Slipper Limpet (Crepidula fornicata) Shells Support In Vitro Osteogenesis of Human Adipose-Derived Stem Cells

Abstract

This study aimed to investigate a cost-effective alternative to man-made calcium phosphate ceramics for treating bone defects. The slipper limpet is an invasive species in European coastal waters, and its shells composed of calcium carbonate could potentially be a cost-effective source of bone graft substitutes. This research analyzed the mantle of the slipper limpet (Crepidula fornicata) shells to enhance in vitro bone formation. Discs machined from the mantle of C. fornicata were analyzed using scanning electron microscopy with energy dispersive spectroscopy (SEM-EDS), X-ray crystallography (XRD), Fourier-transform infrared spectroscopy (FT-IR) and profilometry. Calcium release and bioactivity were also studied. Cell attachment, proliferation, and osteoblastic differentiation (RT-qPCR and alkaline phosphatase activity) were measured in human adipose-derived stem cells grown on the mantle surface. The mantle material was mainly composed of aragonite and showed a sustained Ca2+ release at physiological pH. In addition, apatite formation was observed in simulated body fluid after three weeks, and the materials supported osteoblastic differentiation. Overall, our findings suggest the mantle of C. fornicata shows potential as a material for fabricating bone graft substitutes and structural biomaterials for bone regeneration.