PURPOSE: This study was performed to characterize the effects of zirconia coated with calcium phosphate and hydroxyapatite compared to smooth zirconia after bone marrow-derived osteoblast culture. MATERIALS AND METHODS: Bone marrow-derived osteoblasts were cultured on (1) smooth zirconia, (2) zirconia coated with calcium phosphate (CaP), and (3) zirconia coated with hydroxyapatite (HA). The tetrazolium-based colorimetric assay (MTT test) was used for cell proliferation evaluation. Scanning electron microscopy (SEM) and alkaline phosphatase (ALP) activity was measured to evaluate the cellular morphology and differentiation rate. X-ray photoelectron spectroscopy (XPS) was employed for the analysis of surface chemistry. The genetic expression of the osteoblasts and dissolution behavior of the coatings were observed. Assessment of the significance level of the differences between the groups was done with analysis of variance (ANOVA). RESULTS: From the MTT assay, no significant difference between smooth and surface coated zirconia was found (P>.05). From the SEM image, cells on all three groups of discs were sporadically triangular or spread out in shape with formation of filopodia. From the ALP activity assay, the optical density of osteoblasts on smooth zirconia discs was higher than that on surface treated zirconia discs (P>.05). Most of the genes related to cell adhesion showed similar expression level between smooth and surface treated zirconia. The dissolution rate was higher with CaP than HA coating. CONCLUSION: The attachment and growth behavior of bone-marrow-derived osteoblasts cultured on smooth surface coated zirconia showed comparable results. However, the HA coating showed more time-dependent stability compared to the CaP coating.
Alkaline Phosphatase ; Calcium ; Cell Adhesion ; Cell Proliferation ; Chemistry ; Durapatite ; Microscopy, Electron, Scanning ; Osteoblasts* ; Photoelectron Spectroscopy ; Pseudopodia

NHERF1/EBP50 (Na⁺/H⁺ exchanger regulating factor 1; Ezrin-binding phosphoprotein of 50 kDa) organizes stable protein complexes beneath the apical membrane of polar epithelial cells. By contrast, in cancer cells without any fixed polarity, NHERF1 often localizes in the cytoplasm. The regulation of cytoplasmic NHERF1 and its role in cancer progression remain unclear. In this study, we found that, upon lysophosphatidic acid (LPA) stimulation, cytoplasmic NHERF1 rapidly translocated to the plasma membrane, and subsequently to cortical protrusion structures, of ovarian cancer cells. This movement depended on direct binding of NHERF1 to C-terminally phosphorylated ERM proteins (cpERMs). Moreover, NHERF1 depletion downregulated cpERMs and further impaired cpERM-dependent remodeling of the cell cortex, suggesting reciprocal regulation between these proteins. The LPA-induced protein complex was highly enriched in migratory pseudopodia, whose formation was impaired by overexpression of NHERF1 truncation mutants. Consistent with this, NHERF1 depletion in various types of cancer cells abolished chemotactic cell migration toward a LPA gradient. Taken together, our findings suggest that the high dynamics of cytosolic NHERF1 provide cancer cells with a means of controlling chemotactic migration. This capacity is likely to be essential for ovarian cancer progression in tumor microenvironments containing LPA.
Cell Membrane ; Cell Movement ; Cytoplasm ; Cytosol ; Epithelial Cells ; Membranes ; Ovarian Neoplasms* ; Pseudopodia ; Tumor Microenvironment

STATEMENT OF PROBLEM: A biochemical approach for surface modification has offered an alternative for physicochemical and morphological methods to obtain desirable bone-implant interfaces. PURPOSE: The purpose of the present study was to investigate cell responses to poly (D,L-lactide-co-glycolide) (PLGA)/1alpha, 25-(OH)2D3 coating with reference to cellular proliferation and differentiation in vitro. MATERIAL AND METHODS: 96 titanium discs were fabricated and divided into four groups. Group 1 was anodized under 300 V as control. Group 2, 3 and 4 were anodized then coated with 3 ml PLGA/1alpha, 25-(OH)2D3 solutions. Amount of the solutions were 2 ul, 20 ul and 200ul respectively. The osteoblast-like Human Osteogenic Sarcoma (HOS) cells were seeded and cultured for 1, 3 and 7 days. MTSbased cell proliferation assay and ALPase activity test were carried out. RESULTS: PLGA nanoparticles were observed as fine, smooth and round and HOS cells attached to the anodized surfaces through strand-like and sheet-like filopodia. After 3 days of culture, the dendritic filopodia were exaggerated and sheet-like cytoplasmic projections covered the coated titanium surfaces. After 3 days of culture, all of the groups showed increased cellular proliferation and the lowest proliferation rate was measured on group 2. Higher amount of incorporated 1 alpha, 25-(OH)2D3 (Group 3 and 4) improved cellular proliferation but the differences were not significant statistically (P > .05). But they increased the rate of ALP activities than the control group at day 3 (P < .05). CONCLUSION: Biodegradable PLGA nanoparticles incorporated with vitamin D metabolite positively affected proliferation and differentiation of cells on the anodized titanium surface.
Calcitriol ; Cell Proliferation ; Cytoplasm ; Humans ; Lactic Acid ; Nanoparticles ; Osteosarcoma ; Polyglycolic Acid ; Pseudopodia ; Seeds ; Titanium ; Vitamin D

Fluorouracil Was effective in controlling the in vitro cellular proliferation. When given with cultured human retinal pigment epithelial cell, fluoroirracil decreased the cellular activity of the cultured retinal pigment epithelial cell. The attachment and migratien of the cultured retinal pigment epithelial cell was markedly reduced after repeated treatment with fluorouracil. Electron micrographs of the cultured retinal pigment epithelial cell given with fluorouracil showed marked reduction of cytoplasmic pseudopodia, rupture of plasma membrane, loss of microorganelles and eventual cytolysis. Based on these data, low dose fluorouracil may be an effective inhibitor of the in vitro proliferation of the retinal pigment epithelium.
Cell Membrane ; Cell Proliferation ; Cytoplasm ; Epithelial Cells ; Fluorouracil ; Humans ; Pseudopodia ; Retinal Pigment Epithelium* ; Retinaldehyde* ; Rupture

It is believed that there exists some relationship between the distribution and morphology of intracellular actin and cell adherence. Cells are likely to be deteched when the quantity of actin filament decreases. Actin filaments locate in the fringe of cancer cells and cells cultured in static state, so that these filaments can stretch out and form pseudopodia to adhere to the matrix. When these cells are stimulated their pseudopodia retract so that they can easily be detached from the matrix. When external forces are exerted on cells to adhere and deadhere from the matrix, the morphology and distribution of skeleton actin will change, so as the cells' morphology. The skeleton actins in cells are changed differently to adapt to different external forces which are imposed on the cells. It is obvious that the relationship between the mechanism of cell adhering to the matrix and the morphology & distribution of actins needs more attention.
Actin Cytoskeleton ; metabolism ; Actins ; metabolism ; Cell Adhesion ; Humans ; Neoplasms ; metabolism ; pathology ; Pseudopodia ; metabolism ; Shear Strength

The changes of marginal epithelial cells in corneal wound healing were observed in a rabbit. The randomly assigned three eyes in the rabbits were extracted at first, third, and eighteenth day after full thickness epithelial removal, then observed under the electronmicroscope. At the 1st and 3rd day ,the thickness of the epithelium at the wound margin was reduced at the leading edge. These flattened epithelial cells showed ruffling and folding of the plasma membrane near free edge to form filopodia or lamellipodia processes, extending onto wound surface. Cytoskeletons reorganized and rearranged in leading edge. Basement membrane of the wound was relatively intact, but on which cellular debris were observed, and cell migration undergone and hemidesmosomes developed incompletely. In eighteenth day, basal cell recovered original cylindric shape, cytoskeletons was originally redistributed in cytoplasm after migratory phase, and hemidesmosome developed completely.
Basement Membrane ; Cell Membrane ; Cell Movement ; Cytoplasm ; Cytoskeleton ; Epithelial Cells* ; Epithelium ; Hemidesmosomes ; Pseudopodia ; Rabbits ; Wound Healing* ; Wounds and Injuries*

BACKGROUND: The natural compound curcumin was known to inhibit migration and invasion of glioblastoma (GBM) cells. Fascin, a kind of actin-binding proteins, is correlated with migration and invasion of GBM cells. The purpose of this study was to investigate anti-migration and anti-invasion effects of curcumin via suppression of fascin expression in GBM cells. METHODS: U87 cell line was used as an experimental model of GBM. Fascin was quantified by Western blot analysis. And, the signal transducer and activator of transcription 3 (STAT3), known to play an important role in migration and invasion of tumor cells, were analyzed by sandwich-ELISA. Migration and invasion capacities were assessed by attachment, migration and invasion assays. Cellular morphology was demonstrated by immunofluorescence. RESULTS: At various concentrations of curcumin and exposure times, fascin expression decreased. After temporarily exposure to 10 µM/L curcumin during 6 hours as less invasive concentration and time, fascin expression temporarily decreased at 12 hours (18.4%, p=0.024), and since then recovered. And, the change of phosphrylated STAT3 level also reflected the temporarily decreased pattern of fascin expression at 12 hours (19.7%, p=0.010). Attachment, migration, and invasion capacities consistently decreased at 6, 12, and 24 hours. And, immunofluorescence showed the change of shape and the reduction of filopodia formation in cells. CONCLUSION: Curcumin is likely to suppress the fascin expression in GBM cells, and this might be a possible mechanism for anti-migration and anti-invasion effects of Curcumin via inhibition of STAT3 phosphorylation.
Blotting, Western ; Cell Line ; Curcumin ; Emigration and Immigration ; Fluorescent Antibody Technique ; Glioblastoma ; Microfilament Proteins ; Models, Theoretical ; Phosphorylation ; Pseudopodia ; STAT3 Transcription Factor

BACKGROUND/OBJECTIVES: Re-epithelialization has an important role in skin wound healing. Astaxanthin (ASX), a carotenoid found in crustaceans including shrimp, crab, and salmon, has been widely used for skin protection. Therefore, we investigated the effects of ASX on proliferation and migration of human skin keratinocyte cells and explored the mechanism associated with that migration. MATERIAL/METHOD: HaCaT keratinocyte cells were exposed to 0.25-1 µg/mL of ASX. Proliferation of keratinocytes was analyzed by using MTT assays and flow cytometry. Keratinocyte migration was determined by using a scratch wound-healing assay. A mechanism for regulation of migration was explored via immunocytochemistry and western blot analysis. RESULTS: Our results suggest that ASX produces no significant toxicity in human keratinocyte cells. Cell-cycle analysis on ASX-treated keratinocytes demonstrated a significant increase in keratinocyte cell proliferation at the S phase. In addition, ASX increased keratinocyte motility across the wound space in a time-dependent manner. The mechanism by which ASX increased keratinocyte migration was associated with induction of filopodia and formation of lamellipodia, as well as with increased Cdc42 and Rac1 activation and decreased RhoA activation. CONCLUSIONS: ASX stimulates the migration of keratinocytes through Cdc42, Rac1 activation and RhoA inhibition. ASX has a positive role in the re-epithelialization of wounds. Our results may encourage further in vivo and clinical study into the development of ASX as a potential agent for wound repair.
Blotting, Western ; Carotenoids ; Cell Movement ; Cell Proliferation ; Clinical Study ; Flow Cytometry ; Humans* ; Immunohistochemistry ; Keratinocytes* ; Pseudopodia ; Re-Epithelialization ; S Phase ; Salmon ; Skin* ; Wound Healing ; Wounds and Injuries

The 5 cases of human fetal liver aged from 11 to 23 weeks of gestation were investigated for the ultrastructure of sinusoidal wall by electron microscopy. The endothelial cells deficient in basement membrane formed almost all the part of sinusoidal wall. The cells were continued with neighboring cells by intercellular junction, and overlapped them and exhibited to maintain unfenestrated capillary wall, which was different from those with fenestra in adults. The cells were found to have coated pits on luminal side and several various vesicles in the cytoplasms. The cells were related with transcellular migration of reticulocytes and acidophilic erythroblastes, which penetrated into the endothelial cytoplasm to form transient migrating pore closing after the migration into sinusoidal lumen. The perivascular cells were present at perivascular space and surrounding the sinusoid discontinuously. The Kupffer cells were easily identified with filipodia and lamellipodia and with phagosome of nuclei enucleated from acidophilic erythroblasts.
Adult ; Basement Membrane ; Capillaries ; Cytoplasm ; Endothelial Cells* ; Erythroblasts ; Erythrocytes ; Humans* ; Intercellular Junctions ; Kupffer Cells* ; Liver* ; Microscopy, Electron ; Phagosomes ; Phenobarbital ; Pregnancy ; Pseudopodia ; Reticulocytes

The purpose of this study was to investigate the ability of Acanthamoeba to adhere to the epithelial cells of human cornea. Human corneas, obtained through the eye bank of Catholic Medical Center, were cultured in Optisol solution at 37degreesC. Trophozoites of Acanthamoeba lugdunensis cultured on non-nutrient agar plate were collected to make a suspension in concentration of 1x106/ml. 100microliter of amoeba suspension was added to the epithelial surface of cultured human corneas and each cornea was incubated for 12 hours. Each cornea was examined with scanning and transmission electron microscope. On scanning electron microscopy, trophozoites adhered to each other and to the corneal epithelium, especially to intercellular junction by their extended lobopodia at 12 hour-incubation. On transmission electron microscopy, trophozoites showed limited regions of attachment to the corneal epithelium at 12 hour-incubation, and the attached areas showed desmosome-like structure. Trophozoites adhered to each other by cytoplasmic interdigitation. In conclusion, trophozoites adhere to the corneal epithelial surface by their cytoplasmic processes and their processes appeared to have affinity to intercellular junctions of the corneal epithelium. Attachment regions between corneal epithelium and amoeba were characterized as desmosomelike junctions.
Acanthamoeba* ; Agar ; Amoeba ; Cornea* ; Cytoplasm ; Epithelial Cells ; Epithelium, Corneal* ; Eye Banks ; Humans* ; Intercellular Junctions ; Microscopy, Electron, Scanning ; Microscopy, Electron, Transmission* ; Organ Culture Techniques ; Pseudopodia ; Trophozoites