To fulfill the properties of membrane for guided bone tissue regeneration, chitosan (CS) and calcium phosphates were blended to produce porous hybrid membranes by lyophilization. We synthesized three different calcium phosphates: calcium deficient hydroxyapatite (CDHA), biphasic calcium phosphate (BCP) and β‑tricalcium phosphate (TCP) by a reverse emulsion method followed by calcination, and compared their efficacy on bone regeneration. The CDHA/CS, BCP/CS, and TCP/CS membranes had an interconnected pore structure with porosity of 91-95% and pore size of 102-147 μm. These hybrid membranes could promote the permeability and adhesiveness to bone cells as demonstrated by in-vitro cell culture of primary osteoblast. Particularly, the CDHA/CS and BCP/CS could further increase the cell attachment and differentiation, whereas the BCP/CS and TCP/CS could enhance cell proliferation. Finally, these hybrid membranes were assessed for guided bone regeneration in the critical-size calvarial bone defects created in SD rats. Histological and histomorphometric analyses revealed that the BCP/CS membrane had the most effective bone regeneration compared to the other two hybrid membranes. At three-week post-surgery, the BCP/CS membrane could enhance new bone generation up to 57% of the original bone defect area. The BCP/CS membrane thus has the potential to be applied for guided bone regeneration.
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