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Objective:To explore the long-term effect of hip replacement and vitamin D in treatment of elderly osteoporotic femoral neck fractures and the transfection of bone morphogenetic protein-7 (BMP-7) /25 hydroxy vitamin D3[ (25- (OH) ) -D3] level.Method:Data of 108 elderly osteoporotic femoral neck fracture patients admitted from Jan. 2018 to Jan. 2020 were selected, and they were divided them into the observation group (hip replacement adjuvant vitamin D treatment) and control group (hip replacement treatment) , 54 cases in each group. All subjects were followed up for 1 year to observe the long-term treatment efficacy and SF-36 scores of the two groups of patients. Before treatment and 1, 3, 6, and 12 months after treatment, the differences in the changes in lumbar and hip bone mineral density, hip joint Harris score, BMP-7, and 25- (OH) -D3 levels were compared between the two groups.Results:12 months after treatment, the long-term treatment effect of the observation group was significantly higher than that of the control group ( P<0.05) ; 1, 3, 6, 12 months after treatment, for the lumbar and hip bone mineral density, SF-36 score, hip Harris score, BMP-7, 25- (OH) -D3 levels of the two groups of patients over time, the degree of increase was not equal, but the treatment group had the greater degree of improvement ( P<0.01) . Conclusion:Hip joint replacement and vitamin D have a good long-term effect in treatment of elderly osteoporotic femoral neck fractures, which can significantly increase bone density and improve BMP-7 and 25- (OH) -D3 levels.
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Objective To detect the expression of transforming growth factor-β1 (TGF-β1), p38MAPK and bone morphogenetic protein-7 (BMP-7) protein in the liver specimens of patients with hepatic alveolar echinococcosis, and to investigate the potential role of TGF-β1, p38MAPK and BMP-7 protein in hepatic fibrosis caused by hepatic alveolar echinococcosis. Methods A total of 20 patients with hepatic alveolar echinococcosis were enrolled as study subjects, and hepatic specimens were sampled from the sites within 0.5 cm (Group A) and 0.5 to 1.5 cm from hepatic alveolar echinococcosis lesions (Group B), while normal liver specimens sampled from the sites 2 cm and greater from hepatic alveolar echinococcosis lesions served as controls (Group C). The fibrosis of liver specimens was pathological examined using HE and Masson staining, and the expression of TGF-β1, p38MAPK and BMP-7 protein was quantified in liver tissues using Western blotting. The associations of TGF-β1, p38MAPK and BMP-7 protein expression with hepatic fibrosis were assessed. Results HE staining showed the malaligned structure of hepatocytes and destruction of the structure of hepatic lobules at various degrees in liver specimens in groups A and B, with hepatocyte degeneration, atrophy and necrosis, hyperplasia of fibrous tissues and eosinophilic granulocyte infiltration seen, while no abnormal pathological alterations of liver tissues, normal hepatocyte structure and morphology and uniform size, no malaligned structure of hepatocytes, clear structure of hepatic lobules, no or mild hepatocyte degeneration or necrosis, and no eosinophilic granulocyte infiltration were seen in Group C. Masson staining showed that there was hyperplasia of multiple fibrous connective tissues in the liver portal areas in groups A and B, with fibrosis seen in hepatic lobules, while no obvious pathological changes were seen in Group C. There were significant differences seen in TGF-β1 (P < 0.001), p38MAPK (P < 0.01) and BMP-7 protein (P < 0.05) expression in liver tissues in groups A, B and C, and higher TGF-β1, p38MAPK and BMP-7 protein expression was quantified in groups A and B than in Group C (all P values < 0.05), while greater TGF-β1, p38MAPK and BMP-7 protein expression was detected in Group B than in Group C (all P values < 0.05). The expression of TGF-β1, p38MAPK and BMP-7 protein correlated positively with the severity of hepatic fibrosis (r = 0.866, 0.702 and 0.801, all P values < 0.05), and there were significant differences in TGF-β1 (F = 72.580, P < 0.01), p38MAPK (χ2 = 31.705, P < 0.01) and BMP-7 protein expression (χ2 = 48.388, P < 0.01) among liver tissues with different degrees of fibrosis. The TGF-β1 protein expression correlated positively with p38MAPK and BMP-7 protein expression (r = 0.607 and 0.702, both P values < 0.001), and the BMP-7 protein expression also correlated positively with p38MAPK protein expression (r = 0.456, P < 0.001). Conclusion The interaction among TGF-β1, p38MAPK and BMP-7 jointly participates in the development of hepatic fibrosis induced hepatic alveolar echinococcosis.
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Diabetic nephropathy (DN) is one of the leading causes of mortality in diabetic patients. Long non-coding RNA zinc finger E-box binding homeobox 1 antisense 1 (ZEB1-AS1) plays a crucial role in the development of various diseases, including DN. However, the molecular mechanism of ZEB1-AS1 in DN pathogenesis remains elusive. An in vitro DN model was established by treating HK-2 cells with high glucose (HG). Quantitative polymerase chain reaction (qRT-PCR) was utilized to detect the expression levels of ZEB1-AS1, microRNA-216a-5p (miR-216a-5p), and bone morphogenetic protein 7 (BMP7). Western blot assay was used to evaluate the protein levels of BMP7, epithelial-to-mesenchymal transition (EMT)-related proteins, and fibrosis markers. Additionally, the interaction among ZEB1-AS1, miR-216a-5p, and BMP7 was predicted by MiRcode (http://www.mircode.org) and starBase 2.0 (omics_06102, omicX), and confirmed by luciferase reporter assay. ZEB1-AS1 and BMP7 were down-regulated, while miR-216a-5p was highly expressed in kidney tissues of DN patients. Consistently, HG treatment decreased the levels of ZEB1-AS1 and BMP7, whereas HG increased miR-216a-5p expression in HK-2 cells in a time-dependent manner. ZEB1-AS1 upregulation inhibited HG-induced EMT and fibrogenesis. Furthermore, ZEB1-AS1 directly targeted miR-216a-5p, and overexpression of miR-216a-5p restored the inhibitory effects of ZEB1-AS1 overexpression on EMT and fibrogenesis. BMP7 was negatively targeted by miR-216a-5p. In addition, ZEB1-AS1 suppressed HG-induced EMT and fibrogenesis by regulating miR-216a-5p and BMP-7. lncRNA ZEB1-AS1 inhibited high glucose-induced EMT and fibrogenesis via regulating miR-216a-5p/BMP7 axis in diabetic nephropathy, providing a potential target for DN therapy.
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Diabetic nephropathy (DN) is one of the leading causes of mortality in diabetic patients. Long non-coding RNA zinc finger E-box binding homeobox 1 antisense 1 (ZEB1-AS1) plays a crucial role in the development of various diseases, including DN. However, the molecular mechanism of ZEB1-AS1 in DN pathogenesis remains elusive. An in vitro DN model was established by treating HK-2 cells with high glucose (HG). Quantitative polymerase chain reaction (qRT-PCR) was utilized to detect the expression levels of ZEB1-AS1, microRNA-216a-5p (miR-216a-5p), and bone morphogenetic protein 7 (BMP7). Western blot assay was used to evaluate the protein levels of BMP7, epithelial-to-mesenchymal transition (EMT)-related proteins, and fibrosis markers. Additionally, the interaction among ZEB1-AS1, miR-216a-5p, and BMP7 was predicted by MiRcode (http://www.mircode.org) and starBase 2.0 (omics_06102, omicX), and confirmed by luciferase reporter assay. ZEB1-AS1 and BMP7 were down-regulated, while miR-216a-5p was highly expressed in kidney tissues of DN patients. Consistently, HG treatment decreased the levels of ZEB1-AS1 and BMP7, whereas HG increased miR-216a-5p expression in HK-2 cells in a time-dependent manner. ZEB1-AS1 upregulation inhibited HG-induced EMT and fibrogenesis. Furthermore, ZEB1-AS1 directly targeted miR-216a-5p, and overexpression of miR-216a-5p restored the inhibitory effects of ZEB1-AS1 overexpression on EMT and fibrogenesis. BMP7 was negatively targeted by miR-216a-5p. In addition, ZEB1-AS1 suppressed HG-induced EMT and fibrogenesis by regulating miR-216a-5p and BMP-7. lncRNA ZEB1-AS1 inhibited high glucose-induced EMT and fibrogenesis via regulating miR-216a-5p/BMP7 axis in diabetic nephropathy, providing a potential target for DN therapy.
Subject(s)
Humans , Diabetic Nephropathies/metabolism , Bone Morphogenetic Protein 7/metabolism , Epithelial-Mesenchymal Transition/physiology , RNA, Long Noncoding/physiology , Zinc Finger E-box-Binding Homeobox 1/metabolism , Down-Regulation , Up-Regulation , Cells, Cultured , MicroRNAs/metabolism , Diabetic Nephropathies/genetics , Real-Time Polymerase Chain ReactionABSTRACT
@#Introduction: Chronic kidney disease (CKD) leads to tubular injury, kidney fibrosis and anemia. These conditions are influenced by fibrotic and anti-fibrotic substances, such as Transforming Growth Factor beta-1 (TGF-β1), Hepatic Growth Factor (HGF), and Bone Morphogenic Protein-7 (BMP-7). Yacon is an herbal medicine which has not been elucidated in CKD. This study aimed to investigate the effect of ethanolic extract of Yacon leaves on attenuating renal injury in CKD model in mice. Methods: We performed 5/6 subtotal nephrectomy (SN) in male Swiss-Webster mice (3 months old, 30–40 grams) to induce chronic kidney disease, then the mice were sacrificed at day 14. The mice (n=25) were divided into five groups: one SN group, three groups of SN with administration of Yacon extract, and one group of sham operation (SO, with supplementation of 0.1% aquadest). There were three different doses of ethanolic extract of Yacon leaves: 98 mg/kg BW (SN+YK1), 49 mg/kg BW (SN+YK2), and 24.5 BW mg/kg (SN+YK3). Tubular injury, perivascular and interstitial fibrosis were quantified based on histopathological examination. Reverse-transcriptase PCR (RT-PCR) was performed to quantify HGF and BMP-7. Results: SN group demonstrated CKD with elevation of creatinine level, anemia, tubular injury, glomerulosclerosis, and fibrosis. Yacon extract treatment showed attenuation of injury with lower creatinine level, tubular injury, glomerulosclerosis and fibrosis compared to the SN group. HGF and BMP-7 mRNA expressions were higher in Yacon-treated groups than the SN group. Conclusion: Yacon treatment might ameliorate CKD through reducing fibrosis and increasing expression of anti-fibrotic genes.
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Objective: To investigate whether microRNA-384-5p can interfere with renal fibrosis by regulating BMP7 transcription in renal tubular epithelial cells. Methods: Unilateral ureteral obstructive (UUO) model was established, and fluorescence activated cell sorter (FACS) was used to isolate renal tubular epithelial cells for culture. Plasmid expressing miR-384-5p, antisense miR-384-5p, and blank plasmid were transfected by liposome and injected in UUO mice. Masson staining was used to detect renal fibrosis, and Western blot and real-time PCR were used to detect the expressions of target-genes of miRNA (miR-384-5p, miR-30, miR-92, and miR-128). Results: There was obvious renal fibrosis at 14 day in UUO model group; the expression of BMP7 mRNA in the renal tissue increased obviously while the expression of BMP7 protein was not increased. Although miR-384-5p could not change BMP7 mRNA in the renal tubular epithelial cells, BMP7 protein in the renal tubular epithelial cells that overexpressed miR-384-5p decreased significantly, and BMP7 protein in these cells overexpressing antisense miR-384-5p increased significantly. Immunohistochemistry and quantitative PCR showed that renal tissue fibrosis in UUO mice increased significantly in 14 days, while antisense miR-384-5p could significantly reduce the renal tissue fibrosis. Although antisense-miR-384-5p treatment did not affect the level of BMP7 mRNA, it could significantly increase the expression of BMP7 protein in renal tissue, suggesting that inhibiting miR-384-5p could lessen renal injury in UUO mice. Conclusion: As a key factor regulating the mechanism of renal fibrosis after renal injury, targeting miR-384-5p may become a new method to prevent and treat renal fibrosis.
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Aim To investigate the anti-proliferation effect of honokial on human colon cancer cells HCT116 and the possible mechanism. Methods Crystal stai-ning and flow cytometry assay were introduced to test the proliferation inhibitory and cell cycle arrested effect of honokial on HCT116 cells. Western blot was used to analyse the expression of PCNA and induction effect of apoptosis. Western blot and PCR assay were conducted to assess the change of BMP7. Via exogenous adenovi-ruses of BMP7 and its antibody combined with honoki-al,crystal violet staining and Western blot were used to analyse the effect on HCT116 cells. Results Honoki-al inhibited the growth of HCT116 in a time-and con- centration-dependent way,induced apoptosis and arres-ted at G2 phase; Western blot assay showed honokial up-regulated the level of PCNA and BMP7,and down-regulated the expression of Bcl-2; Western blot result also showed that exogenous adenoviruses of BMP7 en-hanced the effect of honokial on proliferation inhibition and apoptosis, while BMP7 antibody reversed such effects of honokial. Conclusion Honokial can inhibit the proliferation and induce apoptosis on HCT116 cells,which may be mediated by the up-regulation of BMP7.
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Aim To study the relationship between the anti-proliferative effect of resveratrol (Res) and BMP7 on human colon cancer cells and its possible molecular mechanism. Methods The proliferation of HCT116 cells was analyzed with cell proliferation inhibition assay, flow cytometry, Western blot and Annexin V-EGFP staining. CCK-8, PCR and Western blot assay were used to determine the effect of Res on BMP7 in HCT116 cells and the possible molecular mechanism underlying this process. Results Res inhibited the proliferation,arrested cell cycle at S phase and promo-ted apoptosis in HCT116 cells. Res increased the ex-pression of BMP7 mRNA and protein in HCT116 cells. Overexpression of BMP7 enhanced the anti-proliferative effect of Res on HCT116 cells and promoted the Res-induced apoptosis, whereas BMP7-specific antibody significantly attenuated these effects. Res exerted no apparent effect on the phosphorylation of Smad1/5/8, but decreased the phosphorylation of Akt1/2/3 sub-stantially in HCT116 cells. Overexpression of BMP7 enhanced the inhibitory effect of Res on phosphoryla-tion of Akt1/2/3, while BMP7 specific antibody re-duced this effect notably. Res markedly decreased the phosphorylation of PTEN, which could be boosted by BMP7,but attenuated by the BMP7 specific antibody. Conclusions Res can inhibit the proliferation and promote apoptosis of HCT116 cells,and the anti-canc-er activity of Res may be mediated by inactivating PI3K/Akt signaling through up-regulating BMP7 to de-crease the phosphorylation of PTEN partly.
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OBJECTIVE@#To investigate the effect of Yiqi Wenyang Huoxue Huatan Fang (YWHHF) on alleviating hypoxia-hypercarbia pulmonary hypertension by inhibiting endothelial-mesenchymal transition (EndoMT) BMP-7/Smads pathway.@*METHODS@#Fifty male healthy SD rats of clean grede, weighting (180~220) g, were randomly divided into 5 groups (=10):normoxia group (N), hypoxia-hypercarbia group (HH); YWHHF high dose group (YH), middle dose group (YM) and low dose group (YL). The rats in N group were kept in normal oxygen environment, the remaining four groups were intermittently exposed to hypoxia-hypercarbia environment (9%~11% O, 5%~6% CO) for 4 weeks, 6 days a week, 8 hours per day. The rats in YH, YM, YL groups were received YWHHF gavage in a dosageof 0.6, 0.3, 0.15g/kg respectively (3 ml/kg),the rats in N and HH groups were received equal volume of normal saline. After 4 weeks, the mean pulmonary arterial pressure(mPAP) was detected,the right ventricular free wall and left ventricle plus ventricular septum were isolated to determine the right ventricular hypertrophy index. Lung ultrastructural changes were surveyed under an electronic microscopy, the changes of pulmonary artery structure surveyed by immunofluorescence, the mRNA levels of alpha-smooth muscle actin (α-SMA)、platelet endothelial cell adhesion molecule-1 (CD31)、bone morphogenetic protein-7 (BMP-7)、drosophila mothers against decapentaplegic protein1/5/8 (Smad1/5/8) were detected by RT-PCR, and the protein levels of α-SMA、CD31、BMP-7、p-Smad1/5/8 and Smad1/5/8 were detected by Western blot.@*RESULTS@#Compared with N group, mPAP and the right ventricular hypertrophy index were increased,some significant injuries also were discovered under microscopic observation,the mRNA and protein expression of α-SMA was increased, and the mRNA expressions of CD31、BMP-7、Smad1/5/8 were decreased in the other four groups, the protein expressions of CD31、BMP-7、p-Smad1/5/8 were decreased(<0.05). Compared with HH group, the above changes in YH、YM、YL groups were all improved (<0.05).@*CONCLUSIONS@#YWHHF can inhibit EndoMT to alleviate pulmonary hypertension, and the mechanism may be related to the promotion of the expression of BMP-7/Smads pathway.
Subject(s)
Animals , Male , Rats , Hypercapnia , Hypertension, Pulmonary , Hypoxia , Pulmonary Artery , Rats, Sprague-DawleyABSTRACT
Objective:To investigate the effects of bone morphogenetic protein 7 (BMP-7) on the biological properties of osteoblasts exposed to high glucose.Methods:MC3T3 cells were cultured in the high glucose condition and were divided into 4 groups according to BMP-7 concentration (ng/ml):0 (control group),50,100 and 200 ng/mL BMP-7 (test groups).The cell proliferation and ALP activity were examind 1,3,5 and 7 d after exposure.The number of intracellular calcium nodules were observed with Alizarin red staining after osteogenesis induction for 21 days.The F-actin cytoskeleton of MC3T3 was stained with Rhodamine and examined 24 h after culture.The mRNA expression of OCN and Runx2 were quantified by RT-qPCR 48h after exposure to BMP-7.Results:BMP-7 significantly promoted proliferation of MC3T3 cells and enhanced ALP activity(P ＜ 0.05),increased the number and volume of intracellular calcium nodules.In the cells exposed to BMP-7,the osteoblast form was improved and F-actin cytoskeleton started to change with network structure.Compared with control group,the mRNA levels of OCN and Runx2 were up-regulated in BMP-7 groups (P ＜ 0.05).The mRNA levels of OCN and Runx2 were the highest in 200 ng/ml BMP-7 group(P ＜ 0.05).Conclusion:BMP 7 may inhibit the action of the high concentration of glucose on MC3T3 cells.BMP-7 may be benificial to osseointegration of dental implant in patietents with diabetics.
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To describe the relationship between miRNA-21 and renal fibrosis.Methods NRK-52E cells cultured in vitro,then constructed fibrosis model induced by transforming growth factorβ1 ( TGF-β1 ) , then bone morphogenetic protein-7 ( BMP-7 )-containing lentivirus vector was used to transfect the NRK-52E cells to anti-fibrosis, SMAD1, SMAD6, FN1, rno-miR-21-5p gene expression were detected.Results After TGF-β1 treatment, the BMP-7 mRNA and protein expressions were significantly decreased, FN1 mRNA and protein expressions were significantly increased ( P<0.05 ) . After transfected by BMP-7-containing lentivirus vector, BMP-7 mRNA expression significantly increased(P<0.05), FN1, SMAD6 and rno-miRNA-21-5p mRNA decreased, SMAD1 mRNA increased(P<0.05).Conclusion Epithelial-to-mesenchymal transition (EMT) is positively correlated with the miRNA21 and TGF-beta 1, its maybe related to the changes of BMP-7, SMAD1 and SMAD6.
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Objective:To examine the expression of odontoblast related proteins in dental pulp stem cells(DPSCs)induced by BMP-7.Methods:DPSCs were cultured in the common culture medium or medium supplemented with 1 00 ng/ml BMP-7.Electron microscope,CCK8 and immunohistochemical staining were carried out to estimate the cell morphology and differentiation.Results:In-duced by BMP-7,the morphology of DPSCs was not changed,the proliferation of DPSCs was slower than that of the cells without BMP-7 treatment.DPSCs were negative for the expression of DSPP,DMP-7 and ALP.However,DPSCs were found strongly positive for DSPP,DMP-7 and ALP after the induction of BMP-7.Conclusion:BMP-7 induction may promote the differentiation of DPSCs.
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Objective To explore the expression of thrombospondin-1 ( TSP-1 ) and bone morphogenetic protein-7 ( BMP-7 ) in mucus membrcane of chronic rhinosinusitis ( CRS ) and its potential significance in pathogenesis . Methods TSP-1 ,BMP-7 and TGF-β1 were detected by ELISA .Results The expression level of TSP-1 and TGF-β1 increased gradually with the advanced stage of CRSsNP ( P<0.05 ) .The expression of BMP-7 decreased gradu-ally with the advanced stage of CRSsNP ( P<0.05 ) .The expression of TSP-1 and TGF-β1 decreased gradually with the advanced stage of CRSwNP ( P<0.05 ) .The expression level of BMP-7 increased gradually with the ad-vanced stage of CRSwNP ( P<0.05 ) .Conclusions The expressions of TSP-1 and BMP-7 were abnormal in nasal mucosa of CRS , which might be involved in the pathogenesis of CRS .
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This study was conducted in order to verify the effect of different concentrations of BMP-7 in the in vitro survival and development of caprine preantral follicles. Fragments of caprine ovarian cortical tissue were cultured for 1 or 7 days in Minimum Essential Medium (MEM+) supplemented with different concentrations of BMP-7 (1, 10, 50 or 100ng/ml). Non-cultured fragments or those cultured for 1 or 7 days were processed for classical histology and transmission electron microscopy (TEM). Parameters such as follicular survival, activation and growth were evaluated. The results showed that, after 1 or 7 days of culture, the percentage of morphologically normal follicles was significantly reduced in all treatments when compared with fresh control, except at 1ng/ml of BMP-7 for 1 day. In addition, the concentration of 10ng/ml of BMP-7 significantly increases follicular diameter from day 1 to 7 of culture. There was no influence of the other concentrations of BMP-7 regarding to the follicular and oocyte diameter. Ultrastructure studies confirmed follicular integrity after 7 days of culture in 1ng/ml BMP-7. In conclusion, small concentrations of BMP-7 can improve the survival and growth of caprine preantral follicles during in vitro culture.
O presente trabalho foi conduzido de modo a se verificar o efeito de diferentes concentrações da BMP-7 no desenvolvimento in vitro de folículos pré-antrais caprinos. Fragmentos de tecido cortical ovariano caprino foram cultivados por 1 ou 7 dias em Minimum Essential Medium (MEM+) suplementado com diferentes concentrações de BMP-7 (1, 10, 50 ou 100ng/ml). Os fragmentos não cultivados ou aqueles cultivados por 1 ou 7 dias foram processados para histologia clássica e microscopia eletrônica de transmissão (TEM), sendo avaliados parâmetros morfológicos indicativos de viabilidade, ativação e crescimento. Os resultados mostraram que o percentual de folículos morfologicamente normais diminuiu significativamente em todos os tratamentos quando comparados ao controle, exceto na concentração de 1ng/ml por 1 dia de cultivo. Já no D7 todos os tratamentos reduziram significativamente os percentuais de folículos morfologicamente normais. Utilizando 10ng/ml de BMP-7 foi observado um aumento significativo no diâmetro folicular quando comparados os diferentes períodos de cultivo. Não houve influência das demais concentrações de BMP-7 quando avaliados além do diâmetro folicular o diâmetro oocitário. A análise por TEM confirmou a integridade ultra-estrutural nos folículos após 7 dias de cultivo com 1ng/ml de BMP-7 . Em conclusão, o BMP-7 em baixas concentrações pode melhorar a sobrevivência e o crescimento durante o cultivo in vitro de folículos pré-antrais caprinos.
Subject(s)
Animals , Ovary/anatomy & histology , Bone Morphogenetic Proteins/administration & dosage , GoatsABSTRACT
Objective To study the role of bone morphogenetic protein-7 in the osteogenic differentiation of periosteal cellsin vitro. Methods Periosteal cells, obtained from adult tibial periosteum, were cultured by routine method in vitro, and divided into two groups. One group cultured with BMP7 and the supplements of 100 nmol dexametasone, 10 mmol b-glycerophosphate and 50 mg/mL L-ascorbic acid (BMP7 group), the other cultured with the supplements alone as the control (control group). Ultrastructure and morphological changes of periosteal cells were observed by contrast phase microscope and electron microscope. In order to test the expression of markers of osteoblastic differantiation in periosteal cells, involved mineralized node and alkaline phosphatase. Each group was tested at the time of 5 d, 10 d, 15 d, 20 d, respectively, using ALP kit stain and Von Kossa stain with 3 samples at each time. Results The periosteal cells cultured by routine method and induced into osteoblast differentiation with BMP7 were both growing well, in vitro. Microscope observations showed that the periosteal cells were spindle-shaped, well-stacked, transparent and three-dimensional in the early stage, and cube-shaped or puncheon shaped in the mitotic phase, gradually became wide shuttle and irregular shape with a lot secretion in telophase. The positive cells were visible by the ALP kit staining and Von Kossa staining of calcium nodules at 5 d, 10 d, 15 d and 20 d in both groups.A difference of positive rate at each time point was found between BMP7 group and control group at 5 d, 10 d, 15 d, 20 d, and the difference was statistically significant (P ＜ 0.01). Conclusion It displayed well regeneration and osteogenesis ability in the periosteal cell. BMP7 has definite osteo-inductive activity, which can obviously enhance the proliferation and ossifyng differentiation of periosteal cells.
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Objective: To observe polyethylenimine-mediated BMP-7 gene transfection in promoting fracture healing in elderly rats. Methods: Male SD rats (20-month-old) were randomly divided into 3 groups: group A received BMP-7 gene therapy mediated by polyehtylenimine, group B was treated with normal saline and group C was treated with BMP-7 plasmid without polyethylenimine. Right femoral shaft fracture model was established in all rats. Animals in group A received transdermal injection of pcDNA3. 1-BMP-7/PET; those in group B and C received the same volume of normal saline and pcDNA3. 1-BMP-7, respectively. X-ray photography, histological observation (H-E staining), immunohistochemical staining of collagen type I, biomechanical and bone mineral density test were employed to assess the healing of fracture 2, 4, and 8 weeks after treatment. Results: The results of X ray and H-E staining showed no fracture healing in the 3 groups during the 8th week after fracture; however, the growth of coloboma in group A was better than that of the other 2 groups, with partial bone union between the fracture ends. Staining of collagen type I showed deeper, wider staining in group A compared with the other 2 group. Anti-bending intensity and bone mineral density tests showed that the parameters in group A were better than those of the other 2 groups (P<0.05 or 0.01). Conclusion: Polyethylenimine-mediated hBMP-7 gene transfer can effectively promote fracture healing in elderly rats.
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[Objective]To evaluate the inhibiting affection of Ad-BMP-7 to osteoporosis of ovariectomized rats.[Method]Mice model of osteoporosis was made by surgical ovariectomy,and divided into 4 groups:A.group of vertebral body intramedullary injection of normal saline;B.injection of Adovirus(Ad)-GFp;C.injection of Ad;D.injection of Ad+BMP(bone morphogenetic protein),the bone losing of whole body were monitored by measuring dry bone weight after 1 month,the osteoporosic condition of bone trabecular were observed by histology,and bone histomorphometry of vertebral bodies were carried to quatitatively measure the dynamic and static indexes of osteoporosic bone.[Result]The dry bone weight of AdBMP-7 rats werehigher than that of other groups.Under the microscope,the bone trabecula form of the Ad BMP-7 groups were more integrity,however,the bone trabecular of the other groups showed sharpen and ruptured.In the bone histomorphomitry,the numbers of the osteoblasts,the thickness of osteoid of group D were obviously higher than other groups.[Conclusion]Ad BMP-7 injected into the vertebral bodies of ovariectomized rats may secrete BMP-7,and inhibits the osteoporosis of ovariectomized rats in the early stage.The protective effect of this gene was not restricted to bones receiving intramedullary injection of the vector,but occurred in all bones that were evaluated.This proof of concept encourages further development of gene therapy approaches to the treatment of osteoporosis.
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Objective:To construct recombinant retrovirus expressing human bone morphogenetic protein-7 gene BMP7 and to discuss its apoptosis-inducing activities and the mechanism in liver cancer cell line HepG2. Methods:BMP7 gene was amplified and reconstructed with retroviral plasmid pLP-LNCX by loxP homologous recombination,and then the plasmid pLP-LNCX-BMP7 (pLLBMP7) was transferred into packing cell line PT67 and the supernatant was collected to assay viral titer. MTT assay was adopted to observe HepG2 cells amplification. 48h after pLLBMP7 infection agarose electrophoresis and flow cytometry were used to verify apoptosis of tumor cells,and then the expression of BMP7,caspase-3 and bcl-2 proteins were detected by Western blotting. Results:Recombinant retrovirus pLLBMP7 was justified and transformed into PT67 package cell with supernatant viral titer amounted to 5?109 pfu/ml. In MTT assay retrovirus group had no evident difference from controls in cellular inhibition 72h later (35.1% vs. 5.3%,68.5% vs.18.3%,p
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To evaluate availability of the BMP-7 adenovirus (AdBMP-7) as a gene therapy for osteoinduction, we investigated in morphological aspect at 1, 2, 4, 6 weeks after cells injection. Primary cultured human dermal fibroblasts, transduced with AdBMP-7, were injected into gastrocnemius muscle of the nude mice. One week after fibroblasts transplantation new tissue was observed in the muscle. Majority of new tissue was evaluated as cartilage and calcification in the matrix was confirmed by Von Kossa stain as well as electron microscopy. Two weeks after transplantation, spongy bone was built up and adipocytes were observed in intertrabecular spaces. Osteoblasts and osteoclasts were observed in the bony tissue surface. In the result of Von Kossa-Van Gieson stain, osteolysis was dominant in bony trabeculae. Bone marrow was established in 4th weeks with intertrabecular space filled up by hematopoietic cells. At the 6th weeks, the number of trabeculae decreased and thickness of the cortical bone was increased. A great part of bone matrix has laminar structure which run paralleled to surface and which included osteocytes and canaliculi. These data demonstrate that cell mediated AdBMP-7 for gene therapy initiate development of cartilage and calcification of matrix within 1 week and complete bone and bone marrow formation within 4 weeks, so then, could be made practical application for promotion of osteoinduction.
Subject(s)
Animals , Humans , Mice , Adenoviridae , Adipocytes , Bone Marrow , Bone Matrix , Bone Morphogenetic Protein 7 , Cartilage , Fibroblasts , Genes, vif , Genetic Therapy , Mice, Nude , Microscopy, Electron , Muscle, Skeletal , Osteoblasts , Osteoclasts , Osteocytes , OsteolysisABSTRACT
No abstract available.