The aim of this study was to investigate the cytotoxicity of modified nonequilibrium plasma with chlorhexidine digluconate（CHX） on human gingival fibroblasts（HGFs）, and to evaluate the biosecurity of modified none...The aim of this study was to investigate the cytotoxicity of modified nonequilibrium plasma with chlorhexidine digluconate（CHX） on human gingival fibroblasts（HGFs）, and to evaluate the biosecurity of modified nonequilibrium plasma with 2% CHX as a new method of root canal treatment. Tissue samples taken from human gingiva were primarily cultured and passaged. Cells from passages 3–7 were used. HGFs were treated by modified nonequilibrium plasma with 2% CHX for 0 min（control group）, 30 s, 1 min, 1.5 min, 3 min, 5 min, and 10 min, respectively, and then they were incubated for 0, 24, and 48 h. After that, cell counting kit-8（CCK-8） assay was applied to analyze the cytotoxicity of modified nonequilibrium plasma with 2% CHX on HGFs. There was no significant difference between the 0 h group treated with the modified nonequilibrium plasma for 1 min and the control group（P〉0.05）. However, there were significant differences between all the other treated groups and the control group（P〈0.05）. When treated for 1.5 min or shorter, the cell viability was obviously increased; while treated for 3 min or longer, it was obviously reduced. Moreover, when successively cultured for 0, 24, and 48 h, cell viability was decreased at first and then increased in the 3-min-treated and 5-min-treated groups. The modified nonequilibrium plasma with 2% CHX was of no influence on cell viability in 1.5 min treatment, and it could be safely used on root canal treatment.展开更多
The effect of collagen-chitosan membrane with different proportionate collagen and bFGF were investigated for culture human fibroblast. The optimum weight ratio of collagen/chitoson and bFGF were selected. Using cultu...The effect of collagen-chitosan membrane with different proportionate collagen and bFGF were investigated for culture human fibroblast. The optimum weight ratio of collagen/chitoson and bFGF were selected. Using culture human fibroblast technologies and cytotoxicity evaluated in vitro, Cell morphology was observed. Experimental results show that collagen-chitosan with bFGF promoted human fibroblast adhesion and supported cell proliferation for a long time. Furthermore collagen-chitosan membrane obviously degrade after 18d when human fibroblast was exhibited fusion spreading, compacting and stabilize. Cytotoxic to human fibroblast was revealed very low . Collagen- chitosan with bFGF should be useful as a tissue engineering biomaterial scaffold for cell culture.展开更多
This paper sets out to demonstrate that scraping of the flat dorsal surface of human dermis with a scalpel blade and cell plating without centrifugation can lead to the recognition and identification of the individual...This paper sets out to demonstrate that scraping of the flat dorsal surface of human dermis with a scalpel blade and cell plating without centrifugation can lead to the recognition and identification of the individual packing micro pattern of dermal reticular fibroblasts at confluence. The characteristic alignment of papillary and reticular fibroblasts at right angles to each other led to the positive identification of reticular fibroblasts. A non-enzymatic means of sub-culturing (passaging), which yields fully functional, healthy cells with normal, phenotypic morphology is also described. Implications for published subcutaneous wound healing studies are discussed as well as the confluent reticular fibroblast configuration, interpreted as ananatomic site identity code,which may be the address of a specific fibroblast gene pattern expression.展开更多
Fanconi anemia (FA) is a fatal heterogeneous autosomal recessive disorder, characterized by progressive bone marrow failure, congenital defect and cancer predisposition. Cell culture from FA fibroblast (FAF) displays ...Fanconi anemia (FA) is a fatal heterogeneous autosomal recessive disorder, characterized by progressive bone marrow failure, congenital defect and cancer predisposition. Cell culture from FA fibroblast (FAF) displays certain abnormalities as compared to normal human dermal fibroblast (NHDF). This prompted us to investigate the effect of a specific nutrient mixture (NM) containing ascorbic acid, lysine, proline and green tea extract, which has demonstrated a broad spectrum of pharmacological activities, on FAF compared to NHDF. We investigated the in vitro effect of NM on FAF and NHDF cell proliferation by MTT assay, MMPs secretion by zymography, morphology by H&E staining and apoptosis by green caspase assay. FAF (FA-A: PD20, FA-A: PD220) and NHDF were cultured in modified Dulbecco Eagle media. At near confluence, the cells were treated with different concentrations of NM (0, 50, 100, 250, 500 and 1000 μg/ml) in triplicate. The cells were also treated with PMA to induce MMP-9 activity. NM had no effect on FAF cell viability in both cell lines compared to control. In contrast NM exhibited 20% at 50 and 100, 50% at 250, 60% at 500 and 70% toxicity at 1000 μg/ml on NHDF cells. Zymography demonstrated MMP-2 and MMP-9 on PMA stimulation in FAF and NM inhibited the activity of both MMP-2 and MMP-9 in a dose response fashion with total block at 500 μg/ml. In contrast, NHDF exhibited only MMP-2, both active and inactive forms, and NM inhibited their activities in a dose-dependent manner with total block at 1000 μg/ml. H&E staining did not indicate any morphological changes in FAF nor induced apoptosis at higher concentrations, as seen by caspases assay. However, although no morphological changes in NHDF were noted up to NM 100 μg/ml, progressive changes in cell shrinkage, rounding and nuclear condensation, pertaining to apoptosis, were observed at higher concentrations. These changes were consistent with the results from the green caspases apoptosis assay. Our data demonstrate that NM exhibited different responses toward FAF and NHDF. This may in part be due to elevated chromosomal break, deletion and hypersensitivity to cross linking agents, a DNA repair disorder in FAF that is lacking in NHDF.展开更多
Objective:To exploretheserum-freecultureconditionsfordifferentiatingmouseembryonicstemcells(ES cells)intoneuralprecursorcells(NPC)andcomparetheeffectsof humanembryonicfibroblasts(HEF)as thefeederlayer of ES withthatof...Objective:To exploretheserum-freecultureconditionsfordifferentiatingmouseembryonicstemcells(ES cells)intoneuralprecursorcells(NPC)andcomparetheeffectsof humanembryonicfibroblasts(HEF)as thefeederlayer of ES withthatof mouseembryonicfibroblasts(MEF)in vitro.Methods:MouseES cellswereculturedin or notin feederlayer cellsmediumcontainingor notleukemiainhibitoryfactorto suppresstheirdifferentiation.Immunocytochemicalmethod was usedto identifyNPCby detectingnestinantigenandalkalinephosphatase.Results: TheES cellsculturedin HEF werepositiveto alkalinephosphatase.Serum-freemediumallowedthedifferentiationof ES cellsintoNPC.Conclusion:HEFcouldreplaceMEFandkeeptheundifferentiatedconditionof ES cellswithmorebenefits.NPCof highpuritycould be culturedfromEScellsby serum-freeculturemethod.展开更多
The human dermis presents an ongoing problem for regenerative medicine. Current medical management uses various acellular dermal matrices on wound sites. The challenge for scientists is to examine, then to question ac...The human dermis presents an ongoing problem for regenerative medicine. Current medical management uses various acellular dermal matrices on wound sites. The challenge for scientists is to examine, then to question accepted conventional wisdom and to present new concepts. In this paper, Autologous Cell Therapy will be described by using cell culture of autologous dermal fibroblasts and their extracellular matrix as a foundation for rebuilding the dermis in conditioned wound beds. This proposal seems to create a conflict with the medical approach to keeping a wound bed “moist”.展开更多
Oat contains different components that possess antioxidant properties;no study to date has addressed the antioxidant effect of the extract of oat bran on the cellular level.Therefore,the present study focuses on the i...Oat contains different components that possess antioxidant properties;no study to date has addressed the antioxidant effect of the extract of oat bran on the cellular level.Therefore,the present study focuses on the investigation of the protective effect of oat bran extract by enzymatic hydrolysates on human dermal fibroblast injury induced by hydrogen peroxide(H2O2).Kjeldahl determination,phenol-sulfuric acid method,and high-performance liquid chromatography(HPLC) analysis indicated that the enzymatic products of oat bran contain a protein amount of 71.93%,of which 97.43% are peptides with a molecular range from 438.56 to 1 301.01 Da.Assays for 1,1-diphenyl-2-picrylhydrazyl(DPPH) radical scavenging activity indicate that oat peptide-rich extract has a direct and concentration-dependent antioxidant activity.3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide(MTT) colorimetric assay and the TdT-mediated digoxigenin-dUTP nick-end labeling(TUNEL) assay for apoptosis showed that administration of H2O2 in human dermal fibroblasts caused cell damage and apoptosis.Pre-incubation of human dermal fibroblasts with the Oatp for 24 h markedly inhibited human dermal fibroblast injury induced by H2O2,but application oat peptides with H2O2 at same time did not.Pre-treatment of human dermal fibroblasts with Oatp significantly reversed the H2O2-induced decrease of superoxide dismutase(SOD) and the inhibition of malondialdehyde(MDA).The results demonstrate that oat peptides possess antioxidant activity and are effective against H2O2-induced human dermal fibroblast injury by the enhanced activity of SOD and decrease in MDA level.Our results suggest that oat bran will have the potential to be further explored as an antioxidant functional food in the prevention of aging-related skin injury.展开更多
基金supported by grants from the National Natural Science Foundation of China(No.81271189)the Hubei Provincial Science and Technology Support Program of China(No.2015BCE058)
文摘The aim of this study was to investigate the cytotoxicity of modified nonequilibrium plasma with chlorhexidine digluconate（CHX） on human gingival fibroblasts（HGFs）, and to evaluate the biosecurity of modified nonequilibrium plasma with 2% CHX as a new method of root canal treatment. Tissue samples taken from human gingiva were primarily cultured and passaged. Cells from passages 3–7 were used. HGFs were treated by modified nonequilibrium plasma with 2% CHX for 0 min（control group）, 30 s, 1 min, 1.5 min, 3 min, 5 min, and 10 min, respectively, and then they were incubated for 0, 24, and 48 h. After that, cell counting kit-8（CCK-8） assay was applied to analyze the cytotoxicity of modified nonequilibrium plasma with 2% CHX on HGFs. There was no significant difference between the 0 h group treated with the modified nonequilibrium plasma for 1 min and the control group（P〉0.05）. However, there were significant differences between all the other treated groups and the control group（P〈0.05）. When treated for 1.5 min or shorter, the cell viability was obviously increased; while treated for 3 min or longer, it was obviously reduced. Moreover, when successively cultured for 0, 24, and 48 h, cell viability was decreased at first and then increased in the 3-min-treated and 5-min-treated groups. The modified nonequilibrium plasma with 2% CHX was of no influence on cell viability in 1.5 min treatment, and it could be safely used on root canal treatment.
文摘The effect of collagen-chitosan membrane with different proportionate collagen and bFGF were investigated for culture human fibroblast. The optimum weight ratio of collagen/chitoson and bFGF were selected. Using culture human fibroblast technologies and cytotoxicity evaluated in vitro, Cell morphology was observed. Experimental results show that collagen-chitosan with bFGF promoted human fibroblast adhesion and supported cell proliferation for a long time. Furthermore collagen-chitosan membrane obviously degrade after 18d when human fibroblast was exhibited fusion spreading, compacting and stabilize. Cytotoxic to human fibroblast was revealed very low . Collagen- chitosan with bFGF should be useful as a tissue engineering biomaterial scaffold for cell culture.
文摘This paper sets out to demonstrate that scraping of the flat dorsal surface of human dermis with a scalpel blade and cell plating without centrifugation can lead to the recognition and identification of the individual packing micro pattern of dermal reticular fibroblasts at confluence. The characteristic alignment of papillary and reticular fibroblasts at right angles to each other led to the positive identification of reticular fibroblasts. A non-enzymatic means of sub-culturing (passaging), which yields fully functional, healthy cells with normal, phenotypic morphology is also described. Implications for published subcutaneous wound healing studies are discussed as well as the confluent reticular fibroblast configuration, interpreted as ananatomic site identity code,which may be the address of a specific fibroblast gene pattern expression.
文摘Fanconi anemia (FA) is a fatal heterogeneous autosomal recessive disorder, characterized by progressive bone marrow failure, congenital defect and cancer predisposition. Cell culture from FA fibroblast (FAF) displays certain abnormalities as compared to normal human dermal fibroblast (NHDF). This prompted us to investigate the effect of a specific nutrient mixture (NM) containing ascorbic acid, lysine, proline and green tea extract, which has demonstrated a broad spectrum of pharmacological activities, on FAF compared to NHDF. We investigated the in vitro effect of NM on FAF and NHDF cell proliferation by MTT assay, MMPs secretion by zymography, morphology by H&E staining and apoptosis by green caspase assay. FAF (FA-A: PD20, FA-A: PD220) and NHDF were cultured in modified Dulbecco Eagle media. At near confluence, the cells were treated with different concentrations of NM (0, 50, 100, 250, 500 and 1000 μg/ml) in triplicate. The cells were also treated with PMA to induce MMP-9 activity. NM had no effect on FAF cell viability in both cell lines compared to control. In contrast NM exhibited 20% at 50 and 100, 50% at 250, 60% at 500 and 70% toxicity at 1000 μg/ml on NHDF cells. Zymography demonstrated MMP-2 and MMP-9 on PMA stimulation in FAF and NM inhibited the activity of both MMP-2 and MMP-9 in a dose response fashion with total block at 500 μg/ml. In contrast, NHDF exhibited only MMP-2, both active and inactive forms, and NM inhibited their activities in a dose-dependent manner with total block at 1000 μg/ml. H&E staining did not indicate any morphological changes in FAF nor induced apoptosis at higher concentrations, as seen by caspases assay. However, although no morphological changes in NHDF were noted up to NM 100 μg/ml, progressive changes in cell shrinkage, rounding and nuclear condensation, pertaining to apoptosis, were observed at higher concentrations. These changes were consistent with the results from the green caspases apoptosis assay. Our data demonstrate that NM exhibited different responses toward FAF and NHDF. This may in part be due to elevated chromosomal break, deletion and hypersensitivity to cross linking agents, a DNA repair disorder in FAF that is lacking in NHDF.
文摘Objective:To exploretheserum-freecultureconditionsfordifferentiatingmouseembryonicstemcells(ES cells)intoneuralprecursorcells(NPC)andcomparetheeffectsof humanembryonicfibroblasts(HEF)as thefeederlayer of ES withthatof mouseembryonicfibroblasts(MEF)in vitro.Methods:MouseES cellswereculturedin or notin feederlayer cellsmediumcontainingor notleukemiainhibitoryfactorto suppresstheirdifferentiation.Immunocytochemicalmethod was usedto identifyNPCby detectingnestinantigenandalkalinephosphatase.Results: TheES cellsculturedin HEF werepositiveto alkalinephosphatase.Serum-freemediumallowedthedifferentiationof ES cellsintoNPC.Conclusion:HEFcouldreplaceMEFandkeeptheundifferentiatedconditionof ES cellswithmorebenefits.NPCof highpuritycould be culturedfromEScellsby serum-freeculturemethod.
文摘The human dermis presents an ongoing problem for regenerative medicine. Current medical management uses various acellular dermal matrices on wound sites. The challenge for scientists is to examine, then to question accepted conventional wisdom and to present new concepts. In this paper, Autologous Cell Therapy will be described by using cell culture of autologous dermal fibroblasts and their extracellular matrix as a foundation for rebuilding the dermis in conditioned wound beds. This proposal seems to create a conflict with the medical approach to keeping a wound bed “moist”.
文摘Oat contains different components that possess antioxidant properties;no study to date has addressed the antioxidant effect of the extract of oat bran on the cellular level.Therefore,the present study focuses on the investigation of the protective effect of oat bran extract by enzymatic hydrolysates on human dermal fibroblast injury induced by hydrogen peroxide(H2O2).Kjeldahl determination,phenol-sulfuric acid method,and high-performance liquid chromatography(HPLC) analysis indicated that the enzymatic products of oat bran contain a protein amount of 71.93%,of which 97.43% are peptides with a molecular range from 438.56 to 1 301.01 Da.Assays for 1,1-diphenyl-2-picrylhydrazyl(DPPH) radical scavenging activity indicate that oat peptide-rich extract has a direct and concentration-dependent antioxidant activity.3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide(MTT) colorimetric assay and the TdT-mediated digoxigenin-dUTP nick-end labeling(TUNEL) assay for apoptosis showed that administration of H2O2 in human dermal fibroblasts caused cell damage and apoptosis.Pre-incubation of human dermal fibroblasts with the Oatp for 24 h markedly inhibited human dermal fibroblast injury induced by H2O2,but application oat peptides with H2O2 at same time did not.Pre-treatment of human dermal fibroblasts with Oatp significantly reversed the H2O2-induced decrease of superoxide dismutase(SOD) and the inhibition of malondialdehyde(MDA).The results demonstrate that oat peptides possess antioxidant activity and are effective against H2O2-induced human dermal fibroblast injury by the enhanced activity of SOD and decrease in MDA level.Our results suggest that oat bran will have the potential to be further explored as an antioxidant functional food in the prevention of aging-related skin injury.
