伤科接骨片对MC3T3-E1细胞成骨能力的影响
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摘要
目的观察伤科接骨片对MC3T3-E1细胞成骨能力的影响。方法取BALB/c小鼠30只,随机分为药物+细胞组:伤科接骨片及饲料喂养+MC3T3-E1+明胶海绵;细胞组:单纯饲料+MC3T3-E1+明胶海绵;阴性对照组:单纯饲料+生理盐水+明胶海绵。每组10只。在小鼠右侧股部臀大肌下方制成肌袋,药物+细胞组和细胞组用无菌明胶海绵吸附MC3T3-E1细胞悬液、阴性对照组用无菌明胶海绵吸附生理盐水后植入肌袋内。术后3天,药物+细胞组将伤科接骨片(0.35g/片)用蒸馏水配制成浓度为20mg/m L的混悬液,按2.0mg/(g·d)剂量灌胃给药,细胞组和对照组均灌胃给予2.0mg/(g·d)的蒸馏水。分别于建模后第4、8周每组各处死小鼠5只,取材后分别作大体、切片HE染色光镜,以及扫描电镜下观察。结果术后4周,药物+细胞组和细胞组肌袋内均可见新骨出现,术后8周两组均形成松质骨样结构的板层骨,药物+细胞组形成的板层骨结构较细胞组更为成熟,成骨细胞数量更多,胶原纤维分泌量也更多,并有更多的钙化结节形成。阴性对照组内未见有明显骨化现象。结论伤科接骨片可以促进MC3T3-E1细胞在动物体内的成骨作用。
Objective To investigate the effects of Shangke Jiegu Tablet on the the role of MC3T3-E1 cells in osteogenesis promotionin vivo. Methods Thirty healthy BALB/c mice were randomly divided into 3 groups(10 rats each): drug+cell group, cell group, and negative control group. During the animal operation, the muscle pouch was made in the right thigh of the mouse, and the sterile gelatin sponge was used to absorb the MC3T3-E1 cell suspension or saline. Conventionaldiet was given after operation, and 3days later, the above groups were given treatment by gastrogavagerespectively as follows: Shangke Jiegutablets(0.35g/tablet)withdistilled water toa concentration of20mg/m L suspension, and 2.0mg/(g·d) in drug+cell group; 2.0mg/(g·d) of distilled water incellgroup and negative controlgroup. Every 5 mice were executed at 4 and 8 weeks after oral adiministration, and the specimens were harvested and observedwith HE staining and light microscope and scanning electron microscopy. Results Four weeks after operation, the new bone appeared in the drug +cell and cell groups; at 8 weeks, the cancellous bone with a lamellar structure was formed in both groups. In drug+cell group, more mature cancellous bone, collagen secreted by osteoblasts, and calcified nodules were formed compared withcell group. No new bone was observed in negative control group. Conclusion Shangke Jiegu tablet is conducive to the osteogenesis induced by MC3T3-E1 cells in vivo.
Objective To investigate the effects of Shangke Jiegu Tablet on the the role of MC3T3-E1 cells in osteogenesis promotionin vivo. Methods Thirty healthy BALB/c mice were randomly divided into 3 groups(10 rats each): drug+cell group, cell group, and negative control group. During the animal operation, the muscle pouch was made in the right thigh of the mouse, and the sterile gelatin sponge was used to absorb the MC3T3-E1 cell suspension or saline. Conventionaldiet was given after operation, and 3days later, the above groups were given treatment by gastrogavagerespectively as follows: Shangke Jiegutablets(0.35g/tablet)withdistilled water toa concentration of20mg/m L suspension, and 2.0mg/(g·d) in drug+cell group; 2.0mg/(g·d) of distilled water incellgroup and negative controlgroup. Every 5 mice were executed at 4 and 8 weeks after oral adiministration, and the specimens were harvested and observedwith HE staining and light microscope and scanning electron microscopy. Results Four weeks after operation, the new bone appeared in the drug +cell and cell groups; at 8 weeks, the cancellous bone with a lamellar structure was formed in both groups. In drug+cell group, more mature cancellous bone, collagen secreted by osteoblasts, and calcified nodules were formed compared withcell group. No new bone was observed in negative control group. Conclusion Shangke Jiegu tablet is conducive to the osteogenesis induced by MC3T3-E1 cells in vivo.
引文
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[7]黄长联,董海辉,周建光,等.跌打接骨片促进实验性骨折愈合的骨组织形态计量学研究[J].中医药研究,2001,17(4):47-49.
[2]张一,孙立,简月奎,等.双基因活化纳米骨浆的体内成骨[J].中国组织工程研究,2014,18(3):329-334.
[3]Calvert JW,Chua WC,Gharibjanian NA,et al.Osteoblastic phenotype expression of MC3T3-E1 cells cultured on polymer surfaces[J].Plast Reconstr Surg,2005,116(2):567-576.
[4]Chen VJ,Smith LA,Ma PX.Bone regeneration on computerdesigned nano-fibrous scaffolds[J].Biomaterials,2006,27(21):3973-3979.
[5]姜琳,魏玉玲,杨洪平,等.伤科接骨片促进实验性骨折愈合的超微结构观察[J].中医正骨,2000,12(11):3-4.
[6]魏玉玲,王炳南,粱克玉.伤科接骨片对I、II型胶原基因表达的影响[J].中医正骨,1998,10(5):3-5.
[7]黄长联,董海辉,周建光,等.跌打接骨片促进实验性骨折愈合的骨组织形态计量学研究[J].中医药研究,2001,17(4):47-49.