The Experts below are selected from a list of 2535 Experts worldwide ranked by ideXlab platform
Marcio Mateus Beloti - One of the best experts on this subject based on the ideXlab platform.
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titanium with Nanotopography attenuates the osteoclast induced disruption of osteoblast differentiation by regulating histone methylation
Materials Science and Engineering: C, 2021Co-Authors: Rayana L Bighettitrevisan, Adalberto Luiz Rosa, Luciana O Almeida, Larissa Moreira Spinola De Castroraucci, Jonathan A R Gordon, Coralee E Tye, Gary S Stein, Jane B Lian, Janet L Stein, Marcio Mateus BelotiAbstract:Abstract The bone remodeling process is crucial for titanium (Ti) osseointegration and involves the crosstalk between osteoclasts and osteoblasts. Considering the high osteogenic potential of Ti with Nanotopography (Ti Nano) and that osteoclasts inhibit osteoblast differentiation, we hypothesized that Nanotopography attenuate the osteoclast-induced disruption of osteoblast differentiation. Osteoblasts were co-cultured with osteoclasts on Ti Nano and Ti Control and non-co-cultured osteoblasts were used as control. Gene expression analysis using RNAseq showed that osteoclasts downregulated the expression of osteoblast marker genes and upregulated genes related to histone modification and chromatin organization in osteoblasts grown on both Ti surfaces. Osteoclasts also inhibited the mRNA and protein expression of osteoblast markers, and such effect was attenuated by Ti Nano. Also, osteoclasts increased the protein expression of H3K9me2, H3K27me3 and EZH2 in osteoblasts grown on both Ti surfaces. ChIP assay revealed that osteoclasts increased accumulation of H3K27me3 that represses the promoter regions of Runx2 and Alpl in osteoblasts grown on Ti Control, which was reduced by Ti Nano. In conclusion, these data show that despite osteoclast inhibition of osteoblasts grown on both Ti Control and Ti Nano, the Nanotopography attenuates the osteoclast-induced disruption of osteoblast differentiation by preventing the increase of H3K27me3 accumulation that represses the promoter regions of some key osteoblast marker genes. These findings highlight the epigenetic mechanisms triggered by Nanotopography to protect osteoblasts from the deleterious effects of osteoclasts, which modulate the process of bone remodeling and may benefit the osseointegration of Ti implants.
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titanium with Nanotopography induces osteoblast differentiation by regulating endogenous bone morphogenetic protein expression and signaling pathway
Journal of Cellular Biochemistry, 2016Co-Authors: Larissa Moreira Spinola De Castroraucci, Lucas Novaes Teixeira, P T De Oliveira, Mohammad Q Hassan, Emanuela Prado Ferraz, Marcelo S Francischini, Helena Bacha Lopes, Adalberto L Losa, Marcio Mateus BelotiAbstract:We aimed at evaluating the effect of titanium (Ti) with Nanotopography (Nano) on the endogenous expression of BMP-2 and BMP-4 and the relevance of this process to the Nanotopography-induced osteoblast differentiation. MC3T3-E1 cells were grown on Nano and machined (Machined) Ti surfaces and the endogenous BMP-2/4 expression and the effect of BMP receptor BMPR1A silencing in both osteoblast differentiation and expression of genes related to TGF-β/BMP signaling were evaluated. Nano supported higher BMP-2 gene and protein expression and upregulated the osteoblast differentiation compared with Machined Ti surface. The BMPR1A silencing inhibited the osteogenic potential induced by Nano Ti surface as indicated by reduced alkaline phosphatase (ALP), osteocalcin and RUNX2 gene expression, RUNX2 protein expression and ALP activity. In addition, the expression of genes related to TGF-β/BMP signaling was deeply affected by BMPR1A-silenced cells grown on Nano Ti surface. In conclusion, we have demonstrated for the first time that Nanotopography induces osteoblast differentiation, at least in part, by upregulating the endogenous production of BMP-2 and modulating BMP signaling pathway. J. Cell. Biochem. 117: 1718-1726, 2016. © 2015 Wiley Periodicals, Inc.
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Nanotopography directs mesenchymal stem cells to osteoblast lineage through regulation of microrna smad bmp 2 circuit
Journal of Cellular Physiology, 2014Co-Authors: Rogerio Bentes Kato, Adalberto Luiz Rosa, F S De Oliveira, P T De Oliveira, Mohammad Q Hassan, Bhaskar Roy, Emanuela Prado Ferraz, Austin G Kemper, Marcio Mateus BelotiAbstract:The aim of this study was to investigate if chemically produced Nanotopography on titanium (Ti) surface induces osteoblast differentiation of cultured human bone marrow mesenchymal stem cells (hMSCs) by regulating the expression of microRNAs (miRs). It was demonstrated that Ti with Nanotopography induces osteoblast differentiation of hMSCs as evidenced by upregulation of osteoblast specific markers compared with untreated (control) Ti at day 4. At this time-point, miR-sequencing analysis revealed that 20 miRs were upregulated (>twofold) while 20 miRs were downregulated (>threefold) in hMSCs grown on Ti with Nanotopography compared with control Ti. Three miRs, namely miR-4448, -4708, and -4773, which were significantly downregulated (>fivefold) by Ti with Nanotopography affect osteoblast differentiation of hMSCs. These miRs directly target SMAD1 and SMAD4, both key transducers of the bone morphogenetic protein 2 (BMP-2) osteogenic signal, which were upregulated by Ti with Nanotopography. Overexpression of miR-4448, -4708, and 4773 in MC3T3-E1 pre-osteoblasts noticeably inhibited gene and protein expression of SMAD1 and SMAD4 and therefore repressed the gene expression of key bone markers. Additionally, it was observed that the treatment with BMP-2 displayed a higher osteogenic effect on MC3T3-E1 cells grown on Ti with Nanotopography compared with control Ti, suggesting that the BMP-2 signaling pathway was more effective on this surface. Taken together, these results indicate that a complex regulatory network involving a miR-SMAD-BMP-2 circuit governs the osteoblast differentiation induced by Ti with Nanotopography. J. Cell. Physiol. 229: 1690-1696, 2014. © 2014 Wiley Periodicals, Inc.
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Nanotopography drives stem cell fate toward osteoblast differentiation through α1β1 integrin signaling pathway
Journal of Cellular Biochemistry, 2014Co-Authors: Adalberto Luiz Rosa, Rogerio Bentes Kato, L Castro M S Raucci, Lucas Novaes Teixeira, F S De Oliveira, L S Bellesini, P T De Oliveira, Mohammad Q Hassan, Marcio Mateus BelotiAbstract:The aim of our study was to investigate the osteoinductive potential of a titanium (Ti) surface with Nanotopography, using mesenchymal stem cells (MSCs) and the mechanism involved in this phenomenon. Polished Ti discs were chemically treated with H2 SO4 /H2 O2 to yield Nanotopography and rat MSCs were cultured under osteogenic and non-osteogenic conditions on both Nanotopography and untreated polished (control) Ti surfaces. The Nanotopography increased cell proliferation and alkaline phosphatase (Alp) activity and upregulated the gene expression of key bone markers of cells grown under both osteogenic and non-osteogenic conditions. Additionally, the gene expression of α1 and β1 integrins was higher in cells grown on Ti with Nanotopography under non-osteogeneic condition compared with control Ti surface. The higher gene expression of bone markers and Alp activity induced by Ti with Nanotopography was reduced by obtustatin, an α1β1 integrin inhibitor. These results indicate that α1β1 integrin signaling pathway determines the osteoinductive effect of Nanotopography on MSCs. This finding highlights a novel mechanism involved in nanosurface-mediated MSCs fate and may contribute to the development of new surface modifications aiming to accelerate and/or enhance the process of osseointegration.
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Nanotopography drives stem cell fate toward osteoblast differentiation through α1β1 integrin signaling pathway
Journal of Cellular Biochemistry, 2014Co-Authors: Adalberto Luiz Rosa, Rogerio Bentes Kato, L Castro M S Raucci, Lucas Novaes Teixeira, F S De Oliveira, L S Bellesini, P T De Oliveira, Mohammad Q Hassan, Marcio Mateus BelotiAbstract:The aim of our study was to investigate the osteoinductive potential of a titanium (Ti) surface with Nanotopography, using mesenchymal stem cells (MSCs) and the mechanism involved in this phenomenon. Polished Ti discs were chemically treated with H2SO4/H2O2 to yield Nanotopography and rat MSCs were cultured under osteogenic and non-osteogenic conditions on both Nanotopography and untreated polished (control) Ti surfaces. The Nanotopography increased cell proliferation and alkaline phosphatase (Alp) activity and upregulated the gene expression of key bone markers of cells grown under both osteogenic and non-osteogenic conditions. Additionally, the gene expression of α1 and β1 integrins was higher in cells grown on Ti with Nanotopography under non-osteogeneic condition compared with control Ti surface. The higher gene expression of bone markers and Alp activity induced by Ti with Nanotopography was reduced by obtustatin, an α1β1 integrin inhibitor. These results indicate that α1β1 integrin signaling pathway determines the osteoinductive effect of Nanotopography on MSCs. This finding highlights a novel mechanism involved in nanosurface-mediated MSCs fate and may contribute to the development of new surface modifications aiming to accelerate and/or enhance the process of osseointegration. J. Cell. Biochem. 115: 540–548, 2014. © 2013 Wiley Periodicals, Inc.
Adalberto Luiz Rosa - One of the best experts on this subject based on the ideXlab platform.
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titanium with Nanotopography attenuates the osteoclast induced disruption of osteoblast differentiation by regulating histone methylation
Materials Science and Engineering: C, 2021Co-Authors: Rayana L Bighettitrevisan, Adalberto Luiz Rosa, Luciana O Almeida, Larissa Moreira Spinola De Castroraucci, Jonathan A R Gordon, Coralee E Tye, Gary S Stein, Jane B Lian, Janet L Stein, Marcio Mateus BelotiAbstract:Abstract The bone remodeling process is crucial for titanium (Ti) osseointegration and involves the crosstalk between osteoclasts and osteoblasts. Considering the high osteogenic potential of Ti with Nanotopography (Ti Nano) and that osteoclasts inhibit osteoblast differentiation, we hypothesized that Nanotopography attenuate the osteoclast-induced disruption of osteoblast differentiation. Osteoblasts were co-cultured with osteoclasts on Ti Nano and Ti Control and non-co-cultured osteoblasts were used as control. Gene expression analysis using RNAseq showed that osteoclasts downregulated the expression of osteoblast marker genes and upregulated genes related to histone modification and chromatin organization in osteoblasts grown on both Ti surfaces. Osteoclasts also inhibited the mRNA and protein expression of osteoblast markers, and such effect was attenuated by Ti Nano. Also, osteoclasts increased the protein expression of H3K9me2, H3K27me3 and EZH2 in osteoblasts grown on both Ti surfaces. ChIP assay revealed that osteoclasts increased accumulation of H3K27me3 that represses the promoter regions of Runx2 and Alpl in osteoblasts grown on Ti Control, which was reduced by Ti Nano. In conclusion, these data show that despite osteoclast inhibition of osteoblasts grown on both Ti Control and Ti Nano, the Nanotopography attenuates the osteoclast-induced disruption of osteoblast differentiation by preventing the increase of H3K27me3 accumulation that represses the promoter regions of some key osteoblast marker genes. These findings highlight the epigenetic mechanisms triggered by Nanotopography to protect osteoblasts from the deleterious effects of osteoclasts, which modulate the process of bone remodeling and may benefit the osseointegration of Ti implants.
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Nanotopography directs mesenchymal stem cells to osteoblast lineage through regulation of microrna smad bmp 2 circuit
Journal of Cellular Physiology, 2014Co-Authors: Rogerio Bentes Kato, Adalberto Luiz Rosa, F S De Oliveira, P T De Oliveira, Mohammad Q Hassan, Bhaskar Roy, Emanuela Prado Ferraz, Austin G Kemper, Marcio Mateus BelotiAbstract:The aim of this study was to investigate if chemically produced Nanotopography on titanium (Ti) surface induces osteoblast differentiation of cultured human bone marrow mesenchymal stem cells (hMSCs) by regulating the expression of microRNAs (miRs). It was demonstrated that Ti with Nanotopography induces osteoblast differentiation of hMSCs as evidenced by upregulation of osteoblast specific markers compared with untreated (control) Ti at day 4. At this time-point, miR-sequencing analysis revealed that 20 miRs were upregulated (>twofold) while 20 miRs were downregulated (>threefold) in hMSCs grown on Ti with Nanotopography compared with control Ti. Three miRs, namely miR-4448, -4708, and -4773, which were significantly downregulated (>fivefold) by Ti with Nanotopography affect osteoblast differentiation of hMSCs. These miRs directly target SMAD1 and SMAD4, both key transducers of the bone morphogenetic protein 2 (BMP-2) osteogenic signal, which were upregulated by Ti with Nanotopography. Overexpression of miR-4448, -4708, and 4773 in MC3T3-E1 pre-osteoblasts noticeably inhibited gene and protein expression of SMAD1 and SMAD4 and therefore repressed the gene expression of key bone markers. Additionally, it was observed that the treatment with BMP-2 displayed a higher osteogenic effect on MC3T3-E1 cells grown on Ti with Nanotopography compared with control Ti, suggesting that the BMP-2 signaling pathway was more effective on this surface. Taken together, these results indicate that a complex regulatory network involving a miR-SMAD-BMP-2 circuit governs the osteoblast differentiation induced by Ti with Nanotopography. J. Cell. Physiol. 229: 1690-1696, 2014. © 2014 Wiley Periodicals, Inc.
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Nanotopography drives stem cell fate toward osteoblast differentiation through α1β1 integrin signaling pathway
Journal of Cellular Biochemistry, 2014Co-Authors: Adalberto Luiz Rosa, Rogerio Bentes Kato, L Castro M S Raucci, Lucas Novaes Teixeira, F S De Oliveira, L S Bellesini, P T De Oliveira, Mohammad Q Hassan, Marcio Mateus BelotiAbstract:The aim of our study was to investigate the osteoinductive potential of a titanium (Ti) surface with Nanotopography, using mesenchymal stem cells (MSCs) and the mechanism involved in this phenomenon. Polished Ti discs were chemically treated with H2 SO4 /H2 O2 to yield Nanotopography and rat MSCs were cultured under osteogenic and non-osteogenic conditions on both Nanotopography and untreated polished (control) Ti surfaces. The Nanotopography increased cell proliferation and alkaline phosphatase (Alp) activity and upregulated the gene expression of key bone markers of cells grown under both osteogenic and non-osteogenic conditions. Additionally, the gene expression of α1 and β1 integrins was higher in cells grown on Ti with Nanotopography under non-osteogeneic condition compared with control Ti surface. The higher gene expression of bone markers and Alp activity induced by Ti with Nanotopography was reduced by obtustatin, an α1β1 integrin inhibitor. These results indicate that α1β1 integrin signaling pathway determines the osteoinductive effect of Nanotopography on MSCs. This finding highlights a novel mechanism involved in nanosurface-mediated MSCs fate and may contribute to the development of new surface modifications aiming to accelerate and/or enhance the process of osseointegration.
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Nanotopography drives stem cell fate toward osteoblast differentiation through α1β1 integrin signaling pathway
Journal of Cellular Biochemistry, 2014Co-Authors: Adalberto Luiz Rosa, Rogerio Bentes Kato, L Castro M S Raucci, Lucas Novaes Teixeira, F S De Oliveira, L S Bellesini, P T De Oliveira, Mohammad Q Hassan, Marcio Mateus BelotiAbstract:The aim of our study was to investigate the osteoinductive potential of a titanium (Ti) surface with Nanotopography, using mesenchymal stem cells (MSCs) and the mechanism involved in this phenomenon. Polished Ti discs were chemically treated with H2SO4/H2O2 to yield Nanotopography and rat MSCs were cultured under osteogenic and non-osteogenic conditions on both Nanotopography and untreated polished (control) Ti surfaces. The Nanotopography increased cell proliferation and alkaline phosphatase (Alp) activity and upregulated the gene expression of key bone markers of cells grown under both osteogenic and non-osteogenic conditions. Additionally, the gene expression of α1 and β1 integrins was higher in cells grown on Ti with Nanotopography under non-osteogeneic condition compared with control Ti surface. The higher gene expression of bone markers and Alp activity induced by Ti with Nanotopography was reduced by obtustatin, an α1β1 integrin inhibitor. These results indicate that α1β1 integrin signaling pathway determines the osteoinductive effect of Nanotopography on MSCs. This finding highlights a novel mechanism involved in nanosurface-mediated MSCs fate and may contribute to the development of new surface modifications aiming to accelerate and/or enhance the process of osseointegration. J. Cell. Biochem. 115: 540–548, 2014. © 2013 Wiley Periodicals, Inc.
Richard O C Oreffo - One of the best experts on this subject based on the ideXlab platform.
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Image_2_Human Skeletal Stem Cell Response to Multiscale Topography Induced by Large Area Electron Beam Irradiation Surface Treatment.JPEG
2018Co-Authors: Vitali Goriainov, Richard B. Cook, James W. Murray, John C. Walker, Douglas G. Dunlop, Adam T. Clare, Richard O C OreffoAbstract:The healthcare socio-economic environment is irreversibly changing as a consequence of an increasing aging population, consequent functional impairment, and patient quality of life expectations. The increasing complexity of ensuing clinical scenarios compels a critical search for novel musculoskeletal regenerative and replacement strategies. While joint arthroplasty is a highly effective treatment for arthritis and osteoporosis, further innovation and refinement of uncemented implants are essential in order to improve implant integration and reduce implant revision rate. This is critical given financial restraints and the drive to improve cost-effectiveness and quality of life outcomes. Multi-scale modulation of implant surfaces, offers an innovative approach to enhancement in implant performance. In the current study, we have examined the potential of large area electron beam melting to alter the surface Nanotopography in titanium alloy (Ti6Al4V). We evaluated the in vitro osteogenic response of human skeletal stem cells to the resultant Nanotopography, providing evidence of the relationship between the biological response, particularly Collagen type I and Osteocalcin gene activation, and surface nanoroughness. The current studies demonstrate osteogenic gene induction and morphological cell changes to be significantly enhanced on a topography Ra of ~40 nm with clinical implications therein for implant surface treatment and generation.
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Human Skeletal Stem Cell Response to Multiscale Topography Induced by Large Area Electron Beam Irradiation Surface Treatment
'Frontiers Media SA', 2018Co-Authors: Vitali Goriainov, Richard B. Cook, James W. Murray, John C. Walker, Douglas G. Dunlop, Adam T. Clare, Richard O C OreffoAbstract:The healthcare socio-economic environment is irreversibly changing as a consequence of an increasing aging population, consequent functional impairment, and patient quality of life expectations. The increasing complexity of ensuing clinical scenarios compels a critical search for novel musculoskeletal regenerative and replacement strategies. While joint arthroplasty is a highly effective treatment for arthritis and osteoporosis, further innovation and refinement of uncemented implants are essential in order to improve implant integration and reduce implant revision rate. This is critical given financial restraints and the drive to improve cost-effectiveness and quality of life outcomes. Multi-scale modulation of implant surfaces, offers an innovative approach to enhancement in implant performance. In the current study, we have examined the potential of large area electron beam melting to alter the surface Nanotopography in titanium alloy (Ti6Al4V). We evaluated the in vitro osteogenic response of human skeletal stem cells to the resultant Nanotopography, providing evidence of the relationship between the biological response, particularly Collagen type I and Osteocalcin gene activation, and surface nanoroughness. The current studies demonstrate osteogenic gene induction and morphological cell changes to be significantly enhanced on a topography Ra of ~40 nm with clinical implications therein for implant surface treatment and generation
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harnessing Nanotopography and integrin matrix interactions to influence stem cell fate
Nature Materials, 2014Co-Authors: Matthew J Dalby, Nikolaj Gadegaard, Richard O C OreffoAbstract:Stem cells respond to nanoscale surface features, with changes in cell growth and differentiation mediated by alterations in cell adhesion. The interaction of nanotopographical features with integrin receptors in the cells' focal adhesions alters how the cells adhere to materials surfaces, and defines cell fate through changes in both cell biochemistry and cell morphology. In this Review, we discuss how cell adhesions interact with Nanotopography, and we provide insight as to how materials scientists can exploit these interactions to direct stem cell fate and to understand how the behaviour of stem cells in their niche can be controlled. We expect knowledge gained from the study of cell-Nanotopography interactions to accelerate the development of next-generation stem cell culture materials and implant interfaces, and to fuel discovery of stem cell therapeutics to support regenerative therapies.
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nanotopographical control of stem cell differentiation
Journal of Tissue Engineering, 2010Co-Authors: Laura E Mcnamara, Richard O C Oreffo, Rebecca J Mcmurray, Manus J P Biggs, Fahsai Kantawong, Matthew J DalbyAbstract:Stem cells have the capacity to differentiate into various lineages, and the ability to reliably direct stem cell fate determination would have tremendous potential for basic research and clinical therapy. Nanotopography provides a useful tool for guiding differentiation, as the features are more durable than surface chemistry and can be modified in size and shape to suit the desired application. In this paper, Nanotopography is examined as a means to guide differentiation, and its application is described in the context of different subsets of stem cells, with a particular focus on skeletal (mesenchymal) stem cells. To address the mechanistic basis underlying the topographical effects on stem cells, the likely contributions of indirect (biochemical signal-mediated) and direct (force-mediated) mechanotransduction are discussed. Data from proteomic research is also outlined in relation to topography-mediated fate determination, as this approach provides insight into the global molecular changes at the level of the functional effectors.
Kam W Leong - One of the best experts on this subject based on the ideXlab platform.
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Nanotopography as modulator of human mesenchymal stem cell function
Biomaterials, 2012Co-Authors: Karina Kulangara, Yong Yang, Jennifer Yang, Kam W LeongAbstract:Nanotopography changes human mesenchymal stem cells (hMSC) from their shape to their differentiation potential; however little is known about the underlying molecular mechanisms. Here we study the culture of hMSC on polydimethylsiloxane substrates with 350 nm grating topography and investigate the focal adhesion composition and dynamics using biochemical and imaging techniques. Our results show that zyxin protein plays a key role in the hMSC response to Nanotopography. Zyxin expression is downregulated on 350 nm gratings, leading to smaller and more dynamic focal adhesion. Since the association of zyxin with focal adhesions is force-dependent, smaller zyxin-positive adhesion as well as its higher turnover rate suggests that the traction force in focal adhesion on 350 nm topography is decreased. These changes lead to faster and more directional migration on 350 nm gratings. These findings demonstrate that Nanotopography decreases the mechanical forces acting on focal adhesions in hMSC and suggest that force-dependent changes in zyxin protein expression and kinetics underlie the focal adhesion remodeling in response to 350 nm grating topography, resulting in modulation of hMSC function.
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Nanotopography induced changes in focal adhesions cytoskeletal organization and mechanical properties of human mesenchymal stem cells
Biomaterials, 2010Co-Authors: Evelyn K F Yim, Eric M Darling, Karina Kulangara, Farshid Guilak, Kam W LeongAbstract:Abstract The growth of stem cells can be modulated by physical factors such as extracellular matrix Nanotopography. We hypothesize that Nanotopography modulates cell behavior by changing the integrin clustering and focal adhesion (FA) assembly, leading to changes in cytoskeletal organization and cell mechanical properties. Human mesenchymal stem cells (hMSCs) cultured on 350 nm gratings of tissue-culture polystyrene (TCPS) and polydimethylsiloxane (PDMS) showed decreased expression of integrin subunits α2, α6, αV, β2, β3 and β4 compared to the unpatterned controls. On gratings, the elongated hMSCs exhibited an aligned actin cytoskeleton, while on unpatterned controls, spreading cells showed a random but denser actin cytoskeleton network. Expression of cytoskeleton and FA components was also altered by the Nanotopography as reflected in the mechanical properties measured by atomic force microscopy (AFM) indentation. On the rigid TCPS, hMSCs on gratings exhibited lower instantaneous and equilibrium Young's moduli and apparent viscosity. On the softer PDMS, the effects of Nanotopography were not significant. However, hMSCs cultured on PDMS showed lower cell mechanical properties than those on TCPS, regardless of topography. These suggest that both Nanotopography and substrate stiffness could be important in determining mechanical properties, while Nanotopography may be more dominant in determining the organization of the cytoskeleton and FAs.
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Nanotopography induced changes in focal adhesions cytoskeletal organization and mechanical properties of human mesenchymal stem cells
Biomaterials, 2010Co-Authors: Evelyn K F Yim, Eric M Darling, Karina Kulangara, Farshid Guilak, Kam W LeongAbstract:The growth of stem cells can be modulated by physical factors such as extracellular matrix Nanotopography. We hypothesize that Nanotopography modulates cell behavior by changing the integrin clustering and focal adhesion (FA) assembly, leading to changes in cytoskeletal organization and cell mechanical properties. Human mesenchymal stem cells (hMSCs) cultured on 350 nm gratings of tissue-culture polystyrene (TCPS) and polydimethylsiloxane (PDMS) showed decreased expression of integrin subunits alpha2, alpha , alpha V, beta2, beta 3 and beta 4 compared to the unpatterned controls. On gratings, the elongated hMSCs exhibited an aligned actin cytoskeleton, while on unpatterned controls, spreading cells showed a random but denser actin cytoskeleton network. Expression of cytoskeleton and FA components was also altered by the Nanotopography as reflected in the mechanical properties measured by atomic force microscopy (AFM) indentation. On the rigid TCPS, hMSCs on gratings exhibited lower instantaneous and equilibrium Young's moduli and apparent viscosity. On the softer PDMS, the effects of Nanotopography were not significant. However, hMSCs cultured on PDMS showed lower cell mechanical properties than those on TCPS, regardless of topography. These suggest that both Nanotopography and substrate stiffness could be important in determining mechanical properties, while Nanotopography may be more dominant in determining the organization of the cytoskeleton and FAs.
Mohammad Q Hassan - One of the best experts on this subject based on the ideXlab platform.
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titanium with Nanotopography induces osteoblast differentiation by regulating endogenous bone morphogenetic protein expression and signaling pathway
Journal of Cellular Biochemistry, 2016Co-Authors: Larissa Moreira Spinola De Castroraucci, Lucas Novaes Teixeira, P T De Oliveira, Mohammad Q Hassan, Emanuela Prado Ferraz, Marcelo S Francischini, Helena Bacha Lopes, Adalberto L Losa, Marcio Mateus BelotiAbstract:We aimed at evaluating the effect of titanium (Ti) with Nanotopography (Nano) on the endogenous expression of BMP-2 and BMP-4 and the relevance of this process to the Nanotopography-induced osteoblast differentiation. MC3T3-E1 cells were grown on Nano and machined (Machined) Ti surfaces and the endogenous BMP-2/4 expression and the effect of BMP receptor BMPR1A silencing in both osteoblast differentiation and expression of genes related to TGF-β/BMP signaling were evaluated. Nano supported higher BMP-2 gene and protein expression and upregulated the osteoblast differentiation compared with Machined Ti surface. The BMPR1A silencing inhibited the osteogenic potential induced by Nano Ti surface as indicated by reduced alkaline phosphatase (ALP), osteocalcin and RUNX2 gene expression, RUNX2 protein expression and ALP activity. In addition, the expression of genes related to TGF-β/BMP signaling was deeply affected by BMPR1A-silenced cells grown on Nano Ti surface. In conclusion, we have demonstrated for the first time that Nanotopography induces osteoblast differentiation, at least in part, by upregulating the endogenous production of BMP-2 and modulating BMP signaling pathway. J. Cell. Biochem. 117: 1718-1726, 2016. © 2015 Wiley Periodicals, Inc.
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Nanotopography directs mesenchymal stem cells to osteoblast lineage through regulation of microrna smad bmp 2 circuit
Journal of Cellular Physiology, 2014Co-Authors: Rogerio Bentes Kato, Adalberto Luiz Rosa, F S De Oliveira, P T De Oliveira, Mohammad Q Hassan, Bhaskar Roy, Emanuela Prado Ferraz, Austin G Kemper, Marcio Mateus BelotiAbstract:The aim of this study was to investigate if chemically produced Nanotopography on titanium (Ti) surface induces osteoblast differentiation of cultured human bone marrow mesenchymal stem cells (hMSCs) by regulating the expression of microRNAs (miRs). It was demonstrated that Ti with Nanotopography induces osteoblast differentiation of hMSCs as evidenced by upregulation of osteoblast specific markers compared with untreated (control) Ti at day 4. At this time-point, miR-sequencing analysis revealed that 20 miRs were upregulated (>twofold) while 20 miRs were downregulated (>threefold) in hMSCs grown on Ti with Nanotopography compared with control Ti. Three miRs, namely miR-4448, -4708, and -4773, which were significantly downregulated (>fivefold) by Ti with Nanotopography affect osteoblast differentiation of hMSCs. These miRs directly target SMAD1 and SMAD4, both key transducers of the bone morphogenetic protein 2 (BMP-2) osteogenic signal, which were upregulated by Ti with Nanotopography. Overexpression of miR-4448, -4708, and 4773 in MC3T3-E1 pre-osteoblasts noticeably inhibited gene and protein expression of SMAD1 and SMAD4 and therefore repressed the gene expression of key bone markers. Additionally, it was observed that the treatment with BMP-2 displayed a higher osteogenic effect on MC3T3-E1 cells grown on Ti with Nanotopography compared with control Ti, suggesting that the BMP-2 signaling pathway was more effective on this surface. Taken together, these results indicate that a complex regulatory network involving a miR-SMAD-BMP-2 circuit governs the osteoblast differentiation induced by Ti with Nanotopography. J. Cell. Physiol. 229: 1690-1696, 2014. © 2014 Wiley Periodicals, Inc.
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Nanotopography drives stem cell fate toward osteoblast differentiation through α1β1 integrin signaling pathway
Journal of Cellular Biochemistry, 2014Co-Authors: Adalberto Luiz Rosa, Rogerio Bentes Kato, L Castro M S Raucci, Lucas Novaes Teixeira, F S De Oliveira, L S Bellesini, P T De Oliveira, Mohammad Q Hassan, Marcio Mateus BelotiAbstract:The aim of our study was to investigate the osteoinductive potential of a titanium (Ti) surface with Nanotopography, using mesenchymal stem cells (MSCs) and the mechanism involved in this phenomenon. Polished Ti discs were chemically treated with H2 SO4 /H2 O2 to yield Nanotopography and rat MSCs were cultured under osteogenic and non-osteogenic conditions on both Nanotopography and untreated polished (control) Ti surfaces. The Nanotopography increased cell proliferation and alkaline phosphatase (Alp) activity and upregulated the gene expression of key bone markers of cells grown under both osteogenic and non-osteogenic conditions. Additionally, the gene expression of α1 and β1 integrins was higher in cells grown on Ti with Nanotopography under non-osteogeneic condition compared with control Ti surface. The higher gene expression of bone markers and Alp activity induced by Ti with Nanotopography was reduced by obtustatin, an α1β1 integrin inhibitor. These results indicate that α1β1 integrin signaling pathway determines the osteoinductive effect of Nanotopography on MSCs. This finding highlights a novel mechanism involved in nanosurface-mediated MSCs fate and may contribute to the development of new surface modifications aiming to accelerate and/or enhance the process of osseointegration.
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Nanotopography drives stem cell fate toward osteoblast differentiation through α1β1 integrin signaling pathway
Journal of Cellular Biochemistry, 2014Co-Authors: Adalberto Luiz Rosa, Rogerio Bentes Kato, L Castro M S Raucci, Lucas Novaes Teixeira, F S De Oliveira, L S Bellesini, P T De Oliveira, Mohammad Q Hassan, Marcio Mateus BelotiAbstract:The aim of our study was to investigate the osteoinductive potential of a titanium (Ti) surface with Nanotopography, using mesenchymal stem cells (MSCs) and the mechanism involved in this phenomenon. Polished Ti discs were chemically treated with H2SO4/H2O2 to yield Nanotopography and rat MSCs were cultured under osteogenic and non-osteogenic conditions on both Nanotopography and untreated polished (control) Ti surfaces. The Nanotopography increased cell proliferation and alkaline phosphatase (Alp) activity and upregulated the gene expression of key bone markers of cells grown under both osteogenic and non-osteogenic conditions. Additionally, the gene expression of α1 and β1 integrins was higher in cells grown on Ti with Nanotopography under non-osteogeneic condition compared with control Ti surface. The higher gene expression of bone markers and Alp activity induced by Ti with Nanotopography was reduced by obtustatin, an α1β1 integrin inhibitor. These results indicate that α1β1 integrin signaling pathway determines the osteoinductive effect of Nanotopography on MSCs. This finding highlights a novel mechanism involved in nanosurface-mediated MSCs fate and may contribute to the development of new surface modifications aiming to accelerate and/or enhance the process of osseointegration. J. Cell. Biochem. 115: 540–548, 2014. © 2013 Wiley Periodicals, Inc.
