Three-dimensional nanofibrous PCL/gelatin scaffold fabricated using centrifugal force assisted wet electrospinning technique

A three-dimensional (3D) scaffold that enables optimal cell-matrix interactions is essential for developing physiologically relevant neural tissue models. In this study, wet electrospinning was optimized to fabricate nanofibrous PCL/gelatin scaffolds with a well-controlled 3D architecture, using centrifugal force to tune scaffold morphology, porosity, and mechanical properties. The effects of centrifugal force intensity (5000 vs. 10,000 rpm) and application time (5 vs. 10 min) were systematically investigated. Scaffolds fabricated at 5000 rpm exhibited poor structural integrity and were excluded from further analysis. Among scaffolds produced at 10,000 rpm, blends of PCL/gelatin at 70:30 and 60:40 demonstrated excellent porosity (98.1 ± 1.9% and 97.3 ± 1.1%, respectively) and favorable fiber architecture. The 70:30-10 min scaffold achieved the highest tensile strength (57.03 ± 1.50 kPa) and modulus (53.00 ± 2.00 kPa), aligning with the physiological range of neural tissues. MTT assays confirmed robust biocompatibility, with C6 glial cell viability increasing by + 4.30% on the 70:30-10 min scaffold and + 5.88% on the 60:40-10 min scaffold over 14 days. Although the 70:30-5 min scaffold showed the highest proliferation (+ 12.16%), the 10-min variant was selected for detailed morphological evaluation due to its superior mechanical performance and structural uniformity. DAPI and H&E staining further validated the enhanced cell aggregation, ECM deposition, and neural-like morphology within the 70:30-10 min scaffold. These results collectively highlight the critical role of scaffold composition and processing parameters in engineering 3D neural tissue scaffolds, with the optimized 70:30-10 min scaffold emerging as a promising candidate for advanced neural tissue engineering applications.