ABSTRACT
BACKGROUND:Periosteal osteoblasts possess strong reproductive activity,as well as osteoblastic differentiation potential,which is an ideal seed cell if can shorten the culture time.OBJECTIVE:Modified enzymatic digestion was used to culture rabbits'osteoblasts.and to study the adherence and proliferation of osteoblasts on the sudace of sandblasting titanium.MEITHODS:Periostea were harvested from the theanteromedial surface of the proximal tibia of male,Japanese white rabbits,and cultured as follow:①Routine method:Digested with 0.25%trypsinase at 37 ℃ for 30 minutes,followed by digestion with 0.1%type I collagenase at 37 ℃ for 30 minutes,vibration.removed trypsinase and dried.After 2 hours,DMEM containing 15% fetal bovine serums were added.②Modified method:30 minutes culture of type I collagenase was prolonged to 1 hour.The osteoblasts were identified by alkaline phosphatase staining and calcium node staining.The adherence and proliferation of osteoblasts cultured on sandblasting surface were measured by scanning electron microscopy and MTT.RESULTS AND CONCLUSION:Five days after culture.the periosteal steoblasts crawled out from tissues,gathered as monolayer with tdangle or polygon at after 25 days of modified culture.After 1 month of culture,superposition growth of calcium nodus appeared.The cultured cells possessed the morphological characteristic and biological behavior of osteoblasts.which were positive to alkaline phosphorase and calcium node staining.The time of cells cultured with routine method covered flask delayed 12 days than modified method.The osteoblasts were inseted into sandblasting titatium with pseudopodium.However,the adherence and proliferation of osteoblasts cultured on sandblasting surface had no obviously difference between two culture methods.The results suggested that modified enzymatic digestion can sho～en the culture time without effect on adherence and proliferation of osteoblasts.
ABSTRACT
BACKGROUND: Autologous bone, bone substitute materials and guided bone regeneration (GBR) technique can repair jaw defects, but the absorption speed of bone substitute materials and GBR membrane are faster than the formation speed of new bone, therefore, it affects the volume and shape of new bone.OBJECTIVE: To investigate the role of personalized prefabricated titanium template, autologous bone and nano-hydroxyapatite on restoration of maxillary defect in rabbit.METHODS: A total of 18 rabbits were randomly divided into two groups, and maxillary alveolar defect with 10 mm length and 5 mm high was created. The template was implanted in both two groups, and fastened with titanium screws. Autologous and nano-hydroxyapatite were placed into the defect in experimental group; neither autologous bone nor bone substitute materials were implanted into the defect in control group. New bone formation, X-ray findings, and histological changes with HE stain were carded out 4, 8 and 12 weeks postoperatively.RESULTS AND CONCLUSION: The quality of new bone in experimental group was batter than that in control group 4 weeks postoperatively, but the quality of new bone was almost the same 8 and 12 weeks postoperatively. By paired t-test, there was significant difference in new bone density between the experimental group and the control group 4 .weeks after operation (P<0.01), but there was no significant difference in new bone density between the experimental group and the control group 8 and 12 weeks after operation (P > 0.05). Autologous bone and nano-hydroxyapatite can restore the defect of maxillary alveola.Personalized prefabricated titanium template can play an important role of screen membrane and external scaffold in new bone formation, and remain shape of new bone.