Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: UNIPROT:A7KAX9 (grit)
 1,275 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Phenotypic responses of rat calvarial osteoblast-like cells (RCOB) were evaluated on commercially pure titanium (cpTi) surfaces when cultured at high density (5100 cells/mm2). These surfaces were prepared to three different clinically relevant surface preparations (1-micron, 600-grit, and 50-microns-grit sand-blast), followed by sterilization with either ultraviolet light, ethylene oxide, argon plasma-cleaning, or routine clinical autoclaving. Osteocalcin and alkaline phosphatase, but not collagen expression, were significantly affected by surface roughness when these surfaces were altered by argon plasma-cleaning. In general, plasma-cleaned cpTi surfaces demonstrated an inverse relationship between surface roughness and phenotypic markers for a bone-like response. On a per-cell basis, levels of the bone-specific protein, osteocalcin, and the enzymatic activity of alkaline phosphatase were highest on the smooth 1-micron polished surface and lowest on the roughest surfaces for the plasma-cleaned cpTi. Detectable bone cell expression can be altered by clinically relevant surfaces prepared by standard dental implant preparation techniques.
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PMID:Bone cell expression on titanium surfaces is altered by sterilization treatments. 800 33

Surface topography plays a critical role in the interaction of dental implants with adjacent tissues. No statistical differences in oxide composition and surface contamination were observed between 600 grit and polished titanium surfaces. The expression of osteoblast phenotype was enhanced when osteoblast progenitor cells (2T9) were stimulated with bone morphogenetic protein-2 on polished and 600 grit titanium surfaces. Bone morphogenetic protein-2 stimulated phenotypic expression on 600 grit titanium surfaces was marked by prolonged alkaline phosphatase specific activity and more rapid osteocalcin production as compared with the polished titanium surfaces. Because the surface area of the 600 grit titanium surface was shown to be 8 percent greater than that of the polished titanium surface, it is possible that increased surface area played a role in the enhanced expression of the osteoblast phenotype.
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PMID:Surface roughness of titanium on bone morphogenetic protein-2 treated osteoblast cells in vitro. 920 1

Surface roughness affects proliferation, differentiation (alkaline phosphatase and osteocalcin), local factor production (transforming growth factor (TGF beta) and prostaglandin E2 (PGE2)], and response to 1,25-(OH)2D3 (1,25) of MG63 osteoblast-like cells. In this study, we examined whether the effect of surface roughness on MG63 cells is mediated by prostaglandins produced by the cells. Unalloyed titanium (Ti) disks were pretreated with HF/HNO3 (PT) and then machined and acid-etched (MA). Disks were also coarse grit-sandblasted (SB), coarse grit-sandblasted and acid-etched (CA), or plasma-sprayed with Ti particles (PS). The surfaces, from smoothest to roughest, were PT, MA, CA, SB, and PS. MG63 cells were cultured to confluence on the Ti disks in the presence or absence of 10(-7) M indomethacin (Indo), a specific inhibitor of cyclooxygenase activity, resulting in decreased prostaglandin production. When the cells reached confluence, cell number, cell layer alkaline phosphatase specific activity (ALPase), and osteocalcin (OC) and latent TGF beta (LTGF beta) production were determined. In addition, confluent cultures which had been grown in the absence of Indo were exposed to 10(-7) M 1,25, 10(-7) M Indo, or a combination of the two for 24 h. On the rougher surfaces, cell number was decreased and ALPase, OC, and LTGF beta were increased. When indomethacin was present throughout the culture period, the effect of surface roughness on cell number, OC, and LTGF beta was abolished. ALPase was reduced, but surface roughness-dependent effects were still observed. Addition of indomethacin to confluent cultures for 24 h had no effect on any of the parameters examined, with one exception: Cells cultured on MA surfaces exhibited a more differentiated phenotype. 1,25 increased all parameters examined on SB, CA, and PS surfaces. When indomethacin was added with 1,25, the 1,25-dependent effects on cell number and OC and LTGF beta production were abolished; however, ALPase was unaffected. This indicates that bone cell response to systemic hormones may be modified by implant surface roughness. This effect may be mediated, at least in part, by prostaglandins produced by the same cells.
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PMID:Prostaglandins mediate the effects of titanium surface roughness on MG63 osteoblast-like cells and alter cell responsiveness to 1 alpha,25-(OH)2D3. 965 20

When osteoblasts are cultured on surfaces of increasing microroughness, they exhibit decreases in proliferation, increases in differentiation and local factor production, and enhanced response to 1alpha,25(OH)(2)D(3). The cells interact with surfaces through integrins, which signal by the same pathways used by 1alpha,25(OH)(2)D(3), including protein kinase C via phospholipase C and protein kinase A via phospholipase A(2). This provides opportunities for crosstalk that may contribute to the synergistic effects of surface roughness and the vitamin D metabolite. Because these pathways converge at mitogen-activated protein kinase (MAPK), we tested the hypothesis that the extracellular signal-regulated kinase (ERK1/2) subclass of MAPKs mediates the effects of surface roughness and 1alpha,25(OH)(2)D(3). MG63 osteoblast-like osteosarcoma cells were cultured on commercially pure Ti disks with various surface roughnesses: pretreatment (PT; 0.6 microm average roughness [Ra]), coarse grit-blasted and acid-etched (SLA; 4 microm RA), and titanium plasma-sprayed (TPS; 5.2-microm R(a)). At confluence, cells were treated for 24 h with control media or media containing 10(-7) M 1alpha,25(OH)(2)D(3). One-half of the cultures received 1 microm or 10 microm PD98059, a specific inhibitor of the ERK family of MAPKs. PD98059 alone did not affect proliferation, osteocalcin production, or production of transforming growth factor-beta1 or nitric oxide, regardless of the surface roughness. Alkaline phosphatase was reduced by the inhibition of the ERK family kinases on all surfaces to a comparable extent. However, when PD98059 was added to the cultures with 1alpha,25(OH)(2)D(3), the effects of the seco-steroid were blocked, including the synergistic increases seen in MG63 cells cultured on SLA or TPS. These results indicate that ERK1/2 MAPK is required for the maintenance of alkaline phosphatase at control levels and that the effects of 1alpha,25(OH)(2)D(3) are mediated by ERK1/2. However, the effects of surface roughness are not due to the ERK family of MAPKs. This suggests that alternative pathways may be used, including those mediated by other MAPK subclasses.
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PMID:Osteoblast response to titanium surface roughness and 1alpha,25-(OH)(2)D(3) is mediated through the mitogen-activated protein kinase (MAPK) pathway. 1137 60

Osteoblasts exhibit a more differentiated morphology on surfaces with rough microtopographies. Surface effects are often mediated through integrins that bind the RGD motif in cell attachment proteins. Here, we tested the hypothesis that modulating access to RGD binding sites can modify the response of osteoblasts to surface microtopography. MG63 immature osteoblast-like cells were cultured on smooth (Ti sputter-coated Si wafers) and rough (grit blasted/acid etched) Ti surfaces that were modified with adsorbed monomolecular layers of a comb-like graft copolymer, poly-(L-lysine)-g-poly(ethylene glycol) (PLL-g-PEG), to limit nonspecific protein adsorption. PLL-g-PEG coatings were functionalized with varying amounts of an integrin-receptor-binding RGD peptide GCRGYGRGDSPG (PLL-g-PEG/PEG-RGD) or a nonbinding RDG control sequence GCRGYGRDGSPG (PLL-g-PEG/PEG-RDG). Response to PLL-g-PEG alone was compared with response to surfaces on which 2-18% of the polymer sidechains were functionalized with the RGD peptide or the RDG peptide. To examine RGD dose-response, peptide surface concentration was varied between 0 and 6.4 pmol/cm(2). In addition, cells were cultured on uncoated Ti or Ti coated with PLL-g-PEG or PLL-g-PEG/PEG-RGD at an RGD surface concentration of 0.7 pmol/cm(2), and free RGDS was added to the media to block integrin binding. Analyses were performed 24 h after cultures had achieved confluence on the tissue culture plastic surface. Cell number was reduced on smooth Ti compared to plastic or glass and further decreased on surfaces coated with PLL-g-PEG or PLL-g-PEG/PEG-RDG, but was restored to control levels when PLL-g-PEG/PEG-RGD was present. Alkaline phosphatase specific activity and osteocalcin levels were increased on PLL-g-PEG alone or PLL-g-PEG/PEG-RDG, but PLL-g-PEG/PEG-RGD reduced the parameters to control levels. On rough Ti surfaces, cell number was reduced to a greater extent than on smooth Ti. PLL-g-PEG coatings reduced alkaline phosphatase and increased osteocalcin in a manner that was synergistic with surface roughness. The RDG peptide did not alter the PLL-g-PEG effect but the RGD peptide restored these markers to their control levels. PLL-g-PEG coatings also increased TGF-beta1 and PGE(2) in conditioned media of cells cultured on smooth or rough Ti; there was a 20x increase on rough Ti coated with PLL-g-PEG. PLL-g-PEG effects were inhibited dose dependently by addition of the RGD peptide to the surface. Free RGDS did not decrease the effect elicited by PLL-g-PEG surfaces. These unexpected results suggest that PLL-g-PEG may have osteogenic properties, perhaps correlated with effects that alter cell attachment and spreading, and promote a more differentiated morphology.
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PMID:RGD-containing peptide GCRGYGRGDSPG reduces enhancement of osteoblast differentiation by poly(L-lysine)-graft-poly(ethylene glycol)-coated titanium surfaces. 1476 25

Osteoblasts are attachment-dependent cells that interact with their surface through integrin-mediated mechanisms. Their differentiation is regulated by 1,25-dihydroxyvitamin D3 [1alpha,25(OH)(2)D(3)] and is affected by substrate chemistry and microtopography, suggesting that 1alpha,25(OH)(2)D(3) may regulate integrin expression in a surface-specific manner. To test this hypothesis, osteoblast-like human MG63 cells were grown on tissue culture plastic and on grit-blasted and acid-etched titanium disks with a complex microtopography to induce osteoblast differentiation. Expression of alpha(2), alpha(5), alpha(v), beta(1), and beta(3) integrins were quantified by real-time polymerase chain reaction (PCR) as a function of time in culture and treatment with 1alpha,25(OH)(2)D(3). Results were correlated with expression of osteocalcin, a marker of a differentiated osteoblast. Osteocalcin mRNA increased with time and 1alpha,25(OH)(2)D(3) treatment and these changes were greater in cultures on the titanium disks. Integrin expression varied with time in culture and this was also surface dependent. At each time point, beta(1) and alpha(2) mRNAs were greater on titanium than on plastic, whereas alpha(5) expression was reduced and alpha(v),beta(3) expression was unaffected. 1alpha,25(OH)(2)D(3) increased beta(1) mRNA on both surfaces at all time points, but it increased alpha(2) expression only in 8-d cultures. 1alpha,25(OH)(2)D(3) caused reduced alpha(5) expression only in cultures grown on plastic for 8 d, and had no effect on either alpha(v) or beta(3) expression regardless of surface. These results show that integrin expression in human osteoblast-like cells is differentially modulated by 1alpha,25(OH)(2)D(3) in a time-dependent manner that is sensitive to the surface on which the cells are grown.
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PMID:1alpha,25(OH)2D3 regulation of integrin expression is substrate dependent. 1538 91

Microtextured titanium implant surfaces enhance bone formation in vivo and osteoblast phenotypic expression in vitro, but the mechanisms are not understood. To determine the roles of specific microarchitectural features in modulating osteoblast behavior, we used Ti surfaces prepared by electrochemical micromachining as substrates for MG63 osteoblast-like cell culture. Cell response was compared to tissue culture plastic, a sand-blasted with large grit and acid-etched surface with defined mixed microtopography (SLA), polished Ti surfaces, and polished surfaces electrochemically machined through a photoresist pattern to produce cavities with 100, 30 and 10 microm diameters arranged so that the ratio of the microscopic-scale area of the cavities versus the microscopic-scale area of the flat region between the cavities was equal to 1 or 6. Microstructured disks were acid-etched, producing overall sub-micron-scale roughness (Ra=0.7 microm). Cell number, differentiation (alkaline phosphatase; osteocalcin) and local factor levels (TGF-beta1; PGE(2)) varied with microarchitecture. 100 microm cavities favored osteoblast attachment and growth, the sub-micron-scale etch enhanced differentiation and TGF-beta1 production, whereas PGE(2) depended on cavity dimensions but not the sub-micron-scale roughness.
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PMID:Differential regulation of osteoblasts by substrate microstructural features. 1557 58

Increasing bone formation at endosseous titanium implants may be achieved by modification of topographically enhanced surfaces. The aim of this study was to determine the effect of fluoride ion modification of TiO2 grit-blasted, c.p. titanium implants on osteoblastic differentiation and interfacial bone formation by parallel in vitro and in vivo investigations. Human mesenchymal stem cells (Osiris Therapeutics, Inc.) were cultured on TiO2 grit-blasted c.p.titanium disks with and without fluoride ion modification. Cell adhesion, proliferation, and osteoblastic gene expression was measured by scanning electron microscopy, tritiated-thymidine uptake into insoluble DNA, and reverse transcription polymerase chain reaction detection of mRNAs encoding collagen 1, osteopontin, bone sialoprotein, osteocalcin and BMP-2. After 24 h, there were no differences in cell adhesion among the surfaces tested. Fluoride-treated surfaces supported greater proliferation and increased bone sialoprotein and BMP-2 expression. Additionally, 12 TiO2 grit-blasted and 12 fluoride ion modified implants were placed randomly into medial and distal osteotomies prepared in the tibia of 300 g Sprague Dawley rats. After 21 days, the tibiae were harvested and 100 microm ground sections were examined by backscatter scanning electron microscopy. The bone-to-implant contact formed at TiO2 grit-blasted and fluoride-treated versus TiO2 grit-blasted surfaces was 55.45% versus 34.21% (p<0.027), respectively. Fluoride ion modification of the TiO2 grit-blasted surface enhanced osteoblastic differentiation in vitro and interfacial bone formation in vivo. This parallel in vitro and in vivo investigation demonstrates that fluoride ion modification enhanced osteoblastic differentiation and interfacial bone formation. The mechanism(s) by which fluoride ion modification of c.p.titanium enhanced osteoblastic differentiation and osseointegration merit careful investigation.
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PMID:Fluoride modification effects on osteoblast behavior and bone formation at TiO2 grit-blasted c.p. titanium endosseous implants. 1611 91

The aim of this study was to investigate the influence of hydrophobic acid-etched (A) and coarse-blasted large-grit and acid-etched (SLA) surfaces as well as hydrophilic modified acid-etched (modA) and modified coarse-blasted large-grit and acid-etched (modSLA) surfaces on the behavior of MG63 cells grown on these surfaces through determination of cell attachment and cell proliferation, time-lapse microscopy of fluorescence-labeled cells, and determination of gene expression by reverse transcription-polymerase chain reaction (RT-PCR). No significant difference of cell attachment on various titanium surfaces was found. Increased cell proliferation was observed on the A surface and the SLA surface compared with the modA surface and the modSLA surface. After 2 days of incubation, on modSLA and modA surfaces a tendency of formation of cell clusters has been observed, which was most pronounced on modSLA surface. On the A and the SLA surface, cell cluster formation started after longer incubation periods. The expression level of the bone-associated genes (alkaline phosphatase, osteocalcin, type-I-collagen, osteoprotegerin, and glyceraldehyde-3-phosphate-dehydrogenase) detected by RT-PCR was highest on the modSLA surface. In conclusion it has been demonstrated that the modSLA surface results in an enhanced cluster formation of osteoblasts grown on this surface and in an increased expression of key osteogenic regulatory genes in osteoblasts.
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PMID:The initial attachment and subsequent behavior regulation of osteoblasts by dental implant surface modification. 1732 17

The objective of this study was to characterize the physicochemical, dissolution, and osteogenic properties of radio frequency magnetron sputtered dicalcium pyrophosphate/tricalciumphosphate (Pyro/TCP) and hydroxylapatite (HA) coatings. Therefore Pyro/TCP and HA coatings were deposited on grit-blasted titanium discs. The results showed that the deposited coatings were amorphous and changed into a crystalline structure after IR heat-treatment of 550 degrees C for HA and 650 degrees C for Pyro/TCP. Heat-treated HA coatings appeared to be stable during immersion in simulated body fluid (SBF), that is no changes in the XRD pattern were observed. Also, no dissolution of the coating was observed by scanning electron microscopy (SEM). Energy dispersive spectroscopy (EDS) revealed that the Ca/P ratio of the HA coatings remained constant during SBF immersion. On the other hand, the heat-treated Pyro/TCP coatings showed a surface reaction of calcium pyrophosphate into a beta-tricalcium phosphate phase during SBF immersion. This was confirmed by EDS analysis. Rat bone marrow-derived osteoblast-like cells cultured on the heat-treated substrates showed that cell proliferation and differentiation occurred on both types of bioceramic coatings. No significant differences for proliferation and early differentiation were observed between cells cultured on heat-treated Pyro/TCP and HA at individual time points. However, osteocalcin expression, a late marker for osteoblast-like cell differentiation, was significantly increased after 12 days of culture on HA-coatings. These results were confirmed by SEM observations and suggest increased osteogenic properties for HA-coatings over Pyro/TCP-coatings. Additional research is necessary to obtain conclusive evidence on the in vivo osteogenic capacity of Pyro/TCP coatings.
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PMID:Characterization and in vitro evaluation of biphasic calcium pyrophosphate-tricalciumphosphate radio frequency magnetron sputter coatings. 1763 19


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