Hemostasis and Bone Regeneration Using Chitosan/Gelatin-BCP Bi-layer Composite Material

The aim of the study was to determine the hemostatic activity of a composite bi-layered topical hemostat composed of electrospun gelatin loaded with bi-phasic calcium phosphate and chitosan layer and its effect on bone formation. Morphology of the composite hemostat and its individual components were observed using scanning electron microscopy. In vitro biocompatibility of the topical hemostat tested using preosteoblasts cells (MC3t3-E1) showed no adverse toxicity. Confocal microscopy of seeded cells showed good cell adhesion while 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay indicated 15% increased cell proliferation at the end of 1 week culture period. The material’s efficiency as a hemostatic agent was tested by testing blood adsorption capacity and in vivo bone bleeding models. Blood absorption indicates that test sample measuring 14.14 mm 3 becomes fully saturated within 5 minutes of blood contact. Bone bleeding was induced on the frontal plates of rat skulls and samples were applied as either removable topical hemostat or actual degradable hemostat. The effect of the bi-layered hemostat on bone formation was determined through analysis of micro-computed tomography (microCT) and observation of histological sections of extracted bone tissue samples. Results indicate that the bi-layer hemostat was able to halt bleeding within 3 minutes of application on the bleeding site and significantly enhanced bone regeneration. Using the bi-layer material as a degradable hemostat also drastically improved bone regeneration of the 3 mm defect.