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OBJECTIVE:To investigate the effect of Guijiajiao (Colla Carapacis et Plastri,CCP),the glue of tortoise shell,on bone regeneration in zebrafish and male Wistar rats.METHODS:In this study,we applied in vitro and in vivo models,tissue section analysis,and quantitative polymerase chain reaction to assess the effects of CCP on bone repair.MG-63 cells were used in alkaline phosphatase (ALP) activity and mineralization assays.Zebrafish and male Wistar rats were used to evaluate the effects of CCP on bone repair in vivo.RESULTS:A simple preparation of CCP promoted osteogenesis in vivo and in vitro,and promoted MG-63 cell proliferation as well as ALP activity and mineralization.In addition,CCP activated Akt and extracellular signaling-regulated kinase pathways and significantly increased the expression of ossification-related genes and proteins such as runt-related transcription factor-2,osteocalcin,and osteopontin.The in vivo results revealed promotion of osteogenesis by immersing zebrafish in CCP for 72 h.An oral dose of 1.25 g/kg CCP significantly improved skull defects in rats,which was accompanied by an increase in serum ALP levels.CONCLUSION:One of the ingredients of Guilu Erxianjiao (tortoise shell and deer h formula) provides a practical altative therapy for bone regeneration.