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.

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.

OBJECTIVE: To assess the relationship between the prognosis of the patients with diffuse traumatic brain swelling (DTBS) and the changes of the ventricles and the cisterns in CT scans. METHODS: The outcome of the patients with DTBS and the changes of the ventricles and the cisterns in CT scans were studied and analyzed in a group of 268 cases. We focused on the changes of the third ventricle and the basal cistern, age and Glasgow Coma Scale (GCS). RESULTS: Of 268 cases, there were changes of the third ventricle and/or the basal cistern in 124, 65 died. In l8 cases, the third ventricle and the basal cistern were both absent and l6 died (88.9%). The third ventricle changed significantly in 59 cases, 33 died (55.9%), while the basal cistern changed in 47 cases and 16 died (34%). Of the 124 patients with changes of the third ventricle and/or the basal cistern, 26 were children, 8 died; 98 adults, 57 died. CONCLUSIONS: For patients with DTBS, the outcome was in direct correlation with the change of the third ventricle and/or the basal cistern, the change of the third ventricle was much more important in assessment of the outcome than that of basal cisterns. There is no significant difference in, the incidence of DTBS between children and adults while the outcome of children is much better than that of adults. The patients with the changes of the third ventricle and the basal cistern accompanied with lower GCS scores have poor outcome.

Objective To investigate the biocompatibility of Ti-6Al-4V scaffolds fabricated by electron beam melting(EBM).Methods Bone marrow mesenchymal stem cells(BMSC) co-cultured with Ti-6A1-4V specimens fabricated with EBM was prepared as experimental group and the regular cells culture was employed as control.The biocompatibility was detected using CCK-8 and cytoskeleton staining.The osteogenic differentiation ability was assessed using mineralization nodule formation.A 24 mm defect was created on the right mandibular body in 12 beagles.The mandibular defects were repaired with Ti-6A1-4V scaffolds mesh fabricated by EBM.General observation,CT and histology examination was carried out to evaluated the biocompatibility of Ti-6A1-4V scaffolds in vivo.Results CCK-8 result showed the A values of the two groups had no significant difference(P ＞ 0.05).There was no significant difference between the two groups (P＞0.05).Cytoskeletal staining showed that cells were fully stretched out and grew well on T-i6A1-4V specimen.The actin fibers were arranged in parallel and stained uniformly with fluorescent.After osteogenic culture,the quantity of the nodule formation of the experimental group and control group were 5.7±0.7 and 5.1 ± 0.6,respectively(P＞0.05).All animals had tolerated the surgery and healed well.CT examination showed that Ti-6A1-4V scaffolds mesh had good retention with surrounding bone and the continuity of mandible was restored.Histological examination showed that no inflammation reaction or toxity was caused in the soft tissue surrounding the scaffolds and in the liver and kidney after implantation.Ti-6A1-4V scaffolds had good retention with surrounding bone.Conclusions Ti-6Al-4V fabricated with electron beam melting has good biocompatibility.

Objective To evaluate the surface characteristics and cytocompatibility of Ti-6Al-4V alloy fabricated using select laser melting (SLM) and electron beam melting (EBM) technique.Methods Ti-6Al-4V alloy specimens were fabricated with SLM and EBM.A wrought form of Ti-6Al-4V alloy was used as a control.Its properties were evaluated using component analysis,contact angle test,surface roughness,surface topography,cell ultrastructure,cell attachment and proliferation observation,metal ion precipitation examination.Results The roughness of SLM and EBM specimens was suitable for cell attachment but not the best.The character of SLM and EBM specimens was hydrophobic (＞65°).The surface topography of EBM and SLM specimens were similar,but were not the best type for cell attachment.The components of Ti-alloy oxide film were detected in all the specimens.The content of Ti,A1,V ions of EBM,SLM and wrought specimens were very low and did not affect the cell attachment and proliferation.The ultrastructure of cell was normal,and the cytomembrane was intact.The number of cells was similar to each other among the three kinds of specimens and increased obviously with the culture time.Conclusions The results of the study suggested that EBM and SLM Ti-6A1-4V specimens possessed good surface characteristics.However,the surface modification are needed further.

OBJECTIVETo establish a RP-HPLC method for determination of glycosides in Traditional Chinese Medicine Liu-wei Di-huang.

METHODThe samples were analyzed on an ODS column at 30 degrees C, with mobile phase of methanol/water (33:67) at flow rate 1.0 mL.min and detection at wavelength of 236 nm.

RESULTThree major components reached base-line separation and were identified to be mononiside, loganin, paeoniflorin. Respectively for the three components, linear correlations were found between peak areas and concentrations in the ranges of 7.4-60, 7.7-62 mg.L-1 and 8.5-68 mg.L-1, and the recoveries were 98.8%, 98.3%, 99.6%.

CONCLUSIONThe established method is proved to be suitable for simultaneous quantification of three major glycosidic components in Liuwei Dihuang decoction and can be used for evaluation of the quality of Liuwei Dihuang preparations.

Benzoates ; analysis ; Bridged-Ring Compounds ; analysis ; Chromatography, High Pressure Liquid ; methods ; Cornus ; chemistry ; Dioscorea ; chemistry ; Drug Combinations ; Drugs, Chinese Herbal ; analysis ; chemistry ; Glucosides ; analysis ; Iridoids ; analysis ; Monoterpenes ; Plants, Medicinal ; chemistry ; Quality Control ; Rehmannia ; chemistry