rhPTH(1-34)对成骨细胞增殖、Collagen I及Osterix mRNA表达的影响

目的 观察间歇给予重组人甲状旁腺激素(1-34)[rhPTH(1-34)]对体外成骨细胞增殖、I型胶原蛋白(CoHagen I)及Osterix mRNA表达的影响,初步探讨rhPTH(1-34)对体外成骨细胞的作用机制.方法 体外培养新生大鼠成骨细胞,间歇循环给予0、10-11、10-10、10-9、10-8、10-7 M rhPTH(1-34)干预,(24 h为一循环,前12h给药),共2次,用MTT法检测细胞的增殖;RT-PCR法半定量测定成骨细胞Collagen I、Ostefix mRNA的表达.结果 显示rhPTH(1-34)可明显促进成骨细胞的增殖(P<0.05),促进成骨细胞Collagen I和Ostefix mRNA表达(P<0.05),101-9 M增殖、表达最明显,呈剂量依赖关系.结论 rhPTH(1-34)可促进成骨细胞的增殖、分化,可能是通过Collagen I和Ostefix mRNA表达来调节.     

rhPTH(1-34)对成骨细胞增殖、Collagen I及Osterix mRNA表达的影响

目的 观察间歇给予重组人甲状旁腺激素(1-34)[rhPTH(1-34)]对体外成骨细胞增殖、I型胶原蛋白(CoHagen I)及Osterix mRNA表达的影响,初步探讨rhPTH(1-34)对体外成骨细胞的作用机制.方法 体外培养新生大鼠成骨细胞,间歇循环给予0、10-11、10-10、10-9、10-8、10-7 M rhPTH(1-34)干预,(24 h为一循环,前12h给药),共2次,用MTT法检测细胞的增殖;RT-PCR法半定量测定成骨细胞Collagen I、Ostefix mRNA的表达.结果 显示rhPTH(1-34)可明显促进成骨细胞的增殖(P<0.05),促进成骨细胞Collagen I和Ostefix mRNA表达(P<0.05),101-9 M增殖、表达最明显,呈剂量依赖关系.结论 rhPTH(1-34)可促进成骨细胞的增殖、分化,可能是通过Collagen I和Ostefix mRNA表达来调节.     

rhPTH（1-34）对成骨细胞增殖、CollagenⅠ及Osterix mRNA表达的影响

目的观察间歇给予重组人甲状旁腺激素(1-34)[rhPTH(1-34)]对体外成骨细胞增殖、Ⅰ型胶原蛋白(Collagen Ⅰ)及Osterix mRNA表达的影响,初步探讨rhPTH(1-34)对体外成骨细胞的作用机制。方法体外培养新生大鼠成骨细胞,间歇循环给予0、10-11、10-10、10-9、10-8、10-7M rhPTH(1-34)干预,(24 h为一循环,前12h给药),共2次,用MTT法检测细胞的增殖;RT-PCR法半定量测定成骨细胞Collagen Ⅰ、Osterix mRNA的表达。结果显示rhPTH(1-34)可明显促进成骨细胞的增殖(P<0.05),促进成骨细胞Collagen Ⅰ和Osterix mRNA表达(P<0.05),10-9 M增殖、表达最明显,呈剂量依赖关系。结论 rhPTH(1-34)可促进成骨细胞的增殖、分化,可能是通过Collagen Ⅰ和Osterix mRNA表达来调节。     

rhIGF-1对成骨细胞增殖、BMP-2及Cbfa1基因表达的影响

目的通过重组人胰岛素样生长因子(rhIGF-1)对体外成骨细胞增殖、骨形态蛋白-2(BMP-2)及核心结合因子1(Cbfa1)基因表达的影响,探讨rhIGF-1对骨代谢影响的可能机制。方法不同浓度的rhIGF-1(0、10、50、100ng/ml)刺激体外培养的大鼠成骨细胞,采用噻唑蓝(MTT)法检测细胞增殖能力,用半定量RT-PCR法检测成骨细胞BMP-2、Cbfa1基因的表达。结果rhIGF-1可明显促进成骨细胞增殖(P0.05),并促进成骨细胞BMP-2、Cbfa1基因的表达(P0.01),具有浓度依赖性。结论rhIGF-1可促进成骨细胞的增殖、分化及成熟,可能是通过增强BMP-2、Cbfa1基因的表达来实现的。     

联合甲状旁腺激素和普萘洛尔对人成骨细胞增殖及OPG和RANKL mRNA表达的影响

目的 联合甲状旁腺激素(rhPTH 1-34)和普萘洛尔(PRO)处理体外培养的人松质骨源性成骨细胞(HOB),观察其对人成骨细胞增殖及骨保护素(OPO)和核因子KB受体活化因子配体(RANKL )基因表达的影响,探讨其对骨代谢影响的可能机制。方法 (1)以成人骼骨和股骨颈部松质骨为原料,分离培养原代人成骨细胞并对其鉴定。(2)以人成骨细胞为体外实验模型,固定浓度rhPTH1- 34 (50 ng/ml)和不同浓度PRO(0. 1μM,1μM,10μM)分别及联合刺激体外培养的人成骨细胞72 h后,采用CCK-8法检测细胞增殖能力,用实时荧光定量PCR法检测成骨细胞OPG和RANKL基因的表达。结果 rhPTH1-34和PRO单独给药均可促进成骨细胞增殖(P <0.05),在PRO 10μM时成骨细胞OPG基因的表达量增加(P<0.05),且大于RANKL基因的表达量;相反,联合rhPTH 1-34和PRO给药抑制成骨细胞增殖(P<0.05),并抑制成骨细胞OPG和RANKL基因的表达(P<0.05),且随着PRO浓度的增加,OPG和RANKL基因表达均呈下降趋势。结论 rhPTH1- 34和PRO单独给药均可促进成骨细胞增殖,而两者联合给药后却抑制了成骨细胞的增殖,可能是通过调控OPG和RANKL基因的表达来实现的。     

胰岛素样生长因子1对成骨细胞中BMP-2、BMP-7基因表达的影响

目的研究胰岛素样生长因子1(IGF-1)对成骨细胞中BMP-2、BMP-7基因表达的影响,探讨IGF-1促进成骨细胞增殖、分化的分子机理.方法以不同浓度的IGF-1刺激分离培养的大鼠成骨细胞,提取细胞总RNA,RT-PCR扩增BMP-2及BMP-7基因cDNA,同时扩增管家基因βactin作为内对照.扫描PCR产物电泳照片,计算BMP-2/β-actin及BMP-7/-βactin的积分光密度比值,推算BMP-2及BMP-7基因的相对表达水平.结果 IGF-1可以在转录水平增加大鼠成骨细胞中BMP-2及BMP-7基因的表达,并呈明显的剂量依赖关系.结论 IGG-1促进成骨细胞增殖、分化的作用可能是通过增加BMP-2及BMP-7基因的表达来实现的.     

甲状旁腺激素联合辛伐他汀对成骨细胞分化及OPG和RANKL mRNA表达的影响

目的联合甲状旁腺激素(rhPTH1-34)和辛伐他汀(SIM)在体外对乳鼠颅盖骨成骨细胞分化及骨保护素(OPG)和核因子κB受体活化因子配体(RANKL)基因表达的影响。方法以乳鼠成骨细胞为体外试验模型,rhPTH1-34(10-9mol/L)联合不同浓度SIM(10-8、10-7、10-6mol/L)作用于体外培养的乳鼠成骨细胞,采用对硝基苯磷酸盐(PNPP)法测定碱性磷酸酶(ALP)活性;RT-PCR法测定OPG和RANKL基因的表达。结果 rhPTH1-34和SIM单独给药均可促进成骨细胞ALP活性及OPG基因、降低RANKL基因表达(P0.05);两者联合后与SIM单独作用组比较,ALP活性明显增加,并能协同促进OPG、降低RANKL基因表达(P0.05)。结论 rhPTH1-34和SIM联合应用对成骨细胞分化和代谢有协同作用。     

胰岛素样生长因子-1对甲状旁腺激素介导成骨细胞增殖及成骨活性的影响

目的：通过构建乳鼠颅盖骨成骨细胞分离培养与鉴定方法,研究胰岛素样生长因子－1（Insulin－like growth factor－1,IGF－1）及IGF－1联合重组人甲状旁腺激素1－34（recombinant human parathyroid hormone 1－34,rhPTH1－34）对成骨细胞增殖及I型胶原蛋白mRNA表达的影响。方法培养乳鼠成骨细胞,并观察其形态及功能,以原代培养乳鼠成骨细胞为实验模型,分空白组、PTH组、IGF－1组及不同浓度IGF－1作用的PTH介导的成骨细胞组（0、10、50、100 ng／L）。通过不同剂量的胰岛素样生长因子－1（0、10、50、100 ng／L）联合10－9 mol／L重组人甲状旁腺素刺激体外培养的成骨细胞,用噻唑蓝（ MTT）法检测细胞的增殖能力,RT－PCR法检测成骨细胞I型胶原蛋白mRNA的表达。结果 PTH和IGF－1均可促进成骨细胞增殖;IGF－1联合PTH可以促进成骨细胞增殖,且具有剂量依赖性。 PTH联合IGF－1使成骨细胞增殖能力增强、I型胶原蛋白mRNA表达增强。 IGF－1可以促进成骨细胞I型胶原蛋白mRNA的表达,且具有剂量依赖性。 IGF－1可以促进成骨细胞I型胶原蛋白mRNA的表达,且具有剂量依赖性。结论 PTH与IGF－1均可刺激成骨细胞增殖和分化,IGF－1可促进PTH对成骨细胞的分化、增殖。两者合用其作用增强,有协同促进作用。     

低能体外冲击波和低剂量间歇人重组甲状旁腺激素1-34刺激对体外培养大鼠成骨细胞增殖分化的影响

目的探讨低能体外冲击波（ESW）和低剂量间歇人重组甲状旁腺素1-34（rhPTH1-34）对体外培养大鼠成骨细胞（ROBs）增殖和成骨分化的影响。方法分别用不同次数的低能ESW刺激、不同浓度及作用方式的rhPTH1-34刺激，以及ESW和低剂量间歇性rhPTH1-34刺激共同作用于ROBs后，用细胞计数、MTY和流式细胞术细胞周期分析检测ROBs的增殖，用酶标仪检测ALP活性，用免疫组化检测I型胶原表达来观察ROBs成骨分化。结果60～150次0．18mJ／mm^2 ESW刺激、间歇性rhPTH1-34（1×10^-11和1×10^-10 mol／L）刺激以及ESW＋间歇性rhPTH1-34（1×10^-11mol／L）刺激均可显著促进体外培养ROBs细胞增殖和成骨分化（与对照组比较，P〈0．05）；其中60～150次ESW刺激＋间歇性rhPTH1-34（1×10^-11 mol／L）刺激各组作用最强。结论适当的ESW应力刺激和低剂量间歇性rhPTH1-34刺激可显著促进体外培养ROBs细胞增殖和成骨分化，两者联合应用作用最强。     

甲状旁腺素羧基端肽段的表达及其对大鼠成骨细胞增殖的影响

目的：探讨 rhPTH1～84、rhPTH37～84及 hPTH1～34对大鼠成骨细胞增殖的影响。方法分子生物学方法克隆表达 hPTH1～84、hPTH37～84重组蛋白。体外培养的大鼠成骨细胞给予不同浓度的 PTH 作用后,采用噻唑蓝（MTT）法检测细胞增殖。结果获得了浓度约为261．82 mg／L 的 rhPTH1～84和浓度为165．45 mg／L 的rhPTH37～84。10－10～10－7 mol／L 范围的 rhPTH1～84和 hPTH1～34促进大鼠成骨细胞的增殖;rhPTH37～84则抑制大鼠成骨细胞增殖。结论成功获得了高浓度高纯度的 rhPTH1～84及 rhPTH37～84。rhPTH37～84抑制细胞增殖,为进一步研究甲状旁腺激素及其羧基端肽段的功能奠定了基础。     

Prevention of atrophic nonunion development by recombinant human bone morphogenetic protein-7.

Severe periosteal and soft tissue disruption at the time of fracture may result in the formation of an atrophic nonunion. We have developed a reproducible atrophic nonunion in an animal model. The purpose of this study was to evaluate whether the immediate application of recombinant human BMP-7 to the fracture site could rescue the healing process in this nonunion model. A total of 56 three month old Fisher 344 rats were utilized. A 1.25 mm diameter K-wire was inserted into the femur in a retrograde fashion, and a mid-diaphyseal closed transverse fracture was created using a standard three point bending device. To create a nonunion, the fracture site was exposed and 2 mm of the periosteum was cauterized on each side of the fracture. The fracture site was immediately treated with either the application of rhBMP-7 50 microg in 25 microl of rat tail tendon collagen buffer (BMP-7 group), or with 25 microl of rat tail tendon collagen buffer only (Control group). Fracture healing was evaluated with serial radiographs every two weeks for an eight weeks period. Specimens at four and eight weeks were subjected to biomechanical and histological evaluation. None of the Control group healed throughout the eight weeks experimental duration. At four weeks 63% of the BMP-7 group had healed, and all had healed by six weeks. This investigation showed pronounced differences between the BMP-7 group and the Control group both histologically and biomechanically. In conclusion, we have demonstrated that the immediate application of BMP-7 may rescue the fracture healing process and prevent the development of nonunion following severe periosteal disruption.     

骨形态构建蛋白-2在气道高敏感反应小鼠模型肺组织中的表达

目的 观察骨形态构建蛋白-2(bone morphogenetic protein-2,BMP-2)在气道高敏感反应模型小鼠肺组织中的表达. 方法 Balb/C小鼠20只,按随机数字表法分为2组.对照组在0 d和7 d腹腔注射氢氧化铝2 mg(溶于生理盐水0.3 ml),于14、21、22 d用生理盐水雾化吸入,每天1次,每次30min.气道高敏感反应模型组在Od和7 d腹腔注射抗原混悬液(卵清蛋白20μg加氢氧化铝2 mg溶于生理盐水0.3 m1)致敏,于14、21、22 d用20 g/L卵清蛋白雾化吸入,每天1次,每次30 min.通过免疫学、组织学及气道反应性检测气道高敏感反应;Western检测肺组织BMP-2蛋白,荧光定量PCR检测肺组织mRNA含量表达. 结果模型组发生了气道高敏感反应.模型组较对照组肺组织BMP-2蛋白(1.64±0.05 vs 0.85±0.08)及mRNA(1.47±0.19vs 0.45±0.09)表达水平增高,差异有统计学意义(P<0.01).结论 卵清蛋白致敏并激发可产生小鼠气道高敏感反应,BMP-2参与了气道高敏感反应.     

体内负压技术对家兔颅骨修复及BMP-2 mRNA表达水平的影响

[目的]间歇性负压可以促进人骨髓间充质干细胞(bone marrow-derived stroma cells,BMSCs)向成骨细胞定向分化,上调成骨相关基因的表达水平。通过观察体内负压技术对兔颅骨骨孔修复过程及骨形态发生蛋白-2(bone morphogenetic protein-2,BMP-2)表达水平的影响,探讨负压技术在体内对骨组织修复作用。[方法]建立体内负压吸引干预动物模型,负压处2个骨孔为实验组,另外2个骨孔为对照组。对动物进行体内负压吸引技术干预(50 kPa,30 min/次,2次/d,间歇性负压干预1周)后,通过X线、骨密度观察其对家兔颅骨修复过程的影响,荧光定量RT-PCR检测BMP-2 mRNA的表达水平。[结果]术后2、4周实验组骨孔区X线阻射程度分别为(43.14±2.26)、(82.95±2.43),对照组分别为(25.13±2.12)、(51.49±2.37),差异有统计学意义(P<0.05);2周及4周实验组骨密度分别为(0.236±0.012)、(0.282±0.004),对照组分别为(0.174±0.005)、(0.213±0.009),两组相比差异有统计学意义(P<0.05);术后2、4周实验组BMP-2 mRNA的表达水平分别为(0.52±0.11)、(0.73±0.14),对照组分别为(0.29±0.07)、(0.36±0.09),两组相比差异有统计学意义(P<0.05)。[结论]体内负压技术可以上调BMP-2 mRNA表达水平,加速兔颅骨修复的进程,促进骨组织再生。     

Amniotic membrane attenuates heterotopic ossification following high-dose bone morphogenetic protein-2 treatment of segmental bone defects

Treatment of large bone defects with supraphysiological doses of bone morphogenetic protein-2 (BMP-2) has been associated with complications including heterotopic ossification (HO), inflammation, and pain, presumably due to poor spatiotemporal control of BMP-2. We have previously recapitulated extensive HO in our rat femoral segmental defect model by treatment with high-dose BMP-2 (30 μg). Using this model and BMP-2 dose, our objective was to evaluate the utility of a clinically available human amniotic membrane (AM) around the defect space for guided bone regeneration and reduction of HO. We hypothesized that AM surrounding collagen sponge would attenuate heterotopic ossification compared with collagen sponge alone. In vitro, AM retained more BMP-2 than a synthetic poly(ε-caprolactone) membrane through 21 days. In vivo, as hypothesized, the collagen + AM resulted in significantly less heterotopic ossification and correspondingly, lower total bone volume (BV), compared with collagen sponge alone. Although bone formation within the defect was delayed with AM around the defect, by 12 weeks, defect BVs were equivalent. Torsional stiffness was significantly reduced with AM but was equivalent to that of intact bone. Collagen + AM resulted in the formation of dense fibrous tissue and mineralized tissue, while the collagen group contained primarily mineralized tissue surrounded by marrow-like structures. Especially in conjunction with high doses of growth factor delivered via collagen sponge, these findings suggest AM may be effective as an overlay adjacent to bone healing sites to spatially direct bone regeneration and minimize heterotopic ossification.     

In vivo bone formation in fracture repair induced by direct retroviral-based gene therapy with bone morphogenetic protein-4

This study sought to develop an in vivo gene therapy to accelerate the repair of bone fractures. In vivo administration of an engineered viral vector to promote fracture healing represents a potential high-efficacy, low-risk procedure. We selected a murine leukemia virus (MLV)-based retroviral vector, because this vector would be expected to target transgene expression to the proliferating periosteal cells arising shortly after bone fracture. This vector transduced a hybrid gene that consisted of a bone morphogenetic protein (BMP)-4 transgene with the BMP-2 secretory signal to enhance the secretion of mature BMP-4. The MLV vector expressing this BMP-2/4 hybrid gene or β-galactosidase control gene was administered at the lateral side of the fracture periosteum at 1 day after fracture in the rat femoral fracture model. X-ray examination by radiograph and peripheral quantitative computed tomography at 7, 14, and 28 days after fracture revealed a highly significant enhancement of fracture tissue size in the MLV-BMP-2/4-treated fractures compared to the control fractures. The tissue was extensively ossified at 14 and 28 days, and the newly formed bone exhibited normal bone histology. This tissue also exhibited strong immunohistochemical staining of BMP-4. Additional control and MLV-BMP-2/4-treated animals each were monitored for 70 days to determine the fate of the markedly enhanced fracture callus. Radiographs showed that the hard callus had been remodeled and substantial healing at the fracture site had occurred, suggesting that the union of the bone at the fracture site was at least as high in the BMP-4-treated bone as in the control bone. There was no evidence of viral vector infection of extraskeletal tissues, suggesting that this in vivo gene therapy for fracture repair is safe. In summary, we have demonstrated for the first time that a MLV-based retroviral vector is a safe and effective means of introducing a transgene to a fracture site and that this procedure caused an enormous augmentation of fracture bone formation.     

骨形态发生蛋白-2对兔骨髓基质细胞向软骨细胞分化基因表达的影响

目的 观察骨形态发生蛋白-2(BMP-2)基因对兔骨髓基质细胞(MSCs)向软骨细胞分化mRNA表达的影响.方法 从兔骨髓细胞中分离提取BMP-2 mRNA,构建重组BMP-2真核表达质粒,并转染兔MSCs,采用荧光定量逆转录-聚合酶链反应(RT-PCR)方法检测各组MSCs内Ⅱ型胶原和蛋白多糖mRNA表达水平.结果 转染后2、4 d实验组MSCs内的Ⅱ型胶原和蛋白多糖mRNA表达量28.7±4.0、236.7±48.5及26.9±4.3、208.2±36.7,均明显高于转染后2、4 d的空白对照组(16.1±2.8、99.2±24.8及14.6±2.7、111.1±18.9)和空载体对照组(14.6±2.6、85.4±24.7及16.1±2.8、98.0±22.5)(P＜0.05).结论 重组真核表达载体BMP-2质粒能够诱导MSCs向软骨细胞分化.

Abstract：

Objective To observe the effect of bone morphogenetic protein-2 (BMP-2) on gene expression during the differentiation of marrow stromal cells (MSCs) into chondrocytes. Methods The BMP-2 mRNA was extracted from the rabbit bone marrow cells. The recombinant BMP-2 eukaryotic expression plasmids were constructed and transfected into MSCs. The mRNA expression of type Ⅱ collagen and proteoglycan was detected by using quantitative polymerase chain reaction (PGR) technique. Results At 2nd, and 4th day after transfection of plasmid pEGFP-C3-BMP-2 into MSCs, the mRNA expression levels of type Ⅱ collagen and proteoglycan in MSCs of experimental group were significantly higher than controls (P ＜ 0. 05 ). Conclusion Recombinant eukaryotic expression plastmid pEGFP-C3-BMP-2 can induce MSCs differentiation towards chondrocytes.     

Neuroforaminal chondrocyte metaplasia and clustering associated with recombinant bone morphogenetic protein-2 usage in transforaminal lumbar interbody fusion

Background contextRecombinant human bone morphogenetic protein-2 (rhBMP-2) is commonly used to augment posterior and interbody spinal fusion techniques and has many reported side effects. Neuroforaminal heterotopic ossification (HO) is a known cause of postoperative leg pain, but the pathohistologic composition of this material is not well understood.PurposeThe purpose of this article was to report the histologic composition of a case of HO and lumbar radiculopathy after transforaminal lumbar interbody fusion with rhBMP-2.Study design/settingThis is a case report.Patient sampleThis is a single patient case report.Outcome measuresThe outcomes considered were physician-recorded clinical, physiological, and functional measures.MethodsA retrospective review of a single patient was performed. Clinical, radiographic, and pathologic specimens were reviewed and are reported.ResultsA 69-year-old woman presented with low back pain and right leg radicular pain associated with L4–L5 stenosis and a recurrent facet cyst. After attempted nonsurgical care, she underwent an L4–L5 revision decompression with interbody and posterolateral fusions including off-label rhBMP-2. Postoperatively, her symptoms resolved for approximately 7 months but then returned in association with right L4–L5 foraminal HO. The ectopic tissue was notably larger than suggested by preoperative computed tomographic scan. It was decompressed, which then improved her symptoms. Histologic examination of the specimen revealed three discrete tissue types: a nonspecific fibrovascular stroma; immature osteoid and woven bone; and chondrocyte metaplasia with chondrocyte clustering.ConclusionsNeuroforaminal HO formation is a reported side effect associated with the off-label use of rhBMP-2 for posterior lumbar interbody fusion. The mechanism of formation and the composition of this material are not well understood but may involve a chondrocyte differentiation pathway.     

高糖对骨形态发生蛋白-2、胰岛素样生长因子-1基因转染大鼠骨髓基质干细胞增殖的影响

目的 观察高糖环境下骨形态发生蛋白-2(BMP-2)和胰岛素样生长因子-1(IGF-1)基因转染大鼠骨髓基质干细胞(BMSC)后BMSC的增殖.方法 用Ad-BMP-2和Ad-IGF-1转染大鼠BMSC,Wester blot检测蛋白表达.噻唑蓝(MTT)比色法及流式细胞术检测细胞增殖.结果 Western blot检测到转染组细胞中有目的 蛋白表达.MTT结果显示第5天细胞增殖能力达到高峰,5 d光吸收值:A～E组分别为0.324±0.027、0.319±0.017、0.622±0.028、0.626±0.020、0.778±0.031.流式细胞术结果显示A～E组细胞处于增殖期的比重分别为23.92±3.07、23.51±2.11、34.37±6.85、35.04±1.45、42.56±1.15.结论 高糖环境下BMP-2和IGF-1基因转染均能促进BMSC增殖,联合转染对BMSC增殖有协同效应.

Abstract：

Objective To observe the proliferation of bone marrow stromal cell (BMSC) transfected by bone morphogenetic protein-2 (BMP-2) and insulin-like growth factor-1 ( IGF-1 ) gene under high glucose condition.Methods Ad-BMP-2 and Ad-IGF-1 transfected rat BMSC,protein expression of BMSC were detected by Western blotting analysis.Methyl thiazol tetrazolium (MTT) assay and flow cytometry to observe the proliferation potential of BMSC.Results In the Western blotting analysis,positive signal lane of protein was observed in transfected group.MTT assay show that proliferation reached the peak in all groups on the fifth day,and the absorbency values of A to E group were 0.324 ± 0.027,0.319 ± 0.017,0.622 ±0.028,0.626 ± 0.020,0.778 ± 0.031.Flow cytometry show that the proliferative percentage from A to E group were 23.92 ±3.07,23.51 ±2.11,34.37 ±6.85,35.04 ± 1.45,42.56 ± 1.15.Conclusion BMP-2 or IGF-1 can promote the proliferation of BMSC under high glucose condition,but the combined has the synergy effect.