Doymuş Burcu, Peközer Görke Gürel, Önder Sakip
Department of Biomedical Engineering, Yildiz Technical University, Esenler, 34220, Istanbul, Türkiye.
Department of Molecular Biology and Genetics, Istanbul Technical University, Istanbul, Türkiye.
Ann Biomed Eng. 2025 Apr;53(4):980-993. doi: 10.1007/s10439-025-03684-4. Epub 2025 Jan 27.
Titanium (Ti)-based materials are favored for hard tissue applications, yet their bioinertness limits their success. This study hypothesizes that functionalizing Ti materials with chitosan nano/microspheres and calcitriol (VD) will enhance their bioactivity by improving cellular activities and mineralization. To test this, chitosan particles were applied uniformly onto Ti surfaces using electrophoretic deposition (EPD) at 20 V for 3 minutes. VD was then loaded onto the coated surfaces, and the release profile of VD was monitored. Human fetal osteoblastic cells (hFOB) were cultured on the VD-loaded Ti surfaces. Cellular activities such as proliferation, Alkaline phosphatase (ALP) activity, osteogenic gene expression (runt-related transcription factor 2 (Runx2), collagen type 1 (Col I), osteocalcin ( OCn), osteopontin (OP)), and mineralization were assessed. Von Kossa staining was performed to analyze mineralization, and the expression of cell adhesion proteins (N-cadherin (NC), integrin alpha V (IaV), integrin beta 3, (Ib3)) was measured. The results showed that approximately 50% of the VD released over 50 hours. The chitosan coating increased surface roughness three-fold, and this, combined with VD release, resulted in reduced cell proliferation but increased ALP activity, suggesting enhanced differentiation. VD-functionalized Ti surfaces showed statistically significant differences in osteogenic gene expressions, particularly on rougher surfaces. Additionally, the expression of cell adhesion proteins (NC, IaV, Ib3) was upregulated on VD-containing coated surfaces. Von Kossa analysis revealed that surface roughness significantly enhanced mineralization, particularly on VD-free surfaces by day 7, while mineralization on VD-containing bare surfaces started on day 14. These findings demonstrate that VD-loaded chitosan coatings significantly enhance the biocompatibility and bioactivity of Ti-based materials, highlighting their potential for applications in bone regeneration.
钛（Ti）基材料在硬组织应用中备受青睐，但其生物惰性限制了它们的应用成效。本研究假设，用壳聚糖纳米/微球和骨化三醇（VD）对Ti材料进行功能化处理，将通过改善细胞活性和矿化作用来增强其生物活性。为验证这一假设，使用电泳沉积（EPD）在20伏电压下持续3分钟，将壳聚糖颗粒均匀地施加到Ti表面。然后将VD负载到涂层表面，并监测VD的释放情况。将人胎儿成骨细胞（hFOB）培养在负载VD的Ti表面上。评估细胞增殖、碱性磷酸酶（ALP）活性、成骨基因表达（ runt相关转录因子2（Runx2）、I型胶原蛋白（Col I）、骨钙素（OCn）、骨桥蛋白（OP））和矿化等细胞活性。进行冯·科萨染色以分析矿化情况，并测量细胞粘附蛋白（N-钙粘蛋白（NC）、整联蛋白α V（IaV）、整联蛋白β 3（Ib3））的表达。结果表明，约50%的VD在50小时内释放。壳聚糖涂层使表面粗糙度增加了两倍，这与VD释放相结合，导致细胞增殖减少，但ALP活性增加，表明分化增强。VD功能化的Ti表面在成骨基因表达上显示出统计学上的显著差异，尤其是在较粗糙的表面上。此外，含VD的涂层表面上细胞粘附蛋白（NC、IaV、Ib3）的表达上调。冯·科萨分析表明，表面粗糙度显著增强了矿化作用，尤其是在第7天的无VD表面上，而含VD的裸露表面上的矿化作用在第14天开始。这些发现表明，负载VD的壳聚糖涂层显著增强了Ti基材料的生物相容性和生物活性，突出了它们在骨再生应用中的潜力。
