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    Chitosan scaffolds containing boron compound for bone tissue engineering
    (Adana Alparslan Türkeş Bilim ve Teknoloji Üniversitesi, 2025) Al Dham, Oula Alkhalifa ; Çapkın Yurtsever, Merve
    In this study, scaffolds produced with sodium pentaborate pentahydrate (SPP) containing chitosan (Ch) and medical grade (MG) chitosan were obtained by freeze drying method, characterization studies were performed and cell culture studies were carried out to determine their potential in bone tissue engineering applications. SPP was added to the chitosan polymer solution dissolved in 1% (v/v) acetic acid at ratios of 0.125%, 0.25%, 0.5%, 0.75% and 1% (w/w). In order to obtain a 3-dimensional pore structure, the polymer was frozen at -20 oC for 2 days and freeze dried at -80 oC in and tey were stabilized by ethanol or NaOH and freeze dried again. Three different experimental groups were studied in the thesis: free SPP dissolved in cell culture medium, chitosan scaffolds containing SPP at concentrations ranging from 0.125% to 1% (w/w), and medical-grade chitosan scaffolds containing 0.5% and 0.75% (w/w) SPP. Scanning Electron Microscopy (SEM) and ImageJ analysis of the studies with chitosan (Ch) polymer showed that increasing SPP concentration decreased the average pore size. The smallest pore structure was observed in the scaffold containing 1% SPP. Water retention tests revealed that all scaffolds reached swelling equilibrium within 30 minutes, but the swelling capacity decreased as the SPP concentration increased. The highest water uptake capacity was measured in the pure chitosan scaffold and the lowest in the scaffold containing 1% SPP. Four-week biodegradation tests showed greater weight loss in scaffolds with higher SPP content. When the antimicrobial properties of Ch – SPP scaffolds were examined, it was determined that Ch - SPP scaffolds showed a dose-dependent antifungal effect against Aspergillus niger and Candida albicans. When MC3T3-E1 mouse osteoblast cells were cultured on the scaffolds, it was observed that cell viability decreased in all groups in the following days. Considering that cell proliferation was not supported by Ch-SPP scaffolds, the effect of both chitosan type and stabilization method on the proliferation of MC3T3-E1 cells was examined. The results showed that the scaffolds produced with medical grade chitosan (MG) and stabilized with NaOH promoted the proliferation of MC3T3-E1 cells. Therefore, in the ongoing studies, scaffolds containing 0.5%, 0.75% (w/w) SPP with medical grade chitosan were produced, characterized and cell culture studies were carried out. Resazurin viability assay results showed that MCET3-E1 cells adhered to the scaffolds and supported their proliferation. Although metabolic activity decreased in the last days, DAPI staining results revealed increased cell nuclei density at day 10, especially in the MG Ch - 0.5 SPP and MG Ch - 0.75 SPP groups, indicating that cell proliferation supported. SEM analysis results also showed that the cells proliferated and spread intensively, especially in the MG Ch - 0.5 SPP group. In conclusion, SPP-enriched medical grade chitosan scaffolds were found to be antifungal and biocompatible by supporting the proliferation of MC3T3-E1 cells. Among all formulations, the scaffold containing 0.5% (w/w) SPP in NaOH-stabilized medical grade chitosan stands out as the most promising scaffold for bone tissue engineering.