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 47,337 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Effect of surface topography upon cell-adhesion, -orientation and -differentiation was investigated by in vitro study on cellular responses to titanium substratum with different surface roughness. Cell-shape, -function and -differentiation depending upon the surface topography were clarified by use of bone formative group cells (BFGCs) derived from bone marrow of beagle's femur. BFGCs consisted of hematopoietic stem cells (HSC) and osteogenetic stem cells (OSC). Cell differentiation of BFGCs was expressed and promoted by structural changes of cytoskeleton, and cell-organella, which was caused by mechanical stress with cytoplasmic stretching of cell adhesions to the substratum. Phagocytic monocytes of HSC differentiated to osteomediator cells (OMC) by cytoplasmic stretching with cell adhesion to the substratum. The OMC mediated and promoted cell differentiation from OSC to osteoblast through osteoblastic phenotype cell (OBC) by cell-aggregation of nodules with "pile up" phenomenon of OBC onto OMC. The osteogenesis might be performed by coupling work of both cells, OMC originated from monocyte of HSC and OBC originated from OSC, which were explained by SEM, TEM and fluorescent probe investigation on BFGCs on the test plate of cp titanium plates with different topographies. This osteogenetic process was proved by investigating cell proliferation, DNA contents, cell-adhesion, alkaline phosphatase activity and osteocalcine productivity for cells on the titanium plates with different topographies. The study showed increased osteogenic effects for cells cultured on Ti with increased surface roughness. Possible mechanisms were discussed from a biomechanical perspective.
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PMID:In vitro study on bone formation and surface topography from the standpoint of biomechanics. 1574 82

A bioactive glass belonging to the system SiO(2)-CaO-Na(2)O was doped with silver ions by ion exchange in molten salts as well as in aqueous solution. The ion exchange in the solution was done to check if it is possible to prepare an antimicrobial material using a low silver content. The doped glass was characterized by means of X-ray diffraction, SEM observation, EDS analysis, bioactivity test (soaking in a simulated body fluid), leaching test (GFAAS analyses) and cytotoxicity test. It is demonstrated that these surface silver-doped glasses maintain, or even improve, the bioactivity of the starting glass. The measured quantity of released silver into simulated body fluid compares those reported in literature for the antibacterial activity and the non-cytotoxic effect of silver. Cytotoxicity tests were carried out to understand the effect of the doped surfaces on osteogenic cell adhesion and proliferation.
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PMID:Surface characterization of silver-doped bioactive glass. 1579 37

Embryonic stem (ES) cells represent a potentially useful cell source for tissue regeneration. Previously, using factors known to enhance differentiation and mineralization of primary osteoblasts, we were able to generate cell populations enriched with osteoblasts from a murine ES cell source. Dexamethasone was a potent inducer of osteoblast differentiation and the timing of stimulation markedly increased the proportion of osteoblast lineage cells. This study examined whether inorganic stimuli derived from bioactive glasses could affect the differentiation of osteoblasts in an ES-cell based system. Previous work has demonstrated the ability of soluble ions released from bioactive glasses undergoing dissolution in vitro to stimulate gene expression characteristic of a mature phenotype in primary osteoblasts. We report here on the potential of soluble extracts prepared from 58S sol-gel bioactive glass to further enhance lineage-specific differentiation in murine ES cells. Differentiation of ES cells into osteogenic cells was characterized by the formation of multilayered, mineralized nodules. These nodules contained cells expressing the transcription factor runx2/cbfa-1, and deposition of osteocalcin in the extracellular matrix was detected by immunostaining. When differentiating cells were placed in an osteoblast maintenance medium supplemented with soluble extracts prepared from bioactive glass powders, we observed increased formation of mineralized nodules (98 +/- 6%, mean +/- SEM) and alkaline phosphatase activity (56 +/- 14%, mean +/- SEM) in a pattern characteristic of osteoblast differentiation. This effect of the glass extracts exhibited dose dependency, with alkaline phosphatase activity and nodule formation increasing with extract concentrations. Compared with medium supplemented with dexamethasone, which had previously been used to enhance osteoblast lineage derivation, the glass extracts were as effective at inducing formation of mineralized nodules by murine ES cells. When glass extracts were used in combination with dexamethasone, a further increase in the number of nodules was observed (110 +/- 16%; cf. 83 +/- 7% for dexamethasone alone). This study demonstrates the capacity of an entirely inorganic material to stimulate differentiation of ES cells toward a lineage with therapeutic potential in tissue-engineering applications.
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PMID:Enhanced derivation of osteogenic cells from murine embryonic stem cells after treatment with ionic dissolution products of 58S bioactive sol-gel glass. 1586 26

Biomimetic apatites have been reported to promote osteogenic activities in numerous in vivo and in vitro models, but the precise mechanism by which the apatite microenvironment promotes such activities is not well understood. Such mechanistic studies require reproducible model systems that are relevant to tissue engineering practices. Although two-dimensional (2D) apatite-coated polystyrene culture dishes provide practicality and reproducibility, they do not simulate the effects of the three-dimensional (3D) microenvironment and degrading polymeric substrates. A simple 3D model system to address these relevant effects, and its utilization in the investigation of apatite-promoted osteoblastic differentiation in vitro is reported in this paper. Apatite coating was achieved by sequentially immersing poly(lactide-co-glycolide) (PLGA) scaffolds into different simulated body fluids (SBF). SEM, EDX, FTIR, TEM electron diffraction confirmed the apatite coating to comprise of calcium-deficient carbonated hydroxyapatite crystals. While both apatite-coated and non-coated PLGA scaffolds supported MC3T3-E1 attachment, spreading, and proliferation, significant differences in osteoblastic differentiation were observed. Relative to non-coated controls, quantitative real-time PCR revealed significant apatite-associated suppression of alkaline phosphatase (ALP), early upregulation of osteopontin (OPN) at 3 days, and upregulation of osteocalcin (OCN) and bone sialoprotein (BSP) at 4 weeks. In summary, apatite-promoted osteoblastic differentiation can be observed in a 3D model system that is relevant to tissue engineering.
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PMID:In vitro response of MC3T3-E1 pre-osteoblasts within three-dimensional apatite-coated PLGA scaffolds. 1600 21

The aim of this study was to evaluate the osteogenic properties of magnetron sputtered dicalcium pyrophaosphate (DCPP) and hydroxylapatite (HA) coatings. Therefore, DCPP and HA coatings were deposited on grit-blasted titanium discs. The substrates were used as-prepared or received an additional heat treatment which changed the amorphous coating structure to a crystalline structure. Subsequently, rat bone marrow stromal cells were cultured for 1-24 days on the various substrates. DNA and alkaline phosphatase activity was determined after 1, 3, 5, 8, and 12 days of incubation. Osteocalcin expression was evaluated after 8, 12, 16, and 24 days of incubation. Scanning electron microscopical analysis of cell morphology and coating characteristics was done after 8 and 16 days of incubation. All assays were done in duplicate and in each assay all specimens were present in fourfold. Results demonstrated that the cells did not proliferate and differentiate on all amorphous coatings. SEM revealed that the amorphous coatings showed significant dissolution. On the crystalline DCPP and HA coatings an increase in DNA and alkaline phosphatase activity was seen starting at day 8 of incubation. Osteocalcin expression on the crystalline coatings started to increase at day 16 of incubation. SEM showed that the growth and differentiation of the cells was associated with extensive collagen fiber formation and surface mineralization in the form of globular accretions. Further, statistical testing revealed that proliferation and differentiation of the rat bone marrow stromal cells started significantly earlier on the crystalline HA coatings than that on the crystalline DCPP coatings. These results demonstrate that the rat bone marrow stromal cells proliferated and differentiated only on crystalline magnetron sputtered DCPP as well as HA coatings, which warrants the further in vivo analysis of the bone healing supporting properties of these coatings.
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PMID:Growth behavior of rat bone marrow cells on RF magnetron sputtered hydroxyapatite and dicalcium pyrophosphate coatings. 1660 22

During the process of bone formation, titanium (Ti) surface is an important factor in the modulation of osteoblastic function. This study was conducted in order to determine the effects of different Ti surfaces on the biological responses of a human osteoblast-like cell line (MG63). MG63 cells were cultured on smooth (S), sandblasted large-grit and acid etching (SLA), hydroxyapatite (HA), hydroxyfluoride (HF), titanium nitrate (TIN), and diamond-like carbon (DLC) Ti. The morphology of these cells were assessed by SEM. The cDNAs prepared from the total RNAs of the MG63 were hybridized into a human cDNA microarray (1152 elements). The appearances of the surfaces observed by SEM were different on each of the six dental substrate types. The SLA and HA surfaces were determined to be rougher than the others. MG63 cells cultured on SLA and HA exhibited cell-matrix interactions. In the expression of genes involved in osseointegration, several genes, including bone morphogenetic protein, cadherin, integrin, and insulin-like growth factors, were upregulated on the different surfaces. Several genes, including fibroblast growth factor receptor 4, Bcl 2-related protein, and collagen, were downregulated on the different surfaces. The attachment and expression of key osteogenic regulatory genes were enhanced by the surface roughness of the dental materials used.
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PMID:Effect of various implant coatings on biological responses in MG63 using cDNA microarray. 1662 96

Hydroxyapatite (HA) has been widely used as a bone graft substitute. In this study, we investigated whether the addition of osteogenic protein-1 (OP-1) further enhanced the weak osteoinductive properties of hydroxyapatite when loaded with human mesenchymal stem cells (h-MSCs). Over a 14 day period, cell proliferation in both groups was assessed qualitatively using SEM and quantitatively using alamar blue assay. Cell differentiation was also evaluated by measurement of ALP activity, which was expressed against total DNA. HA/MSC loaded with OP-1 demonstrated a statistically significant increase (p<0.001) in cell proliferation at all time points in comparison to unloaded samples. ALP activity per DNA was also significantly enhanced (p<0.001) in loaded samples when compared to unloaded controls. SEM demonstrated increased cellular attachment and proliferation into HA pores at all time points in the loaded samples. Our study suggests that the osteoinductive potential of HA can be improved in vitro by the combined incorporation of MSCs and OP-1.
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PMID:Enhancing the osteoinductive properties of hydroxyapatite by the addition of human mesenchymal stem cells, and recombinant human osteogenic protein-1 (BMP-7) in vitro. 1696 59

Nanofibers have recently gained substantial interest for potential applications in tissue engineering. The objective of this study was to determine whether electrospun nanofibers accommodate the viability, growth, and differentiation of human mesenchymal stem cells (hMSCs) as well as their osteogenic (hMSC-Ob) and chondrogenic (hMSC-Ch) derivatives. Poly(d,l-lactide-co-glycolide) (PLGA) beads with a PLA:PGA ratio of 85:15 were electrospun into non-woven fibers with an average diameter of 760+/-210 nm. The average Young's modulus of electrospun PLGA nanofibers was 42+/-26 kPa, per nanoindentation with atomic force microscopy (AFM). Human MSCs were seeded 1-4 weeks at a density of 2 x 10(6)cells/mL in PLGA nanofiber sheets. After 2 week culture on PLGA nanofiber scaffold, hMSCs remained as precursors upon immunoblotting with hKL12 antibody. SEM taken up to 7 days after cell seeding revealed that hMSCs, hMSC-Ob and hMSC-Ch apparently attached to PLGA nanofibers. The overwhelming majority of hMSCs was viable and proliferating in PLGA nanofiber scaffolds up to the tested 14 days, as assayed live/dead tests, DNA assay and BrdU. In a separate experiment, hMSCs seeded in PLGA nanofiber scaffolds were differentiated into chodrogenic and osteogenic cells. Histological assays revealed that hMSCs continuously differentiated into chondrogenic cells and osteogenic cells after 2 week incubation in PLGA nanofibers. Taken together, these data represent an original investigation of continuous differentiation of hMSCs into chondrogenic and osteogenic cells in PLGA nanofiber scaffold. Consistent with previous work, these findings also suggest that nanofibers may serve as accommodative milieu for not only hMSCs, but also as a 3D carrier vehicle for lineage specific cells.
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PMID:Continuing differentiation of human mesenchymal stem cells and induced chondrogenic and osteogenic lineages in electrospun PLGA nanofiber scaffold. 1701 Apr 25

The purpose of this study was to evaluate the effect of osteogenic differentiation of bone marrow stromal stem cells (BMSCs) on bone formation in a novel interconnected porous calcium hydroxyapatite (IP-CHA). BMSCs/IP-CHA composites, as a cell-hybrid artificial bone, were made by injecting BMSCs solution into IP-CHA scaffolds. To induce osteogenic differentiation, BMSCs/IP-CHA composites were subcultured for three, seven, 10, and 14 days. At the end of each subculture period, BMSCs/IP-CHA composites were examined by SEM and ALP staining. BMSCs/IP-CHA composites of different osteogenic groups of subculture were also placed into bone sockets in the right femur of beagle dogs. After four weeks, same placement procedure was done in the left femur. BMSCs/IP-CHA subcultured for 10 and 14 days were ALP-positive as opposed to those of three and seven days. At four weeks after placement, bone formation was superior at the 10- and 14-day subculture groups. Based on the results obtained, it was suggested that osteogenic differentiation periods with 10 and 14 days of subculture for BMSCs/IP-CHA as a cell-hybrid artificial bone were beneficial in promoting bone formation.
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PMID:Development of cell-hybrid artificial bone: effect of osteogenic differentiation of bone marrow stromal stem cells on bone formation with newly developed interconnected porous calcium hydroxyapatite. 1762 30

The aim of the study was to assess the suitability of different Ti-6Al-4V surfaces produced by the electron beam melting (EBM) process as matrices for attachment, proliferation, and differentiation of human fetal osteoblasts (hFOB 1.19). Human osteoblasts were cultured in vitro on smooth and rough-textured Ti-6Al-4V alloy disks. By means of cell number and vitality and SEM micrographs cell attachment and proliferation were observed. The differentiation rate was examined by using quantitative real-time PCR analysis for the gene expression of alkaline phosphatase (ALP), type I collagen (Coll-I), bone sialoprotein (BSP) and osteocalcin (OC). After 3 days of incubation there was a significant higher vitality (p < 0.02) and proliferation (p < 0.02) of hFOB cells on smooth surfaces (R(a) = 0.077 microm) and compact surfaces with adherent partly molten titanium particles on the surface (R(a) </= 24.9 microm). On these samples cells spread over almost the whole surface. On porous surfaces with higher R(a) values, cell proliferation was reduced significantly. Quantitative real-time PCR analysis showed that the expression of osteogenic differentiation markers was not influenced by surface characteristics. Gene expression did not differ more than twofold for the different samples. Compact titanium samples with adherent partly molten titanium particles on the surface (R(a) </= 24.9 microm) fabricated by the EBM process turned out to be best suited for cell proliferation, while highly rough surfaces (R(a) >/= 56.9 microm) reduced proliferation of hFOB cells. Surface characteristics of titanium can easily be changed by EBM in order to further improve proliferation.
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PMID:Effects of topographical surface modifications of electron beam melted Ti-6Al-4V titanium on human fetal osteoblasts. 1768 9


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