Composite Spheroid-Laden Bilayer Hydrogel for Engineering Three-Dimensional Osteochondral Tissue

球体 自愈水凝胶 组织工程 生物医学工程 明胶 软骨发生 化学 粘附 间充质干细胞 材料科学 生物加工 细胞粘附 生物物理学 细胞生物学 复合材料 高分子化学 体外 生物 医学 生物化学
作者
Jin‐Kyu Lee,Eun-Jin Lee,Seung Jae Huh,Jeon Il Kang,Kyung Min Park,Hayeon Byun,Sang‐Min Lee,Eunhyung Kim,Heungsoo Shin
出处
期刊:Tissue Engineering Part A [Mary Ann Liebert]
卷期号:30 (5-6): 225-243
标识
DOI:10.1089/ten.tea.2023.0299
摘要

A combination of hydrogels and stem cell spheroids has been used to engineer three-dimensional (3D) osteochondral tissue, but precise zonal control directing cell fate within the hydrogel remains a challenge. In this study, we developed a composite spheroid-laden bilayer hydrogel to imitate osteochondral tissue by spatially controlled differentiation of human adipose-derived stem cells. Meticulous optimization of the spheroid-size and mechanical strength of gelatin methacryloyl (GelMA) hydrogel enables the cells to homogeneously sprout within the hydrogel. Moreover, fibers immobilizing transforming growth factor beta-1 (TGF-β1) or bone morphogenetic protein-2 (BMP-2) were incorporated within the spheroids, which induced chondrogenic or osteogenic differentiation of cells in general media, respectively. The spheroids-filled GelMA solution was crosslinked to create the bilayer hydrogel, which demonstrated a strong interfacial adhesion between the two layers. The cell sprouting enhanced the adhesion of each hydrogel, demonstrated by increase in tensile strength from 4.8 ± 0.4 to 6.9 ± 1.2 MPa after 14 days of culture. Importantly, the spatially confined delivery of BMP-2 within the spheroids increased mineral deposition and more than threefold enhanced osteogenic genes of cells in the bone layer while the cells induced by TGF-β1 signals were apparently differentiated into chondrocytes within the cartilage layer. The results suggest that our composite spheroid-laden hydrogel could be used for the biofabrication of osteochondral tissue, which can be applied to engineer other complex tissues by delivery of appropriate biomolecules.
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