甲状旁腺激素(1-34)联合介孔生物活性玻璃促进大鼠脊柱后外侧融合  

Local administration of PTH(1-34)combined with mesoporous bioactive glass scaffolds to promote posterolateral spinal fusion in a rat model

作　　者：梁博[1] 黄晶焕 李晓林[1] 徐建广[1] LIANG Bo;HUANG Jinghuan;LI Xiaolin;XU Jianguang(Department of Orthopedic Surgery,Shanghai Jiao Tong University Affiliated Sixth People′s Hospital,Shanghai 200233,China)

机构地区：[1]上海交通大学附属第六人民医院骨科,200233

出　　处：《国际骨科学杂志》2018年第4期255-262,共8页International Journal of Orthopaedics

摘　　要：目的探究以介孔生物活性玻璃(MBG)为载体搭载甲状旁腺激素(PTH)(1-34)制备支架材料,局部应用,促进大鼠脊柱后外侧融合的效用及相关机制。方法通过改进的溶胶-凝胶-聚氨酯泡沫模板法制备MBG材料,加载不同浓度(0.1 mg、0.5 mg)PTH(1-34)制备PTH(1-34)-MBG支架。将支架材料分为3组:0.5 mg PTH组[含0.5 mg PTH(1-34)-MBG的支架]、0.1 mg PTH组[含0.1 mg PTH(1-34)-MBG的支架]和对照组[不含PTH(1-34)的MBG支架]。将各组支架材料与大鼠骨髓间充质干细胞(rBMSC)共同培养。培养7d时通过扫描电镜观察其促进rBMSC黏附的能力,1、3、7 d时使用CCK-8法检测细胞黏附与增殖情况,培养7 d和14 d时检测rBMSC碱性磷酸酶(ALP)活性和成骨相关基因[骨特异性转录因子(Runx)2、骨钙素(OCN)和Ⅰ型胶原(COLⅠ)]的表达情况。将18只SD大鼠随机均分为3组(n=6),分别将3组支架材料植入SD大鼠L4~L5双侧横突间进行脊柱后外侧融合,于术后12周取实验大鼠腰椎标本,通过Micro-CT和手动触摸检查融合情况。结果培养7d时,0.1 mg PTH组和0.5 mg PTH组黏附的rBMSC胞质伸展程度均高于对照组。CCK-8检测显示,培养3d和7d时,0.1 mg PTH组和0.5 mg PTH组吸光度值均高于对照组(P均<0.05)。培养7d和14 d时,0.1 mg PTH组和0.5 mg PTH组的ALP活性均高于对照组(P均<0.05)。培养7d和14 d时,0.1 mg PTH组和0.5 mg PTH组的Runx2、OCN和COLⅠ表达均显著高于对照组(P均<0.05)。术后12周,Micro-CT检查显示,0.1 mg PTH组和0.5 mg PTH组大鼠均为脊柱成功融合,对照组融合失败。手动触摸检查示,0.5 mg PTH组牢固融合6例(100%),0.1 mg PTH组牢固融合4例(66.7%),对照组融合失败。结论以MBG为载体,搭载PTH(1-34)局部应用,可成功促进大鼠的脊柱后外侧融合。Objective To investigate the efficacy and the mechanism of the local use of PTH（1-34）released from mesoporous bioactive glass（MBG）in the treatment of the posterolateral spinal fusion in SD rats.Methods Using MBG scaffolds as a control,the PTH（1-34）-MBG scaffolds with different concentrations of PTH（1-34）were co-cultured with rat bone marrow mesenchymal stem cells（rBMSCs）.The scaffolds were allocated to three groups：0.5 mg PTH（1-34）-MBG scaffold group,0.1 mg PTH（1-34）-MBG scaffold group and MBG scaffold alone group.Cell attachment was evaluated by using scanning electron microscopy（SEM）after 7 days,and cell proliferation was evaluated by means of cell counting kit-8 assay（CCK-8）on day 1,3 and 7.Moreover,osteogenic differentiation was evaluated through alkaline phosphatase activity as well as the expression of osteogenic gene,including Runx-2,OCN and COL-1 on day 7 and 9.Furthermore,18 SD rats were randomly divided into 3 groups,and PTH（1-34）-MBG scaffolds were implanted at the transverse processes of L4-L5 lumbar vertebrae.Twelve weeks after the implantation,lumbar specimen was harvested,and manual palpation and Micro-CT examination were used to evaluate the results of fusion.Results PTH（1-34）-MBG scaffolds group demonstrated better cell attachment than the control group after 7 days of co-culture.Cells in the experimental groups showed better spreading patterns than control group.At day 3 and 7,CCK-8 analysis showed that the OD value in the PTH（1-34）-MBG scaffolds groups was significantly higher than that in the control group（P〈0.05）.At day7 and14,ALP activity in both the 0.5 mg PTH（1-34）-MBG scaffold group and the 0.1 mg PTH（1-34）-MBG scaffold group was better than that in the control group（P〈0.05）,and the expression of osteogenic gene Runx-2,OCNand COLⅠ was significantly higher in the experimental groups than that in the control group（P〈0.05）.Twelve weeks after the implantation of PTH（1-34）-MBG scaffolds,manual palpation and Micro

关 键 词：骨组织工程 甲状旁腺激素 支架材料 脊柱融合 

分 类 号：R687[医药卫生—骨科学]