Mesenchymal stem cells(MSCs)originate from many sources,including the bone marrow and adipose tissue,and differentiate into various cell types,such as osteoblasts and adipocytes.Recent studies on MSCs have revealed th...Mesenchymal stem cells(MSCs)originate from many sources,including the bone marrow and adipose tissue,and differentiate into various cell types,such as osteoblasts and adipocytes.Recent studies on MSCs have revealed that many transcription factors and signaling pathways control osteogenic development.Osteogenesis is the process by which new bones are formed;it also aids in bone remodeling.Wnt/β-catenin and bone morphogenetic protein(BMP)signaling pathways are involved in many cellular processes and considered to be essential for life.Wnt/β-catenin and BMPs are important for bone formation in mammalian development and various regulatory activities in the body.Recent studies have indicated that these two signaling pathways contribute to osteogenic differen-tiation.Active Wnt signaling pathway promotes osteogenesis by activating the downstream targets of the BMP signaling pathway.Here,we briefly review the molecular processes underlying the crosstalk between these two pathways and explain their participation in osteogenic differentiation,emphasizing the canonical pathways.This review also discusses the crosstalk mechanisms of Wnt/BMP signaling with Notch-and extracellular-regulated kinases in osteogenic differentiation and bone development.展开更多
The periodontal ligament-derived mesenchymal stem cell is regarded as a source of adult stem cells due to its multipotency. However, the proof of chondrogenic potential of the cells is scarce. Therefore, we investigat...The periodontal ligament-derived mesenchymal stem cell is regarded as a source of adult stem cells due to its multipotency. However, the proof of chondrogenic potential of the cells is scarce. Therefore, we investigated the chondrogenic differentiation capacity of periodontal ligament derived mesenchymal stem cells induced by transforming growth factor （TGF）-β3 and bone morphogenetic protein （BMP）-6. After isolation of periodontal ligament stem cells （PDLSCs） from human periodontal ligament, the cells were cultured in Dulbecco＇s modified Eagle＇s medium （DMEM） with 20% fetal bovine serum （FBS）. A mechanical force initiated chondrogenic differentiation of the cells. For chondrogenic differentiation, 10 μg ·L-1 TGF-β3 or 100 μg ·L-1 BMP-6 and the combination treating group for synergistic effect of the growth factors. We analyzed the PDLSCs by fluorescence-activated cell sorting and chondrogenesis were evaluated by glycosaminoglycans assay, histology, immunohistochemistry and genetic analysis. PDLSCs showed mesenchymal stem cell properties proved by FACS analysis. Glycosaminoglycans contents were increased 217% by TGF-β3 and 220% by BMP-6. The synergetic effect of TGF-β3 and BMP-6 were shown up to 281% compared to control. The combination treatment increased Sox9, aggrecan and collagen II expression compared with not only controls, but also TGF-β3 or BMP-6 single treatment dramatically. The histological analysis also indicated the chondrogenic differentiation of PDLSCs in our conditions. The results of the present study demonstrate the potential of the dental stem cell as a valuable cell source for chondrogenesis, which may be applicable for regeneration of cartilage and bone fracture in the field of cell therapy.展开更多
Half of altogether 60 cylindrical implant devices made of titanium-aluminum-vanadium alloy (Ti-6Al-4V) were plasma-sprayed with a hydroxyapatite-coating and the other half had a corundum blasted porous surface. 15 imp...Half of altogether 60 cylindrical implant devices made of titanium-aluminum-vanadium alloy (Ti-6Al-4V) were plasma-sprayed with a hydroxyapatite-coating and the other half had a corundum blasted porous surface. 15 implants of each group of the titanium test implants were coated with 230 μg porcine, high-purified BMP-3-precipitate per implant. In each case a BMP-3-coated and an uncoated control-device were implanted into the femoral part of the patellofemoral joint of the right and left leg of 30 adult giant rabbits. Histomorphological and histomorphometrical we found in both groups with BMP-3-coated test devices an improved osteointegration. Statistical evaluation using the t-test for matched samples showed 5 weeks after surgery a significant higher volume of new formed bone of the BMP-3-coated corundum-blasted or hydroxyapatite-coated Ti-6Al-4V test devices compared to the non-coated controls of the same type (p<0.01, t-test for matched samples). In both implant groups with BMP-coating a synergetic effect was verifiable although the bone ongrowth in the hydroxyapatite coated implants was more extensive than in the corundum blasted implants. Light microscopy demonstrated osteointegration without connective tissue membrane around the surface of the implants. Our results indicate that composite metal implants, as used in endoprosthetics and implantology, are suitable carriers for BMP-3 and improved fixation of the implants can be achieved. The hydroxyapatite surface is superior to the corundum-blasted surface with regards to the observed parameters because of its pronounced bioactivity and its osteoconductive characteristics.展开更多
The extracellular matrix-associated bone morphogenetic proteins(BMPs) govern a plethora of biological processes. The BMPs are members of the transforming growth factor-β protein superfamily, and they actively partici...The extracellular matrix-associated bone morphogenetic proteins(BMPs) govern a plethora of biological processes. The BMPs are members of the transforming growth factor-β protein superfamily, and they actively participate to kidney development, digit and limb formation, angiogenesis, tissue fibrosis and tumor development. Since their discovery, they have attracted attention for their fascinating perspectives in the regenerative medicine and tissue engineering fields. BMPs have been employed in many preclinical and clinical studies exploring their chondrogenic or osteoinductive potential in several animal model defects and in human diseases. During years of research in particular two BMPs, BMP2 and BMP7 have gained the podium for their use in the treatment of various cartilage and bone defects. In particular they have been recently approved for employment in non-union fractures as adjunct therapies. On the other hand, thanks to their potentialities in biomedical applications, there is a growing interest in studying the biology of mesenchymal stem cell(MSC), the rules underneath their differentiation abilities, and to test their true abilities in tissue engineering. In fact, the specific differentiation of MSCs into targeted celltype lineages for transplantation is a primary goal of the regenerative medicine. This review provides an overview on the current knowledge of BMP roles and signaling in MSC biology and differentiation capacities. In particular the article focuses on the potential clinical use of BMPs and MSCs concomitantly, in cartilage and bone tissue repair.展开更多
BACKGROUND Liver fibrosis is a refractory disease whose persistence can eventually induce cirrhosis or even liver cancer.Early liver fibrosis is reversible by intervention.As a member of the transforming growth factor...BACKGROUND Liver fibrosis is a refractory disease whose persistence can eventually induce cirrhosis or even liver cancer.Early liver fibrosis is reversible by intervention.As a member of the transforming growth factor-beta(TGF-β)superfamily,bone morphogenetic protein 7(BMP7)has anti-liver fibrosis functions.However,little is known about BMP7 expression changes and its potential regulatory mechanism as well as the relationship between BMP7 and TGF-βduring liver fibrosis.In addition,the mechanism underlying the anti-liver fibrosis function of BMP7 needs to be further explored.AIM To investigate changes in the dynamic expression of BMP7 during liver fibrosis,interactions between BMP7 and TGF-β1,and possible mechanisms underlying the anti-liver fibrosis function of BMP7.METHODS Changes in BMP7 expression during liver fibrosis and the interaction between BMP7 and TGF-β1 in mice were observed.Exogenous BMP7 was used to treat mouse primary hepatic stellate cells(HSCs)to observe its effect on activation,migration,and proliferation of HSCs and explore the possible mechanism underlying the anti-liver fibrosis function of BMP7.Mice with liver fibrosis received exogenous BMP7 intervention to observe improvement of liver fibrosis by using Masson’s trichrome staining and detecting the expression of the HSC activation indicator alpha-smooth muscle actin(α-SMA)and the collagen formation associated protein type I collagen(Col I).Changes in the dynamic expression of BMP7 during liver fibrosis in the human body were further observed.RESULTS In the process of liver fibrosis induced by carbon tetrachloride(CCl4)in mice,BMP7 protein expression first increased,followed by a decrease;there was a similar trend in the human body.This process was accompanied by a sustained increase in TGF-β1 protein expression.In vitro experiment results showed that TGF-β1 inhibited BMP7 expression in a time-and dose-dependent manner.In contrast,high doses of exogenous BMP7 inhibited TGF-β1-induced activation,migration,and proliferation of HSCs;this inhibitory effect was associated with upregulation of pSmad1/5/8 and downregulation of phosphorylation of Smad3 and p38 by BMP7.In vivo experiment results showed that exogenous BMP7 improved liver fibrosis in mice.CONCLUSION During liver fibrosis,BMP7 protein expression first increases and then decreases.This changing trend is associated with inhibition of BMP7 expression by sustained upregulation of TGF-β1 in a time-and dose-dependent manner.Exogenous BMP7 could selectively regulate TGF-β/Smad pathway-associated factors to inhibit activation,migration,and proliferation of HSCs and exert antiliver fibrosis functions.Exogenous BMP7 has the potential to be used as an antiliver fibrosis drug.展开更多
AIM:To observe the effect of Danshao Huaxian capsule(DHC)on the expression of Gremlin and bone morphogenetic protein-7(BMP-7)in the liver of hepatic fibrosis rats.METHODS:A total of 75 male Wistar rats were randomly d...AIM:To observe the effect of Danshao Huaxian capsule(DHC)on the expression of Gremlin and bone morphogenetic protein-7(BMP-7)in the liver of hepatic fibrosis rats.METHODS:A total of 75 male Wistar rats were randomly divided into a normal control group(A),a CCl4-induced hepatic fibrosis model group(B),a natural recovery group(C),a low-dose DHC-treated group(D),and a high-dose DHC-treated group(E),with 15 rats in each group.Liver fibrosis was induced by subcutaneous injections of carbon tetrachloride(CCl4)and a highlipid/low-protein diet for 8 wk,except for the rats in group A.Then,the rats in the two DHC-treated groups were administered 0.5 and 1.0 g/kg DHC by gastrogavage once per day for 8 successive weeks,respectively.By the end of the experiment,the level of transforming growth factorβ1(TGF-β1)in the liver homogenate was determined by an enzyme-linked immunosorbent assay.The mRNA and protein expression of Gremlin and BMP-7 in the liver tissue was determined by reversetranscription polymerase chain reaction,an immunohistochemical assay,and Western blot analysis.RESULTS:Compared with group A,the level of TGF-β1and the mRNA and protein expression of Gremlin were significantly higher in group B(TGF-β1:736.30±24.40μg/g vs 284.20±18.32μg/g,P<0.01;mRNA of Gremlin:80.40±5.46 vs 49.83±4.20,P<0.01;positive protein expression rate of Gremlin:38.46%±1.70%vs 3.83%±0.88%,P<0.01;relative protein expression of Gremlin:2.81±0.24 vs 0.24±0.06,P<0.01),and the mRNA and protein expression of BMP-7was significantly lower in group B(mRNA:54.00±4.34vs 93.99±7.03,P<0.01;positive protein expression rate:28.97%±3.14%vs 58.29%±6.02,P<0.01;relative protein expression:0.48±0.31 vs 1.05±0.12,P<0.01).Compared with groups B and C,the degree of hepatic fibrosis was significantly improved,and the level of TGF-β1 and the mRNA and protein expression of Gremlin were significantly lowered in the two DHCtreated groups(TGF-β1:523.14±21.29μg/g,441.86±23.18μg/g vs 736.30±24.40μg/g,651.13±15.75μg/g,P<0.01;mRNA of Gremlin:64.86±2.83,55.82±5.39 vs 80.40±5.46,70.37±4.01,P<0.01;positive protein expression rate of Gremlin:20.78%±1.60%,17.43%±2.02%vs 38.46%±1.70%,29.50%±2.64%,P<0.01;relative protein expression of Gremlin:1.95±0.26,1.65±0.20 vs 2.81±0.24,2.22±0.63,P<0.01),and the mRNA and protein expression of BMP-7 was higher in the two DHC-treated groups(mRNA:73.52±4.56,81.78±5.38 vs 54.00±4.34,62.28±4.51,P<0.01;positive protein expression rate:41.44%±4.77%,47.49%±4.59%vs28.97%±3.14%,35.85%±3.50%,P<0.01;relative protein expression:0.71±0.06,0.81±0.07 vs 0.48±CONCLUSION:The therapeutic mechanism of DHC forhepatic fibrosis in rats may be associated with inhibitionof the expression of Gremlin and up-regulation of the expression of BMP-7.展开更多
AIM To present the incidence of heterotopic ossification after the use of recombinant human bone morphogenetic protein-7(rhB MP-7) for the treatment of nonunions.METHODS Bone morphogenetic proteins(BMPs) promote bone ...AIM To present the incidence of heterotopic ossification after the use of recombinant human bone morphogenetic protein-7(rhB MP-7) for the treatment of nonunions.METHODS Bone morphogenetic proteins(BMPs) promote bone formation by auto-induction. Recombinant human BMP-7 in combination with bone grafts was used in 84 patients for the treatment of long bone nonunions. All patients were evaluated radiographicaly for the development of heterotopic ossification during the standard assessment for the nonunion healing. In all patients(80.9%) with radiographic signs of heterotopic ossification, a CT scan was performed. Nonunion site palpation and ROM evaluation of the adjacent jointswere also carried out. Factors related to the patient(age, gender), the nonunion(location, size, chronicity, number of previous procedures, infection, surrounding tissues condition) and the surgical procedure(graft and fixation type, amount of rhB MP-7) were correlated with the development of heterotopic ossification and statistical analysis with Pearsons χ~2 test was performed.RESULTS Eighty point nine percent of the nonunions treated with rh BMP-7, healed with no need for further procedures. Heterotopic bone formation occurred in 15 of 84 patients(17.8%) and it was apparent in the routine radiologi-cal evaluation of the nonunion site, in a mean time of 5.5 mo after the rh BMP-7 application(range 3-12). The heterotopic ossification was located at the femur in 8 cases, at the tibia in 6, and at the humerus in οne patient. In 4 patients a palpable mass was present and only in one patient, with a para-articular knee nonunion treated with rhB MP-7, the size of heterotopic ossification affected the knee range of motion. All the patients with heterotopic ossification were male. Statistical analysis proved that patient's gender was the only important factor for the development of heterotopic ossification(P = 0.007). CONCLUSION Heterotopic ossification after the use of rh BMP-7 in nonunions was common but it did not compromise the final clinical outcome in most cases, and affected only male patients.展开更多
AIM: To investigate the influence of bone morphogenetic protein type IA receptor [BMPR-IA(ALK3)] conditional knockout in lens on expression of bone morphogenetic protein 4(BMP4) in lens during the development of the v...AIM: To investigate the influence of bone morphogenetic protein type IA receptor [BMPR-IA(ALK3)] conditional knockout in lens on expression of bone morphogenetic protein 4(BMP4) in lens during the development of the vertebrate eye.METHODS: Cre-positive mice were mated with Crenegative mice to generate 50% Cre-positive(conditional knockout, CKO) 4 embryos, 8 eyes and 50% Cre-negative offspring(wild type, WT) 4 embryos, 8 eyes. The embryos were fixed in 4% paraformaldehyde, embedded in paraffin, and sectioned to a thickness of 4 μm.Removal of paraffin wax and dehydrating for sections,and then the procedure of in situ hybridization was processed, BMP4 MK1784-m(BOSTER) was used, and observed the expression of BMP4 in the lens in experimental group and control group. We selected SPSS11.0 software for statistical analysis, P<0.05 showed that the difference was statistically significant.· RESULTS: Four embryos of each genotype were examined, totally we had 8 embryos, 16 eyes. We got the uniform outcomes in all the embryos. We found ALK3 was required during lens growing, but was not essential for the formation of lens. We observed that the expression of BMP4 in the lens was significantly reduced in all 8 ALK3 CKO lens, BMP4 expression was normal in all the 8 WT lens, P <0.01. This phenomenon became increasingly visible in accordance with embryo development. The most apparent alteration was present at stage E15.5.CONCLUSION: ALK3 is essential for lens growth. The influence of ALK3 on the expression of BMP4 is present during the development of mice lens.展开更多
AIM: To investigate the antifibrotic effects of bone morphogenetic protein-7 (BMP-7) on Schistosoma japonicum (S. japonicum )-induced hepatic fibrosis in BALB/C mice. METHODS: Sixty BALB/C mice were randomly divided i...AIM: To investigate the antifibrotic effects of bone morphogenetic protein-7 (BMP-7) on Schistosoma japonicum (S. japonicum )-induced hepatic fibrosis in BALB/C mice. METHODS: Sixty BALB/C mice were randomly divided into three groups, including a control group (group A, n = 20), model group (group B, n = 20) and BMP-7 treated group (group C, n = 20). The mice in group B and group C were abdominally infected with S. japonicum cercariae to induce a schistosomal hepatic fibrosis model. The mice in group C were administered human recombinant BMP-7. Liver samples were extracted from mice sacrificed at 9 and 15 wk after modeling. Hepatic histopathological changes were assessed using Masson's staining. Transforming growth factor-beta 1 (TGF-β1), alpha-smooth muscle actin (α-SMA), phosphorylated Smad2/3 (pSmad2/3) and Smad7 protein levels and localization were measured by Western blotting and immunohistochemistry, respectively, and their mRNA expressions were detected by reverse transcriptionpolymerase chain reaction (RT-PCR). RESULTS: The schistosomal hepatic fibrosis mouse model was successfully established, as the livers of mice in group B and group C showed varying degrees of typical schistosomal hepatopathologic changes such as egg granuloma and collagen deposition. The degree of collagen deposition in group C was higher than that in group A (week 9: 22.95±6.66vs 2.02±0.76; week 15: 12.84±4.36 vs 1.74±0.80; P<0.05), but significantly lower than that in group B (week 9: 22.95±6.66 vs 34.43±6.96; week 15: 12.84±4.36 vs 18.90±5.07;P<0.05) at both time points. According to immunohistochemistry data, the expressions of α-SMA, TGF-β1 and pSmad2/3 protein in group C were higher than those in group A (α-SMA: week 9: 21.24±5.73 vs 0.33±0.20; week 15: 12.42±4.88 vs 0.34±0.27; TGF-β1: week 9: 37.00±13.74 vs 3.73±2.14; week 15: 16.71±9.80 vs 3.08±2.35; pSmad2/3: week 9: 12.92±4.81 vs 0.83±0.48; week 15: 7.87±4.09 vs 0.90±0.45; P<0.05), but significantly lower than those in group B (α-SMA: week 9: 21.24±5.73 vs 34.39±5.74; week 15: 12.42±4.88 vs 25.90±7.01; TGF-β1: week 9: 37.00±13.74 vs 55.66±14.88; week 15: 16.71±9.80 vs 37.10±12.51; pSmad2/3: week 9: 12.92±4.81 vs 19.41±6.87; week 15: 7.87±4.09vs 13.00±4.98;P<0.05) at both time points; the expression of Smad7 protein in group B was higher than that in group A and group C at week 9 (8.46±3.95 vs 1.00±0.40 and 8.46±3.95 vs 0.77±0.42; P<0.05), while there were no differences in Smad7 expression between the three groups at week 15 (1.09±0.38 vs 0.97±0.42 vs 0.89±0.39; P>0.05). Although minor discrepancies were observed, the results of RT-PCR and Western blotting were mainly consistentwith the immunohistochemical results. CONCLUSION: Exogenous BMP-7 significantly decreased the degree of hepatic fibrosis in both the acute and chronic stages of hepato-schistosomiasis, and the regulatory mechanism may involve the TGF-β/Smad signaling pathway.展开更多
Bone morphogenetic proteins(BMPs)are a family of potent,multifunctional growth factors belonging to transforming growth factor-(TGF-).They are highly conservative in structures.Over 20 members of BMPs with varying fun...Bone morphogenetic proteins(BMPs)are a family of potent,multifunctional growth factors belonging to transforming growth factor-(TGF-).They are highly conservative in structures.Over 20 members of BMPs with varying functions such as embryogenesis,skeletal formation,hematopoiesis and neurogenesis have been identified in human body.BMPs are unique growth factors that can induce the formation of bone tissue individually.BMPs can induce the differentiation of bone marrow mesenchymal stem cells into osteoblastic lineage and promote the proliferation of osteoblasts and chondrocytes.BMPs stimulate the target cells by specific membrane-bound receptors and signal transduced through mothers against decapentaplegic(Smads)and mitogen activated protein kinase(MAPK)pathways.It has been demonstrated that BMP-2,BMP-4,BMP-6,BMP-7,and BMP-9 play an important role in bone formation.This article focuses on the molecular characterization of BMPs family members,mechanism of osteogenesis promotion,related signal pathways of osteogenic function,relationships between structure and osteogenetic activity,and the interactions among family members at bone formation.展开更多
AIM: To characterize the expression and dynamic changes of bone morphogenetic protein (BMP)-2 in hepatocytes in the regenerating liver in rats after partial hepatectomy (PH), and examine the effects of BMP-2 on prolif...AIM: To characterize the expression and dynamic changes of bone morphogenetic protein (BMP)-2 in hepatocytes in the regenerating liver in rats after partial hepatectomy (PH), and examine the effects of BMP-2 on proliferation of human Huh7 hepatoma cells. METHODS: Fifty-four adult male Wistar rats were randomly divided into three groups: A normal control (NC) group, a partial hepatectomized (PH) group and a sham operated (SO) group. To study the effect of liver regeneration on BMP-2 expression, rats were sacrificed before and at different time points after PH or the sham intervention (6, 12, 24 and 48 h). For each time point, six rats were used in parallel. Expression and distribution of BMP-2 protein were determined in regenerating liver tissue by Western blot analysis and immunohistochemistry. Effects of BMP-2 on cell proliferation of human Huh7 hepatoma cell line were assessed using an MTT assay.RESULTS: In the normal liver strong BMP-2 expression was observed around the central and portal veins. The expression of BMP-2 decreased rapidly as measured by both immunohistochemistry and Western blot analysis. This decrease was at a maximum of 3.22 fold after 12 h and returned to normal levels at 48 h after PH. No significant changes in BMP-2 immunoreactivity were observed in the SO group. BMP-2 inhibited serum induced Huh7 cell proliferation.CONCLUSION: BMP-2 is expressed in normal adult rat liver and negatively regulates hepatocyte proliferation. The observed down regulation of BMP-2 following partial hepatectomy suggests that such down regulation may be necessary for hepatocyte proliferation.展开更多
AIM:To explore ex vivo the role of bone morphogenetic protein-4(BMP-4) and transforming growth factorbeta1(TGF-β1) in acute valvular response to fluid shear stress(FSS) abnormalities.METHODS:Porcine valve leaflets we...AIM:To explore ex vivo the role of bone morphogenetic protein-4(BMP-4) and transforming growth factorbeta1(TGF-β1) in acute valvular response to fluid shear stress(FSS) abnormalities.METHODS:Porcine valve leaflets were subjected ex vivo to physiologic FSS,supra-physiologic FSS magnitude at normal frequency and supra-physiologic FSS frequency at normal magnitude for 48 h in a double-sided cone-and-plate bioreactor filled with standard culture medium. The role of BMP-4 and TGF-β1 in the valvular response was investigated by promoting or inhibiting the downstream action of those cytokines via culture medium supplementation with BMP-4 or the BMP antagonist noggin,and TGF-β1 or the TGF-β1 inhibitor SB-431542,respectively. Fresh porcine leaflets were used as controls. Each experimental group consisted of six leaflet samples. Immunostaining and immunoblotting were performed to assess endothelial activation in terms of intercellular adhesion molecule-1 and vascular cell adhesion molecule-1 expressions,paracrine signaling in terms of BMP-4 and TGF-β1 expressions and extracellular matrix(ECM) remodeling in terms of cathepsin L,cathepsin S,metalloproteinases(MMP)-2 and MMP-9 expressions. Immunostained images were quantified by normalizing the intensities of positively stained regions by the number of cells in each image while immunoblots were quantified by densitometry. R E S U LT S :Regardless of the culture medium,physiologic FSS maintained valvular homeostasis. Tissue exposure to supra-physiologic FSS magnitude in standard medium stimulated paracrine signaling(TGF-β1:467% ± 22% vs 100% ± 6% in freshcontrols,BMP-4:258% ± 22% vs 100% ± 4% in fresh controls; P < 0.05) and ECM degradation(MMP-2:941% ± 90% vs 100% ± 19% in fresh controls,MMP-9:1219% ± 190% vs 100% ± 16% in fresh controls,cathepsin L:1187% ± 175% vs 100% ± 12% in fresh controls,cathepsin S:603% ± 88% vs 100% ± 13% in fresh controls; P < 0.05),while BMP-4 supplementation also promoted fibrosa activation and TGF-β1 inhibition reduced MMP-9 expression to the native tissue level(MMP-9:308% ± 153% with TGF-β1 inhibition vs 100% ± 16% in fresh control; P > 0.05). Supra-physiologic FSS frequency had no effect on endothelial activation and paracrine signaling regardless of the culture medium but TGF-β1 silencing attenuated FSS-induced ECM degradation via MMP-9 downregulation(MMP-9:302% ± 182% vs 100% ± 42% in fresh controls; P > 0.05).CONCLUSION:Valvular tissue is sensitive to FSS abnormalities. The TGF-β1 inhibitor SB-431542 is a potential candidate molecule for attenuating the effects of FSS abnormalities on valvular remodeling.展开更多
The advent of recombinant DNA technology has substantially increased the intra-operative utilization of biologic augmentation in spine surgery over the past several years after the Food and Drug Administration approva...The advent of recombinant DNA technology has substantially increased the intra-operative utilization of biologic augmentation in spine surgery over the past several years after the Food and Drug Administration approval of the bone morphogenetic protein(BMP) class of molecules for indications in the lumbar spine. Much less is known about the potential benefits and risks of the "off-label" use of BMP in the cervical spine. The history and relevant literature pertaining to the use of the "off-label" implantation of the BMP class of molecules in the anterior or posterior cervical spine are reviewed and discussed. Early prospective studies of BMP-2 implantation in anterior cervical spine constructs showed encouraging results. Later retrospective studies reported potentially "life threatening complications" resulting in a 2007 public health advisory by the FDA. Limited data regarding BMP-7 in anterior cervical surgery was available with one group reporting a 2.4% early(< 30 d) complication rate(brachialgia and dysphagia). BMP use in the decompressed posterior cervical spine may result in neurologic or wound compromise according to several retrospective reports, however, controlled use has been reported to increase fusion rates in select complex and pediatric patients. There were no cases of de novo neoplasia related to BMP implantation in the cervical spine. BMP-2 use in anterior cervical spine surgery has been associated with a high early complication rate. Definitive recommendations for BMP-7 use in anterior cervical spine surgery cannot be made with current clinical data. According to limited reports, select complex patients who are considered "high risk" for pseudoarthrosis undergoing posterior cervical or occipitocervical arthrodesis or children with congenital or traumatic conditions may be candidates for "off-label" use of BMP in the context of appropriate informed decision making. At the present time, there are no highlevel clinical studies on the outcomes and complication rates of BMP implantation in the cervical spine.展开更多
To study the cartilage differentiation of mouse mesenchymal stem cells(MSCs) induced by cartilage-derived morphogenetic proteins-2 in vitro,the MSCs were isolated from mouse bone marrow and cultured in vitro.The cells...To study the cartilage differentiation of mouse mesenchymal stem cells(MSCs) induced by cartilage-derived morphogenetic proteins-2 in vitro,the MSCs were isolated from mouse bone marrow and cultured in vitro.The cells in passage 3 were induced into chondrogenic differentiation with different concentrations of recombinant human cartilage-derived morphogenetic proteins-2(0,10,20,50 and 100 ng/mL).After 14 days of induction,morphology of cells was observed under phase-contrast microscope.Collagen Ⅱ mRNA and protein were examined with RT-PCR,Western blotting and immunocytochemistry respectively and the sulfate glycosaminoglycan was measured by Alcian blue staining.RT-PCR showed that CDMP-2 could promote expression of collagen Ⅱ mRNA in an dose-dependant manner,especially at the concentration of 50 ng/mL and 100 ng/mL.Immunocytochemistry and Western blotting revealed a similar change.Alcian blue staining exhibited deposition of typical cartilage extracellular matrix.Our results suggest that mouse bone marrow mesencymal stem cells can differentiate into chondrogenic phonotype with the induction of CDMP-2 in vitro,which provides a basis for further research on the role of CDMP-2 in chondrogenesis.展开更多
Objective To evaluate the effects of retinoic acid(RA)on expression of bone morphogenetic protein 7(BMP-7)in rat fetus with cleft palate,and the effects of RA on proliferation and apoptosis of osteoblasts.Methods All-...Objective To evaluate the effects of retinoic acid(RA)on expression of bone morphogenetic protein 7(BMP-7)in rat fetus with cleft palate,and the effects of RA on proliferation and apoptosis of osteoblasts.Methods All-trans RA(ATRA)was used to induce congenital cleft palate in Wistar rat.BMP-7 mRNA expression in maxillary bone tissue of fetal rats was measured by Northern blotting analysis.Flow cytometry and MTT assay were used to measure the apoptosis and proliferation of ATRA-treated MC-3T3-E1 cells.BMP-7 mRNA and protein expressions in ATRA-treated MC-3T3-E1 cells were detected by RT-PCR and Western blotting analysis.Results ATRA could induce cleft palate of rat fetus.The incidence rate of cleft palate induced by 100 mg/kg ATRA(45.5%)was significantly higher than 50 mg/kg ATRA(12.5%,P<0.05).BMP-7 mRNA expression decreased in maxillary bone tissue of rat fetus with cleft palate.MC-3T3-E1 cells proliferation treated with 1×10-6 mol/LATRA decreased by 60%,the cell apoptosis increased by 2 times.BMP-7 mRNA and protein levels in MC-3T3-E1 cells treated with 1×10-6 mol/L ATRA decreased by 60% and 80%,respectively,compared with ATRA-untreated cells(P<0.05).Conclusions BMP-7 may play an important role in embryonic palate development.RA may possess the ability to down-regulate cell proliferation through regulation of BMP-7 gene expression.展开更多
Bone morphogenesis proteins(BMPs) are multi-functional growth factors. They are expressed in retina,retinal pigment epithelium(RPE) and sclera and serve as a regulator in the growth and development of the eye. This ar...Bone morphogenesis proteins(BMPs) are multi-functional growth factors. They are expressed in retina,retinal pigment epithelium(RPE) and sclera and serve as a regulator in the growth and development of the eye. This article reviewed the chondrogenic potency of the sclera,biochemical and pathological changes of myopic scleral tissue and the differentiation of chondrogenesis by BMP-2. We proposed the hypothesis that BMP-2 can regulate differentiate of scleral fibroblasts and affect the development of myopia.展开更多
An experimental model of femoral muscular pouch in 20 mice was adopted. The expression of VEGF was examined by in situ hybridization method and immunohistochemical method in bone morphogenetic protein-2 induced osteog...An experimental model of femoral muscular pouch in 20 mice was adopted. The expression of VEGF was examined by in situ hybridization method and immunohistochemical method in bone morphogenetic protein-2 induced osteogenesis. The experimental results demonstrated that the expression signals of VEGF mRNA and VEGF appeared in cytoplasm during condensation of mesenchymal cell. As the mesenchymal cells differentiated into precartilage, the expression signals decreased in mesenchymal cells, but increased in chondrocytes and kept getting denser in the process of cartilage maturity. The peak expression of VEGF mRNA and VEGF in the experimental group appeared on the 14th day, accompanied by numerous hypertrophic chondrocytes. When mature cartilage calcified and new bone trabecula formed, the expression of VEGF mRNA and VEGF decreased in chondrocytes, but still expressed moderately in the osteoblasts and osteocytes. Signals of VEGF mRNA and VEGF can not be detected in the control groups.展开更多
基金Indian Council of Medical Research,2020-0282/SCR/ADHOC-BMSDepartment of Science and Technology,India,DST/INSPIRE Fellowship:2021/IF210073.
文摘Mesenchymal stem cells(MSCs)originate from many sources,including the bone marrow and adipose tissue,and differentiate into various cell types,such as osteoblasts and adipocytes.Recent studies on MSCs have revealed that many transcription factors and signaling pathways control osteogenic development.Osteogenesis is the process by which new bones are formed;it also aids in bone remodeling.Wnt/β-catenin and bone morphogenetic protein(BMP)signaling pathways are involved in many cellular processes and considered to be essential for life.Wnt/β-catenin and BMPs are important for bone formation in mammalian development and various regulatory activities in the body.Recent studies have indicated that these two signaling pathways contribute to osteogenic differen-tiation.Active Wnt signaling pathway promotes osteogenesis by activating the downstream targets of the BMP signaling pathway.Here,we briefly review the molecular processes underlying the crosstalk between these two pathways and explain their participation in osteogenic differentiation,emphasizing the canonical pathways.This review also discusses the crosstalk mechanisms of Wnt/BMP signaling with Notch-and extracellular-regulated kinases in osteogenic differentiation and bone development.
基金supported by the Bio & Medical Technology Development Program of the National Research Foundation(NRF) funded by the Korean government(MEST)(No.860-20110087)
文摘The periodontal ligament-derived mesenchymal stem cell is regarded as a source of adult stem cells due to its multipotency. However, the proof of chondrogenic potential of the cells is scarce. Therefore, we investigated the chondrogenic differentiation capacity of periodontal ligament derived mesenchymal stem cells induced by transforming growth factor （TGF）-β3 and bone morphogenetic protein （BMP）-6. After isolation of periodontal ligament stem cells （PDLSCs） from human periodontal ligament, the cells were cultured in Dulbecco＇s modified Eagle＇s medium （DMEM） with 20% fetal bovine serum （FBS）. A mechanical force initiated chondrogenic differentiation of the cells. For chondrogenic differentiation, 10 μg ·L-1 TGF-β3 or 100 μg ·L-1 BMP-6 and the combination treating group for synergistic effect of the growth factors. We analyzed the PDLSCs by fluorescence-activated cell sorting and chondrogenesis were evaluated by glycosaminoglycans assay, histology, immunohistochemistry and genetic analysis. PDLSCs showed mesenchymal stem cell properties proved by FACS analysis. Glycosaminoglycans contents were increased 217% by TGF-β3 and 220% by BMP-6. The synergetic effect of TGF-β3 and BMP-6 were shown up to 281% compared to control. The combination treatment increased Sox9, aggrecan and collagen II expression compared with not only controls, but also TGF-β3 or BMP-6 single treatment dramatically. The histological analysis also indicated the chondrogenic differentiation of PDLSCs in our conditions. The results of the present study demonstrate the potential of the dental stem cell as a valuable cell source for chondrogenesis, which may be applicable for regeneration of cartilage and bone fracture in the field of cell therapy.
文摘Half of altogether 60 cylindrical implant devices made of titanium-aluminum-vanadium alloy (Ti-6Al-4V) were plasma-sprayed with a hydroxyapatite-coating and the other half had a corundum blasted porous surface. 15 implants of each group of the titanium test implants were coated with 230 μg porcine, high-purified BMP-3-precipitate per implant. In each case a BMP-3-coated and an uncoated control-device were implanted into the femoral part of the patellofemoral joint of the right and left leg of 30 adult giant rabbits. Histomorphological and histomorphometrical we found in both groups with BMP-3-coated test devices an improved osteointegration. Statistical evaluation using the t-test for matched samples showed 5 weeks after surgery a significant higher volume of new formed bone of the BMP-3-coated corundum-blasted or hydroxyapatite-coated Ti-6Al-4V test devices compared to the non-coated controls of the same type (p<0.01, t-test for matched samples). In both implant groups with BMP-coating a synergetic effect was verifiable although the bone ongrowth in the hydroxyapatite coated implants was more extensive than in the corundum blasted implants. Light microscopy demonstrated osteointegration without connective tissue membrane around the surface of the implants. Our results indicate that composite metal implants, as used in endoprosthetics and implantology, are suitable carriers for BMP-3 and improved fixation of the implants can be achieved. The hydroxyapatite surface is superior to the corundum-blasted surface with regards to the observed parameters because of its pronounced bioactivity and its osteoconductive characteristics.
文摘The extracellular matrix-associated bone morphogenetic proteins(BMPs) govern a plethora of biological processes. The BMPs are members of the transforming growth factor-β protein superfamily, and they actively participate to kidney development, digit and limb formation, angiogenesis, tissue fibrosis and tumor development. Since their discovery, they have attracted attention for their fascinating perspectives in the regenerative medicine and tissue engineering fields. BMPs have been employed in many preclinical and clinical studies exploring their chondrogenic or osteoinductive potential in several animal model defects and in human diseases. During years of research in particular two BMPs, BMP2 and BMP7 have gained the podium for their use in the treatment of various cartilage and bone defects. In particular they have been recently approved for employment in non-union fractures as adjunct therapies. On the other hand, thanks to their potentialities in biomedical applications, there is a growing interest in studying the biology of mesenchymal stem cell(MSC), the rules underneath their differentiation abilities, and to test their true abilities in tissue engineering. In fact, the specific differentiation of MSCs into targeted celltype lineages for transplantation is a primary goal of the regenerative medicine. This review provides an overview on the current knowledge of BMP roles and signaling in MSC biology and differentiation capacities. In particular the article focuses on the potential clinical use of BMPs and MSCs concomitantly, in cartilage and bone tissue repair.
基金Supported by the National Natural Science Foundation of China,No.81560104 and No.81860115
文摘BACKGROUND Liver fibrosis is a refractory disease whose persistence can eventually induce cirrhosis or even liver cancer.Early liver fibrosis is reversible by intervention.As a member of the transforming growth factor-beta(TGF-β)superfamily,bone morphogenetic protein 7(BMP7)has anti-liver fibrosis functions.However,little is known about BMP7 expression changes and its potential regulatory mechanism as well as the relationship between BMP7 and TGF-βduring liver fibrosis.In addition,the mechanism underlying the anti-liver fibrosis function of BMP7 needs to be further explored.AIM To investigate changes in the dynamic expression of BMP7 during liver fibrosis,interactions between BMP7 and TGF-β1,and possible mechanisms underlying the anti-liver fibrosis function of BMP7.METHODS Changes in BMP7 expression during liver fibrosis and the interaction between BMP7 and TGF-β1 in mice were observed.Exogenous BMP7 was used to treat mouse primary hepatic stellate cells(HSCs)to observe its effect on activation,migration,and proliferation of HSCs and explore the possible mechanism underlying the anti-liver fibrosis function of BMP7.Mice with liver fibrosis received exogenous BMP7 intervention to observe improvement of liver fibrosis by using Masson’s trichrome staining and detecting the expression of the HSC activation indicator alpha-smooth muscle actin(α-SMA)and the collagen formation associated protein type I collagen(Col I).Changes in the dynamic expression of BMP7 during liver fibrosis in the human body were further observed.RESULTS In the process of liver fibrosis induced by carbon tetrachloride(CCl4)in mice,BMP7 protein expression first increased,followed by a decrease;there was a similar trend in the human body.This process was accompanied by a sustained increase in TGF-β1 protein expression.In vitro experiment results showed that TGF-β1 inhibited BMP7 expression in a time-and dose-dependent manner.In contrast,high doses of exogenous BMP7 inhibited TGF-β1-induced activation,migration,and proliferation of HSCs;this inhibitory effect was associated with upregulation of pSmad1/5/8 and downregulation of phosphorylation of Smad3 and p38 by BMP7.In vivo experiment results showed that exogenous BMP7 improved liver fibrosis in mice.CONCLUSION During liver fibrosis,BMP7 protein expression first increases and then decreases.This changing trend is associated with inhibition of BMP7 expression by sustained upregulation of TGF-β1 in a time-and dose-dependent manner.Exogenous BMP7 could selectively regulate TGF-β/Smad pathway-associated factors to inhibit activation,migration,and proliferation of HSCs and exert antiliver fibrosis functions.Exogenous BMP7 has the potential to be used as an antiliver fibrosis drug.
基金Supported by The Science and Technology Project of Guizhou Province,China,No.SZ[2008]3049
文摘AIM:To observe the effect of Danshao Huaxian capsule(DHC)on the expression of Gremlin and bone morphogenetic protein-7(BMP-7)in the liver of hepatic fibrosis rats.METHODS:A total of 75 male Wistar rats were randomly divided into a normal control group(A),a CCl4-induced hepatic fibrosis model group(B),a natural recovery group(C),a low-dose DHC-treated group(D),and a high-dose DHC-treated group(E),with 15 rats in each group.Liver fibrosis was induced by subcutaneous injections of carbon tetrachloride(CCl4)and a highlipid/low-protein diet for 8 wk,except for the rats in group A.Then,the rats in the two DHC-treated groups were administered 0.5 and 1.0 g/kg DHC by gastrogavage once per day for 8 successive weeks,respectively.By the end of the experiment,the level of transforming growth factorβ1(TGF-β1)in the liver homogenate was determined by an enzyme-linked immunosorbent assay.The mRNA and protein expression of Gremlin and BMP-7 in the liver tissue was determined by reversetranscription polymerase chain reaction,an immunohistochemical assay,and Western blot analysis.RESULTS:Compared with group A,the level of TGF-β1and the mRNA and protein expression of Gremlin were significantly higher in group B(TGF-β1:736.30±24.40μg/g vs 284.20±18.32μg/g,P<0.01;mRNA of Gremlin:80.40±5.46 vs 49.83±4.20,P<0.01;positive protein expression rate of Gremlin:38.46%±1.70%vs 3.83%±0.88%,P<0.01;relative protein expression of Gremlin:2.81±0.24 vs 0.24±0.06,P<0.01),and the mRNA and protein expression of BMP-7was significantly lower in group B(mRNA:54.00±4.34vs 93.99±7.03,P<0.01;positive protein expression rate:28.97%±3.14%vs 58.29%±6.02,P<0.01;relative protein expression:0.48±0.31 vs 1.05±0.12,P<0.01).Compared with groups B and C,the degree of hepatic fibrosis was significantly improved,and the level of TGF-β1 and the mRNA and protein expression of Gremlin were significantly lowered in the two DHCtreated groups(TGF-β1:523.14±21.29μg/g,441.86±23.18μg/g vs 736.30±24.40μg/g,651.13±15.75μg/g,P<0.01;mRNA of Gremlin:64.86±2.83,55.82±5.39 vs 80.40±5.46,70.37±4.01,P<0.01;positive protein expression rate of Gremlin:20.78%±1.60%,17.43%±2.02%vs 38.46%±1.70%,29.50%±2.64%,P<0.01;relative protein expression of Gremlin:1.95±0.26,1.65±0.20 vs 2.81±0.24,2.22±0.63,P<0.01),and the mRNA and protein expression of BMP-7 was higher in the two DHC-treated groups(mRNA:73.52±4.56,81.78±5.38 vs 54.00±4.34,62.28±4.51,P<0.01;positive protein expression rate:41.44%±4.77%,47.49%±4.59%vs28.97%±3.14%,35.85%±3.50%,P<0.01;relative protein expression:0.71±0.06,0.81±0.07 vs 0.48±CONCLUSION:The therapeutic mechanism of DHC forhepatic fibrosis in rats may be associated with inhibitionof the expression of Gremlin and up-regulation of the expression of BMP-7.
基金The European Union(European Social Fund-ESF)Greek national funds through the Operational Program "Education and Lifelong Learning" of the National Strategic Reference Framework(NSRF)-Research Funding Program:Heracleitus Ⅱ
文摘AIM To present the incidence of heterotopic ossification after the use of recombinant human bone morphogenetic protein-7(rhB MP-7) for the treatment of nonunions.METHODS Bone morphogenetic proteins(BMPs) promote bone formation by auto-induction. Recombinant human BMP-7 in combination with bone grafts was used in 84 patients for the treatment of long bone nonunions. All patients were evaluated radiographicaly for the development of heterotopic ossification during the standard assessment for the nonunion healing. In all patients(80.9%) with radiographic signs of heterotopic ossification, a CT scan was performed. Nonunion site palpation and ROM evaluation of the adjacent jointswere also carried out. Factors related to the patient(age, gender), the nonunion(location, size, chronicity, number of previous procedures, infection, surrounding tissues condition) and the surgical procedure(graft and fixation type, amount of rhB MP-7) were correlated with the development of heterotopic ossification and statistical analysis with Pearsons χ~2 test was performed.RESULTS Eighty point nine percent of the nonunions treated with rh BMP-7, healed with no need for further procedures. Heterotopic bone formation occurred in 15 of 84 patients(17.8%) and it was apparent in the routine radiologi-cal evaluation of the nonunion site, in a mean time of 5.5 mo after the rh BMP-7 application(range 3-12). The heterotopic ossification was located at the femur in 8 cases, at the tibia in 6, and at the humerus in οne patient. In 4 patients a palpable mass was present and only in one patient, with a para-articular knee nonunion treated with rhB MP-7, the size of heterotopic ossification affected the knee range of motion. All the patients with heterotopic ossification were male. Statistical analysis proved that patient's gender was the only important factor for the development of heterotopic ossification(P = 0.007). CONCLUSION Heterotopic ossification after the use of rh BMP-7 in nonunions was common but it did not compromise the final clinical outcome in most cases, and affected only male patients.
基金Supported by National Natural Science Foundation of China(No.30872836)Natural Science Foundation of Liaoning Province,China(No.201102054)
文摘AIM: To investigate the influence of bone morphogenetic protein type IA receptor [BMPR-IA(ALK3)] conditional knockout in lens on expression of bone morphogenetic protein 4(BMP4) in lens during the development of the vertebrate eye.METHODS: Cre-positive mice were mated with Crenegative mice to generate 50% Cre-positive(conditional knockout, CKO) 4 embryos, 8 eyes and 50% Cre-negative offspring(wild type, WT) 4 embryos, 8 eyes. The embryos were fixed in 4% paraformaldehyde, embedded in paraffin, and sectioned to a thickness of 4 μm.Removal of paraffin wax and dehydrating for sections,and then the procedure of in situ hybridization was processed, BMP4 MK1784-m(BOSTER) was used, and observed the expression of BMP4 in the lens in experimental group and control group. We selected SPSS11.0 software for statistical analysis, P<0.05 showed that the difference was statistically significant.· RESULTS: Four embryos of each genotype were examined, totally we had 8 embryos, 16 eyes. We got the uniform outcomes in all the embryos. We found ALK3 was required during lens growing, but was not essential for the formation of lens. We observed that the expression of BMP4 in the lens was significantly reduced in all 8 ALK3 CKO lens, BMP4 expression was normal in all the 8 WT lens, P <0.01. This phenomenon became increasingly visible in accordance with embryo development. The most apparent alteration was present at stage E15.5.CONCLUSION: ALK3 is essential for lens growth. The influence of ALK3 on the expression of BMP4 is present during the development of mice lens.
文摘AIM: To investigate the antifibrotic effects of bone morphogenetic protein-7 (BMP-7) on Schistosoma japonicum (S. japonicum )-induced hepatic fibrosis in BALB/C mice. METHODS: Sixty BALB/C mice were randomly divided into three groups, including a control group (group A, n = 20), model group (group B, n = 20) and BMP-7 treated group (group C, n = 20). The mice in group B and group C were abdominally infected with S. japonicum cercariae to induce a schistosomal hepatic fibrosis model. The mice in group C were administered human recombinant BMP-7. Liver samples were extracted from mice sacrificed at 9 and 15 wk after modeling. Hepatic histopathological changes were assessed using Masson's staining. Transforming growth factor-beta 1 (TGF-β1), alpha-smooth muscle actin (α-SMA), phosphorylated Smad2/3 (pSmad2/3) and Smad7 protein levels and localization were measured by Western blotting and immunohistochemistry, respectively, and their mRNA expressions were detected by reverse transcriptionpolymerase chain reaction (RT-PCR). RESULTS: The schistosomal hepatic fibrosis mouse model was successfully established, as the livers of mice in group B and group C showed varying degrees of typical schistosomal hepatopathologic changes such as egg granuloma and collagen deposition. The degree of collagen deposition in group C was higher than that in group A (week 9: 22.95±6.66vs 2.02±0.76; week 15: 12.84±4.36 vs 1.74±0.80; P<0.05), but significantly lower than that in group B (week 9: 22.95±6.66 vs 34.43±6.96; week 15: 12.84±4.36 vs 18.90±5.07;P<0.05) at both time points. According to immunohistochemistry data, the expressions of α-SMA, TGF-β1 and pSmad2/3 protein in group C were higher than those in group A (α-SMA: week 9: 21.24±5.73 vs 0.33±0.20; week 15: 12.42±4.88 vs 0.34±0.27; TGF-β1: week 9: 37.00±13.74 vs 3.73±2.14; week 15: 16.71±9.80 vs 3.08±2.35; pSmad2/3: week 9: 12.92±4.81 vs 0.83±0.48; week 15: 7.87±4.09 vs 0.90±0.45; P<0.05), but significantly lower than those in group B (α-SMA: week 9: 21.24±5.73 vs 34.39±5.74; week 15: 12.42±4.88 vs 25.90±7.01; TGF-β1: week 9: 37.00±13.74 vs 55.66±14.88; week 15: 16.71±9.80 vs 37.10±12.51; pSmad2/3: week 9: 12.92±4.81 vs 19.41±6.87; week 15: 7.87±4.09vs 13.00±4.98;P<0.05) at both time points; the expression of Smad7 protein in group B was higher than that in group A and group C at week 9 (8.46±3.95 vs 1.00±0.40 and 8.46±3.95 vs 0.77±0.42; P<0.05), while there were no differences in Smad7 expression between the three groups at week 15 (1.09±0.38 vs 0.97±0.42 vs 0.89±0.39; P>0.05). Although minor discrepancies were observed, the results of RT-PCR and Western blotting were mainly consistentwith the immunohistochemical results. CONCLUSION: Exogenous BMP-7 significantly decreased the degree of hepatic fibrosis in both the acute and chronic stages of hepato-schistosomiasis, and the regulatory mechanism may involve the TGF-β/Smad signaling pathway.
基金This work was supported by National Natural Science Foundation Funding(3110131631371805)Program for New Century Excellent Talents in University of Ministry of Education of China(NCET-11-0796)and Heilongjiang Province Postdoctoral Science Foundation.
文摘Bone morphogenetic proteins(BMPs)are a family of potent,multifunctional growth factors belonging to transforming growth factor-(TGF-).They are highly conservative in structures.Over 20 members of BMPs with varying functions such as embryogenesis,skeletal formation,hematopoiesis and neurogenesis have been identified in human body.BMPs are unique growth factors that can induce the formation of bone tissue individually.BMPs can induce the differentiation of bone marrow mesenchymal stem cells into osteoblastic lineage and promote the proliferation of osteoblasts and chondrocytes.BMPs stimulate the target cells by specific membrane-bound receptors and signal transduced through mothers against decapentaplegic(Smads)and mitogen activated protein kinase(MAPK)pathways.It has been demonstrated that BMP-2,BMP-4,BMP-6,BMP-7,and BMP-9 play an important role in bone formation.This article focuses on the molecular characterization of BMPs family members,mechanism of osteogenesis promotion,related signal pathways of osteogenic function,relationships between structure and osteogenetic activity,and the interactions among family members at bone formation.
基金Supported by a grant from the Research Funds for Returned Overseas Scholars of Shanxi Province, China, No. 200568
文摘AIM: To characterize the expression and dynamic changes of bone morphogenetic protein (BMP)-2 in hepatocytes in the regenerating liver in rats after partial hepatectomy (PH), and examine the effects of BMP-2 on proliferation of human Huh7 hepatoma cells. METHODS: Fifty-four adult male Wistar rats were randomly divided into three groups: A normal control (NC) group, a partial hepatectomized (PH) group and a sham operated (SO) group. To study the effect of liver regeneration on BMP-2 expression, rats were sacrificed before and at different time points after PH or the sham intervention (6, 12, 24 and 48 h). For each time point, six rats were used in parallel. Expression and distribution of BMP-2 protein were determined in regenerating liver tissue by Western blot analysis and immunohistochemistry. Effects of BMP-2 on cell proliferation of human Huh7 hepatoma cell line were assessed using an MTT assay.RESULTS: In the normal liver strong BMP-2 expression was observed around the central and portal veins. The expression of BMP-2 decreased rapidly as measured by both immunohistochemistry and Western blot analysis. This decrease was at a maximum of 3.22 fold after 12 h and returned to normal levels at 48 h after PH. No significant changes in BMP-2 immunoreactivity were observed in the SO group. BMP-2 inhibited serum induced Huh7 cell proliferation.CONCLUSION: BMP-2 is expressed in normal adult rat liver and negatively regulates hepatocyte proliferation. The observed down regulation of BMP-2 following partial hepatectomy suggests that such down regulation may be necessary for hepatocyte proliferation.
基金Supported by American Heart Association Scientist Development Grant,No.11SDG7600103
文摘AIM:To explore ex vivo the role of bone morphogenetic protein-4(BMP-4) and transforming growth factorbeta1(TGF-β1) in acute valvular response to fluid shear stress(FSS) abnormalities.METHODS:Porcine valve leaflets were subjected ex vivo to physiologic FSS,supra-physiologic FSS magnitude at normal frequency and supra-physiologic FSS frequency at normal magnitude for 48 h in a double-sided cone-and-plate bioreactor filled with standard culture medium. The role of BMP-4 and TGF-β1 in the valvular response was investigated by promoting or inhibiting the downstream action of those cytokines via culture medium supplementation with BMP-4 or the BMP antagonist noggin,and TGF-β1 or the TGF-β1 inhibitor SB-431542,respectively. Fresh porcine leaflets were used as controls. Each experimental group consisted of six leaflet samples. Immunostaining and immunoblotting were performed to assess endothelial activation in terms of intercellular adhesion molecule-1 and vascular cell adhesion molecule-1 expressions,paracrine signaling in terms of BMP-4 and TGF-β1 expressions and extracellular matrix(ECM) remodeling in terms of cathepsin L,cathepsin S,metalloproteinases(MMP)-2 and MMP-9 expressions. Immunostained images were quantified by normalizing the intensities of positively stained regions by the number of cells in each image while immunoblots were quantified by densitometry. R E S U LT S :Regardless of the culture medium,physiologic FSS maintained valvular homeostasis. Tissue exposure to supra-physiologic FSS magnitude in standard medium stimulated paracrine signaling(TGF-β1:467% ± 22% vs 100% ± 6% in freshcontrols,BMP-4:258% ± 22% vs 100% ± 4% in fresh controls; P < 0.05) and ECM degradation(MMP-2:941% ± 90% vs 100% ± 19% in fresh controls,MMP-9:1219% ± 190% vs 100% ± 16% in fresh controls,cathepsin L:1187% ± 175% vs 100% ± 12% in fresh controls,cathepsin S:603% ± 88% vs 100% ± 13% in fresh controls; P < 0.05),while BMP-4 supplementation also promoted fibrosa activation and TGF-β1 inhibition reduced MMP-9 expression to the native tissue level(MMP-9:308% ± 153% with TGF-β1 inhibition vs 100% ± 16% in fresh control; P > 0.05). Supra-physiologic FSS frequency had no effect on endothelial activation and paracrine signaling regardless of the culture medium but TGF-β1 silencing attenuated FSS-induced ECM degradation via MMP-9 downregulation(MMP-9:302% ± 182% vs 100% ± 42% in fresh controls; P > 0.05).CONCLUSION:Valvular tissue is sensitive to FSS abnormalities. The TGF-β1 inhibitor SB-431542 is a potential candidate molecule for attenuating the effects of FSS abnormalities on valvular remodeling.
文摘The advent of recombinant DNA technology has substantially increased the intra-operative utilization of biologic augmentation in spine surgery over the past several years after the Food and Drug Administration approval of the bone morphogenetic protein(BMP) class of molecules for indications in the lumbar spine. Much less is known about the potential benefits and risks of the "off-label" use of BMP in the cervical spine. The history and relevant literature pertaining to the use of the "off-label" implantation of the BMP class of molecules in the anterior or posterior cervical spine are reviewed and discussed. Early prospective studies of BMP-2 implantation in anterior cervical spine constructs showed encouraging results. Later retrospective studies reported potentially "life threatening complications" resulting in a 2007 public health advisory by the FDA. Limited data regarding BMP-7 in anterior cervical surgery was available with one group reporting a 2.4% early(< 30 d) complication rate(brachialgia and dysphagia). BMP use in the decompressed posterior cervical spine may result in neurologic or wound compromise according to several retrospective reports, however, controlled use has been reported to increase fusion rates in select complex and pediatric patients. There were no cases of de novo neoplasia related to BMP implantation in the cervical spine. BMP-2 use in anterior cervical spine surgery has been associated with a high early complication rate. Definitive recommendations for BMP-7 use in anterior cervical spine surgery cannot be made with current clinical data. According to limited reports, select complex patients who are considered "high risk" for pseudoarthrosis undergoing posterior cervical or occipitocervical arthrodesis or children with congenital or traumatic conditions may be candidates for "off-label" use of BMP in the context of appropriate informed decision making. At the present time, there are no highlevel clinical studies on the outcomes and complication rates of BMP implantation in the cervical spine.
基金This project was supported by a grant from the National Natural Sciences Foundation of China (No 30471753)
文摘To study the cartilage differentiation of mouse mesenchymal stem cells(MSCs) induced by cartilage-derived morphogenetic proteins-2 in vitro,the MSCs were isolated from mouse bone marrow and cultured in vitro.The cells in passage 3 were induced into chondrogenic differentiation with different concentrations of recombinant human cartilage-derived morphogenetic proteins-2(0,10,20,50 and 100 ng/mL).After 14 days of induction,morphology of cells was observed under phase-contrast microscope.Collagen Ⅱ mRNA and protein were examined with RT-PCR,Western blotting and immunocytochemistry respectively and the sulfate glycosaminoglycan was measured by Alcian blue staining.RT-PCR showed that CDMP-2 could promote expression of collagen Ⅱ mRNA in an dose-dependant manner,especially at the concentration of 50 ng/mL and 100 ng/mL.Immunocytochemistry and Western blotting revealed a similar change.Alcian blue staining exhibited deposition of typical cartilage extracellular matrix.Our results suggest that mouse bone marrow mesencymal stem cells can differentiate into chondrogenic phonotype with the induction of CDMP-2 in vitro,which provides a basis for further research on the role of CDMP-2 in chondrogenesis.
基金Supported by National Natural Science Foundation of China(30500414)Scientific Research Project in Department of Education of Liaoning Province(05L508,20061010)
文摘Objective To evaluate the effects of retinoic acid(RA)on expression of bone morphogenetic protein 7(BMP-7)in rat fetus with cleft palate,and the effects of RA on proliferation and apoptosis of osteoblasts.Methods All-trans RA(ATRA)was used to induce congenital cleft palate in Wistar rat.BMP-7 mRNA expression in maxillary bone tissue of fetal rats was measured by Northern blotting analysis.Flow cytometry and MTT assay were used to measure the apoptosis and proliferation of ATRA-treated MC-3T3-E1 cells.BMP-7 mRNA and protein expressions in ATRA-treated MC-3T3-E1 cells were detected by RT-PCR and Western blotting analysis.Results ATRA could induce cleft palate of rat fetus.The incidence rate of cleft palate induced by 100 mg/kg ATRA(45.5%)was significantly higher than 50 mg/kg ATRA(12.5%,P<0.05).BMP-7 mRNA expression decreased in maxillary bone tissue of rat fetus with cleft palate.MC-3T3-E1 cells proliferation treated with 1×10-6 mol/LATRA decreased by 60%,the cell apoptosis increased by 2 times.BMP-7 mRNA and protein levels in MC-3T3-E1 cells treated with 1×10-6 mol/L ATRA decreased by 60% and 80%,respectively,compared with ATRA-untreated cells(P<0.05).Conclusions BMP-7 may play an important role in embryonic palate development.RA may possess the ability to down-regulate cell proliferation through regulation of BMP-7 gene expression.
基金Supported by National Natural Science Foundation of China(No.81070753)Natural Science Foundation of Guangdong Province,China(No.10251008901000025)
文摘Bone morphogenesis proteins(BMPs) are multi-functional growth factors. They are expressed in retina,retinal pigment epithelium(RPE) and sclera and serve as a regulator in the growth and development of the eye. This article reviewed the chondrogenic potency of the sclera,biochemical and pathological changes of myopic scleral tissue and the differentiation of chondrogenesis by BMP-2. We proposed the hypothesis that BMP-2 can regulate differentiate of scleral fibroblasts and affect the development of myopia.
文摘An experimental model of femoral muscular pouch in 20 mice was adopted. The expression of VEGF was examined by in situ hybridization method and immunohistochemical method in bone morphogenetic protein-2 induced osteogenesis. The experimental results demonstrated that the expression signals of VEGF mRNA and VEGF appeared in cytoplasm during condensation of mesenchymal cell. As the mesenchymal cells differentiated into precartilage, the expression signals decreased in mesenchymal cells, but increased in chondrocytes and kept getting denser in the process of cartilage maturity. The peak expression of VEGF mRNA and VEGF in the experimental group appeared on the 14th day, accompanied by numerous hypertrophic chondrocytes. When mature cartilage calcified and new bone trabecula formed, the expression of VEGF mRNA and VEGF decreased in chondrocytes, but still expressed moderately in the osteoblasts and osteocytes. Signals of VEGF mRNA and VEGF can not be detected in the control groups.
