Enhancing 3D poly[ɛ]caprolactone scaffold properties through alkaline hydrolysis for improved chondrocyte culture: Morphological, physicochemical, and biocompatibility evaluations

Abstract

This study investigates the impact of alkaline hydrolysis on the morphology, physicochemical properties, and mechanical strength of poly[ε]caprolactone (PCL) scaffolds fabricated using a melt-based electrohydrodynamic process. We systematically analyzed how a brief, 5-min alkaline hydrolysis treatment influenced scaffold characteristics compared to longer durations (15 and 30 min). Results demonstrated that the short-duration treatment significantly increased fiber diameter and reduced pore size while modifying the scaffold's surface chemistry. Importantly, the 5-min treatment optimized the water contact angle to 55–65°, enhancing hydrophilicity which facilitated superior cell attachment and proliferation of the C28/I2 immortalized human chondrocyte cell line. Mechanical testing revealed that while alkaline hydrolysis decreased the scaffold's Young's modulus, a 5-min treatment maintained sufficient mechanical integrity for potential tissue engineering applications, in contrast to the more substantial degradation observed with longer treatments.Biocompatibility evaluations showed that the scaffolds, particularly those treated for 5 min, supported cell viability and proliferation without inducing cytotoxic effects. The molecular analysis indicated a favorable upregulation of chondrogenic genes such as SOX9 and COL2A1, and a downregulation of COL1A1, suggesting potential benefits for cartilage tissue engineering. These findings highlight that precise control over the duration of alkaline hydrolysis can significantly enhance PCL scaffold properties, suggesting a scalable and cost-effective method for scaffold optimization in regenerative medicine applications. The results pave the way for further in vivo studies to validate these promising in vitro findings and explore the clinical applicability of the optimized scaffolds.