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Establishment and identification of fibroblast clones expressing human bone morphogenetic protein 2
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作者 JuanWang WeibinSun +1 位作者 ChunLu GuixiaTang 《Journal of Nanjing Medical University》 2005年第1期51-54,共4页
Objective:To establish fibroblasts stably expressing human bone morphogenetic protein 2(hBMP2). Methods:Eukaryonic expression vector(pcDNA3.1-B2) was transduced into NIH3T3 cells using SofastTM, a new generation catio... Objective:To establish fibroblasts stably expressing human bone morphogenetic protein 2(hBMP2). Methods:Eukaryonic expression vector(pcDNA3.1-B2) was transduced into NIH3T3 cells using SofastTM, a new generation cationic polymer gene transfection reagent. The positive cell clones were selected with G418. The stable transfection and expression of BMP2 in the NIH3T3 cells were determined by RT-PCR and immunohistochemical stain. Results: BMP2 mRNA was transcripted and expressed in the transfected NIH3T3 cells. Conclusion: With positive compound transfection, outside human BMP2 gene can be successfully transducted into NIH3T3 cells, which is the key step to induce periodontal cells to osseous phenotypes. 展开更多
关键词 bone morphogenetic protein 2 gene transfection fibroblastic cells
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EFFECTS OF TRANSFORMING GROWTH FACTOR β AND RECOMBINANT HUMAN BONE MORPHOGENETIC PROTEIN 2 ON HUMAN PERIODONTAL LIGAMENT FIBROBLASTS
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作者 司晓辉 刘正 《Journal of Shanghai Second Medical University(Foreign Language Edition)》 2001年第1期36-40,共5页
Objective To evaluate the effects of transforming growth factor β(TGF-β) and recombinant human bone morphogenetic protein 2 (rhBMP2) on human periodontal ligament fibroblasts (HPDLFs). Methods HPDLFs were done prima... Objective To evaluate the effects of transforming growth factor β(TGF-β) and recombinant human bone morphogenetic protein 2 (rhBMP2) on human periodontal ligament fibroblasts (HPDLFs). Methods HPDLFs were done primary culture to detect the distinct concentrations of TGF-P and rhBMF2 on its proliferation, alkaline phosphatase (ALP) activity, osteocalcin (OC) synthesis and formation of the minerali-zed nodules, respectively. Results TGF-β (5~100ng/ml) significantly stimulated the proliferation of HPDLFs. The ALP activity of HPDLFs was evaluated evidently by 5ng/ml TGF-β. TGF-β( 0. 5 ~ 100ng/ml) had no effects on OC synthesis and formation of the mineralized nodules of HPDLFs. rhBMP2 (0. 25~2mg/ ml) had no remarkable effect on the proliferation of HPDLFs. The ALP activity, OC synthesis and forma-tion of the mineralized nodules of HPDLFs were significantly stimulated by 0. 5~ 2mg /ml rhBMP2. Conclusion The effects of TGF-β and rhBMP2 on HPDLFs are dose-dependent. TGF-P can stimulate HPDLFs to express the early marker of osteoblastic phenotype, and it lacks the ability to promote maturation of the osteogenic phenotype. rhBMP2 can not only stimulate the expression but also promote the maturation of osteoblas-tic phenotype of HPDLFs. 展开更多
关键词 transforming growth factor Precombinant human bone morphogenetic protein 2human periodontal ligament fibroblastsalkaline phosphataseosteocalcin mineralization
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Construction of bicistronic green fluorescent protein labeled pSELECT GFPzeo human bone morphogenetic protein 2 eukaryotic expression vector
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作者 黄洪超 《外科研究与新技术》 2011年第2期91-91,共1页
Objective To construct green fluorescent protein (GFP)-labeled pSELECT-GFP zeohBMP2 eukaryotic expression vector.Methods The encoding fragment of hBMP2 gene was obtained from a recombinant plasmid pcDNA3.1/CT-hBMP2 by... Objective To construct green fluorescent protein (GFP)-labeled pSELECT-GFP zeohBMP2 eukaryotic expression vector.Methods The encoding fragment of hBMP2 gene was obtained from a recombinant plasmid pcDNA3.1/CT-hBMP2 by using polymerase 展开更多
关键词 PCR GFP Construction of bicistronic green fluorescent protein labeled pSELECT GFPzeo human bone morphogenetic protein 2 eukaryotic expression vector GENE
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Acupotomy ameliorates subchondral bone absorption and mechanical properties in rabbits with knee osteoarthritis by regulating bone morphogenetic protein 2-Smad1 pathway 被引量:2
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作者 Chen Xilin GUO Yan +8 位作者 LU Juan QIN Luxue HU Tingyao ZENG Xin WANG Xinyue ZHANG Anran ZHUANG Yuxin ZHONG Honggang GUO Changqing 《Journal of Traditional Chinese Medicine》 SCIE CSCD 2023年第4期734-743,共10页
OBJECTIVE:To investigate the effects of acupotomy on the subchondral bone absorption and mechanical properties in rabbits with knee osteoarthritis(KOA).METHODS:The rabbits were divided into blank control,model,acupoto... OBJECTIVE:To investigate the effects of acupotomy on the subchondral bone absorption and mechanical properties in rabbits with knee osteoarthritis(KOA).METHODS:The rabbits were divided into blank control,model,acupotomy and electroacupuncture(EA)groups,with 12 rabbits in each.Modified Videman's method was used to prepare KOA model.The acupotomy and EA group were given indicated intervention for 3 weeks.The behavior of rabbits in each group was recorded.Subsequently,cartilage–subchondral bone units were obtained and morphological changes were observed by optical microscope and micro computed tomography.Compression test was used to detect the mechanical properties of subchondral bone,Western blot and real-time polymerase chain reaction(RT-PCR)were applied to detect the expression of bone morphogenetic protein 2-Smad1(BMP2-Smad1)pathway in subchondral bone.RESULTS:Compared with the control group,rabbits in the KOA group showed lameness,knee pain,and cartilage degradation;the subchondral bone showed active resorption,the mechanical properties decreased significantly and the BMP2-Smad1 pathway downregulated significantly.Both acupotomy and EA intervention could increase the thickness of trabecular bone(Tb.Th),the bone volume fraction(BV/TV)and the thickness of subchondral bone plate,reduce the separation of trabecular bone(Tb.Sp),improve the maximum load and elastic modulus of subchondral bone,and effectively delay cartilage degeneration in KOA rabbits.This process may be achieved through upregulation the related proteins of BMP2-Smad1 pathway.The maximum load and elastic modulus of subchondral bone in the acupotomy group were slightly better than those in the EA group.CONCLUSIONS:Acupotomy could effectively protect cartilage by inhibiting abnormal bone resorption and improving mechanical properties of subchondral bone thorough the related proteins of BMP2-Smad1 pathway in KOA rabbits. 展开更多
关键词 acupuncture therapy OSTEOARTHRITIS knee subchondral bone mechanical properties bone morphogenetic protein 2 Smad1 protein
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Bone morphogenetic protein 2 promotes transforming growth factor β3-induced chondrogenesis of human osteoarthritic synovium-derived stem cells 被引量:19
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作者 RUI Yun-feng DU Lin +5 位作者 WANG You WANG Yang LUI Pauline po-yee TANG Ting-ting CHAN Kai-ming DAI Ke-rong 《Chinese Medical Journal》 SCIE CAS CSCD 2010年第21期3040-3048,共9页
Background Synovium-derived stem cells (SDSCs) with higher chondrogenic potential are attracting considerable attention as a cell source for cartilage regeneration. We investigated the effect of bone morphogenetic p... Background Synovium-derived stem cells (SDSCs) with higher chondrogenic potential are attracting considerable attention as a cell source for cartilage regeneration. We investigated the effect of bone morphogenetic protein 2 (BMP-2) on transforming growth factor beta3 (TGF-β3)-induced chondrogenesis of SDSCs isolated from human osteoarthritic synovium in a pellet culture system. Methods The clonogenicity, stem cell marker expression and multi-differentiation potential of isolated SDSCs were determined by colony forming unit assay, flow cytometry and specific staining including alizarin red S, Oil red O and alcian blue staining, respectively. SDSCs pellet was cultured in chondrogenic medium with or without TGF-β3 or/and BMP-2. At day 21, the diameter and the weight of the pellets were measured. Chondrogenic differentiation of SDSCs was evaluated by Safranin O staining, immunohistochemical staining of collagen type Ⅱ, sulfated glycosaminoglycan (sGAG) synthesis and mRNA expression of collagen type Ⅱ, aggrecan, SOX9, link-protein, collagen type X and BMP receptor Ⅱ. Results Cells isolated under the optimized culturing density (104/60 cm2) showed clonogenicity and multi-differentiation potential. These cells were positive (〉99%) for CD44, CD90, CD105 and negative (〈10%) for CD34 and CD71. SDSCs differentiated to a chondrocytic phenotype in chondrogenic medium containing TGF-β3 with or without BMP-2. Safranin O staining of the extracellular matrix was positive and the expression of collagen type Ⅱ was detected. Cell pellets treated with TGF-β3 and BMP-2 were larger in diameter and weight, produced more sGAGs, and expressed higher levels of collagen type Ⅱ and other chondrogenic markers, except COL10A1, than medium with TGF-β3 alone. Conclusions SDSCs could be isolated from human osteoarthritic synovium. Supplementation with BMP-2 significantly promoted the in vitro TGF-β3-induced chondrogenic differentiation of SDSCs. 展开更多
关键词 CHONDROGENESIS synovial membrane mesenchymal stem cell OSTEOARTHRITIS bone morphogenetic protein 2 transforming growth factor beta3
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Osteogenic potential of the human bone morphogenetic protein 2 gene activated nanobone putty 被引量:10
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作者 TIAN Xiao-bin SUN Li +3 位作者 YANG Shu-hua ZHANG Yu-kun HU Ru-yin FU De-hao 《Chinese Medical Journal》 SCIE CAS CSCD 2008年第8期745-751,共7页
Background Nanobone putty is an injectable and bioresorbable bone substitute. The neutral-pH putty resembles hard bone tissue, does not contain polymers or plasticizers, and is self-setting and nearly isothermic, prop... Background Nanobone putty is an injectable and bioresorbable bone substitute. The neutral-pH putty resembles hard bone tissue, does not contain polymers or plasticizers, and is self-setting and nearly isothermic, properties which are helpful for the adhesion, proliferation, and function of bone cells. The aim of this study was to investigate the osteogenic potential of human bone morphogenetic protein 2 (hBMP2) gene activated nanobone putty in inducing ectopic bone formation, and the effects of the hBMP2 gene activated nanobone putty on repairing bone defects. Methods Twenty four Kunming mice were randomly divided into two groups. The nanobone putty + hBMP2 plasmid was injected into the right thigh muscle pouches of the mice (experiment side). The nanobone putty + blank plasmid or nanobone putty was injected into the left thigh muscle pouches of the group 1 (control side 1) or group 2 (control side 2), respectively. The effects of ectopic bone formation were evaluated by radiography, histology, and molecular biology analysis at 2 and 4 weeks after operation. Bilateral 15 mm radial defects were made in forty-eight rabbits. These rabbits were randomly divided into three groups: Group A, nanobone putty + hBMP2 plasmid; Group B, putty + blank plasmid; Group C, nanobone putty only. Six rabbits with left radial defects served as blank controls. The effect of bone repairing was evaluated by radiography, histology, molecular biology, and biomechanical analysis at 4, 8, and 12 weeks after operation. Results The tissue from the experimental side of the mice expressed hBMP2. Obvious cartilage and island-distributed immature bone formation in implants of the experiment side were observed at 2 weeks after operation, and massive mature bone observed at 4 weeks. No bone formation was observed in the control side of the mice. The ALP activity in the experiment side of the mice was higher than that in the control side. The tissue of Group A rabbits expressed hBMP2 protein and higher ALP level. The new bone formation rate and antibending strength of group A was significantly higher than those of group B and C. The defects in blank control were not healed. Conclusions The hBMP2 gene activated nanobone putty exhibited osteoinductive ability, and had a better bone defect repair capabilitv than that of nanobone putty only. 展开更多
关键词 bone morphogenetic protein 2 gene activated matrix nanobone putty nanocrystalline hydroxyapatite gene therapy
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Construction and expression of a bicistronic vector containing human bone morphogenetic protein 2 and vascular endothelial growth factor-165 genes in vitro 被引量:3
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作者 TIAN Xiao-bin SUN Li +3 位作者 ZHANG Yu-kun GAO Yong FU De-hao YANG Shu-hua 《Chinese Medical Journal》 SCIE CAS CSCD 2009年第4期471-473,共3页
It has been well documented that bone morphogenetic .proteins (BMPs), a group of proteins belonging to the transforming growth factor-β (TGFβ) superfamily, can induce bone formation, both in vivo and in vitro. B... It has been well documented that bone morphogenetic .proteins (BMPs), a group of proteins belonging to the transforming growth factor-β (TGFβ) superfamily, can induce bone formation, both in vivo and in vitro. Bone morphogenetic protein-2 (BMP2) is a potent osteoinductive factor and is being evaluated as a bone growth inducer for orthopedic applications.1 Vascular endothelial growth factor (VEGF), the best-characterized angiogenic factor, 展开更多
关键词 bone morphogenetic protein 2 vascular endothelial growth factor-165 CO-EXPRESSION bicistronic vector
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Effects of secretive bone morphogenetic protein 2 induced by gene transfection on the biological changes of NIH3T3 cells 被引量:3
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作者 SUN Wei-bin WANG Juan +1 位作者 LU Chun TANG Gui-xia 《Chinese Medical Journal》 SCIE CAS CSCD 2005年第20期1703-1709,共7页
Background Bone morphogenetic proteins (BMPs), which belong to the transforming growth factor beta superfamily, are powerful regulators of cartilage and bone formation. This study investigated the biological changes... Background Bone morphogenetic proteins (BMPs), which belong to the transforming growth factor beta superfamily, are powerful regulators of cartilage and bone formation. This study investigated the biological changes of NIH3T3 ceils incubated with secretive BMP2 that was induced by gene transfection through transwell.Methods Eukaryonic expression vector (pcDNA3. 1-B2 ) was transfered into NIH333 cells with Sofast^TM, a positive compound transfection agent. The positive cell clones were selected with G418. The cytoplasmic and extracellular expressions of BMP2 were determined by immunohistochemical stain and enzyme-linked immunosorbent assay. NIH333 ceils were co-cultured with hBMP2 gene transfecting ceils through transwell, and the ultrastructure, alkaline phosphatase activity and the expression of osteocalcin (the marker of osteogenetic differentiation) changes were observed.Results There were cytoplasmic and extracellular expressions of BMP2 in transfecting NIH3T3 ceils. The ultrastructural changes, the high activity of alkaline phosphatase and the positive stain of osteocalcin suggested the osteogenetic differentiation tendency of NIH3T3 cells co-cultured with transfecting NIH3T3 cells.Conclusion Secretive BMP2 that is induced by gene transfection could promote the osteogenetic differentiation of fibroblast ceils. 展开更多
关键词 guided periodontal tissue regeneration bone morphogenetic protein 2 ultrastructure ·alkaline phosphatase OSTEOCALCIN
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Effects of Modified Qing'e Pill(加味青娥丸) on Expression of Adiponectin,Bone Morphogenetic Protein 2 and Coagulation-Related Factors in Patients with Nontraumatic Osteonecrosis of Femoral Head 被引量:7
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作者 LI Cheng-gang SHEN Lin +3 位作者 YANG Yan-Ping XU Xiao-Juan SHUAI Bo MA Chen 《Chinese Journal of Integrative Medicine》 SCIE CAS CSCD 2017年第3期183-189,共7页
Objective: To observe the regulation of Chinese herbal medicine, Modified Qing'e Pill(加味青娥丸, MQEP), on the expression of adiponectin, bone morphogenetic protein 2(BMP2), osteoprotegerin(OPG) and other pot... Objective: To observe the regulation of Chinese herbal medicine, Modified Qing'e Pill(加味青娥丸, MQEP), on the expression of adiponectin, bone morphogenetic protein 2(BMP2), osteoprotegerin(OPG) and other potentially relevant risk factors in patients with nontraumatic osteonecrosis of the femoral head(ONFH). Methods: A total of 96 patients with nontraumatic ONFH were unequal randomly divided into treatment group(60 cases) and control group(36 cases). The treatment group were treated with MQEP while the control group were treated with simulated pills. Both groups were given caltrate D. Six months were taken as a treatment course. Patients were followed up every 2 months. The levels of plasma adiponectin, BMP2, OPG, von Willebrand factor(vWF), von Willebrand factor cleaving protease(vWF-cp), plasminogen activator inhibitor 1(PAI-1), tissue plasminogen activator(tPA), C-reactive protein(CRP), blood rheology, bone mineral density(BMD) of the femoral head and Harris Hip Score were measured before and after treatment. Results: After 6 months of treatment, compared with the control group, patients in the treatment group had significantly higher adiponectin and BMP2 levels(P〈0.01 and P=0.013, respectively), lower vWF, PAI-1 and CRP levels(P=0.019, P〈0.01 and P〈0.01, respectively), and lower blood rheology parameters. BMD of the femoral neck, triangle area and Harris Hip Score in the treatment group were significantly higher than those in the control group. Moreover, plasma adiponectin showed a positive association with BMP2(r=0.231, P=0.003) and a negative association with PAI-1(r=–0.159, P〈0.05). Conclusions: MQEP may play a protective role against nontraumatic ONFH by increasing the expression of adiponectin, regulating bone metabolism and improving the hypercoagulation state, which may provide an experimental base for its clinical effects. 展开更多
关键词 osteonecrosis femoral head Chinese herbal medicine adiponectin bone morphogenetic protein 2 plasminogen activator inhibitor 1
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SPECIFIC BINDING OF HUMAN BONEMORPHOGENETIC PROTEIN (2A) WITH MOUSEOSTEOBLASTIC CELLS
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作者 刘新平 陈苏民 +2 位作者 陈南春 高磊 赵忠良 《Chinese Medical Sciences Journal》 CAS CSCD 1996年第2期97-99,共3页
Human bone morphogenetic protein 2A (hBMP2A) cDNA terminal 567 nucleotides were cloned and expressed in a phage display vector pCSM21. Human BMP2A C-terminal peptide displayed on the surface of the phage can bind spec... Human bone morphogenetic protein 2A (hBMP2A) cDNA terminal 567 nucleotides were cloned and expressed in a phage display vector pCSM21. Human BMP2A C-terminal peptide displayed on the surface of the phage can bind specifically to the surface of mouse osteoblastic cell (MC3T3) membrane. ELISA assay showed a positive signal of the binding by using antibody against M13 phage gene 8 protein. After labeling with 3HTdR,the counts of the binding groups were 3 to 10 times higher than the control groups. It suggests that the surface of MC3T3 cells exist the receptor for hBMP2A. 展开更多
关键词 human bone morphogenetic protein 2A ELISA 3HTdR incorporation
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CORAL AS A CARRIER FOR RECOMBINANT HUMAN BONE MORPHOGENETIC PROTEIN-2
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作者 张森林 毛天球 +1 位作者 孟昭业 王会信 《Chinese Medical Sciences Journal》 CAS CSCD 1999年第2期125-128,共4页
By combining coral with recombinant human bone morphogenetic protein-2 (rhBMP-2), rhBMP-2/coral composite was obtained in this study. Following implantation of the composite into the muscle pouches of mice, cartilage ... By combining coral with recombinant human bone morphogenetic protein-2 (rhBMP-2), rhBMP-2/coral composite was obtained in this study. Following implantation of the composite into the muscle pouches of mice, cartilage growth was induced in the pores or on the surface of the implants at one week, woven bone at three week and lamellar bone with bone marrow at six week, and coral was absorbed partially. The induced formation of endochondral bone was time-related and rhBMP-2 dose-related. The results of this study indicate that the composite possesses a superior ability of osteogenesis, and coral acts as one of the most suitable rhBMP-2 slowrelease carriers currently available. The composite will be a new type of bone substitute to be used in orthopaedics and maxillofacial surgery. 展开更多
关键词 recombinant human bone morphogenetic protein 2 OSTEOINDUCTION CARRIER
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Human bone morphogenetic protein-2 gene transfer induces human mesenchymal stem cell proliferation and differentiation in vitro
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作者 李军 范清宇 +3 位作者 钱济先 马保安 周勇 张明华 《Journal of Medical Colleges of PLA(China)》 CAS 2004年第2期115-119,共5页
Objective: To identify eukaryotic expression vector of human bone morphogenetic protein 2 pcDNA3/BMP2, verify its expression in transfected human mesenchymal stem cells (hMSCs) and the effect on hMSCs differentiation.... Objective: To identify eukaryotic expression vector of human bone morphogenetic protein 2 pcDNA3/BMP2, verify its expression in transfected human mesenchymal stem cells (hMSCs) and the effect on hMSCs differentiation. Methods: The BMP2 gene was cloned into a eukaryotic expression vector pcDNA3. Transfected the recombinant into hMSCs by liposome. Immunnohistochemistry and in situ hybridization methods were used to identify the expression of BMP2 mRNA and protein; ALP and Von Kossa stains were performed to identify the BMP2 gene differentiated effect on the hMSCs. Results: The pcDNA3/BMP2 fragments were as large as theory. BMP2 mRNA and protein were expressed and synthesized both in 48 h and 4 weeks after transfection, the ALP and Ca deposit exhibition, which marked the osteogenic lineage of hMSCs, were enhanced and sped. Conclusion: Transfection of pcDNA3/BMP2 is able to provide transient and persistent expression in hMSCs, and promote the MSCs differentiation to osteogenic lineage. 展开更多
关键词 bone morphogenetic protein(BMP2) TRANSFECTION human mesenchymal stem cells (hMSCs) osteogenic differentiation
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Ectopic Bone Formation in vivo Induced by a Novel Synthetic Peptide Derived from BMP-2 Using Porous Collagen Scaffolds 被引量:11
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作者 袁泉 郭晓东 《Journal of Wuhan University of Technology(Materials Science)》 SCIE EI CAS 2007年第4期701-705,共5页
To investigate the osteoinductive and ectopicly osteogenic effects of a novel peptide P24 derived from bone morphogenetic protein 2 (BMP2), biodegradable collagen scaffolds (CS) were used to load BMP-2-derived pep... To investigate the osteoinductive and ectopicly osteogenic effects of a novel peptide P24 derived from bone morphogenetic protein 2 (BMP2), biodegradable collagen scaffolds (CS) were used to load BMP-2-derived peptide solutions with different concentrations (0.4 mg peptide/CS, 0.1 mg peptide/CS and pure CS, respectively), and the implants were implanted into muscular pockets on the back of Wistar rats. Radiographs and histological analysis were performed to evaluate the ectopic bone effects. Active ectopic bone formation was seen in both groups containing the peptide at different concentration (0.4 mg and 0.1 mg), whereas no bone formation and only fibrous tissue was seen in the pure CS group. The new bone formation induced by the peptide P24 displayed a dose-dependent and time-dependent efficiency. The new bone formation in the 0.4 mg peptide/CS group significantly increased than that of the 0.1 mg peptide/CS group. This novel BMP-2-derived peptide had excellent osteoinductive and ectopicly osteogenic properties which were similar to those of BMP2. 展开更多
关键词 bone tissue engineering biomimetic material bone morphogenetic protein 2 OSTEOINDUCTION PEPTIDE
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CO-TRANSFECTION OF RAT BONE MARROW MESENCHYMAL STEM CELLS WITH HUMAN BMP2 AND VEGF165 GENES 被引量:1
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作者 蒋佳 范存义 曾炳芳 《Journal of Shanghai Second Medical University(Foreign Language Edition)》 2009年第1期12-18,共7页
Objective To explore the feasibility and efficacy of lentivirus-mediated co-transfection of rat bone marrow mesenchymal stem cells (MSCs) with human vascular endothelial growth factor 165 (hVEGFI65) gene and human... Objective To explore the feasibility and efficacy of lentivirus-mediated co-transfection of rat bone marrow mesenchymal stem cells (MSCs) with human vascular endothelial growth factor 165 (hVEGFI65) gene and human bone morphogenetic protein 2 (hBMP2) gene. Methods The hVEGF165 and hBMP2 cDNAs were obtained from human osteosarcoma cell line MG63 and cloned into lentiviral expression vectors designed to co-express the copepod green fluorescent protein (copGFP). The expression lentivector and packaging Plasmid Mix were co-transferred to 293TN cells, which produced the lentivirus carrying hVEGF165 (Lv-VEGF) or hBMP2 ( Lv-BMP) , respectively. MSCs of Wistar rats were co-transfected with Lv-BMP and Lv-VEGF (BMP + VEGF group), or each alone (BMP group and VEGF group), or with no virus ( Control group). The mRNA and protein expressions of hVEGF165 and hBMP2 genes in each group were detected by real-time PCR and enzyme linked immunosorbent assay (ELISA). Results Lentiviral expression vectors carrying hVEGF165 or hBMP2 were correctly constructed and confirmed by restriction endonucleses analysis and DNA sequencing analysis. A transfer efficiency up to 90% was archieved in all the transfected groups detected by the fraction of fluorescent cells using fluorescent microscopy. From the results generated by real-time PCR and ELISA, VEGF165 and BMP2 genes were co-expressed in BMP + VEGF group. No significant difference of BMP2 expression was detected between BMP + VEGF and BMP groups ( P 〉 0. 05). Similarly, there was no significant difference of VEGF165 expression between BMP + VEGF and VEGF groups ( P 〉 0. 05). Conclusion VEGF165 and BMP2 genes were successfully co-expressed in MSCs by lentivirus-mediated co-transfection, which provided a further foundation for the combined gene therapy of bone regeneration. 展开更多
关键词 mesenchymal stem cells vascular endothelial growth factor lentiviral vector bone morphogenetic protein 2 gene therapy co-transfection
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Effect of nano-hydroxyapatite/collagen composite and bone morphogenetic protein-2 on lumbar intertransverse fusion in rabbits 被引量:11
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作者 孙天胜 关凯 +5 位作者 时述山 朱兵 郑永军 崔福斋 张伟 廖素三 《Chinese Journal of Traumatology》 CAS 2004年第1期18-24,共7页
Objective: To investigate the effect of nano hydroxyapatite/collagen (nHA/collagen) composite as a graft extender and enhancer when combined with recombinant human bone morphogenetic protein 2 (rhBMP 2) on lumbar inte... Objective: To investigate the effect of nano hydroxyapatite/collagen (nHA/collagen) composite as a graft extender and enhancer when combined with recombinant human bone morphogenetic protein 2 (rhBMP 2) on lumbar intertransverse fusion in rabbits. Methods: Sixty four adult female New Zealand white rabbits, aged 1 year and weighing 3.5 4.5 kg, underwent similar posterolateral intertransverse process arthrodesis and were randomly divided into 4 groups based on different grafts: autogenous cancellous bone alone (ACB group), nHA/collagen alone (HAC group), half autogenous cancellous bone and half nHA/collagen (ACB+HAC group) and nHA/collagen combined with rhBMP 2 (HAC+BMP group). The fusion masses were analyzed by manual palpation, radiography, biomechanical testing and histological examination. Results: Fusion was observed in 4 cases in the 6th week and in 5 cases in the 10th week after surgery in ACB group. No case showed fusion in HAC group. In ACB+HAC group, there was fusion in 3 cases in the 6th week and in 4 cases in the 10th week after surgery. In HAC+BMP group, fusion in 1 case was found in the 4th week, in 5 cases in the 6th week and in 6 cases in the 10th week after surgery. It suggested that ACB, ACB+HAC and HAC+BMP groups showed similar fusion ratio and mechanical strength in the 6th and 10th week after surgery. According to the microstructure analysis of the samples, nHA/collagen had no negative effect when implanted together with ilium autograft. In HAC+BMP group, new bone like tissue was observed in the 2nd week postoperatively, and nearly all of the implanted composites were replaced by mature bone matrix and new bones in 10th week postoperatively. Conclusions: The nHA/collagen, especially combined with rhBMP 2, is a promising bone substitute, for it has quick biodegradation, fine bone bending ability, and high osteoconductivity on posterolateral spinal fusion in rabbits. 展开更多
关键词 Lumbar vertebrae Spinal fusion Bone morphogenetic protein 2 Nano hydroxyapatite/collagen composite
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Co-delivery of rhBMP-2 and zoledronic acid using calcium sulfate/hydroxyapatite carrier as a bioactive bone substitute to enhance and accelerate spinal fusion
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作者 Xinggui Tian Corina Vater +10 位作者 Deepak Bushan Raina Lisa Findeisen Lucas-Maximilian Matuszewski Magnus Tägil Lars Lidgren Anja Winkler Robert Gottwald Niels Modler Klaus-Dieter Schaser Alexander C.Disch Stefan Zwingenberger 《Bioactive Materials》 SCIE CSCD 2024年第6期256-271,共16页
Recombinant human bone morphogenetic protein-2(rhBMP-2)has been FDA-approved for lumbar fusion,but supraphysiologic initial burst release due to suboptimal carrier and late excess bone resorption caused by osteoclast ... Recombinant human bone morphogenetic protein-2(rhBMP-2)has been FDA-approved for lumbar fusion,but supraphysiologic initial burst release due to suboptimal carrier and late excess bone resorption caused by osteoclast activation have limited its clinical usage.One strategy to mitigate the pro-osteoclast side effect of rhBMP-2 is to give systemic bisphosphonates,but it presents challenges with systemic side effects and low local bioavailability.The aim of this in vivo study was to analyze if posterolateral spinal fusion(PLF)could be improved by utilizing a calcium sulfate/hydroxyapatite(CaS/HA)carrier co-delivering rhBMP-2 and zoledronic acid(ZA).Six groups were allocated(CaS/HA,CaS/HA+BMP-2,CaS/HA+systemic ZA,CaS/HA+local ZA,CaS/HA+BMP-2+systemic ZA,and CaS/HA+BMP-2+local ZA).10-week-old male Wistar rats,were randomly assigned to undergo L4-L5 PLF with implantation of group-dependent scaffolds.At 3 and 6 weeks,the animals were euthanized for radiography,μCT,histological staining,or biomechanical testing to evaluate spinal fusion.The results demonstrated that the CaS/HA biomaterial alone or in combination with local or systemic ZA didn’t support PLF.However,the delivery of rhBMP-2 significantly promoted PLF.Combining systemic ZA with rhBMP-2 didn’t enhance spinal fusion.Notably,the co-delivery of rhBMP-2 and ZA using the CaS/HA carrier significantly enhanced and accelerated PLF,without inhibiting systemic bone turnover,and potentially reduced the dose of rhBMP-2.Together,the treatment regimen of CaS/HA biomaterial co-delivering rhBMP-2 and ZA could potentially be a safe and cost-effective off-the-shelf bioactive bone substitute to enhance spinal fusion. 展开更多
关键词 Calcium sulfate/hydroxyapatite Bone morphogenetic protein 2 BISPHOSPHONATE Spinal fusion Bone substitute
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A segmental defect adaptation of the mouse closed femur fracture model for the analysis of severely impaired bone healing
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作者 Amandeep Kaur Subburaman Mohan Charles H.Rundle 《Animal Models and Experimental Medicine》 CSCD 2020年第2期130-139,共10页
Objective: To better characterize nonunion endochondral bone healing and evaluate novel therapeutic approaches for critical size defect healing in clinically challenging bone repair, a segmental defect model of bone i... Objective: To better characterize nonunion endochondral bone healing and evaluate novel therapeutic approaches for critical size defect healing in clinically challenging bone repair, a segmental defect model of bone injury was adapted from the threepoint bending closed fracture technique in the murine femur.Methods: The mouse femur was surgically stabilized with an intramedullary threaded rod with plastic spacers and the defect adjusted to different sizes. Healing of the different defects was analyzed by radiology and histology to 8 weeks postsurgery. To determine whether this model was effective for evaluating the benefits of molecular therapy, BMP-2 was applied to the defect and healing then examined.Results: Intramedullary spacers were effective in maintaining the defect. Callus bone formation was initiated but was arrested at defect sizes of 2.5 mm and above, with no more progress in callus bone development evident to 8 weeks healing. Cartilage development in a critical size defect attenuated very early in healing without bone development, in contrast to the closed femur fracture healing, where callus cartilage was replaced by bone. BMP-2 therapy promoted osteogenesis of the resident cells of the defect, but there was no further callus development to indicate that healing to pre-surgery bone structure was successful.Conclusions: This segmental defect adaptation of the closed femur fracture model of murine bone repair severely impairs callus development and bone healing, reflecting a challenging bone injury. It is adjustable and can be compared to the closed fracture model to ascertain healing deficiencies and the efficacy of therapeutic approaches. 展开更多
关键词 bone fractures bone morphogenetic protein 2 intramedullary fracture fixation ununited fractures
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Immunoadsorption therapy for Klinefelter syndrome with antiphospholipid syndrome in a patient:A case report
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作者 You Song Yong-Zhen Xiao +1 位作者 Cheng Wang Rong Du 《World Journal of Clinical Cases》 SCIE 2022年第19期6580-6586,共7页
BACKGROUND Klinefelter syndrome(KS) is a genetic disease of male sex chromosome malformations that affects sperm production and reduces testosterone production. It has been reported that there is currently more than 1... BACKGROUND Klinefelter syndrome(KS) is a genetic disease of male sex chromosome malformations that affects sperm production and reduces testosterone production. It has been reported that there is currently more than 10 cases of KS combined with antiphospholipid syndrome(APS).CASE SUMMARY Here, we describe a 31-year-old male patient with chromosome 47, XXY type, who suffered deep vein thrombosis of the lower limbs accompanied by abnormal antiphospholipid antibody, lupus anticoagulant and factor VⅢ. After treatment with immunoadsorption therapy, glucocorticoids, cyclophosphamide, intravenous immunoglobulin and anticoagulant therapy, the patient showed dramatic symptomatic improvement. During the follow-up, the patient did not develop any new thrombotic events.CONCLUSION Immunoadsorption combined with glucocorticoid and cyclophosphamide shock comprehensive treatment has achieved significant results for patients with KS combined with antiphospholipid syndrome. 展开更多
关键词 Klinefelter syndrome Antiphospholipid syndrome IMMUNOADSORPTION GLUCOCORTICOID CYCLOPHOSPHAMIDE Bone morphogenetic protein receptor type-2 Case report
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CLONING AND SEQUENCING OF MATURE FRAGMENT OF HUMAN BMP4 GENE
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作者 王欣璐 刘淼 +2 位作者 杨广夫 王全颍 杨广笑 《Academic Journal of Xi'an Jiaotong University》 2000年第2期155-159,共5页
Objective To study the cloning and sequencing of mature fragment of human bone morphogenetic protein 4 gene. Methods The template DNA was obtained from the human osteosarcoma cell line U2OS. By using RT PCR method, th... Objective To study the cloning and sequencing of mature fragment of human bone morphogenetic protein 4 gene. Methods The template DNA was obtained from the human osteosarcoma cell line U2OS. By using RT PCR method, the cDNA coding for the mature fragment of BMP 4 was amplified, cloned into the vector pUC19, and sequenced by Sanger Dideoxy mediated Chain Termination method. Results The mature fragment of BMP4 cDNA was obtained by RT PCR and determined by sequencing. Through the computer search on Genebank, the analysis showed that the homology of nucleotides and amino acids between cDNA of rhBMP4 mature fragment of this study and the published sequence was 99%. Sequence analysis showed that there were two differences, one was at base 1154(201): G→C, which had no influence on the corresponding amino acids(Val). Another was at base1222(269):C→T, the mutation at the base 1222 had the change of Ala to Val. Conclusion The mature fragment of BMP4 gene has been cloned. The results will be of great significance in treatment of skeletal injuries and diseases. 展开更多
关键词 recombinant human bone morphogenetic protein 4(rhBMP4(2b)) molecular clonging sequencin
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Rhizoma Drynariae-derived nanovesicles reverse osteoporosis by potentiating osteogenic differentiation of human bone marrow mesenchymal stem cells via targeting ERα signaling
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作者 Qing Zhao Junjie Feng +11 位作者 Fubin Liu Qianxin Liang Manlin Xie Jiaming Dong Yanfang Zou Jiali Ye Guilong Liu Yue Cao Zhaodi Guo Hongzhi Qiao Lei Zheng Kewei Zhao 《Acta Pharmaceutica Sinica B》 SCIE CAS CSCD 2024年第5期2210-2227,共18页
Although various anti-osteoporosis drugs are available,the limitations of these therapies,including drug resistance and collateral responses,require the development of novel anti-osteoporosis agents.Rhizoma Drynariae ... Although various anti-osteoporosis drugs are available,the limitations of these therapies,including drug resistance and collateral responses,require the development of novel anti-osteoporosis agents.Rhizoma Drynariae displays a promising anti-osteoporosis effect,while the effective component and mechanism remain unclear.Here,we revealed the therapeutic potential of Rhizoma Drynariae-derived nanovesicles(RDNVs)for postmenopausal osteoporosis and demonstrated that RDNVs potentiated osteogenic differentiation of human bone marrow mesenchymal stem cells(hBMSCs)by targeting estrogen receptor-alpha(ERα).RDNVs,a natural product isolated from fresh Rhizoma Drynariae root juice by differential ultracentrifugation,exhibited potent bone tissue-targeting activity and anti-osteoporosis efficacy in an ovariectomized mouse model.RDNVs,effectively internalized by hBMSCs,enhanced proliferation and ERαexpression levels of hBMSC,and promoted osteogenic differentiation and bone formation.Mechanistically,via the ERαsignaling pathway,RDNVs facilitated mRNA and protein expression of bone morphogenetic protein 2 and runt-related transcription factor 2 in hBMSCs,which are involved in regulating osteogenic differentiation.Further analysis revealed that naringin,existing in RDNVs,was the active component targeting ERαin the osteogenic effect.Taken together,our study identified that naringin in RDNVs displays exciting bone tissue-targeting activity to reverse osteoporosis by promoting hBMSCs proliferation and osteogenic differentiation through estrogen-like effects. 展开更多
关键词 Rhizoma Drynariae-derived nanovesicles Bone marrow mesenchymal stem cells Bone targeting Osteogenic differentiation ERαsignaling NARINGIN Bone morphogenetic protein 2 Runt-related transcription factor 2
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