Department of Chemical and Materials Engineering, Chang Gung University, Kwei-San, Taoyuan 33302, Taiwan.
Department of Plastic and Reconstructive Surgery, Chang Gung Memorial Hospital at Keelung, Keelung 20401, Taiwan; Department of Plastic and Reconstructive Surgery and Craniofacial Research Center, Chang Gung Memorial Hospital at Linkou, Chang Gung University School of Medicine, Kwei-San, Taoyuan 33305, Taiwan.
Int J Biol Macromol. 2023 Dec 31;253(Pt 1):126528. doi: 10.1016/j.ijbiomac.2023.126528. Epub 2023 Aug 24.
In this study, we fabricate squeezable cryogel microbeads as injectable scaffolds for minimum invasive delivery of chondrocytes for cartilage tissue engineering applications. The microbeads with different glucosamine concentrations were prepared by combining the water-in-oil emulsion and cryogelation through crosslinking of gelatin with glutaraldehyde in the presence of glucosamine. The physicochemical characterization results show the successful preparation of cryogel microbeads with uniform shape and size, high porosity, large pore size, high water uptake capacity, and good injectability. In vitro analysis indicates proliferation, migration, and differentiated phenotype of rabbit chondrocytes in the cryogel scaffolds. The seeded chondrocytes in the cryogel scaffold can be delivered by injecting through an 18G needle to fully retain the cell viability. Furthermore, the incorporation of glucosamine in the cryogel promoted the differentiated phenotype of chondrocytes in a dose-dependent manner, from cartilage-specific gene expression and protein production. The in vivo study by injecting the cryogel microbeads into the subcutaneous pockets of nude mice indicates good retention ability as well as good biocompatibility and suitable biodegradability of the cryogel scaffold. Furthermore, the injected chondrocyte/cryogel microbead constructs can form ectopic functional neocartilage tissues following subcutaneous implantation in 21 days, as evidenced by histological and immunohistochemical analysis.
在这项研究中，我们制备了可挤压的水凝胶微球作为注射式支架，用于最小侵入式递送软骨细胞，以应用于软骨组织工程。通过在存在葡糖胺的情况下使用戊二醛使明胶交联，将具有不同葡糖胺浓度的微球通过水包油乳液和凝胶化结合制备。理化特性分析结果表明成功制备了具有均匀形状和尺寸、高孔隙率、大孔径、高吸水性和良好可注射性的水凝胶微球。体外分析表明兔软骨细胞在水凝胶支架中增殖、迁移和分化。通过 18G 针注射可将接种在水凝胶支架中的软骨细胞完全保留其活力。此外，葡糖胺的掺入以剂量依赖的方式促进软骨细胞的分化表型，从软骨特异性基因表达和蛋白产物方面得到证实。将水凝胶微球注射到裸鼠的皮下囊中进行的体内研究表明，水凝胶支架具有良好的保留能力、良好的生物相容性和合适的生物降解性。此外，在 21 天的皮下植入后，注射的软骨细胞/水凝胶微球构建体可以形成异位功能性新软骨组织，这可通过组织学和免疫组织化学分析得到证明。
