1.Prevention and treatment of experimental orthopaedic infection with a fibrin sealant cefazoline sodium implant
Bowen LIN ; Zhongshi XU ; Deming XIAO ; Weifan LI ; Li ZHANG
Chinese Journal of Trauma 1990;0(03):-
Objective To investigate the effect of fibrin sealant cefazoline sodium implant on prevention and treatment of orthopaedic infection. Methods The fibrin sealant antibiotic implant was made by 10 ml (100 mg/ml) profibrin (Human) mixed with 2.0 g cefazoline sodium, after which 5 ml (200 PE/ml) prothrombin complex concentrate (Human) was added. A total of 24 female New Zealand white rabbits were recruited in the study and divided into three groups, ie, Group A that was implanted with the fibrin sealant antibiotic implant, Group B that was implanted with antibiotic and Group C that was as control. The model of bone infection was made by injecting 0.2 ml(1?10~6 CFU/ml) staphylococcus aureus into the bone hole in tibia of two legs. The histologic and radiologic bone changes were evaluated 2, 4 and 8 weeks after therapy. And in vitro release test was performed. Results There was not evidence of osteomyelitis in Group A, but the osteomyelitis was observed in Groups B and C. Concentration of cefazoline sodium about (1 043.94?0.20) ?g per piece was released from fibrin sealant cefazoline sodium implant within the first 24 hours by in vitro diffusion test but felt to (7.21?0.02) ?g per piece at the 35th day. Conclusion The fibrin sealant cefazoline sodium implant is simple to make and effective in preventing and treating orthopaedic infection as well as in improving repair of the bone defects.
2.Osteogenic differentiation induced by bone morphogenetic protein 2 and long non-coding RNA AK007000
Yan GAO ; Chen CHENG ; Jing LI ; Yue ZHANG ; Weifan XIAO ; Qiuhui PAN
Chinese Journal of Tissue Engineering Research 2014;(15):2297-2302
BACKGROUND:Long non-coding RNA (lncRNA) regulates a series of physiological processes and it is considered to play important roles in the gene regulation of development, differentiation and metabolism. MC3T3-E1, C2C12 and C3H10T1/2 cells are able to differentiate into different celllineages, such as bone cells and muscle cells, and they can be used in the study of musculoskeletal diseases.
OBJECTIVE:To study the role of lncRNA in osteogenic differentiation induced by bone morphogenetic protein 2.
METHODS:Osteogenic differentiation of MC3T3-E1, C2C12 and C3H10T1/2 cells was induced by bone morphogenetic protein 2, and microarray expression profiling of lncRNA was undertaken in osteogenic differentiation. LncRNA simultaneous changes in three cells were found out. The siRNA interference of the lncRNA was used to study its effects on the osteogenic differentiation induced by bone morphogenetic protein 2. Real-time PCR and alkaline phosphatase staining were applied to detect osteogenesis related indicators.
RESULTS AND CONCLUSION:In the process of osteogenic differentiation induced by bone morphogenetic protein 2, osteogenic differentiation indicators were increased, while myogenic differentiation indicator myogenin was reduced. LncRNA AK007000 was screened out to play a role in osteogenic differentiation induced by bone morphogenetic protein 2. Knockdown of lncRNA AK007000 decreased the expression of osteogenic differentiation indicators, while increased the expression of myogenin. Therefore, AK007000 may play a role in promoting osteogenic differentiation and inhibiting myogenic differentiation.
3.Bone morphogenetic protein 2-induced C2C12 and MC3T3-E1 osteoblast differentiation and autophagy
Yan GAO ; Chen CHENG ; Jing LI ; Yue ZHANG ; Weifan XIAO ; Qiuhui PAN
Chinese Journal of Tissue Engineering Research 2014;(20):3236-3241
BACKGROUND:A series of studies indicate that autophagy is closely linked with differentiation. Bone morphogenetic protein 2 (BMP-2) is the classical pathway for C2C12 and MC3T3-E1 osteoblast differentiation, and the ideal model to study osteogenic differentiation process.
OBJECTIVE:To observe the relationship between autophagy and BMP-2-induced celllines C2C12, MC3T3-E1 osteoblast differentiation.
METHODS:Real-Time PCR was applied to detect osteogenic differentiation and autophagy related index after C2C12 and MC3T3-E1 were induced with BMP-2 (100μg/L) for 72 hours. The osteogenic index alkaline phosphatase in BMP-2-induced MC3T3-E1 and C2C12 cultured with different concentrations of 3-methyladenine (0, 1, 5, 10 mmol/L) was determined with alkaline phosphatase staining. Western blot analysis was applied to detect LC3-I/II expression levels in C2C12 and MC3T3-E1 induced with BMP-2 for different time points (0, 12, 24, 48, 72, 96 hours).
RESULTS AND CONCLUSION:The autophagy-related mRNA and protein expression showed an increasing tendency and autophagy-related protein LC3 levels was increased, which was associated with the time, during the BMP-2-induced celllines C2C12, MC3T3-E1 osteoblast differentiation. Meanwhile, alkaline phosphatase expression levels were inhibited by autophagy in the process of osteogenic differentiation. Therefore, there is a close relationship between autophagy and the BMP-2-induced celllines C2C12, MC3T3-E1 osteoblast differentiation.
4.Prevention of epidural adhesion after laminectomy by a salvia miltiorrhiza radix-gel
Ji CHEN ; Deming XIAO ; Hongtu YANG ; Bowen LIN ; Wanxin ZHEN ; Weifan LI ; Meng Lü
Chinese Journal of Tissue Engineering Research 2007;11(18):3647-3652
BACKGROUND:Different methods and biomaterials have been applied in animal experiments and clinical practice to prevent the formation of epidural scars,Biodegradable and sticky semi-fluid gels are the most often used material.Salvia miltforrhiza radix (SMR) and carbomer have been clinically confirmed to be the safe and effective drugs and gel agents. OBJECTIVE:To observe the effect of SMR-gel on preventing epidural adhesion after laminectomy.DESIGN:A complete randomized grouping design, a controlled experiment. SETTING:Department of Orthopaedics,Shenzhen People's Hospital (Second Clinical College of Jinan University). MATERIALS:Thirty-six healthy pure New Zealand rabbits were used,either male of female,clean degree,2-3 years of age. They were randomly divided into four groups with 9 rabbits in each group:blank control group,gel contro group, HA group and SMR-gel group. Carbomer934 powder (Shanghai People's Pharmaceutical Factory, batch number: 20000510) , hyaluronic acid (HA) [Shandong Bausch & Lomb Freda Pharaceutical, Co., Ltd.,No.H10960136,2 mL (20 mg)].METHODS:The experiments were carried out in the animal laboratory of Shenzhen People's Hospital from April 2002 to August 2003.①Preparing SMR-gel:SMR was prepared into extract powder.Carbomer934 powder was added by water for dissolving and swelling and stayed overnight,then SMR-gel was prepared by dipping with triethanolamine,adding with SMR extract powder (2 g),then adding with purified water till 100.0 g and stirring uniformly.②The rabbits were anesthetized. and the lamina of vertebra was totally resected at L3 and L6 (reserving superior and Inferior articular processes).then defects of 10 mm×5 mm were made to expose the dura mater.The vertebral defects were added with 1 mL carbomer gel, 1 mL HA (20 g/L) and 1 mL SMR-gel in the gel control group,HA group and SMR-gel group respectively.whereas nothing was added in the blank control group.③Gross samples:Three rabbits were killed 4,6 and 8 weeks postoperatively in each group.vertebraI ventral fascia were stripped to remove the spinal segments (L3,L6) for operation completely,and totally 24 samples for each time.One sample was selected in each group 4 weeks postoperatively. and the samples were observed under H-600 transmission electron microscope (Hitachi). ④The adhesion compactness of scar tissue with dura mater was evaluated in the 24 samples of the 4 groups at 8 weeks postoperatively:There were 4 grades:No obvious adhesion between dural sac and scar tissue for grade O:Extensive and compact adhesion between dural sac and scar tissue. impossible blunt dissection between dural sac and scar tissue.incomplete dural sac after sharp dissection for grade Ⅲ.Each spinal segment was cut into 4 parts equally,and all were prepared into sections and stained,then the thickness of epidural scar was determined with Tiger2000 image analyzer. ⑤The rank sum test was used in the scar adhesion compactness grading evaluated with naked eyes,whereas analysis of variance.and two-two comparison were used in analyzing the thickness of epidural scar.P<0.05 was considered as significant difference.MAIN OUTCOME MEASURES:①Results of gross scar adhesion compactness grading at 8 weeks and comparison of the thickness of epidural scar at 4.6 and 8 weeks;②Results of gross observation,histological examination and ultrastructure.RESULTS: All the 36 rabbits were involved in the analysis of results. ①Results of gross observation and pathohistological examination:There was compact adhesion at each time point in the blank control group,part adhesion in the gel control group and HA group, and no obvious adhesion in the SMR-gel group.②Results of quantitative analysis:The rabbits with lower scores of scar adhesion compactness grading In the blank control group,gel control group and HA group were obviously fewer than those in the SMR-gel group (W=45-52,P<0.05-0.01).The scar thickness at 4 and 8 weeks in the SMR-gel group was obviously less than that in the other 3 groups(F=128.657,152.246,80.891,P<0.01).③Results of observation under transmission electron microscope:The proliferation of fibroblasts at 4 week was active in the blank control group,gel control group and HA group,but inactive in the SMR-gel group.CONCLUSION:①SMR can inhibit the fibroblasts to proliferate,differentiate and synthetize into secretory collagens,and then inhibit the formation of epidural scar adhesion.②HA can be absorbed by organs very early,which reduces its role in preventing adhesion.Whereas carbomer gel can stay longer, and it plays a role in inhibiting and blocking adhesion in the whole process of wound repairing.