The effect of sterilization methods on biological activity of fibronectin on the surface of biomaterials was elaborated in the present study. Sterile protein- modified biomaterials were fabricated by microfilter filtration and UV irradiation, respectively. UV irradiation altered the conformation of surface- adsorbed fibronectin and further affected the attachment, morphology and biological function of endothelial cells. However, microfilter filtration did not to change the normal conformation of fibronectin, or the proliferation and biological function of endothelial cells, indicating that microfilter filtration sterilization is the most suitable method for protein-substrate.
Cell Adhesion ; radiation effects ; Fibronectins ; radiation effects ; Filtration ; Prostheses and Implants ; microbiology ; Sterilization ; methods ; Ultraviolet Rays

OBJECTIVETo evaluate the adhesion, proliferation and differentiation of osteoblast-like cells on the ultraviolet (UV)-treated titanium in different storing environment, and to find a way to enhance the bioactivity of titanium and to prevent its age-related degradation.

METHODSAcid-etched titanium disks stored under ambient conditions for 4 weeks and treated with UV light for 48 h.Then disks were divided into three groups and placed in a sealed container for 0 h (no-stored,NO group) , 4 weeks (air-stored, AS group) or in a sealed container filled with nitrogen for 4 weeks (nitrogen-stored,NS group) respectively. A group of UV-untreated titanium served as negative control (NC group).The surface morphology was evaluated using scanning electron microscopy (SEM), and hydrophilicity of disks were measured using contact angle measuring device. Cell counting kit-8 was used to measure the cell adhesion and proliferation. Cell differentiation was evaluated by testing alkaline phosphatase (ALP) activity using ALP reagent kit.

RESULTSThere was no difference in surface topography among groups.Contact angels in NS group [(67.70 ± 3.59)°] and NO group [(0.70 ± 0.28)°] were smaller than the others (P < 0.05). Cell adhesion in NS group at 2 h and 4 h point was (0.237 ± 0.006) and (0.578 ± 0.039), respectively, and proliferation at 3 d and 5 d point was (0.743 ± 0.026) and (1.548 ± 0.046) respectively, which were significantly higher than those in AS group [(0.158 ± 0.036), (0.400 ± 0.010), (0.499 ± 0.019) and (1.174 ± 0.062)] and in NC group [(0.161 ± 0.024), (0.390 ± 0.011), (0.508 ± 0.015) and (1.209 ± 0.025)] at the same time point (P < 0.05). How ever the results mention above in the NS group were lower than those in the NO group (P < 0.05). No difference were found between data from the AS group and NS group (P > 0.05). Osteoblast-like cells had an abundant spread on NS and NO group during 2 h incubation, but did not exactly spread on AS and NC group after incubation for 4 h. No difference were found in ALP among groups.

CONCLUSIONSUV treatment can enhance bioactivity of titanium, and nitrogen storage can slow down its biological aging.

Alkaline Phosphatase ; metabolism ; Animals ; Biocompatible Materials ; chemistry ; Cell Adhesion ; radiation effects ; Cell Differentiation ; radiation effects ; Cell Proliferation ; radiation effects ; Cells, Cultured ; Mice ; Microscopy, Electron, Scanning ; Nitrogen ; chemistry ; Osteoblasts ; cytology ; metabolism ; Surface Properties ; Titanium ; chemistry ; radiation effects ; Ultraviolet Rays

OBJECTIVETo observe the effects of calcium phosphate cement/eucommia ulmoides extracts compound (CPC/EUE) on cell proliferation and adhesion of MC3T3E1 osteoblasts in vitro.

METHODSCultured MC3T3E1 osteoblasts were divided into 5 groups: blank control, CPC/EUE 0, 0.152, 0.304 and 0.608 mg/ml. 3H-TdR and scan electronic microscopy (SEM) were used to observe the cell growth, proliferation and adhesion.

RESULTThe CPC/EUE compound enhanced the cell growth, proliferation and adhesion. The concentrations of 0.152 mg/ml, 0.304 mg/ml were more significant in enhancing the cell proliferation rate (P<0.05).

CONCLUSIONCPC/EUE can promote the cell growth, proliferation and adhesion of the osteoblasts.

Alkaline Phosphatase ; metabolism ; Bone Cements ; Calcium Phosphates ; pharmacology ; Cell Adhesion ; radiation effects ; Cell Differentiation ; radiation effects ; Cell Proliferation ; radiation effects ; Cells, Cultured ; Gonadotropins ; metabolism ; Humans ; Osteoblasts ; cytology ; drug effects ; Tissue Adhesions

Animals ; Catalysis ; Cell Adhesion ; drug effects ; radiation effects ; Humans ; Hydrophobic and Hydrophilic Interactions ; Osseointegration ; drug effects ; radiation effects ; Osteoblasts ; cytology ; drug effects ; radiation effects ; Osteogenesis ; drug effects ; radiation effects ; Photochemical Processes ; Surface Properties ; radiation effects ; Time Factors ; Titanium ; chemistry ; pharmacology ; radiation effects ; Ultraviolet Rays

OBJECTIVETo explore the changes in the expressions of the tight junction related protein occludin and junctional adhesion molecule-1 (JAM-1) of the blood-testis barrier and their significance in rats after microwave radiation.

METHODSEighty male Wistar rats were exposed to microwave radiation with average power density of 0, 10, 30 and 100 mW/cm2 for five minutes, and dynamic changes in the expressions of testicular occludin and JAM-1 were observed by Western blot and image analysis at 6 h, 1 d, 3 d, 7 d and 14 d after the radiation.

RESULTSThere was a significant down-regulation in the expression of the occludin protein at 3 - 7 d, 6 h - 7 d and 6 h - 14 d (P < 0. 05), as well as in that of JAM-1 at 3 - 7 d, 1 - 7 d and 1-14 d (P < 0.05) after exposure to 10, 30 and 100 mW/cm2 microwave radiation.

CONCLUSIONThe decreased protein expressions of occludin and JAM-1 may play an important role in the microwave radiation induced-damage to the blood-testis barrier.

Animals ; Blood-Testis Barrier ; metabolism ; radiation effects ; Cell Adhesion Molecules ; metabolism ; Down-Regulation ; Male ; Membrane Proteins ; metabolism ; Microwaves ; Occludin ; Rats ; Rats, Wistar ; Testis ; metabolism ; radiation effects

In order to determine the effect of ultraviolet radiation (UVR) on the cell adhesion molecules expressed in human dermal microvascular endothelial cells (HDMEC), the cells were exposed to varying UVR doses and the cell surface was examined for expression of intercellular cell adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule-1 (VCAM- 1), and E-selectin. The effect of UVB irradiation on the binding of T lymphocytes to HDMEC was also examined. UVA irradiation did not affect the surface expression of ICAM-1, VCAM-1, or E-selectin on the HDMEC. However, following UVB exposure, ELISA demonstrated a significant increase in the baseline ICAM-1 cell surface expression on the HDMEC. However, no induction of either E-selectin or VCAM-1 was noted. UVB also significantly augmented ICAM-1 induction by IL-1 alpha and TNF- alpha. VCAM-1 was induced by stimulating HDMEC with IL-1alpha following a UVB irradiation dose of 100 mJ/cm2. Flow cytometric analysis of the HDMEC stimulated with IL-1 alpha for 24h demonstrated that 12% of the cells expressed VCAM-1 but either IL-1 alpha or UVB irradiation alone failed to induce VCAM-1 expression. Enhancement of T cell-HDMEC binding by IL-1 alpha; or TNF- alpha treatment was not significantly affected after UVB irradiation. This study demonstrated that UVB irradiation can alter ICAM-1 and VCAM-1 expression on the HDMEC surface and that augmentation of ICAM-1 expression and the IL-1 alpha -dependent induction of VCAM-1 following UVB exposure might be important steps in the pathogenesis of sunburn.
Cell Adhesion/radiation effects ; Cell Adhesion Molecules/*metabolism ; Cells, Cultured ; Endothelium, Vascular/cytology/*physiology ; Human ; Microcirculation ; Skin/*blood supply ; T-Lymphocytes/*physiology ; *Ultraviolet Rays

The present paper is aimed to investigate the effects of pulse electrical stimulation on mutual adhesion of vascular endothelial cell and endothelial progenitor cell (EPC). EPC was induced from periphery blood, labeled with fluorescence dye and then co-cultured with vascular endothelial cell. With a fixed electric voltage and frequency of 5V and 5Hz, respectively, the co-culture system was continually stimulated for 24h under different pulse width, 1, 3, 6 and 9ms. After pulse stimulation, fluorescence intensity of adherent labeled EPC was measured and converted to fluorescence ratio. Compared to that in the control group, fluorescence ratio of 3 ms and 6 ms group were significantly larger, while that in the 9 ms group was lower. The peak fluorescence ratio value was appeared at 6 ms group. It is indicated that suitable pulse electrical stimulation could benefit the adhesion of endothelial cell and EPC. All these results provide a new theoretical basis about why electrical stimulation could contribute to neovascularization.
Adult ; Cell Adhesion ; radiation effects ; Cells, Cultured ; Coculture Techniques ; Electric Stimulation ; Female ; Hematopoietic Stem Cells ; cytology ; radiation effects ; Human Umbilical Vein Endothelial Cells ; cytology ; radiation effects ; Humans ; Male ; Young Adult

To evaluate the effect of Ligustrazine on the expression of VEGF in bone marrow stromal cells (BMSCs) of radiation injured mice and to explore the effect of VEGF on the recovery of hematopoiesis and the mechanism of signal transduction, the protein expression of VEGF, focal adhesion kinase (FAK) and mitogen-activated protein kinase (MAPK) in BMSCs were assayed by Western blot, the cell cycle and apoptosis rate of BMSCs were tested by flow cytometry. The effect of Ligustrazine on the hematopoiesis was evaluated at the same time. The results showed that the protein expression of VEGF in BMSCs was decreased significantly after irradiation and increased slowly with the time. The value in Ligustrazine-treated group almost reached normal level, but it remained lower than that in control group on day 14. The changes of phosphorylated FAK and MAPK protein expression had the same tendency. After (60)Co gamma-irradiation, the BMSCs were arrested in G0-G1 phase and apoptosis rate increased; these values recovered slowly with the time and remained higher than that in normal control group on day 14. The recovery of these values in Ligustrazine-treated group was sooner than that in irradiated control group, and they almost reached to the normal levels on day 14. It is concluded that irradiation could inhibit the expression of VEGF in BMSCs and induce apoptosis. The Ligustrazine promotes the recovery of bone marrow microenvironment probably by increasing the expression of phosphorylated FAK and MAPK in BMSCs.
Animals ; Apoptosis ; drug effects ; Bone Marrow Cells ; drug effects ; metabolism ; Cell Cycle ; drug effects ; Cell Division ; drug effects ; Focal Adhesion Kinase 1 ; Focal Adhesion Protein-Tyrosine Kinases ; Mice ; Protein-Tyrosine Kinases ; analysis ; Pyrazines ; pharmacology ; Radiation Injuries, Experimental ; drug therapy ; metabolism ; Radiation-Protective Agents ; pharmacology ; Stromal Cells ; drug effects ; metabolism ; Vascular Endothelial Growth Factor A ; analysis

NF-kappa B promotes cell survival against external stress such as radiation. We examined whether NF-kappa B decoy transfection enhances the antiproliferative effects of radiation on vascular smooth muscle cells (VSMCs) in vitro. The irradiation induced activation or nuclear translocation of NF-kappa B p65 in VSMCs was confirmed by immunofluorescence. NF-kB decoy transfection resulted in inhibition of the radiation-induced NF-kB activation in VSMCs and the subsequent reduction of transcription and translocation of ICAM, iNOS, and TNF-alpha, downstream molecules under the control of NF-kappa B. By using MTT assay, NF-kappa B decoy augmented the antiproliferative effects of radiation, where the effect of low dose radiation (2 and 8-Gy) of the cells transfected with NF-kappa B decoy was equivalent to the high dose (16-Gy) irradiated non-transfected cells at 48 h after irradiation: 1.06+/-0.16, 1.11+/-0.22, 1.20+/-0.25, respectively. The decrease in proliferation and survival of the radiation treated cells by flow cytometry analysis showed that NF-kappa B inhibition did not show any additive effects on the cell cycle of the irradiated VSMCs, while apoptosis was significantly increased after NF-kappa B decoy transfection in the irradiated VSMCs (apoptosis fraction: 13.33+/-2.08% vs. 26.29+/-7.43%, for radiation only vs. radiation+NF-kappa B decoy transfection, P < 0.05). In addition, at 48 h, NF-kappa B decoy transfection dose dependently (10 mM vs. 20 mM) inhibited proliferation of 16Gy-irradiated VSMCs, and showed greater antiproliferative efficacy than 100 mM sulfasalazine, a specific NF-kappa B inhibitor. These results indicate that NF-kappa B inhibition reduces proliferation and survival of irradiated VSMCs, likely by increased apoptosis rather than additive cell cycle arrest and suggest the possibility of adjunctive gene therapy using NF-kappa B decoy to improve efficacy and to decrease the adverse effects of intracoronary radiation therapy.
Animals ; Aorta/cytology/radiation effects ; *Apoptosis ; Cell Cycle/physiology/radiation effects ; Cell Proliferation/radiation effects ; Cells, Cultured ; Gamma Rays ; Intercellular Adhesion Molecule-1/metabolism ; Male ; Muscle, Smooth, Vascular/cytology/physiology/*radiation effects ; Myocytes, Smooth Muscle/cytology/radiation effects ; NF-kappa B/*antagonists & inhibitors/metabolism ; Nitric-Oxide Synthase/metabolism ; Protein Transport ; Rats ; Rats, Sprague-Dawley ; Research Support, Non-U.S. Gov't ; Transcription, Genetic ; Transfection ; Tumor Necrosis Factor-alpha/metabolism

OBJECTIVETo study the effects of ultraviolet (UV) light-treatment on the physicochemical properties and bioactivity of micro-arc oxidation (MAO) titanium surfaces in vitro.

METHODSThe pure titanium were prepared using MAO. MAO titanium samples were treated with 15 W bactericidal lamp UVC [λ = (250 ± 20) nm] or 15 W mercury lamp [λ = (360 ± 20) nm] for 24 h under ambient conditions. Three sample groups were prepared: MAO, UVA treated after MAO (MAO + UVA), UVC treated after MAO (MAO + UVC). The surface physicochemical properties were evaluated using scanning electron microscopy (SEM), contact angle measuring device, energy dispersive X-ray spectrometer (EDX), and X-ray diffraction (XRD). Bicinchoninic acid (BCA) based colorimetric detection was used to quantify the percentage of albumin adsorption after 2 h, 6 h, and 24 h incubation on the titanium surfaces. The rates of MG-63 cells attached to each group titanium surfaces were calculated by nucleus immunofluorescence using Hoechst 33342 after 1 h, 2 h, and 4 h incubation. SEM was used to observe cell morphology on titanium surfaces in each group.

RESULTSNo obvious differences in surface topography, TiO(2) crystal and elemental composition were detected on titanium surfaces with or without UV treatment. Statistically significant difference in contact angles among MAO + UVC group (65.34 ± 1.16)°, MAO + UVA group (44.64 ± 1.28)°, and MAO group (3.41 ± 0.48)° were found (P < 0.001). The percentage of albumin adsorption reached the plateau after 2 h incubation on MAO + UVC titanium surfaces (48.16 ± 1.24)%, which was higher than those in MAO [(8.22 ± 2.99)%] and MAO + UVA groups [(5.29 ± 2.27)%, P < 0.001]. The rates of cells attached to the surfaces of MAO + UVC titanium was greater than that on MAO surfaces and MAO + UVA surfaces after 1 h [(40.71 ± 4.08)%], 2 h [(53.72 ± 2.38)%], 4 h [(70.32 ± 2.85)%] incubation (P < 0.05). The MAO + UVC surfaces remarkably enhanced the spread of MG-63 cells, however, there was no significant difference between the group of MAO and MAO + UVA.

CONCLUSIONSPretreatment of micro-arc oxidation titanium with UVC light considerably improved the surface bioactivity to MG-63 cells, which showed an increase in cellular attachment and spread.

Absorption ; Albumins ; metabolism ; Cell Adhesion ; Cell Line, Tumor ; Humans ; Microscopy, Electron, Scanning ; Osteosarcoma ; pathology ; Oxidation-Reduction ; Spectrometry, X-Ray Emission ; Surface Properties ; Titanium ; chemistry ; radiation effects ; Ultraviolet Rays ; X-Ray Diffraction