共查询到20条相似文献,搜索用时 15 毫秒
1.
Once thought to provide only structural support to tissues by acting as a scaffold to which cells bind, it is now widely recognized that the extracellular matrix (ECM) provides instructive signals that dictate cell behavior. Recently we demonstrated that mechanical cues intrinsic to the ECM directly regulate the behavior of pre-osteoblastic MC3T3-E1 cells. We hypothesized that one possible mechanism by which ECM compliance exerts its influence on osteogenesis is by modulating the mitogen-activated protein kinase (MAPK) pathway. To address this hypothesis, the differentiation of MC3T3-E1 cells cultured on poly(ethylene glycol) (PEG)-based model substrates with tunable mechanical properties was assessed. Alkaline phosphatase (ALP) levels at days 7 and 14 were found to be significantly higher in cells grown on stiffer substrates (423.9 kPa hydrogels and rigid tissue culture polystyrene (TCPS) control) than on a soft hydrogel (13.7 kPa). Osteocalcin (OCN) and bone sialoprotein (BSP) gene expression levels followed a similar trend. In parallel, MAPK activity was significantly higher in cells cultured on stiffer substrates at both time points. Inhibiting this activation pharmacologically, using PD98059, resulted in significantly lower ALP levels, OCN, and BSP gene expression levels on the hydrogels. Interestingly, the effectiveness of PD98059 was itself dependent on substrate stiffness, with marked inhibition of MAPK phosphorylation in cells grown on compliant hydrogels but insignificant reduction in cells grown on TCPS. Together, these data confirm a role for MAPK in the regulation of osteogenic differentiation by ECM compliance. 相似文献
2.
Bone cells in vivo exist in direct contact with extracellular matrix, which regulates their basic biological processes including metabolism, development, growth and differentiation. Thus, the in vitro activity of cells cultured on tissue culture treated plastic could be different from the activity of cells cultured on their natural substrate. We selected MC3T3-E1 pre-osteoblastic cells to study the effect of extracellular matrix on cell proliferation because these cells undergo a progressive developmental sequence of proliferation and differentiation. MC3T3-E1 cells were cultured on plastic or plastic coated with ECM, fibronectin, collagen type I, BSA or poly l-lysine and their ability to proliferate was assessed by incorporation of [3H]dT or by enumeration of cells. Our results show that (1) ECM inhibits incorporation of [3H]dT by MC3T3-E1 cells; (2) collagen type I, but not BSA, poly l-lysine or fibronectin also inhibits incorporation of [3H]dT; (3) the level of ECM inhibition of [3H]dT incorporation is directly related to the number of cells cultured, but unrelated to the cell cycle distribution or endogenous thymidine content; (4) the kinetic profile of [3H]dT uptake suggest that ECM inhibits transport of [3H]dT from the extracellular medium, and (5) cell counts are similar in cultures whether cells are grown on plastic or ECM. These results suggest that decreased incorporation of [3H]dT by cells cultured on ECM is not reflective of bone cell proliferation. 相似文献
3.
Roman Thaler Heidrun Karlic Silvia Spitzer Klaus Klaushofer Franz Varga 《Apoptosis : an international journal on programmed cell death》2010,15(6):728-737
The extracellular matrix (ECM) of bone consists mainly of collagen type I, which induces osteoblastic differentiation and
prevents apoptosis. Fas induces apoptosis in cells improperly adhering to ECM. Recently, it was described that Fas expression
is modulated by epigenetic DNA methylation. Mouse MC3T3-E1 pre-osteoblastic cells were cultured either on collagen coated
or on uncoated culture dishes for control. mRNA was isolated and gene expression was analyzed by quantitative RT–PCR. Furthermore,
we measured global and specific DNA methylation. Compared to controls, cells cultured on collagen-coated dishes increased
the expression of Runx2 and OCN indicating differentiation of pre-osteoblastic cells. Additionally, collagen up-regulated
cyclin-A2 and down-regulated Fas expression suggesting increased cell multiplication. Furthermore, the expression of Dnmt1
and Hells, key mediators of the DNA-methylation process, was increased. As a consequence, we demonstrate that global DNA methylation
and specific methylation of the Fas promoter was higher in MC3T3-E1 cells cultured on collagen when compared to controls.
Investigation of signal transduction pathways by mean of inhibitors suggests that focal adhesion kinase, MAP- and Jun-kinases
and AP-1 are involved in this process. In summary, we demonstrate that ECM prevents activation of Fas by epigenetic DNA-methylation. 相似文献
4.
Involvement of ligand occupancy in Insulin-like growth factor-I (IGF-I) induced cell growth in osteoblast like MC3T3-E1 cells 总被引:1,自引:0,他引:1
Growth factors and matrix proteins regulate the proliferation and differentiation of osteoblasts. The insulin-like growth factor (IGF) system comprises IGF-I, IGF-II, and six high-affinity IGF-binding proteins (IGFBPs). IGFs stimulate cell growth in many types of tissue; IGF-binding proteins regulate cellular actions and can affect cell growth. IGF-I is involved in differentiation, proliferation, and matrix formation in osteoblasts; IGFBP-5 is associated with the extracellular matrix (ECM) and can potentiate the actions of IGF-I. We investigated the effect of ECM proteins on the responses of MC3T3-E1 osteoblast cells to IGF-I and IGFBP-5. In addition, because extracellular signal-regulated kinases 1 and 2 (Erk 1/2) affect cell growth, we evaluated the effects of IGFBP-5 on Erk 1/2 phosphorylation in MC3T3-E1 cells. IGF-I caused an increase in IGFBP-5 expression in cultured MC3T3-E1 cells, and IGF-I plus IGFBP-5 significantly increased cell growth. Likewise, the addition of IGF-I and IGFBP-5 to cultured MC3T3-E1 cells increased the synthesis of the ECM proteins osteopontin (OPN) and thrombospondin-1 (TSP-1), which can bind to alphaVbeta3 integrin receptors on the cell surface. By contrast, the addition of an antibody against ECM proteins inhibited the effects of OPN and TSP-1 on IGFBP-5 expression. The stimulatory effect of IGFBP-5 was mediated via Erk 1/2 activation. These data suggest that IGFBP-5 regulates Erk 1/2 phosphorylation in cultured MC3T3-E1 cells via ECM proteins that may ultimately stimulate the growth of osteoblasts. We determined whether occupation of the alphaVbeta3 integrin receptor affects IGF-I receptor (IGF-IR)-mediated signaling and function in MC3T3-E1 osteoblast cells. Occupation of the alphaVbeta3 integrin receptor with ECM proteins induced IGF-I-stimulated IGF-IR phosphorylation. Conversely, in the presence of the alphaVbeta3-specific disintegrin echistatin, IGF-I-stimulated IGF-IR activation was inhibited. IGF-I-stimulated IGF-IR phosphorylation was accompanied by IRS-1 phosphorylation and MAPK activation. However, these effects were attenuated by echistatin. Thus, occupancy of the alphaVbeta3 disintegrin receptor modulates IGF-I-induced IGF-IR activation and IGF-IR-mediated function in MC 3T3-E1 osteoblasts. 相似文献
5.
Dun Hong Hai-Xiao Chen Hai-Qiang Yu Carrie Wang Hai-Teng Deng Ren-Shan Ge 《Experimental cell research》2010,316(14):2291-1051
Bone remodeling relies on a dynamic balance between bone formation and resorption, mediated by osteoblasts and osteoclasts, respectively. Under certain stimuli, osteoprogenitor cells may differentiate into premature osteoblasts and further into mature osteoblasts. This process is marked by increased alkaline phosphatase (ALP) activity and mineralized nodule formation. In this study, we induced osteoblast differentiation in mouse osteoprogenitor MC3T3-E1 cells and divided the process into three stages. In the first stage (day 3), the MC3T3-E1 cell under osteoblast differentiation did not express ALP or deposit a mineralized nodule. In the second stage, the MC3T3-E1 cell expressed ALP but did not form a mineralized nodule. In the third stage, the MC3T3-E1 cell had ALP activity and formed mineralized nodules. In the present study, we focused on morphological and proteomic changes of MC3T3-E1 cells in the early stage of osteoblast differentiation — a period when premature osteoblasts transform into mature osteoblasts. We found that mean cell area and mean stress fiber density were increased in this stage due to enhanced cell spreading and decreased cell proliferation. We further analyzed the proteins in the signaling pathway of regulation of the cytoskeleton using a proteomic approach and found upregulation of IQGAP1, gelsolin, moesin, radixin, and Cfl1. After analyzing the focal adhesion signaling pathway, we found the upregulation of FLNA, LAMA1, LAMA5, COL1A1, COL3A1, COL4A6, and COL5A2 as well as the downregulation of COL4A1, COL4A2, and COL4A4. In conclusion, the signaling pathway of regulation of the cytoskeleton and focal adhesion play critical roles in regulating cell spreading and actin skeleton formation in the early stage of osteoblast differentiation. 相似文献
6.
Claudia Tanja Mierke 《Cell biochemistry and biophysics》2009,53(3):115-126
Vinculin couples as a focal adhesion protein the extracellular matrix (ECM) through integrins to the actomyosin cytoskeleton.
During the last years vinculin has become the focus of cell mechanical measurements and a key protein regulating the transmission
of contractile forces. In earlier reports vinculin has been described as an inhibitor of cell migration on planar substrates,
because knock-out of vinculin in F9 mouse embryonic carcinoma cells and mouse embryonic fibroblasts showed increased cell
motility on 2D substrates. The role of vinculin in cell invasion through a 3D extracellular matrix is still fragmentarily
investigated. This review presents vinculin in its role as a regulator of cellular mechanical functions. Contractile force
generation is reduced when vinculin is absent, or enhanced when vinculin is present. Moreover, the generation of contractile
forces is a prerequisite for cell invasion through a dense 3D ECM, where the pore-size is smaller than the diameter of the
cell nucleus (<2 μm). Measurements of cell’s biophysical properties will be presented. In summary, vinculin’s leading role
among focal adhesion proteins in regulating the mechanical properties of cells will be discussed. 相似文献
7.
D. V. Shtansky I. Yu. Zhitnyak I. A. Bashkova Yu. S. Pogozhev A. N. Sheveiko N. A. Gloushankova 《Biochemistry (Moscow) Supplemental Series A: Membrane and Cell Biology》2010,4(3):272-276
The influence of elemental composition and surface topography of substrates on the adhesion, proliferation and early differentiation
stages of mouse osteoblasts, line MC3T3-E1, cultured on the surface of titanium and multicomponent bioactive nanostructured
films (MuBiNaFs) Ti-Ca-(P)-C-O-N and Ti-Ca-Si-C-O-N has been studied. Osteoblasts spread both on the surface of uncoated titanium
samples and on the samples coated with the films and had more elongated and irregular shape than the cells cultured on control
glass substrate. Immunofluorescence study has shown that osteoblasts formed straight actin bundles associated with focal contacts.
Osteoblasts on titanium films formed fewer focal contacts than the cells cultured on Ti-Ca-(P)-C-O-N films. The sandblast
treatment did not affect the distribution of the focal contacts and actin structures within the cells. Osteoblasts actively
divided on the surface of Ti-Ca-(P)-C-O-N and Ti-Ca-Si-C-O-N films and alteration of surface topography did not affect their
growth rate. The quantitative colorimetrical test on alkaline phosphatase activity has shown that modification of surface
topography does not affect the level of osteoblastic differentiation. Thus, the elemental composition plays an important role
in the interaction of osteoblasts with the surface of films, while the modification of the surface topography in the range
of S
q from 0.4 to 1 μm does not influence adhesion, proliferation, and differentiation of osteoblasts. 相似文献
8.
9.
Breast cancer frequently metastasizes to bone, resulting in osteolytic lesions. These lesions, formed by activated osteoclasts, cause pain, an increased susceptibility to fractures, and hypercalcemia. It has been shown that breast cancer cells communicate with osteoblasts and subsequently stimulate osteoclast activity; however, little research has focused on understanding the interaction between breast cancer cells and osteoblasts. We recently reported that conditioned medium from MDA-MB-231 breast cancer cells inhibited the differentiation of MC3T3-E1 osteoblasts through the secretion of transforming growth factor beta (TGFbeta). In addition, the breast cancer conditioned medium altered MC3T3-E1 morphology, the pattern of actin stress fibers, and reduced focal adhesion plaques. In the current study, we identified the mechanism used by MDA-MB-231 cells to cause these effects. When MC3T3-E1 osteoblasts were cultured with MDA-MB-231 conditioned medium preincubated with neutralizing antibodies to platelet derived growth factor (PDGF), insulin-like growth factorII (IGFII), and TGFbeta, focal adhesion plaques and actin stress fiber formation were restored. These cytokines were further found to signal through PI3Kinase and Rac. In conclusion, TGFbeta, PDGF, and IGFII might be good therapeutic targets for treating breast cancer-induced osteolytic lesions. 相似文献
10.
Mingzhi Luo Peili Yu Yang Jin Zhili Qian Yue Wang Jingjing Li Peng Shang Linhong Deng 《Molecular & cellular biomechanics : MCB》2016,13(2):137-157
Random positioning machine (RPM) and diamagnetic levitation are two essential ground-based methods used to stimulate the effect of microgravity in space life science research. However, the force fields generated by these two methods are fundamentally different, as RPM generates a dynamic force field acting on the surface in contact with supporting substrate, whereas diamagnetic levitation generates a static force field acting on the whole body volume of the object (e.g. cell). Surprisingly, it is hardly studied whether these two fundamentally different force fields would cause different responses in mammalian cells. Thus we exposed cultured MC3T3-E1 osteoblasts to either dynamically stimulated effect of microgravity (d-µg) with RPM or statically stimulated effect of microgravity (s-µg) with diamagnetic levitation, respectively, for 3 h. Subsequently, the cells were examined for changes in cell morphology, cytoskeleton (CSK) structure and Ca2+ signaling. The results show that compared to the condition of normal gravity (1g), both d-µg and s-µg resulted in decrease of cell area and disruption of the microfilaments and microtubules in MC3T3-E1 cells, but cells under d-µg were more smooth and round while those under s-µg exhibited more protrusions. The decrease of cell area and disruption of microfilaments and microtubules induced by d-µg but not s-µg were rescued by inhibition of the stretch-activated channel by gadolinium chloride (Gd). Inhibition of calmodulin (CaM) by inhibitor, W-7, promoted the effects of s-µg on cell area and CSK filaments, but inhibition of calmodulin-dependent protein kinase (CaMK) by inhibitor, KN-93, weakened d-µg-induced effects on cell area and cytoskeleton. In addition, both d-µg and s-µg decreased the CaM expression and CaMKⅡ activity in MC3T3-E1 cells. Furthermore, s-µg resulted in decrease of the intracellular free Ca2+ concentration ([Ca2+]i) in MC3T3-E1 cells, which was reversed by disrupting microfilaments with cytochalasin B (CytB). Instead, d-µg induced increase of [Ca2+]i, which was inhibited by Gd. Taken together these data suggest that dynamic and static stimulated microgravity cause different responses in MC3T3-E1 cells. The dynamic force field acts on stretch-activated channels to induce microfilaments disruption and Ca2+ influx in MC3T3-E1 cells whereas the static force field directly induces microfilament disruption, which in turn decreases the [Ca2+]i in MC3T3-E1 cells. Such findings may have important implications to better understanding microgravity related cellular events and their applications. 相似文献
11.
目的:研究高浓度葡萄糖抑制MC3T3-E1细胞成骨分化的机理。方法:建立MC3T3-E1细胞成骨分化诱导体系,观察不同浓度葡萄糖(5.5mM和22mM)对MC3T3-E1细胞成骨分化的影响;用不同浓度的p38 MAPK抑制剂Fr167653(0.1μM、1.0μM和10μM)进行药物干预,观察MC3T3-E1细胞在22mM葡萄糖浓度下成骨分化的变化情况。通过钙含量检测、Real time PCR检测相关分化的变化;用Western Blot方法检测MC3T3-E1细胞分化过程中p38 MAPK磷酸化状态、TXNIP表达水平的变化;使用胰岛素二硫键还原法检测细胞内TRX活性水平;使用活性氧检测试剂盒检测细胞内自由氧生成水平。结果:体外诱导条件下,高浓度(22mM)葡萄糖通过升高p38 MAPK磷酸化水平,上调TXNIP表达水平,同时降低TRX活性,使细胞内自由氧生成增加,抑制MC3T3-E1细胞的成骨分化;Fr167653通过抑制p38 MAPK磷酸化,下调TXNIP表达同时升高TRX活性,抑制细胞内自由氧生成,解除高浓度葡萄糖对细胞成骨分化的抑制作用。结论:高浓度葡萄糖通过p38 MAPK-TXNIP/TRX-ROS信号通路抑制MC3T3-E1细胞成骨分化。 相似文献
12.
To investigate the bioactivities of soybean, which act on bone metabolism, we studied the effect of a soybean ethanol extract on the activity of osteoblast MC3T3-E1 cells. Soy extract (0.01-0.1 g/l) dose-dependently increased survival (P<0.05) and DNA synthesis (P<0.05) of MC3T3-E1 cells. In addition, soy extract (0.05 g/l) increased alkaline phosphatase activity (P<0.05) and collagen synthesis (P<0.05) of MC3T3-E1 cells. Moreover, the anti-estrogen tamoxifen eliminated the stimulation of MC3T3-E1 cells on the proliferation, ALP activity and collagen synthesis by soy extract, indicating that the main action of the soy extract on osteoblastic MC3T3-E1 cells is similar to that of estrogen effects. Treatment with soy extract prevented apoptosis, as assessed by a one-step sandwich immunoassay and DNA gel electrophoresis studies. This effect may be associated with the activation of the estrogen receptor, since we observed soy extract-mediated survival against apoptosis was blocked by the estrogen receptor antagonist tamoxifen in cells, further supporting a receptor-mediated mechanism of cell survival. These results suggest that osteoblast function is promoted by soy extract and that the estrogen receptor is involved in the response, thereby playing an important role in bone remodeling. In conclusion, soy extract has a direct stimulatory effect on bone formation in cultured osteoblastic cell in vitro. Presumably, dietary soy products are useful in the prevention of osteoporosis. 相似文献
13.
Osteoblastic phenotype expression of MC3T3-E1 cultured on electrospun polycaprolactone fiber mats filled with hydroxyapatite nanoparticles 总被引:4,自引:0,他引:4
Electrospun (e-spun) fiber mats of polycaprolactone (PCL; Mn = 80 000 g mol-1) with or without the presence of hydroxyapatite (HAp) nanoparticles (at 1% w/v based on the volume of the PCL solution) were successfully fabricated. The potential for use of these e-spun fiber mats as bone scaffolds was assessed by mouse calvaria-derived pre-osteoblastic cells, MC3T3-E1, in terms of attachment, proliferation, differentiation, and mineralization. Despite the lower number of cells attached at early time points, both the fibrous scaffolds supported the proliferation of MC3T3-E1 at similar levels to tissue-culture polystyrene plate (TCPS), with the cells growing on the PCL/HAp fiber mat (i.e., PCL/HAp-FS) showing the greatest proliferation rate on day 3 after the initial attachment period of 16 h. Alkaline phosphatase (ALP) activity of the cells grown on TCPS was the greatest on day 3 after cell culturing, while that of the cells grown on PCL/HAp-FS reached a maximum on day 5. On the other hand, the ALP activity of the cells grown on the neat PCL fiber mat (i.e., PCL-FS) was the lowest at any given time point. MC3T3-E1 cultured on the surface of PCL/HAp-FS expressed the greatest amount of osteocalcin (OC) gene on day 14 after cell culturing and OC protein on day 21 after cell culturing, respectively, when compared with those cultured on the surfaces of PCL-FS and TCPS. This corresponded to the greatest extent of mineralization for the cells grown on the surface of PCL/HAp-FS on day 21, followed by that for the cells grown on PCL-FS and TCPS, respectively. 相似文献
14.
Koizumi T Hikiji H Shin WS Takato T Fukuda S Abe T Koshikiya N Iwasawa K Toyo-oka T 《The Journal of biological chemistry》2003,278(8):6433-6439
A two-dimensional intracellular Ca(2+) ([Ca(2+)](i)) imaging system was used to examine the relationship between [Ca(2+)](i) handling and the proliferation of MC3T3-E1 osteoblast-like cells. The resting [Ca(2+)](i) level in densely cultured cells was 1.5 times higher than the [Ca(2+)](i) level in sparsely cultured cells or in other cell types (mouse fibroblasts, rat vascular smooth muscle cells, and bovine endothelial cells). A high resting [Ca(2+)](i) level may be specific for MC3T3-E1 cells. MC3T3-E1 cells were stimulated with ATP (10 microM), caffeine (10 mM), thapsigargin (1 microM), or ionomycin (10 microM), and the effect on the [Ca(2+)](i) level of MC3T3-E1 cells was studied. The percentage of responding cells and the degree of [Ca(2+)](i) elevation were high in the sparsely cultured cells and low in densely cultured cells. The rank order for the percentage of responding cells and magnitude of the Ca(2+) response to the stimuli was ionomycin > thapsigargin = ATP > caffeine and suggests the existence of differences among the various [Ca(2+)](i) channels. All Ca(2+) responses in the sparsely cultured MC3T3-E1 cells, unlike in other cell types, disappeared after the cells reached confluence. Heptanol treatment of densely cultured cells restored the Ca(2+) response, suggesting that cell-cell contact is involved with the confluence-dependent disappearance of the Ca(2+) response. Immunohistological analysis of type 1 inositol trisphosphate receptors and electron microscopy showed distinct expression of inositol trisphosphate receptor proteins and smooth-surfaced endoplasmic reticulum in sparsely cultured cells but reduced levels in densely cultured cells. These results indicate that the underlying basis of confluence-dependent [Ca(2+)](i) regulation is down-regulation of smooth-surfaced endoplasmic reticulum by cell-cell contacts. 相似文献
15.
细胞外基对组织细胞起支持、保护、营养作用,对细胞的增殖、分化有重要影响,在细胞和组织工程中,应该充分考虑细胞外基质的作用。本研究首先脱去培养板中融合培养的原代小鼠心肌成纤维细胞和成骨细胞,获得两种体外形成的细胞外基质包被的培养板,其中成骨细胞细胞外基质中含有骨形成蛋白2。然后将MC3T3-E1成骨前体细胞接种在这种培养板中,发现成纤维细胞胞外基质包被的培养板中的细胞增殖活性最高,而成骨细胞胞外基质包被的培养板中细胞的碱性磷酸酶活性、骨形成蛋白2和骨桥蛋白的相对蛋白表达量最高,细胞外钙沉积量比其他组高1倍左右。结果表明:包被在培养板上的这两种细胞外基质有不同的生物活性,成纤维细胞胞外基质可促进成骨前体细胞增殖,成骨细胞胞外基质可促进成骨前体细胞骨向分化。 相似文献
16.
YASUYOSHI TORII KIYOTAKA HITOMI YASUKO YAMAGISHI NORIHIRO TSUKAGOSHI 《Cell biology international》1996,20(7):459-464
MC3T3-E1 cells grown with ascorbic acid express sequentially osteoblastic marker proteins such as alkaline phosphatase (ALPase) and then form a mineralized extracellular matrix (ECM) as a consequence of osteoblastic differentiation. To explore the functional roles of ALPase in the process of osteoblastic maturation, an inducible expression vector for antisense ALPase RNA was constructed and stably transfected into MC3T3-E1 cells. The expression of antisense ALPase RNA in the differentiated MC3T3-E1 transfectants reduced markedly the ALPase activity, which resulted in a significant decrease in the deposition of minerals upon prolonged culture. These findings demonstrated directly that ALPase participated in the mineralizationof ECM. 相似文献
17.
Alignment and proliferation of MC3T3-E1 osteoblasts in microgrooved silicone substrata subjected to cyclic stretching 总被引:1,自引:0,他引:1
Previous studies have shown that many types of cells align in microgrooves in static cultures. However, whether cells remain aligned and also proliferate in microgrooves under stretching conditions has not been determined. We grew MC3T3-E1 osteoblasts in deformable silicone dishes containing microgrooves oriented in the stretch direction. We found that with or without 4% stretching, cells aligned in microgrooves of all sizes, with the groove and ridge widths ranged from 1 to 6microm, but the same groove depth of about 1.6microm. In addition, actin cytoskeleton and nuclei became highly aligned in the microgrooves with and without 4% cyclic stretching. To further examine whether MC3T3-E1 osteoblasts proliferate in microgrooves with cyclic stretching, we grew the cells in six-well silicone dishes containing microgrooves in three wells and smooth surfaces in other three wells. After 4% cyclic stretching for 3, 4, and 7 days, we found that cell numbers in the microgrooves were not significantly different (p>0.05) from those on the smooth surface (p>0.05). Taken together, these results show that MC3T3-E1 osteoblasts can align and proliferate in microgrooves with 4% cyclic stretching. We suggest that the silicone microgrooves can be a useful tool to study the phenotype of MC3T3-E1 osteoblasts under controlled substrate strains. The silicone microgrooves can also be useful for delivering defined substrate strains to other adherent cells in cultures. 相似文献
18.
The GRGDS (Gly-Arg-Gly-Asp-Ser) peptide has intermediate affinity to alphaVbeta3 and alphaIIbbeta3, which are the integrins most reported to be involved in bone function. In this study, biomimetic chitosan films modified with GRGDS peptide were prepared and were used as a substrate for the in vitro culture of MC3T3-E1 cells in order to investigate the effect of GRGDS modification on MC3T3-E1 cell behavior. The results of electron spectroscopy for chemical analysis (ESCA), attenuated total reflection-Fourier transform infrared spectra (ATR-FTIR), and amino acid analysis (AAA) demonstrated that the chitosan films were successfully modified with GRGDS peptides and that the surface density of the immobilized GRGDS was on the order of 10(-9) mol/cm2. The immobilization of the GRGDS sequence on chitosan as well as the peptide concentration play a significant role in MC3T3-E1 cell behavior. MC3T3-E1 cell attachment, proliferation, migration, differentiation, and mineralization were remarkably greater on GRGDS-coupled chitosan than on unmodified chitosan. Besides, the degree of acceleration of these biological processes was found to be dependent on peptide density. Competitive inhibition of MC3T3-E1 cell attachment using soluble GRGDS peptides indicated that the interaction of MC3T3-E1 cells with the surface of the materials was ligand-specific. Cytoskeleton organization in the fully spread MC3T3-E1 cells was highly obvious on GRGDS-coupled chitosan when compared to the lack of actin fibers noted in the round MC3T3-E1 cells on unmodified chitosan. These results suggest that MC3T3-E1 cell function can be modulated, in a peptide density-dependent manner, by the immobilization of GRGDS peptide on chitosan used for scaffold-based bone tissue engineering. 相似文献
19.
MC3T3-E1 cells demonstrate a lag in osteogenic development when seeded onto Poly(beta-hydroxybutyrate-co-beta-hydroxyvalerate) (PHBV), a biomaterial with substantial potential for bone tissue repair. To determine if this was due to the priority of extracellular matrix (ECM) remodelling over other developmental processes, gene expression levels of proteins involved in the production, maintenance and turnover of the ECM were compared between cells grown on PHBV and tissue culture plastic (TCP) 24 h after seeding. When grown on PHBV, MC3T3-E1 cells up-regulated proteins such as the matrix metalloproteinases and down-regulated the expression of proteins such as collagens that are involved in cell-substrate interactions, but in later-stage processes. The results also suggest that proteins such as fibronectin and aggrecan, and particularly osteopontin, may be more suitable candidates for PHBV functionalization for optimal MC3T3-E1 cell growth than proteins like osteonectin, periostin, vitronectin or collagen. This study confirms the importance of understanding the specific response of therapeutically-relevant cells, such as human stem cells, to candidate biomaterial surfaces in order to achieve optimal regenerative therapies. 相似文献
20.
Diabetic bone disease is associated with increased oxidative damage and 2-deoxy-d-ribose (dRib) is used to induce oxidative damage similar to that observed in diabetics. To determine if hesperetin (3′,5,7-trihydroxy-4-methoxyflavanone)
could influence osteoblast dysfunction induced by dRib, osteoblastic MC3T3-E1 cells were treated with dRib and hesperetin.
Then, markers of osteoblast function and oxidative damage were examined. Hesperetin (10−7–10−5 M) caused a significant elevation of alkaline phosphatase (ALP) activity, collagen content, and total antioxidant potential
of MC3T3-E1 cells in the presence of 20 mM dRib (p < 0.05). Moreover, hesperetin (10−7 M) decreased cellular protein carbonyl (PCO), advanced oxidation protein products (AOPP), and malondialdehyde (MDA) contents
of osteoblastic MC3T3-E1 cells in the presence of 20 mM dRib. These results demonstrate that hesperetin attenuates dRib-induced
damage, suggesting that hesperetin may be a useful dietary supplement for minimizing oxidative injury in diabetes related
bone diseases. 相似文献