-D analogs of the extracellular matrix (ECM) of the skin. Bacterial cellulose (BC) has great importance in skin tissue engineering because of its resemblance to ECM and its biocompatibility. The lack of 3-D microporosity and limited biodegradation capacity has restricted its application as a scaffold for skin tissue engineering. Controlled 3-D microporosity of BC via surface modification techniques are required for potential tissue engineering applications.Methods: Freeze-drying is an ex-situ surface modification technique for making macroporous BC scaffolds (MBCSs). This study proposed a new approach to the freeze-drying method for the arrangement of the pore size of MBCSs specifically for the human keratinocyte cell line (KER-CT). Different concentrations of MBCS (0.25%, 0.50%, and 0.75%) were prepared and the KER-CT cell viability was detected via 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) assay.Result: The results of this study indicated that the KER-CT cells were able to proliferate all of the concentrations of MBCS, and the best cell viability value was observed with 0.75% MBCS. The results were supported by FESEM and light microscopic observations.Conclusion: These findings suggested that 0.75% MBCS might be of use in epidermal tissue engineering applications

Perifosine and vitamin D combination induces apoptotic and non-apoptotic cell death in endometrial cancer cells

Endometrial cancer is the most common cancer of the female reproductive system. Combination treatment with specific agents has been widely used as a targeted therapy for cancer. In this study, we aimed to investigate the anti-proliferative and apoptotic effects of varying concentrations of perifosine and vitamin D on the human endometrial cancer cell line (HEC-1A). HEC-1A cells were exposed to perifosine (10 μM, 30 μM), vitamin D (50 nM, 200 nM) and combinations of both for 48 h and 72 h. Monitoring of cell proliferation in a time-dependent manner was performed with the xCELLigence RTCA DP system. The levels of BCL2, BAX and P53 mRNA expression were examined using RT-qPCR. Apoptosis was determined using Annexin V, which were followed by flow cytometry analysis. Ultra-structural morphology of cells was analyzed by transmission electron microscopy (TEM) for 72 h. The anti-proliferative and apoptotic effects of the perifosine+vitamin D combination (30 μM + 200 nM at 48 h and 10 μM + 200 nM at 72 h) on HEC-1A cells were higher than in perifosine and vitamin D alone. It was observed that perifosine has increased the expression of BAX mRNA in HEC-1A cells in a dose-dependent manner. While perifosine+vitamin D combinations increased P53 mRNA expression in HEC-1A cells we did not find any significant change in BCL2, BAX mRNA expression levels. In TEM examinations of HEC-1A cells, perifosine appeared to lead autophagic cell death, whereas vitamin D caused paraptosis-like cell death and combination of perifosine+vitamin D caused apoptotic and non-apoptotic (paraptotic, autophagic and necrotic) cell death. Therefore, it is considered that the combination of both drugs in the treatment of endometrial cancer might be an alternative and effective treatment option through activating the apoptotic and non-apoptotic cell death mechanisms in cancer cells