Baghaei Saman, Azarpira Negar, Paknahad Maryam, Amani Ali Mohammad, Dortaj Hengameh, Koohpeyma Farhad, Sajad Daneshi Seyyed, Vafa Ehsan, Vaez Ahmad, Lavaee Fatemeh, Tayebi Lobat
Student Research Committee, School of Dentistry, Shiraz University of Medical Sciences, Shiraz, Iran.
Craniofacial and Cleft Research Center, Isfahan University of Medical Sciences, Isfahan, Iran.
Res Sq. 2025 Aug 26:rs.3.rs-7159849. doi: 10.21203/rs.3.rs-7159849/v1.
Critical-sized bone defects present significant clinical challenges due to inadequate vascularization and scaffold integration. This study developed a multifunctional 3D-printed polycaprolactone (PCL)-gelatin (Gel) scaffold reinforced with Bioglass particles (BGPs) or copper dopped BGPs (CuBGPs) to synergistically enhance angiogenesis and bone regeneration in rat model. The scaffolds were fabricated by infiltrating gelatin solutions containing BGPs or CuBGPs into the pores of 3D-printed PCL matrices, followed by freeze-drying. Comprehensive characterization of PCL-gel, PCL-gel-BGPs, and PCL-gel-CuBGPs scaffolds revealed optimal porosity (58.76±5.20, 53.27±11.83, and 54.5±7.61%), contact angle (74.53 ±6.6, 71.76±2.65, and 69.89±4.14), and controlled degradation (44.65±4.73, 47.93±2.51, and 50.58±5.68). MTT study demonstrated dose-dependent enhancement of cell proliferation, with BGPs and CuBGPs significantly improving mesenchymal stem cells (MSCs) viability by day 5. In vivo experiments in rat calvarial defects showed that Cu containing scaffolds promoted greater new bone volume compared to other groups at 12 weeks. Histological and immunohistochemical analyses confirmed robust angiogenesis and woven bone formation, with CuBGPs achieving the highest vasculature. This study provides a detailed and reproducible framework for Cu-doped scaffold fabrication, characterization, and application in critical-sized defect regeneration.
临界尺寸骨缺损由于血管化不足和支架整合问题而带来重大临床挑战。本研究开发了一种用生物玻璃颗粒（BGPs）或铜掺杂生物玻璃颗粒（CuBGPs）增强的多功能3D打印聚己内酯（PCL）-明胶（Gel）支架，以协同增强大鼠模型中的血管生成和骨再生。通过将含有BGPs或CuBGPs的明胶溶液渗透到3D打印的PCL基质的孔隙中，然后冷冻干燥来制造支架。对PCL-凝胶、PCL-凝胶-BGPs和PCL-凝胶-CuBGPs支架的全面表征显示出最佳孔隙率（58.76±5.20、53.27±11.83和54.5±7.61%）、接触角（74.53±6.6、71.76±2.65和69.89±4.14）以及可控降解性（44.65±4.73、47.93±2.51和50.58±5.68）。MTT研究表明细胞增殖呈剂量依赖性增强，到第5天，BGPs和CuBGPs显著提高了间充质干细胞（MSCs）的活力。大鼠颅骨缺损的体内实验表明，在12周时，含铜支架比其他组促进了更大的新骨体积。组织学和免疫组织化学分析证实了强大的血管生成和编织骨形成，CuBGPs的血管生成量最高。本研究为铜掺杂支架的制造、表征及其在临界尺寸缺损再生中的应用提供了一个详细且可重复的框架。
