Enhanced hemostasis and alveolar bone regeneration induced by sandwich like egg shell membrane-laponite-chitosan...

Persistent hemorrhage and alveolar bone resorption following tooth extraction can complicate subsequent dental implant interventions, highlighting the need for scaffolds with dual hemostatic and osteogenic properties. In this study, we developed a novel sandwich-like sponge composed of alginate, chitosan, laponite nanoparticles, and soluble eggshell membrane (ALCS). The bioactive ALCS sponge exhibited exceptional hemostatic efficacy in a rat tooth extraction injury model. In vitro cellular assessments demonstrated that the ALCS biomaterial, when encompassing blood clots, significantly elevated the expression of bone-associated RUNX2 and OCN proteins, increased alkaline phosphatase (ALP) activity, and promoted mineralized extracellular matrix deposition by bone marrow stem cells. In vivo bone regeneration, evaluated using CBCT imaging and histological analysis, revealed that ALCS led to a marked improvement in osteogenesis. It achieved a bone volume to tissue volume ratio (BV/TV) of 80 %, significantly higher than that of the commercial Gelita® sponge (68 %) and the untreated control group (60 %) (P-value <0.01). The ALCS scaffold also demonstrated strong clot-forming capacity by promoting platelet activation and accelerating the coagulation cascade. Moreover, the incorporation of osteoinductive components supported stem cell attachment and proliferation, thereby enhancing osteogenic differentiation. These combined features highlight the scaffold's potential as an effective treatment strategy for the clinical management of post-extraction wounds.