Additively Manufactured 3D Clamp-Culture System for the Investigation of Material-Cell Interactions in Multi-Material Hybrid Scaffolds for Musculoskeletal Tissue Defects

The emergence of hybrid scaffolds, blending biomaterials with diverse properties, offers promise in musculoskeletal tissue engineering. However, a need for in vitro platforms investigating biological behavior and the interplay of different load-bearing and colonizable synthetic bone substitute materials remains. Herein, we present a novel, in-house producible, and scalable clamp culture system designed for facile in vitro analysis of interactions between biomaterials, hydrogels, and cells. The system, constructed here from an exemplary 3D-printable polymer and photopolymerizable hydrogel using a widely available benchtop 3D printer, ensures mechanical stability and protection for the embedded hydrogel via its double-clamp structure, facilitating various analytical methods while preserving culture integrity. Hybrid clamp cultures were additively manufactured from polylactic acid, filled with a bone precursor cell-laden methacrylate gelatin hydrogel, cultured for 14 days, and analyzed for cell viability, mineralization, and osseous differentiation. Results indicate no adverse effects on osteogenic differentiation or mineralization compared to conventional droplet cultures, with enhanced cell viability and simplified handling and downstream analysis. This system demonstrates the potential for robust experimentation in tissue engineering and is adaptable to various plate formats, and thus highly suitable for the investigation of biomaterial-cell interactions and the development of implants for musculoskeletal tissue defects.