目的探讨外源性降钙素基因相关肽(calcitonin gene related peptide,CGRP)对糖尿病大鼠骨膜微血管病变和膜性成骨的影响。方法建立糖尿病大鼠模型,外源性CGRP静脉注射,随机分为对照组(CON)、糖尿病组(DM)、CGRP干预组(CGRP),分别在5w、...目的探讨外源性降钙素基因相关肽(calcitonin gene related peptide,CGRP)对糖尿病大鼠骨膜微血管病变和膜性成骨的影响。方法建立糖尿病大鼠模型,外源性CGRP静脉注射,随机分为对照组(CON)、糖尿病组(DM)、CGRP干预组(CGRP),分别在5w、10w后观察各组大鼠骨膜微组织结构及组织计量学测定;墨汁灌注观测骨膜微血管单位面积。结果DM1骨祖细胞数较CON均增大(P<0.01),DM2骨膜厚度等均明显小于CON组(P<0.01);微血管单位面积增大,但渗透性大。CGRP骨膜厚度较DM1增多(P<0.01)。CGRP2较DM2骨膜厚度、骨祖细胞数均增大(P<0.01),微血管连续性好。结论外源性CGRP可改善糖尿病大鼠骨膜的微循环损伤,促进膜性成骨对糖尿病大鼠骨质疏松症起到修复作用。展开更多
目的探讨dickkopf1(DKK1)单克隆抗体逆转多发性骨髓瘤上清液在体外抑制成骨分化的现象,为以DKK1基因为靶点的骨髓瘤骨病(myeloma bone disease MBD)的靶向治疗研究奠定基础。方法在骨髓瘤上清液抑制成骨分化的体系中加入0.1、1和10ng/ml...目的探讨dickkopf1(DKK1)单克隆抗体逆转多发性骨髓瘤上清液在体外抑制成骨分化的现象,为以DKK1基因为靶点的骨髓瘤骨病(myeloma bone disease MBD)的靶向治疗研究奠定基础。方法在骨髓瘤上清液抑制成骨分化的体系中加入0.1、1和10ng/ml DKK1单克隆抗体分别培养5天后检测各项成骨分化指标,如碱性磷酸酶(ALP)染色结节计数、碱性磷酸酶(ALP)吸光度测定以及RT-PCR方法检测碱性磷酸酶(ALP)、骨钙素(OC)和核心结合因子1(Cbfa1)的mRNA水平。结果10ng/ml DKK1的高浓度单克隆抗体组与上清液抑制组相比较ALP染色结节计数(P<0.01)和ALP吸光度测定(P<0.01)以及ALP(P<0.05)和cbfa1(P<0.01)的mRNA都有显著意义的升高。结论 DKK1单克隆抗体在体外能够逆转骨髓瘤细胞上清液所致的成骨分化受抑制状态,骨髓瘤细胞能够分泌可溶性DKK1抑制成骨祖细胞分化。展开更多
The use of periosteum-derived progenitor cells （PCs） combined with bioresorbable materials is an attractive approach for tissue engineering. The aim of this study was to characterize the osteogenic differentiation o...The use of periosteum-derived progenitor cells （PCs） combined with bioresorbable materials is an attractive approach for tissue engineering. The aim of this study was to characterize the osteogenic differentiation of PC in 3-dimensional （3D） poly-lactic-co-glycolic acid （PLGA） fleeces cultured in medium containing allogeneic human serum. PCs were isolated and expanded in monolayer culture. Expanded cells of passage 3 were seeded into PLGA constructs and cultured in osteogenic medium for a maximum period of 28 d. Morphological, histological and cell viability analyses of three-dimensionally cultured PCs were performed to elucidate osseous synthesis and deposition of a calcified matrix. Furthermore, the mRNA expression of type Ⅰ collagen, osteocalcin and osteonectin was semi-quantitively evaluated by real-time reverse transcriptase-polymerase chain reaction （RT-PCR）. The fibrin gel immobilization technique provided homogeneous PCs distribution in 3D PLGA constructs. Live-dead staining indicated a high viability rate of PCs inside the PLGA scaffolds. Secreted nodules ofneo-bone tissue formation and the presence of matrix mineralization were confirmed by positive yon Kossa staining. The osteogenic differentiation of PCs was further demonstrated by the detection of type I collagen, osteocalcin and osteonectin gene expression. The results of this study support the concept that this tissue engineering method presents a promising method for creation of new bone in vivo.展开更多
AIM： To study the effect of Spatholobus suberectus Dunn on the prolilferation and hematonic mechanism of Spatholobus suberectus Dunn. Methods： The techniques of culture of hematopoietic cell and hematopoietic growth...AIM： To study the effect of Spatholobus suberectus Dunn on the prolilferation and hematonic mechanism of Spatholobus suberectus Dunn. Methods： The techniques of culture of hematopoietic cell and hematopoietic growth factor （HGF） assay were used. The method of semi-solid culture with methylcellulose of CFU-GM, CFU-E, BFU-E, CFU-Meg was adopted in bone marrow depressed mice which treated with Spatholobus suberectus Dunn for a long time. Results： Spatholobus suberectus Dunn could obviously promote the proliferation of bone morrow cells and spleen lymphocytes in healthy and anaemic mice. The cuhure medium of spleen cell, macrophage, lung and skeletal muscle treated with Spatholobus suberectus Dunn had much stronger stimulating effects on hematopoietic cells. The numbers of CFU-GM, CFU-E, BFU-E, CFU-Meg in bone marrow depressed mice were raised distinctly under the control of Spatholobus suberectus Dunn as compared with those of contrast group. Conclusions： Spatholobus suberectus Dunn may enhance hematopoiesis by stimulating directly and/or indirectly stroma cell in hematopoietic inductive microenvironment and muscle tissue to secrete some HGF （Epo, GM-CSF, IL, and MK-CSF）. It can promote the proliferation and differentiation of hematopoietic cells in anaemic mice. This is one of the biological mechanisms for hematonic effect of Spatholobus suberectus Dunn.展开更多
文摘目的探讨外源性降钙素基因相关肽(calcitonin gene related peptide,CGRP)对糖尿病大鼠骨膜微血管病变和膜性成骨的影响。方法建立糖尿病大鼠模型,外源性CGRP静脉注射,随机分为对照组(CON)、糖尿病组(DM)、CGRP干预组(CGRP),分别在5w、10w后观察各组大鼠骨膜微组织结构及组织计量学测定;墨汁灌注观测骨膜微血管单位面积。结果DM1骨祖细胞数较CON均增大(P<0.01),DM2骨膜厚度等均明显小于CON组(P<0.01);微血管单位面积增大,但渗透性大。CGRP骨膜厚度较DM1增多(P<0.01)。CGRP2较DM2骨膜厚度、骨祖细胞数均增大(P<0.01),微血管连续性好。结论外源性CGRP可改善糖尿病大鼠骨膜的微循环损伤,促进膜性成骨对糖尿病大鼠骨质疏松症起到修复作用。
基金Project supported by the Investitionsbank Berlin (IBB), Germany (No. 10020666) and the Science and Technology Bureau of ZhejiangProvince, China (No. 991110052)
文摘The use of periosteum-derived progenitor cells （PCs） combined with bioresorbable materials is an attractive approach for tissue engineering. The aim of this study was to characterize the osteogenic differentiation of PC in 3-dimensional （3D） poly-lactic-co-glycolic acid （PLGA） fleeces cultured in medium containing allogeneic human serum. PCs were isolated and expanded in monolayer culture. Expanded cells of passage 3 were seeded into PLGA constructs and cultured in osteogenic medium for a maximum period of 28 d. Morphological, histological and cell viability analyses of three-dimensionally cultured PCs were performed to elucidate osseous synthesis and deposition of a calcified matrix. Furthermore, the mRNA expression of type Ⅰ collagen, osteocalcin and osteonectin was semi-quantitively evaluated by real-time reverse transcriptase-polymerase chain reaction （RT-PCR）. The fibrin gel immobilization technique provided homogeneous PCs distribution in 3D PLGA constructs. Live-dead staining indicated a high viability rate of PCs inside the PLGA scaffolds. Secreted nodules ofneo-bone tissue formation and the presence of matrix mineralization were confirmed by positive yon Kossa staining. The osteogenic differentiation of PCs was further demonstrated by the detection of type I collagen, osteocalcin and osteonectin gene expression. The results of this study support the concept that this tissue engineering method presents a promising method for creation of new bone in vivo.
文摘AIM： To study the effect of Spatholobus suberectus Dunn on the prolilferation and hematonic mechanism of Spatholobus suberectus Dunn. Methods： The techniques of culture of hematopoietic cell and hematopoietic growth factor （HGF） assay were used. The method of semi-solid culture with methylcellulose of CFU-GM, CFU-E, BFU-E, CFU-Meg was adopted in bone marrow depressed mice which treated with Spatholobus suberectus Dunn for a long time. Results： Spatholobus suberectus Dunn could obviously promote the proliferation of bone morrow cells and spleen lymphocytes in healthy and anaemic mice. The cuhure medium of spleen cell, macrophage, lung and skeletal muscle treated with Spatholobus suberectus Dunn had much stronger stimulating effects on hematopoietic cells. The numbers of CFU-GM, CFU-E, BFU-E, CFU-Meg in bone marrow depressed mice were raised distinctly under the control of Spatholobus suberectus Dunn as compared with those of contrast group. Conclusions： Spatholobus suberectus Dunn may enhance hematopoiesis by stimulating directly and/or indirectly stroma cell in hematopoietic inductive microenvironment and muscle tissue to secrete some HGF （Epo, GM-CSF, IL, and MK-CSF）. It can promote the proliferation and differentiation of hematopoietic cells in anaemic mice. This is one of the biological mechanisms for hematonic effect of Spatholobus suberectus Dunn.
