摘要
Gelatin methacrylamide porous microcarriers (GelMA PMS) often fails to achieve complete interconnected pore structures and encounters local structural destruction, spherical distortion and limited cell proliferation rate. This study introduced an innovative dual ice crystal strategy, which enabled staged ice crystal growth before and after photo-crosslinking, along with the low-temperature gelation of GelMA. GelMA PMS with stable spherical structure and controllable interconnected pore structures (12 to 42 μm) were successfully fabricated. Various aspects of cell morphology, viability, proliferation and phenotypic markers were thoroughly investigated to assess the effects of different interconnected-pore structures of GelMA PMS on various cells. Results indicated that PMS60 (−60 °C) promoted bone marrow mesenchymal stem cells (BMSCs) adhesion ratio (91.69 %) and high proliferative activity. PMS196 (−196 °C) facilitated the adhesion and proliferation of human umbilical vein endothelial cells (HUVECs) and mouse renal tubular epithelial cells (mRTECs), as evidenced by amplification times of 7.47 and 8.17, respectively. It is worth mentioning that BMSCs cultured in PMS60 demonstrated a higher differentiation capacity and expression of phenotypic markers, with CD105/CD90 positive rates exceeding 99.54 %. Additionally, the stemness genes Nanog and Sox2 were upregulated by 2.48- and 2.31-fold, respectively. HUVECs in PMS196 exhibited enhanced the vascularization potential, forming well-organized capillary-like structures and more extensive interconnected tubular networks. Meanwhile, mRTECs in PMS196 showed a reduction in epithelial-mesenchymal transition (EMT), along with a significant downregulation of Vimentin/E-cadherin, which was reduced by 2.20-fold compared to cells cultured in Cytodex-3. Overall, the GelMA PMS fabricated in this study advances the development of cell therapy.
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