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1.
This study characterized human umbilical vein endothelial cell (HUVEC) adhesion, proliferation, and gene expression on bilayered polyelectrolyte coatings composed of an outermost layer of glycosaminoglycans (hyaluronan, heparin, or chondroitin sulfate), with an underlying layer of poly-L-lysine or chitosan. The proportion of cells that adhered to the various polyelectrolyte coatings after 1 and 2 h incubations was quantified by the WST-8 assay. Interchanging poly-L-lysine with chitosan resulted in significant differences in cellular adhesion to the outermost glycosaminoglycan layer after 1 h, but these differences became insignificant after 2 h. The proliferation of HUVEC on the various bilayered polyelectrolyte coatings over 10 days was characterized using the WST-8 assay. Regardless of whether the underlying layer was poly-L-lysine or chitosan, HUVEC proliferation on the hyaluronan outermost layer was significantly less than on heparin or chondroitin sulfate. Additionally, it was observed that there was more proliferation with poly-L-lysine as the underlying layer, compared to chitosan. Subsequently, real-time polymerase chain reaction was used to analyze the expression of seven genes related to adhesion, migration, and endothelial function (VWF, VEGFR, VEGFA, endoglin, integrin-α5, ICAM1, and ICAM2) by HUVEC cultured on the various bilayered polyelectrolyte coatings for 3 days. With poly-L-lysine as the underlying layer, biologically significant differences (greater than twofold) in the expression of VWF, VEGFR, VEGFA, endoglin, and ICAM1 were observed among the three glycosaminoglycans. With chitosan as the underlying layer, all three glycosaminoglycans displayed biologically significant differences in the expression of VWF and VEGFR compared to the chitosan control. CT-HA displayed the highest level of expression of VWF, whereas expression levels of VEGFR were almost similar among the three glycosaminoglycans.  相似文献   

2.
Nanotechnology is a rapidly growing field having potential applications in many areas. Nanoparticles (NPs) have been studied for cell toxicity, immunotoxicity, and genotoxicity. Tetrazolium-based assays such as MTT, MTS, and WST-1 are used to determine cell viability. Cell inflammatory response induced by NPs is checked by measuring inflammatory biomarkers, such as IL-8, IL-6, and tumor necrosis factor, using ELISA. Lactate dehydrogenase (LDH) assay is used for cell membrane integrity. Different types of cell cultures, including cancer cell lines have been employed as in vitro toxicity models. It has been generally agreed that NPs interfere with either assay materials or with detection systems. So far, toxicity data generated by employing such models are conflicting and inconsistent. Therefore, on the basis of available experimental models, it may be difficult to judge and list some of the more valuable NPs as more toxic to biological systems and vice versa. Considering the potential applications of NPs in many fields and the growing apprehensions of FDA about the toxic potential of nanoproducts, it is the need of the hour to look for new internationally agreed free of bias toxicological models by focusing more on in vivo studies. Key Words: Cytotoxicity, in vitro, Metal nanoparticles, Toxicology, Review  相似文献   

3.
Protein and polysaccharide was the most important extracellular matrix in dermal tissue. In this study, Silk fibroin (SF) / hyaluronic acid (HA) blend films mimicking the dermal tissue components were prepared and investigated. The results indicated that HA and SF has a good miscibility, HA interfered with SF to form crystal structure. By using EDC as cross-linker, effective cross-linking function on SF and HA macromolecules was reacted, the water solubility of the blend films decreased obviously after being cross-linked by EDC. The existence of EDC could promote SF to form Silk I structure. L929 cells were seeded on these blend films and showed normal attachment morphology. Cell-matrix interactions established by newly formed extracellular matrix were observed after 5 days in culture. The MTT assay showed that cell proliferation on the SF/HA blend films were enhanced significantly compared with that on the SF and HA films. These new 2D SF/HA blend films provided a favorable microenvironment for the proliferation of L929 cells and hold a potential for dermal tissue regeneration.  相似文献   

4.
Selvan ST 《Biointerphases》2010,5(3):FA110-FA115
Fluorescent quantum dots (e.g., CdSe-ZnS) and magnetic nanoparticles (e.g., Fe(2)O(3) or Fe(3)O(4)) are two important candidate systems that have been emerging as potential probes for bioimaging applications. This review focuses on the development of silica-coated inorganic probes (optical and magnetic) that are originated mainly from the author's laboratory for bioimaging applications. The recent developments in the synthesis of rare earth nanoparticles for multimodality imaging are also delineated.  相似文献   

5.
Two strains of Azadinium poporum, one from the Korean West coast and the other from the North Sea, were mass cultured for isolation of new azaspiracids. Approximately 0.9 mg of pure AZA-36 (1) and 1.3 mg of pure AZA-37 (2) were isolated from the Korean (870 L) and North Sea (120 L) strains, respectively. The structures were determined to be 3-hydroxy-8-methyl-39-demethyl-azaspiracid-1 (1) and 3-hydroxy-7,8-dihydro-39-demethyl-azaspiracid-1 (2) by 1H- and 13C-NMR. Using the Jurkat T lymphocyte cell toxicity assay, (1) and (2) were found to be 6- and 3-fold less toxic than AZA-1, respectively.  相似文献   

6.
It is becoming increasingly evident that certain phytochemicals possess cancer chemopreventive properties. In this study, the anti-proliferative activity of plant extracts from olive (Olea europaea L.) leaves was tested on human leukemic cell line (Jurkat). Cytotoxicity of various concentrations of plant extracts was examined and the IC50 was determined. Olive leaf extracts showed concentration-dependent anti-proliferative effect as determined by the WST-1 proliferation kit and [3H]-thymidine incorporation method. To study whether cell death was due to apoptosis, cells were stained with Annexin V-FITC and PI and the expression of important regulatory proteins (Bcl-2, Bax, and p53) involved in apoptosis were examined by Western blot. The antioxidant activity of olive leaves (SC50 = 0.1 mg dry weight) was studied using the DPPH scavenging method. Present findings suggest that olive leaves extracts exhibit anti-proliferative effect on leukemic cells by inducing apoptosis.  相似文献   

7.
野生稻原生质体培养与植株再生   总被引:7,自引:0,他引:7  
三个野生稻(O. rufipogon,O. glumaepatula 和O. latifolia)成熟胚,经愈伤组织诱导和悬浮细胞培养,分离原生质体,利用简化原生质体培养基(SPCM),结合琼脂糖包埋,并附加看护细胞培养液,这三个野生稻原生质体均得到了成功培养,其中两个(O. rufipogon和O. glumaepatula )分化了绿色植株。看护细胞液在野生稻原生质体培养中,可促进细胞分裂,提早细胞分裂始期,细胞分裂频率(3.2%)和植板率(19.0%)分别比未加看护细胞液的培养方法高出13.3%和6.2%。看护细胞液可大大减少野生稻愈伤组织在液体中培养始期的死亡率,提高建立其悬浮细胞的成功率。原生质体克隆在适当的后培养基上进行培养,可以显著改良其结构,明显促进绿苗分化。尤其是L3培养基诱导原生质体克隆形成胚性愈伤组织或类似胚状体结构的频率达32.1%,显著高于MS和N6培养基,从而导致较高绿苗分化频率。  相似文献   

8.
通过田间试验研究硼砂(B,Na2B4O7?10H2O)、Ulexite granular(UB,NaCaB5O9?8H2O)、Etibor-48(EB, Na2B4O7?5H2O)和Colemanite(CB, Ca2B6O11?5H2O)四种硼肥在花生-油菜轮作中的肥效及其后效对作物产量的影响。结果表明,按正常施硼量(纯硼用量1.70 kg/hm2)基施硼砂、UB、EB和CB四种硼肥,花生和油菜产量均有不同程度的提高;两倍施硼量(纯硼用量3.40 kg/hm2)与正常施硼量相比,不同硼肥处理作物增产率均有不同程度降低,其中土壤有效硼含量0.49 mg/kg时,作物增产率并不随施硼量的继续增加而升高。基施四种硼肥,三季作物收获后土壤有效硼含量均高于0.5 mg/kg,秸秆硼含量与作物产量也显著高于不施硼处理。基施硼肥可以改善土壤有效硼状况,具有较强缓释性的硼肥UB和CB在第二季和第三季具有较好的增产效果。在农业生产上可以根据土壤有效硼状况和硼肥本身的缓效性作为基施长效硼肥合理推广利用。  相似文献   

9.
研究沉香叶黄酮稳定性及对H2O2诱导HepG2细胞氧化损伤的保护作用。通过测定溶液吸光值,探讨温度、光照、金属离子、pH值对沉香叶黄酮稳定性的影响。建立H2O2诱导HepG2细胞氧化损伤模型,将细胞分为正常组、模型组以及沉香叶黄酮低、中、高剂量组,采用水溶性四唑盐法检测细胞增殖率,2′,7′-二氢二氯荧光黄双乙酸钠法(DCFH-DA)测定细胞内活性氧(ROS)水平,生化试剂盒检测细胞乳酸脱氢酶(LDH)释放量、丙二醛(MDA)含量、细胞内总超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-Px)和过氧化氢酶(CAT)活性。结果表明,沉香叶黄酮在温度70 ℃以下,pH 3~7范围内,Na+、K+、Mg2+、Ca2+金属离子中,黑暗环境与节能灯光中较稳定;能显著抑制H2O2造成的细胞死亡,降低胞内ROS与MDA的生成,减轻LDH向细胞外扩散程度,提高SOD、CAT、GSH-Px活性。沉香叶黄酮在低温、弱酸、部分金属离子、弱光条件下具有一定稳定性;对H2O2诱导HepG2氧化应激损伤具有保护作用,其作用可能与调节细胞氧化还原系统、清除胞内活性氧水平、提高细胞内抗氧化酶系活性有关。  相似文献   

10.
Opuntia humifusa, a member of the Cactaceae family widely distributed in the southern regions of the Korean peninsula, has potential bioactive functions and medicinal benefits. In the present study, we investigated the effect of hexane, ethyl acetate extracts and water partitioned fraction of O. humifusa on proliferation, G1 arrest and apoptosis in U87MG human glioblastoma cells. Glioblastoma cellular proliferation was evaluated using the MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay, and the effects of O. humifusa partitioned extracts on cell cycle and apoptosis were analyzed by flow cytometry. Our results revealed that when U87MG cells were treated with hexane extracts and water partitioned fraction of O. humifusa, the number of viable cells decreased in a concentration-dependent manner. In addition, water partitioned fractions of O. humifusa induced G1 arrest and non-apoptotic cell death as well as significant increases in ROS production in U87MG cells. In conclusion, water partitioned fractions of O. humifusa induce G1 arrest and inhibit U87MG human glioblastoma cell proliferation.  相似文献   

11.
As a biomaterial, besides excellent biocompatibility and biodegradability, suitable macropores and pores structure should be provided to guide cell extension and migration. In present study, the silk fibroin (SF) scaffold with uniaxial channels was prepared by directional temperature field freezing technique. The average pore diameter, pore density and porosity of the scaffold with oriented channels are ~128.7 µm, ~158 mm?2 and ~91.4 %, respectively. By controlling of the temperature gradient direction, the oriented multichannels of the scaffolds were formed in longitudinal easily. In process of the scaffolds fabrication, the directional growth of ice crystal could shear and draft to the silk fibroin molecule segments, which resulted in the new crystal nucleus formation in new zone and increase of β-sheet components in the scaffolds. In vitro, L929 cells were seeded on the scaffolds with oriented channels to evaluate the effect on cell behavior. Cell viability, adhesion and morphology were determined by methyl thiazolyl tetrazolium, confocal microscope and scanning electron microscope. The results showed that the cells anchored on the oriented channels, spread along the direction of the channels and hold a higher viability on the scaffolds with oriented channels. These new oriented multichannel scaffold could guide the adhesion and proliferation of L929 cells, which hold a potential in tissue engineering.  相似文献   

12.
Three new phomoxanthone compounds, phomolactonexanthones A (1), B (2) and deacetylphomoxanthone C (3), along with five known phomoxanthones, including dicerandrol A (4), dicerandrol B (5), dicerandrol (6), deacetylphomoxanthone B (7) and penexanthone A (8), were isolated in the metabolites of the fungus Phomopsis sp. HNY29-2B, which was isolated from the mangrove plants. The structures of compounds 1–3 were established on the basis of spectroscopic analysis. All compounds were evaluated against four human cancer cell lines including human breast MDA-MB-435, human colon HCT-116, human lung Calu-3 and human liver Huh7 by MTT assay. The compounds 4, 5, 7 and 8 showed cyctotoxic activities against tested cancer cell lines (IC50 < 10 μM).  相似文献   

13.
为研究茶多酚(TP)与油菜花粉提取物对前列腺癌细胞生长抑制效果的差异,应用细胞培养技术、MTT显色试验测定(IC50)法、荧光显微镜观察癌细胞形态变化。结果表明,除花粉醇溶性提取物(B)对前列腺癌细胞没有明显抑制作用外,花粉水溶性提取物(A)及A(B)与TP的等比例混合物均有显著的抑制作用,A、A+TP、B+TP和TP的IC50依次为1470、752、82、45 μg/mL,B+TP对PC-3的生长均有抑制作用,且成剂量效应和时间效应关系,经B+TP处理的PC-3癌细胞,用荧光显微镜可以看到癌细胞的凋亡小体。  相似文献   

14.
Glioblastoma, the most common and aggressive type of brain tumors, has devastatingly proliferative and invasive characteristics. The need for finding a novel and specific drug target is urgent as the current approaches have limited therapeutic effects in treating glioblastoma. Xyloketal B is a marine compound obtained from mangrove fungus Xylaria sp. (No. 2508) from the South China Sea, and has displayed antioxidant activity and protective effects on endothelial and neuronal oxidative injuries. In this study, we used a glioblastoma U251 cell line to (1) explore the effects of xyloketal B on cell viability, proliferation, and migration; and (2) investigate the underlying molecular mechanisms and signaling pathways. MTT assay, colony formation, wound healing, western blot, and patch clamp techniques were employed. We found that xyloketal B reduced cell viability, proliferation, and migration of U251 cells. In addition, xyloketal B decreased p-Akt and p-ERK1/2 protein expressions. Furthermore, xyloketal B blocked TRPM7 currents in HEK-293 cells overexpressing TRPM7. These effects were confirmed by using a TRPM7 inhibitor, carvacrol, in a parallel experiment. Our findings indicate that TRPM7-regulated PI3K/Akt and MEK/ERK signaling is involved in anti-proliferation and migration effects of xyloketal B on U251 cells, providing in vitro evidence for the marine compound xyloketal B to be a potential drug for treating glioblastoma.  相似文献   

15.
In this study, chitosan nanoparticles (HF-CD NPs) were synthesized by an ionic gelation method using negatively charged carboxymethyl-β-cyclodextrin and positively charged 2-hydroxypropyltrimethyl ammonium chloride chitosan bearing folic acid. The surface morphology of HF-CD NPs was spherical or oval, and they possessed relatively small particle size (192 ± 8 nm) and positive zeta potential (+20 ± 2 mV). Meanwhile, doxorubicin (Dox) was selected as model drug to investigate the prepared nanoparticles’ potential to serve as a drug delivery carrier. The drug loading efficiency of drug-loaded nanoparticles (HF-Dox-CD NPs) was 31.25%. In vitro release profiles showed that Dox release of nanoparticles represented a pH-sensitive sustained and controlled release characteristic. At the same time, the antioxidant activity of nanoparticles was measured, and chitosan nanoparticles possessed good antioxidant activity and could inhibit the lipid peroxidation inside the cell and avoid material infection. Notably, CCK-8 assay testified that the nanoparticles were safe drug carriers and significantly enhanced the antitumor activity of Dox. The nanoparticles possessed good antioxidant activity, pH-sensitive sustained controlled release, enhanced antitumor activity, and could be expected to serve as a drug carrier in future with broad application prospects.  相似文献   

16.
宋家乐 《茶叶科学》2012,32(6):539-542
探讨茯砖茶乙醇提取物(FTE)对H2O2诱发猪肾近曲小管LLC-PK1上皮细胞氧化损伤的保护作用.以不同质量浓度(10~200 μg/mL)的FTE预培养LLC-PK1细胞24 h后,换用含500 μmol/L H2O2的DMEM细胞培养液继续培养4h建立细胞氧化损伤模型.MTT法检测细胞生存率,硫代巴比妥酸比色法测定细胞内丙二醛(malondialdehyde,MDA)含量,比色法测定细胞内过氧化氢酶(catalase,CAT),超氧化物歧化酶(superoxide dismutase,SOD)和谷胱甘肽过氧化物酶(glutathione peroxidase,GSH-px)含量.经不同浓度FTE预处理24 h后,受损细胞的生存率上升,细胞内MDA生成量减少.同时,细胞内主要抗氧化酶(CAT、SOD和GSH-px)含量也较对照组增加,并呈剂量效应关系.结果提示,预先经FTE的保护可有效地对抗H2O2诱发的LLC-PK1细胞氧化损伤.  相似文献   

17.
In this study, we synthesed two kind of TiO2 nanomaterial (nanoparticles and nanofiber) for photocatalitic degradation of methyl orange (MO) as pollutant. TiO2 nanoparticles were synthesized by sol-gel technique using titanium (IV) isopropoxide as precursor. Polyvinyl acetate (PVAc)/TiO2 hybrid nanofibers were fabricated by combining sol-gel process with electrospinning technology, which consisted of PVAc as organic segment and TiO2 as inorganic part. Crystalline phase of TiO2 nanomaterials was investigated by X-ray diffraction (XRD). The XRD results show that the TiO2 nanomaterials crystallize in anatase with some rutile phase and these consist of titanium dioxide nano-crystals. The surface structures of TiO2 nanomaterials were examined using scanning electron microscopy (SEM). SEM scanning revealed that the nanoparticle and nanofibrous structure was formed. Fourier transform infrared spectroscopy (FTIR) was employed to analyze the chemical structures of the PVAc/TiO2 hybrid nanofibers. The FTIR analysis indicated the newly formed associated hydrogen bond because of the hybrid effect between PVAc and TiO2 sol. Finally, The photooxidative decomposition of methylene blue by using the titania nanomaterials was examined and compared.  相似文献   

18.
Background: Kinesin spindle protein (KSP) plays a critical role in mitosis. Inhibition of KSP function leads to cell cycle arrest at mitosis and ultimately to cell death. The aim of this study was to suppress KSP expression by specific small-interfering RNA (siRNA) in Hep3B cells and evaluate its anti-tumor activity. Methods: Three siRNA targeting KSP (KSP-siRNA #1-3) and one mismatched-siRNA (Cont-siRNA) were transfected into cells. Subsequently, KSP mRNA and protein levels, cell proliferation, and apoptosis were examined in both Hep3B cells and THLE-3 cells. In addition, the chemosensitivity of KSP-siRNA-treated Hep3B cells with doxorubicin was also investigated using cell proliferation and clonogenic survival assays. Results: The expression of endogenous KSP at both mRNA and protein levels in Hep3B cells was higher than in THLE-3 cells. In Hep3B cells, KSP-siRNA #2 showed a further downregulation of KSP as compared to KSP-siRNA #1 or KSP-siRNA #3. It also exhibited greater suppression of cell proliferation and induction of apoptosis than KSP-siRNA #1 or KSP-siRNA #3; this could be explained by the significant downregulation of cyclin D1, Bcl-2, and survivin. In contrast, KSP-siRNAs had no or lower effects on KSP expression, cell proliferation and apoptosis in THLE-3 cells. We also noticed that KSP-siRNA transfection could increase chemosensitivity to doxorubicin in Hep3B cells, even at low doses compared to control. Conclusion: Reducing the expression level of KSP, combined with drug treatment, yields promising results for eradicating hepatocellular carcinoma (HCC) cells in vitro. This study opens a new direction for liver cancer treatment. Key Words: Apoptosis, Chemosensitivity, Doxorubicin, Hepatocellular carcinoma (HCC) cells, Kinesin spindle protein (KSP)  相似文献   

19.
20.
Methacrylated gelatin films with in situ-generated TiO2 nanoparticles containing varying weight percentages of gelatin (0 %, 0.5 %, 1 %, 2 % and 4 %) were successfully prepared as novel biomaterials. 1H-NMR spectroscopy confirmed their methacrylation with a 79 % degree of substitution. TiO2 nanoparticles were uniformly distributed in the films with the average particle size increasing from 85 to 130 nm in proportion to an increase in TiO2 concentration from 0.5 to 4 wt%. The water absorption of various gelatin methacrylamide/TiO2 films was in the range of 471–758 %, which was enough to prevent wound beds from exudates accumulation. And in vitro degradation test in PBS showed that the three-dimensional structure of all samples basically remained unchanged although more than or nearly half the mass of specimens decreased after 4 weeks’ degradation, and the pH levels of all sample solutions were maintained in an adequate range of 6.5–7.4 for cell and tissue growth during the whole process. The antibacterial activities of the films against E. coli and S. aureus were measured via a shake flask test and demonstrated good performance after the importation of TiO2 nanoparticles. Cytotoxicity testing revealed that all films had no cytotoxicity and showed favorable adherence in the presence of L929 cells. The results suggest that hybrid hydrogel films hold potential for antibacterial wound dressing and tissue engineering scaffold applications.  相似文献   

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