The optimization of pepsin processing time to balance the mechanical and biological properties of xenogeneic bone

Xenogeneic bone showed great prospects to treat large bone defects due to its bionic composition and structure, but the immunogenicity limited its wide applications. Previously, we developed a pepsin treating method to eliminate the immunogenicity of xenogeneic bone. In this study, we further investigated the effect of pepsin processing time on the biological and mechanical properties. The results indicated that increased pepsin treating time impaired the mechanical properties of xenogeneic bone. And MC3T3-E1 cells showed enhanced adhesion ability, as well as increased production of alkaline phosphatase and calcium nodulus production on the xenogeneic bone processed by pepsin for 24 hr (P24), as compared with xenogeneic bone processed by pepsin for 30 hr (P30) and 36 hr (P36). In addition, we found no significant inflammatory responses after implanting different xenogeneic bone into the intermuscular site of rats. These results suggested that xenogeneic bone processed by pepsin for 24 hr may be a preferable choice when using the xenogeneic bone as biomaterials for further researches.