Development of a 3D collagen type I hydrogel scaffold for modeling hyperglycemia-induced spinal neuronal damage and...

This study aimed to develop a three-dimensional (3D) collagen type I hydrogel scaffold for modeling spinal neuron culture under high-glucose conditions, thereby simulating environments such as diabetic neuropathy and maternal diabetes. Hydrogels were fabricated using rat tail collagen type I. Porosity was assessed via scanning electron microscopy (SEM), and viscoelastic properties were evaluated using amplitude sweep testing with a rheometer. Rat embryonic spinal neurons were cultured in 2D and 3D hydrogel models. Neuronal morphology was assessed using Sholl analysis. Cell viability and cytotoxicity under glucose stress were evaluated using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and live/dead imaging. Curcumin was delivered via β-cyclodextrin metal-organic frameworks (β-CD-MOFs). Phosphatase and Tensin Homolog (PTEN) gene expression was analyzed by real-time polymerase chain reaction (PCR). Scanning electron microscopy (SEM) analysis revealed an average pore size of 57.96 μm. The hydrogel's elastic modulus ranged from 138.56 to 178.25 Pa, with tan δ values between 0.124 and 0.180, indicating a predominantly elastic nature. Viability assays confirmed hydrogel biocompatibility. Neurons cultured in 3D exhibited significantly greater neurite outgrowth and branching complexity. High-glucose exposure caused dose-dependent reductions in cell viability. Treatment with 6.8 μM curcumin via β-CD-MOFs improved viability in both 2D and 3D models. Glucose exposure significantly upregulated PTEN expression, which was attenuated by curcumin treatment. The developed collagen type I hydrogel scaffold provides suitable porosity, mechanical compliance, and biocompatibility for 3D spinal neuron culture. It serves as a robust platform for investigating neuronal responses to hyperglycemia and potential therapeutic agents such as curcumin.