铜掺杂硼硅酸盐生物活性玻璃复合水凝胶的 构建及其促口腔黏膜血管化修复的体外研究

Construction of Copper-Doped Borosilicate Bioactive Glass Composite Hydrogel and Its in Vitro Study on Promoting Vascularized Repair of Oral Mucosa

摘要: 【目的】：针对口腔黏膜创面血管化重建不足的临床瓶颈，本研究构建铜掺杂硼硅酸盐生物活性玻璃/海藻酸钠复合水凝胶(Cu-BBG/SA)，并系统评价其体外生物学性能。【方法】：采用溶胶–凝胶法制备Cu-BBG纳米颗粒，与海藻酸钠经CaCl₂离子交联复合成型，得到Cu-BBG/SA水凝胶。通过扫描电子显微镜(SEM)、能量色散X射线光谱(EDS)及傅里叶变换红外光谱(FTIR)对材料进行结构、元素与化学键合表征；以人脐静脉内皮细胞(HUVECs)为体外模型，通过CCK-8法、活/死细胞染色、溶血实验、划痕实验及成管实验系统评价材料的生物相容性及促血管化功能。【结果】：表征结果显示，Cu-BBG颗粒均匀嵌布于SA三维多孔网络中，B、Si、Cu、Ca等元素空间分布均一；FTIR证实复合体系中COO⁻特征峰的规律性红移与O-H峰宽化，提示两相间存在金属离子–羧基配位相互作用与界面氢键的化学层面结合。细胞毒性筛选确定SA适宜浸提浓度为0.1 g/mL、Cu-BBG为50 μg/mL，各组HUVECs细胞活力均≥95%，溶血率低于2%，符合ISO 10993标准。体外功能评价中，Cu-BBG/SA组24 h划痕愈合率(约60%)、成管节点数(约190个)、分支数(约140个)及管腔总长度均显著优于各单组分组(P < 0.05)，呈现明确的有机–无机协同促血管生成效应。【结论】：Cu-BBG/SA复合水凝胶兼具优良生物安全性与体外促血管化功能，为新型口腔黏膜创面修复材料的研发提供了实验依据。

Abstract: [Objective]: To address the clinical challenge of insufficient vascular reconstruction in oral mucosal wounds, this study constructed a copper-doped borosilicate bioactive glass/sodium alginate composite hydrogel (Cu-BBG/SA) and systematically evaluated its in vitro biological performance. [Methods]: Cu-BBG nanoparticles were synthesized via the sol-gel method and composited with sodium alginate through CaCl₂ ionic crosslinking to obtain the Cu-BBG/SA hydrogel. Structural, elemental, and chemical bonding characterization was performed using scanning electron microscopy (SEM), energy-dispersive X-ray spectroscopy (EDS), and Fourier-transform infrared spectroscopy (FTIR). Human umbilical vein endothelial cells (HUVECs) were employed as an in vitro model, and the biocompatibility and pro-angiogenic function of the materials were evaluated via CCK-8 assay, live/dead cell staining, hemolysis test, scratch assay, and tube formation assay. [Results]: Characterization results demonstrated that Cu-BBG particles were uniformly embedded within the three-dimensional porous SA network, with homogeneous spatial distribution of B, Si, Cu, and Ca elements. FTIR analysis confirmed a systematic red shift of the COO⁻ characteristic peaks and broadening of the O-H peak in the composite system, suggesting chemical-level interactions between the two phases in the form of metal ion-carboxylate coordination and interfacial hydrogen bonding. Cytotoxicity screening established optimal extract concentrations of 0.1 g/mL for SA and 50 μg/mL for Cu-BBG, under which HUVECs viability remained ≥ 95% across all groups, with hemolysis rates below 2%, meeting ISO 10993 standards. In vitro functional evaluation showed that the Cu-BBG/SA group significantly outperformed individual component groups in 24 h scratch healing rate (approximately 60%), tube formation junction count (approximately 190), branch count (approximately 140), and total tube length (P < 0.05), demonstrating a distinct organic-inorganic synergistic pro-angiogenic effect. [Conclusion]: The Cu-BBG/SA composite hydrogel exhibits excellent biosafety and in vitro pro-angiogenic function, providing experimental evidence for the development of novel oral mucosal wound repair materials.