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The PHBV nonofibrous membrane fabricated by electron-spinning method for tissue engineered bone regeneration scaffold was evaulated in terms of cellular prolieration and cryopreservation efficiency. The rat calvarial periosteum derived primary cells were cultured with PHBV nanofibrous membranes and analyzed the cellular proliferation and differention fashion and cryopreservation potential by in vitro MTT assay as well as ALP staining and Alizarine red staining with or without cryopreservation for 2 weeks. The rat calvarial periosteum derived primary cells cultured with PHBV nonofibrous membrane showed favorable proliferation and alkaline phosphatase activity with numerous mineral nodule formation regardless of cryopreservation, even though its efficiency was slightly decreased in cryopreserved condition. These findings suggest that PHBV nanofibrous membrane can be applicable as an efficient cell engineered membrane for guided bone regeneration or scaffold for tissue engineered bone regeneration.