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1.
Rev. Assoc. Med. Bras. (1992) ; 68(2): 159-164, Feb. 2022. tab, graf
Article in English | LILACS | ID: biblio-1365364

ABSTRACT

SUMMARY OBJECTIVE: The objective of this study was to explore the molecular mechanism underlying the occurrence of benign bile duct stricture and the target of low-dose paclitaxel in the prevention of benign bile duct stricture. METHODS: Under the stimulation of transforming growth factor beta 1, the expression of collagen type I and connective tissue growth factor were detected on isolated primary fibroblasts. The phosphorylation levels of JNK and Smad2L were detected using Western blot. The effect of low-dose paclitaxel on the transforming growth factor beta 1-induced inhibition of type I collagen and connective tissue growth factor expression and JNK and Smad2L phosphorylation was also observed. RESULTS: Transforming growth factor beta 1 induced the secretion of type I collagen and connective tissue growth factor as well as JNK phosphorylation in biliary fibroblasts. The JNK inhibitor or siRNA-Smad2 inhibited the transforming growth factor beta 1-induced secretion of type I collagen and connective tissue growth factor. Low-dose paclitaxel inhibited the expression of type I collagen induced by transforming growth factor beta 1 and may inhibit the secretion of collagen in biliary fibroblasts. CONCLUSION: The activation of JNK/Smad2L induced by transforming growth factor beta 1 is involved in the occurrence of benign bile duct stricture that is mediated by the overexpression of type I collagen and connective tissue growth factor, and low-dose paclitaxel may inhibit the phosphorylation of JNK/Smad2L.


Subject(s)
Humans , Paclitaxel/pharmacology , Collagen , MAP Kinase Signaling System , Collagen Type I/metabolism , Collagen Type I/pharmacology , Smad2 Protein , Fibroblasts/metabolism
2.
Electron. j. biotechnol ; 52: 67-75, July. 2021. tab, graf, ilus
Article in English | LILACS | ID: biblio-1283594

ABSTRACT

BACKGROUND: Adipogenesis and fibrogenesis can be considered as a competitive process in muscle, which may affect the intramuscular fat deposition. The CCAAT/enhancer-binding protein beta (C/EBPb) plays an important role in adipogenesis, which is well-characterized in mice, but little known in bovine so far. RESULTS: In this study, real-time qPCR revealed that the level of C/EBPb was increased during the developmental stages of bovine and adipogenesis process of preadipocytes. Overexpression of C/EBPb promoted bovine fibroblast proliferation through mitotic clonal expansion (MCE), a necessary process for initiating adipogenesis, by significantly downregulating levels of p21 and p27 (p < 0.01). Also, the PPARc expression was inhibited during the MCE stage (p < 0.01). 31.28% of transfected fibroblasts adopted lipid-laden adipocyte morphology after 8 d. Real-time qPCR showed that C/EBPb activated the transcription of early stage adipogenesis markers C/EBPa and PPARc. Expression of ACCa, FASN, FABP4 and LPL was also significantly upregulated, while the expression of LEPR was weakened. CONCLUSIONS: It was concluded C/EBPb can convert bovine fibroblasts into adipocytes without hormone induction by initiating the MCE process and promoting adipogenic genes expression, which may provide new insights into the potential functions of C/EBPb in regulating intramuscular fat deposition in beef cattle.


Subject(s)
Cattle/metabolism , Adipocytes/metabolism , CCAAT-Enhancer-Binding Protein-beta/metabolism , Fibroblasts/metabolism , Adipose Tissue/metabolism , Clone Cells , Cell Proliferation , Adipogenesis , Real-Time Polymerase Chain Reaction , Mitosis , Muscles
3.
Rev. bras. oftalmol ; 80(1): 17-20, jan.-fev. 2021. tab
Article in English | LILACS | ID: biblio-1251318

ABSTRACT

ABSTRACT Objective: To observe clinically, in rabbits, the side effects of topical injection of subconjunctival cyclophosphamide, studying its role as an antifibrotic drug. Methods: Prospective study in 20 albino rabbits of New Zealand race. All rabbits were treated with cyclophosphamide, 10mg/ml in a volume of 0.3 ml, in the left eye through subconjunctival injection. They were evaluated for 1, 7, 30, and 60 days after the procedure. All the animals were examined for the detection of ocular reactions such as necrosis, hyperemia, chemosis, secretion, opacity, and iritis. Other side effects as changes in the behavior, in the feed, and the water consumption were also evaluated. Results: It was observed that from the 20 rabbits studied, three rabbits (15%) showed side effects only at the 24 hours analysis. One rabbit (5%) presented hyperemia, one rabbit (5%) had hyperemia associated with iritis, and one rabbit (5%) presented hyperemia associated with secretion. These reactions were not observed at 1, 7, 30, and 60 days. Conclusion: Cyclophosphamide subconjunctival injection induces minor side effects on the conjunctiva of rabbits such as hyperemia, associated with iritis and secretion.


RESUMO Objetivo: Observar clinicamente os efeitos colaterais de injeção subconjuntival de ciclofosfamida, pensando em sua ação como um agente antifibrótico. Métodos: Estudo prospectivo realizado com 20 coelhos albinos da raça Nova Zelândia. Todos os coelhos foram submetidos a 0,3 ml de injeção subconjuntival de ciclofosfamida 10mg/ml no olho esquerdo e foram avaliados de acordo com os efeitos locais no primeiro dia após a injeção, 7, 30 e 60 dias. Foram examinados para detecção de reações oculares como necrose, hiperemia, quemose, secreção, opacidade corneana, irite além de alterações comportamentais e variação no consumo de água e alimentação. Resultados: Dos 20 coelhos estudados, apenas 3 apresentaram reações oculares e somente na leitura de 24 horas. Um coelho (5%) apresentou hiperemia, 1 coelho (5%) apresentou hiperemia associada a presença de irite e 1 coelho (5%) apresentou hiperemia associada a presença de secreção. As reações não foram mais observadas durante os exames de 7, 30 e 60 dias. Conclusão: A ciclofosfamida subconjuntival causou poucos efeitos colaterais na conjuntiva dos coelhos. Os únicos efeitos encontrados foram hiperemia, irite e secreção.


Subject(s)
Animals , Rabbits , Fibrosis/prevention & control , Conjunctiva/drug effects , Cyclophosphamide/adverse effects , Cyclophosphamide/pharmacology , Wound Healing/drug effects , Prospective Studies , Mitomycin/pharmacology , Cyclophosphamide/administration & dosage , Injections, Intraocular , Fibroblasts/drug effects , Fibroblasts/metabolism , Slit Lamp Microscopy
4.
Braz. j. med. biol. res ; 54(10): e11156, 2021. graf
Article in English | LILACS | ID: biblio-1285646

ABSTRACT

The objective of this study was to investigate the effect of human esophageal fibroblast-derived exosomal miR-21 on cisplatin sensitivity against esophageal squamous EC9706 cells. EC9706 cells were co-cultured indirectly with human esophageal fibroblasts (HEF) or miR-21 mimics transfected-HEF in the transwell system. The exosomes in HEF-culture conditioned medium were extracted by differential ultracentrifugation. EC9706 cells were co-cultured with HEF-derived exosomes directly. The cisplatin sensitivity against EC9706 cells was revealed via half maximal inhibitory concentration (IC50) values using MTT assay. The expressions of miR-21, programmed cell death 4 (PDCD4) mRNA, and gene of phosphate and tension homology deleted on chromosome ten (PTEN) mRNA were determined by qRT-PCR. The changes of the protein level were detected using western blot assay. IC50 values of cisplatin against EC9706 cells were increased after EC9706 cells were co-cultured with either HEF or exosomes derived from miR-21 mimics-transfected HEF. Following the increased level of miR-21, the mRNA expression and protein levels of PTEN and PDCD4 were decreased in EC9706 cells. The cisplatin sensitivity to EC9706 cells was reduced by HEF-derived exosomal miR-21 through targeting PTEN and PDCD4. This study suggested that non-tumor cells in the tumor micro-environment increased the tumor anti-chemotherapy effects through their exosomes.


Subject(s)
Humans , Esophageal Neoplasms/genetics , Esophageal Neoplasms/drug therapy , Carcinoma , MicroRNAs/genetics , Cisplatin/pharmacology , RNA-Binding Proteins , Apoptosis , Cell Line, Tumor , Cell Proliferation , Apoptosis Regulatory Proteins/metabolism , Tumor Microenvironment , Fibroblasts/metabolism
5.
Article in Chinese | WPRIM | ID: wpr-879937

ABSTRACT

OBJECTIVE@#To investigate the role of IL-17A in promoting the activation of lung fibroblasts and the secretion of chemokine CXCL12, and to explore the possible mechanism.@*METHODS@#Lung tissues of BALB/c mice were collected after intraperitoneal injection of recombinant mouse IL-17A (rmIL-17A). Real-time RT-PCR and Western blotting were used to detect the expression levels of α-smooth muscle actin (α-SMA) and collagen I in lung tissues, and immunohistochemical staining and real-time RT-PCR were used to determine the expression of CXCL12. Normal mouse primary lung fibroblasts were isolated and cultured, and identified by immunofluorescence staining with optical microscopy. Cells and supernatant of culture medium were collected after stimulation with rmIL-17A at different concentrations. mRNA levels of α-SMA, collagen I, and CXCL12 in the cells were determined by real-time RT-PCR, and the levels of collagen I and CXCL12 in the supernatant of culture medium were determined by ELISA.@*RESULTS@#The mRNA and protein levels of α-SMA and collagen I in the lung tissue of mice injected with rmIL-17A were significantly increased compared with the control group (all @*CONCLUSIONS@#s: IL-17A can promote the activation of lung fibroblasts and translation into myofibroblast. The secretion of collagen is increased, which promote the deposition of extracullular matrix, and leads to the occurrence and development of lung fibrosis. CXCL12, a chemokine secreted by activated fibroblasts, may be involved in this process.


Subject(s)
Actins/genetics , Animals , Cells, Cultured , Chemokine CXCL12/metabolism , Fibroblasts/metabolism , Interleukin-17/pharmacology , Lung/metabolism , Mice , Mice, Inbred BALB C
6.
Braz. oral res. (Online) ; 34: e013, 2020. graf
Article in English | LILACS | ID: biblio-1089379

ABSTRACT

Abstract This study evaluated the effect of a cyclopentenone-type PG, 15-Deoxy-Δ12,14-PG J2 (15d-PGJ2), and lectin (ScLL) on the viability of human gingival fibroblasts (HGFs), and on IL-6 and TGFβ-1 release by these fibroblasts, stimulated with lipopolysaccharide (LPS). HGFs were stimulated with LPS 10 μg/ml and treated with 15d-PGJ2 1 and 2 μg/ml, and ScLL 2 and 5 μg/ml, for 1 and 3h, and then evaluated for viability by MTT assay. Supernatant was collected to detect IL-6 and TGFβ-1 release, by ELISA. Positive control was cells kept in Dulbecco's Modified Eagle's Medium, and negative control was those kept in LPS. Data were analyzed by ANOVA and Dunnett's test (α = 0.05). No significant difference was found in viability among experimental groups at 1h (p > 0.05). Percentage of ScLL 5 µg/ml viable cells was similar to that of positive control at evaluated periods (p > 0.05), whereas the other groups had lower levels than the positive control (p < 0.05). IL-6 release was statistically higher for ScLL 5 μg/ml and 15d-PGJ2 2 µg/ml at 1h, compared with the other treated groups and positive control (p < 0.05). No significant differences were found among the groups at 3h (p > 0.05), except for ScLL 2 µg/ml and 15d-PGJ2 1 µg/ml, which showed lower IL-6 release compared with that of negative control (p < 0.05). No significant difference was found among the groups for TGFβ-1 release (p > 0.05). Results indicated that ScLL 5 μg/ml did not interfere in viability, and ScLL 2 µg/ml and 15d-PGJ2 1 µg/ml demonstrated reduced IL-6 release. Tested substances had no effect on TGFβ-1 release.


Subject(s)
Humans , Prostaglandin D2/analogs & derivatives , Lipopolysaccharides/pharmacology , Interleukin-6/metabolism , Plant Lectins/pharmacology , Transforming Growth Factor beta1/metabolism , Fibroblasts/drug effects , Fibroblasts/metabolism , Reference Values , Time Factors , Enzyme-Linked Immunosorbent Assay , Prostaglandin D2/pharmacology , Cell Survival/drug effects , Cells, Cultured , Analysis of Variance , Statistics, Nonparametric , Transforming Growth Factor beta1/drug effects , Gingiva/cytology
7.
Braz. j. med. biol. res ; 52(9): e8551, 2019. graf
Article in English | LILACS | ID: biblio-1019565

ABSTRACT

Fibroblasts are a highly heterogeneous population of cells, being found in a large number of different tissues. These cells produce the extracellular matrix, which is essential to preserve structural integrity of connective tissues. Fibroblasts are frequently engaged in migration and remodeling, exerting traction forces in the extracellular matrix, which is crucial for matrix deposition and wound healing. In addition, previous studies performed on primary myoblasts suggest that the E3 ligase MuRF2 might function as a cytoskeleton adaptor. Here, we hypothesized that MuRF2 also plays a functional role in skeletal muscle fibroblasts. We found that skeletal muscle fibroblasts express MuRF2 and its siRNA knock-down promoted decreased fibroblast migration, cell border accumulation of polymerized actin, and down-regulation of the phospho-Akt expression. Our results indicated that MuRF2 was necessary to maintain the actin cytoskeleton functionality in skeletal muscle fibroblasts via Akt activity and exerted an important role in extracellular matrix remodeling in the skeletal muscle tissue.


Subject(s)
Animals , Rats , Cell Differentiation/physiology , Muscle, Skeletal/physiology , Ubiquitin-Protein Ligases/physiology , Cell Proliferation/physiology , Fibroblasts/physiology , Muscle Proteins/physiology , Blotting, Western , Fluorescent Antibody Technique , Muscle, Skeletal/metabolism , Ubiquitin-Protein Ligases/metabolism , Fibroblasts/metabolism , Muscle Proteins/metabolism
8.
Biol. Res ; 52: 23, 2019. graf
Article in English | LILACS | ID: biblio-1011425

ABSTRACT

BACKGROUND: Conjunctival filtering bleb scar formation is the main reason for the failure of glaucoma filtration surgery. Cytoglobin (Cygb) has been reported to play an important role in extracellular matrix (ECM) remodeling, fibrosis and tissue damage repairing. This study aimed to investigate the role of Cygb in anti-scarring during excessive conjunctival wound healing after glaucoma filtration surgery. METHODS: Cygb was overexpressed in human tenon fibroblasts (hTFs) by transfecting hTFs with lentiviral particles encoding pLenti6.2-FLAG-Cygb. Changes in the mRNA and protein levels of fibronectin, collagen I, collagen III, TGF-ß1, and HIF1α were determined by RT-PCR and western blotting respectively. RESULTS: After Cygb overexpression, hTFs displayed no significant changes in visual appearance and cell counts compared to controls. Whereas, Cygb overexpression significantly decreased the mRNA and protein expression levels of collagen I, collagen III and fibronectin compared with control (p < 0.01). There was also a statistically significant decrease in the mRNA and protein levels of TGF-ß1 and HIF-1α in hTFs with overexpressed Cygb compared with control group (p < 0.05). CONCLUSION: Our study provided evidence that overexpression of Cygb decreased the expression levels of fibronectin, collagen I, collagen III, TGF-ß1 and HIF-1α in hTFs. Therefore, therapies targeting Cygb expression in hTFs may pave a new way for clinicians to solve the problem of post-glaucoma surgery scarring.


Subject(s)
Humans , Extracellular Matrix/metabolism , Tenon Capsule/metabolism , Fibroblasts/metabolism , Cytoglobin/metabolism , RNA, Messenger/analysis , Collagen/analysis , Fibronectins/analysis , Vascular Endothelial Growth Factor A/metabolism , Extracellular Matrix/drug effects , Cytoglobin/pharmacology
9.
Acta cir. bras ; 33(2): 144-155, Feb. 2018. graf
Article in English | LILACS | ID: biblio-886256

ABSTRACT

Abstract Purpose: To evaluate the efficacy of the application of the human amniotic membrane (HAM) on the inflammatory process, fibroblast proliferation, formation of collagenand reduction of skin wound areas in rats. Methods: Thirty six rats were submitted to a surgical injury induction and divided into two groups (n = 18): group C (control) and T (treated with the HAM). The macroscopic evolution in the wound area and the histological characteristics of the skin samples were evaluated. Results: The regression of the wound area was greater in group T. The histological analysis revealed a significant reduction (p < 0.05) in the inflammatory infiltrate in group T at all experimental periods compared with that in the control group. Furthermore, the group T presented a significant increase in the proliferation of fibroblasts at 14 and 21 days compared with group C (p < 0.05). Regarding the deposition of mature collagen fibers, there was an increase in the replacement of type III collagen by type I collagen in group T (p < 0.05). Conclusion: Treatment with the HAM reduced the healing time as well as the inflammatory responses, increased the proliferation of fibroblasts, and induced a higher concentration of mature collagen fibers.


Subject(s)
Humans , Animals , Male , Rats , Skin/injuries , Wound Healing/physiology , Biological Dressings , Collagen/pharmacology , Amnion/transplantation , Skin/pathology , Wound Healing/drug effects , Random Allocation , Rats, Wistar , Collagen Type I/metabolism , Collagen Type I/pharmacology , Collagen Type III/metabolism , Collagen Type III/pharmacology , Disease Models, Animal , Fibroblasts/metabolism , Fibroblasts/pathology , Amnion/chemistry , Inflammation/metabolism
10.
Yonsei Medical Journal ; : 85-91, 2018.
Article in English | WPRIM | ID: wpr-742500

ABSTRACT

PURPOSE: Ascorbic acid has been reported to have an adipogenic effect on 3T3-L1 preadipocytes, while evidence also suggests that ascorbic acid reduces body weight in humans. In this study, we tested the effects of ascorbic acid on adipogenesis and the balance of lipid accumulation in ovariectomized rats, in addition to long-term culture of differentiated 3T3-L1 adipocytes. MATERIALS AND METHODS: Murine 3T3-L1 fibroblasts and ovariectomized rats were treated with ascorbic acid at various time points. In vitro adipogenesis was analyzed by Oil Red O staining, and in vivo body fat was measured by a body composition analyzer using nuclear magnetic resonance. RESULTS: When ascorbic acid was applied during an early time point in 3T3-L1 preadipocyte differentiation and after bilateral ovariectomy (OVX) in rats, adipogenesis and fat mass gain significantly increased, respectively. However, lipid accumulation in well-differentiated 3T3-L1 adipocytes showed a significant reduction when ascorbic acid was applied after differentiation (10 days after induction). Also, oral ascorbic acid administration 4 weeks after OVX in rats significantly reduced both body weight and subcutaneous fat layer. In comparison to the results of ascorbic acid, which is a well-known cofactor for an enzyme of collagen synthesis, and the antioxidant ramalin, a potent antioxidant but not a cofactor, showed only a lipolytic effect in well-differentiated 3T3-L1 adipocytes, not an adipogenic effect. CONCLUSION: Taking these results into account, we concluded that ascorbic acid has both an adipogenic effect as a cofactor of an enzymatic process and a lipolytic effect as an antioxidant.


Subject(s)
3T3-L1 Cells , Adipocytes/drug effects , Adipocytes/metabolism , Adipogenesis/drug effects , Animals , Antioxidants/pharmacology , Ascorbic Acid/pharmacology , Body Composition/drug effects , Body Weight/drug effects , Cell Differentiation/drug effects , Female , Fibroblasts/drug effects , Fibroblasts/metabolism , Lipolysis/drug effects , Mice , Ovariectomy , Rats, Sprague-Dawley
11.
Acta cir. bras ; 32(5): 359-368, May 2017. tab, graf
Article in English | LILACS | ID: biblio-837709

ABSTRACT

Abstract Purpose: To evaluate the changes of caveolin-1 in lung fibroblasts in newborn Wistar rats when exposed to hyperoxic conditions, as well as lung fibroblasts cell cycle. Methods: One hundred newborn Wistar rats were randomly divided (50 rats/group) into experimental and control groups, exposed to hyperoxic conditions or normal air, respectively. The fraction of inspired oxygen (FiO2) in the experimental group was 90%, whereas this value was 21% in the control group. Lung fibroblasts were collected on days 3, 7, and 14 of the experiment. Caveolin-1 expression dynamics in lung fibroblasts was assayed in each group by immunofluorescence and Western blot analyses. Flow cytometry (FCM) was used to assess the proportions of lung fibroblasts at different stages of the cell cycle. Results: On day 3, no significant difference in caveolin-1 expression was observed between the hyperoxic and control groups; however, on days 7 and 14, caveolin-1 expression was significantly lower in the hyperoxic group than in the control (P<0.05). No apparent differences were observed in caveolin-1 expression in the control group at the different time points. Using FCM analysis, we showed that the proportion of lung fibroblasts in G0/G1 phase in the hyperoxic group decreased compared to that of the control group on day 7, while the proportion of S-phase cells increased (P<0.05). These differences were more significant when the groups were compared on day 14 (P<0.01). Conclusion: After seven days the exposure to hyperoxic conditions, lung fibroblasts proliferated and caveolin-1 expression decreased.


Subject(s)
Animals , Female , Cell Proliferation , Caveolin 1/metabolism , Fibroblasts/metabolism , Lung/metabolism , Lung Diseases/metabolism , Oxygen/pharmacology , Random Allocation , Cell Cycle , Cells, Cultured , Chronic Disease , Rats, Wistar , Hyperoxia , Models, Animal , Caveolin 1/pharmacology , Fibroblasts/cytology , Fibroblasts/drug effects , Lung/cytology , Lung/drug effects , Lung Diseases/classification , Lung Diseases/chemically induced , Animals, Newborn
12.
Biol. Res ; 50: 22, 2017. graf
Article in English | LILACS | ID: biblio-950873

ABSTRACT

BACKGROUND: Hypertrophic scarring (HS) is a severe disease, and results from unusual wound healing. Col1A1 could promote the hypertrophic scar formation, and the expression of Col1A1 in HS tissue was markedly higher than that in the normal. In present study, we aimed to identify miRNAs as post-transcriptional regulators of Col1A1 in HS. METHODS: MicroRNA-98 was selected as the key miRNA comprised in HS. The mRNA levels of miR-98 in HS tissues and the matched normal skin tissues were determined by qRT-PCR. MTT and flow cytometry were used to determine the influence of miR-98 on cell proliferation and apoptosis of HSFBs, respectively. Col1A1 was found to be the target gene of miR-98 using luciferase reporter assay. Luciferase assay was performed to determine the relative luciferase activity in mimic NC, miR-98 mimic, inhibitor NC and miR-98 inhibitor with Col1A13'-UTR wt or Col1A13'-UTR mt reporter plasmids. The protein expression of Col1A1 in HSFBs after transfection with mimic NC, miR-98 mimic, inhibitor NC and miR-98 inhibitor were determined by western blotting. RESULTS: The mRNA level of miR-98 in HS tissues was much lower than that in the control. Transfection of HSFBs with a miR-98 mimic reduced the cell viability of HSFBs and increased the apoptosis portion of HSFBs, while inhibition of miR-98 increased cell viability and decreased apoptosis portion of HSFBs. miR-98 inhibitor increased the relative luciferase activity significantly when cotransfected with the Col1A1-UTR reporter plasmid, while the mutant reporter plasmid abolished the miR-98 inhibitor-mediated increase in luciferase activity. Western blotting revealed that overex-pression of miR-98 decreased the expression of Col1A1. CONCLUSIONS: Overexpression of miR-98 repressed the proliferation of HSFBs by targeting Col1A1.


Subject(s)
Humans , RNA Processing, Post-Transcriptional/genetics , Apoptosis/genetics , Collagen Type I/metabolism , MicroRNAs/genetics , Fibroblasts/metabolism , Case-Control Studies , Down-Regulation , Cicatrix, Hypertrophic/genetics , Cicatrix, Hypertrophic/metabolism , Collagen Type I/genetics , MicroRNAs/metabolism , Cell Proliferation
13.
Int. j. odontostomatol. (Print) ; 10(3): 449-454, dic. 2016. ilus
Article in Spanish | LILACS | ID: biblio-840994

ABSTRACT

El objetivo de este estudio fue evaluar el efecto de la radiación ultravioleta (UV) B sobre la expresión del factor de crecimiento transformante (TGF) ß1 por fibroblastos de mucosa oral, con el objetivo de dilucidar si este tipo celular puede contribuir a la expresión de TGFß1 en bermellón labial sobreexpuesto a la radiación UV. Se obtuvieron cultivos primarios de fibroblastos desde explantes de mucosa bucal, los que fueron irradiados con una dosis única de luz UVB (60 mJ/cm2). Se midió proliferación celular con el método MTT, y la expresión de TGFß1, a nivel de ARN mensajero (normalizado a GAPDH) por RT-PCR y a nivel de proteína mediante inmunofluorescencia. Se observó una disminución de la proliferación celular de los fibroblastos de mucosa oral a las 24 hrs post-irradiación en relación a los fibroblastos no irradiados (P<0,05, Mann Whitney). No se encontraron diferencias entre los fibroblastos control y los irradiados en la expresión de TGFß-1 ni a nivel de mensajero (0,5 y 6 h post-irradiación), ni de proteína (24 h post-irradiación). Los resultados sugieren que los fibroblastos de mucosa oral presentan una disminución de su proliferación en respuesta a una dosis única de radiación UVB, sin que se afecte la expresión de TGFß-1, la que fue similar a los fibroblastos no irradiados. Esto sugiere que los fibroblastos contribuirían a la producción de TGFß-1 en respuesta a la exposición crónica a UVB del bermellón labial.


The objective of this study was to characterize the effect of Ultraviolet (UV) B irradiation on the expression of transforming growth factor (TGF) ß1 by oral mucosa fibroblasts, in order to assess if these cells contribute to the production of TGFß-1 in UV-irradiated lip vermillion. Primary cultures of fibroblasts were obtained from oral mucosa explants, and were irradiated with a single dose of UVB light (60 mJ/cm2). The effects of UVB radiation on cell proliferation was evaluated by the MTT method. The effects of UVB on the expression of TGF-ß1 was analyzed by RT-PCR (normalized to GAPDH) and by immunofluorescence. The results showed a decrease in the proliferation of UVB-irradiated fibroblasts as compared to controls at 24h post-irradiation (p<0.05). No variations in the expression of TGFß1, both at the mRNA and protein level, were observed between control and UVB-irradiated fibroblasts during the first 24 h after irradiation. Oral mucosa fibroblasts have reduced proliferation in response to a single dose of UVB, but their expression of TGFß1 was not affected. This suggests that oral mucosa fibroblasts may contribute to the production of TGFß1 in the lip vermillion independent of UVB exposure.


Subject(s)
Humans , Mouth Mucosa/metabolism , Mouth Mucosa/radiation effects , Transforming Growth Factor beta/radiation effects , Ultraviolet Rays , Cell Proliferation , Fibroblasts/metabolism , Transforming Growth Factor beta/metabolism
14.
An. bras. dermatol ; 91(5): 589-594, Sept.-Oct. 2016. tab, graf
Article in English | LILACS | ID: biblio-827757

ABSTRACT

Abstract: Background: The morphological similarities between fibrous papules of the face and multiple sporadic oral fibromas were mentioned long ago and a relationship between them has been reported in the literature. Objective: The aim of this study was to evaluate the participation of mast cells, elastin and collagen in a series of oral fibromas and fibrous papules of the face in order to better understand the possible role of these factors in fibrosis and the formation of these lesions. Methods: Thirty cases of oral fibroma involving the buccal mucosa and 30 cases of fibrous papules of the face were selected. Tissue samples were submitted to picrosirius red staining and immunohistochemistry using anti-elastin and anti-tryptase antibodies. Results: The percentage of tryptase-positive mast cells and expression of elastin were higher in cases of fibrous papules of the face (p < 0.05). In contrast, a higher intensity of collagen deposition was observed in oral fibromas. The results showed mast cell accumulation and higher elastin synthesis in fibrous papules of the face, and mast cell accumulation with higher collagen fiber synthesis in oral fibromas. Conclusion: These findings support the hypothesis that mast cells influence the development and growth of these lesions through different mechanisms.


Subject(s)
Humans , Facial Dermatoses/pathology , Fibroma/pathology , Fibrosis/metabolism , Immunohistochemistry , Collagen/metabolism , Elastin/metabolism , Tryptases/metabolism , Facial Dermatoses/metabolism , Fibroblasts/metabolism , Fibroma/metabolism , Mast Cells/metabolism , Mouth Mucosa/metabolism
15.
Int. j. odontostomatol. (Print) ; 10(2): 237-242, ago. 2016. ilus
Article in English | LILACS | ID: lil-794482

ABSTRACT

The objective of this study was to determine the effects of coating nanoparticles of titanium dioxide (TiO2 NPs) and irradiation -UV on plates of titanium (Ti) for the adhesion and proliferation of human gingival fibroblasts (HGF). A total of 15 Ti plates were divided into three groups (n = 5); (i) control Ti, (ii) experimental: Ti+TiO2 NPs, (iii) experimental: Ti+TiO2 NPs+UV. The plates were analyzed with atomic force microscopy (AFM) and the roughness (Ra and Rmax) was determined. UV irradiation was performed for 20 min. HGF were subcultured in DMEM+10 % fetal bovine serum (FBS) at 37 °C with 5 % CO2. 2x106 cells/mL were inoculated on the plates and incubated for 1 h and washed with phosphate buffer saline (PBS). In the case of cell proliferation, cells were incubated for further 24 h more. Cell viability was determined with the MTT method, the formazan was dissolved with dimethylsulfoxide (DMSO) and analyzed at 540 nm. Experiments were performed of three independent experiments and data were analyzed by Kruskall-Wallis and multiple comparison of Mann-Whitney test. The surface topography of samples corresponded as follow: Ti (Ra= 0,492 µm y Rms= 0.640 µm), Ti+NPs TiO2, (Ra= 0.55 µm y Rms= 0.714 µm), respectively. The coating with TiO2 NPs significantly (p <0.05) increased the adhesion and proliferation of HGF compared with the group. The modification of Ti plates by coated with TiO2 NPs significantly increased adhesion and proliferation of HGF with the formation of a hydrophilic surface which favors the humectancy. This treatment may be reported here convenient to accelerate osseointegration of dental implants based titanium.


El objetivo fue determinar los efectos del recubrimiento con nanopartículas de dióxido de titanio (TiO2 NPs) e irradiación UV sobre placas de titanio (Ti) para la adhesión y proliferación de fibroblastos gingivales humanos (FGH). Un total de 15 placas de Ti se dividieron en tres grupos (n= 5); (i) control Ti, (ii) experimental Ti+NPs TiO2, (iii) experimental: Ti+NPs TiO2+UV. Las placas fueron analizadas en microscopía de fuerza atómica (MFA) y se determinó la rugosidad (Ra y Rmax). La irradiación con UV se realizó durante 20 min. FGH fueron subcultivados en DMEM+10 % de suero fetal bovino a 37 °C con 5 % de CO2. 2x106 células/mL fueron inoculadas sobre las placas e incubadas durante 1 h, se lavaron con solución salina de buffer fosfato. En el caso de la proliferación celular, las células se incubaron por 24 h más. La viabilidad celular se determinó con el método de MTT, el formazan fue disuelto con dimetilsulfoxido y se analizó a 540 nm. Los experimentos se realizaron a partir de tres experimentos independientes y los datos se analizaron por Kruskall-Wallis y por comparación múltiple de Mann-Whitney. La topografía de la superficie de las muestras correspondio de la siguiente manera: Ti (Ra= 0,492 µm y Rms= 0,640 µm), Ti+NPs TiO2, (Ra= 0,55 µm y Rms= 0,714 µm), respectivamente. El recubrimiento con NPs TiO2 aumentó significativamente la adhesión y proliferación de HGF en comparación con el grupo de Ti control (p <0,05). La modificación de la superficie de las placas de Ti recubiertas con NPs TiO2 aumentó significativamente la adhesión y proliferación de HGF con la formación de una superficie hidrófila que favorece la humectancia. Este tratamiento aquí informado tal vez sea un método conveniente para acelerar el proceso de la osteointegración de los implantes dentales a base de titanio.


Subject(s)
Humans , Cell Proliferation/physiology , Fibroblasts/metabolism , Gingiva/metabolism , Titanium , Ultraviolet Rays , Cell Adhesion , Nanoparticles
16.
Yonsei Medical Journal ; : 557-564, 2016.
Article in English | WPRIM | ID: wpr-52546

ABSTRACT

PURPOSE: Periostin mediates critical steps in gastric cancer and is involved in various signaling pathways. However, the roles of periostin in promoting gastric cancer metastasis are not clear. The aim of this study was to investigate the relevance between periostin expression and gastric cancer progression and the role of stress-related hormones in the regulation of cancer development and progression. MATERIALS AND METHODS: Normal, cancerous and metastatic gastric tissues were collected from patients diagnosed with advanced gastric cancer. The in vivo expression of periostin was evaluated by in situ hybridization and immunofluorescent staining. Meanwhile, human gastric adenocarcinoma cell lines MKN-45 and BGC-803 were used to detect the in vitro expression of periostin by using quantitative real-time polymerase chain reaction (PCR) and western blotting. RESULTS: Periostin is expressed in the stroma of the primary gastric tumors and metastases, but not in normal gastric tissue. In addition, we observed that periostin is located mainly in pericryptal fibroblasts, but not in the tumor cells, and strongly correlated to the expression of α-smooth muscle actin (SMA). Furthermore, the distribution patterns of periostin were broader as the clinical staging of tumors progressed. We also identified a role of stress-related signaling in promoting cancer development and progression, and found that isoprenaline upregulated expression levels of periostin in gastric cancer cells. CONCLUSION: These findings suggest that the distribution pattern of periostin was broader as the clinical staging of the tumor progressed and found that isoprenaline upregulated expression levels of periostin in gastric cancer cells.


Subject(s)
Adenocarcinoma/metabolism , Adrenergic beta-Agonists/pharmacology , Aged , Blotting, Western , Cell Adhesion Molecules/drug effects , Cell Line, Tumor , Fibroblasts/metabolism , Gene Expression Regulation, Neoplastic/drug effects , Humans , Isoproterenol/pharmacology , Male , Neoplasm Staging , RNA, Messenger/genetics , Real-Time Polymerase Chain Reaction , Signal Transduction , Stomach/metabolism , Stomach Neoplasms/metabolism , Up-Regulation
17.
Braz. j. med. biol. res ; 49(4): e4324, 2016. tab, graf
Article in English | LILACS | ID: biblio-951663

ABSTRACT

The effects of interleukin-10 (IL-10) and glucose on mRNA and protein expression of osteoprotegerin (OPG), and its ligand, receptor activator of nuclear factor-κB ligand (RANKL), were investigated in human periodontal ligament fibroblasts (HPDLFs). Primary HPDLFs were treated with different concentrations of IL-10 (0, 1, 10, 25, 50, and 100 ng/mL) or glucose (0, 5.5, 10, 20, 30, and 40 mmol/L). Changes in mRNA and protein expression were examined using the reverse-transcription polymerase chain reaction (RT-PCR) and Western blot analysis, respectively. After IL-10 treatment, mRNA and protein levels of OPG were increased, while mRNA and protein levels of RANKL were decreased (P<0.05), both in a concentration-dependent manner. Glucose stimulation had the opposite concentration-dependent effect to that of IL-10 on OPG and RANKL expression. IL-10 upregulated OPG expression and downregulated RANKL expression, whereas high glucose upregulated RANKL and downregulated OPG in HDPLFs. Abnormal levels of IL-10 and glucose may contribute to the pathogenesis of periodontal disease.


Subject(s)
Humans , Periodontal Ligament/drug effects , Interleukin-10/pharmacology , RANK Ligand/metabolism , Osteoprotegerin/metabolism , Fibroblasts/drug effects , Glucose/pharmacology , Periodontal Ligament/cytology , Time Factors , RNA, Messenger/analysis , Down-Regulation , Up-Regulation , Cells, Cultured , Blotting, Western , Analysis of Variance , Reverse Transcriptase Polymerase Chain Reaction , Fibroblasts/metabolism
18.
Braz. oral res. (Online) ; 30(1): e109, 2016. tab, graf
Article in English | LILACS | ID: biblio-952054

ABSTRACT

Abstract Multicystic and unicystic ameloblastomas are benign odontogenic tumors that present distinct biological behavior. The investigation of stem cells has become an important branch of tumor biology, with several studies addressing the possible role of these cells in tumor growth, angiogenesis, progression, infiltration and invasiveness. This study evaluated the immunohistochemical expression of CD90(Thy-1) and P75NTR stem cell markers in multicystic and unicystic ameloblastomas. Seventeen (17) samples of ameloblastomas (multicystic, n = 10; unicystic, n = 7) were submitted to immunohistochemical reactions and graded semi-quantitatively. The Kolmogorov-Smirnov test was used to verify possible differences in CD90 and P75NTR expressions between multicystic and unicystic ameloblastomas (p < 0.05). CD90 immunostaining was observed in all multicystic ameloblastoma specimens (n = 10), in the cytoplasm of the fibroblasts and vascular endothelial cells of the tumor stroma, near the neoplastic odontogenic epithelia. The staining of stromal CD90 was significantly higher in multicystic than in unicystic ameloblastomas (p = 0.003). Nuclear P75NTR immunostaining was observed in all ameloblastoma specimens. A significant difference was seen in the epithelial staining of P75NTR between multicystic and unicystic types (p = 0.007). The increased expression of CD90 and P75NTR found in multicystic ameloblastomas suggests a behavioral biological difference between multicystic and unicystic ameloblastomas, as well as a difference in ameloblastoma development.


Subject(s)
Humans , Male , Female , Adolescent , Adult , Young Adult , Neoplastic Stem Cells/metabolism , Ameloblastoma/metabolism , Mandibular Neoplasms/metabolism , Biomarkers, Tumor/metabolism , Receptors, Nerve Growth Factor/metabolism , Thy-1 Antigens/metabolism , Nerve Tissue Proteins/metabolism , Neoplastic Stem Cells/pathology , Immunohistochemistry , Ameloblastoma/pathology , Mandibular Neoplasms/pathology , Paraffin Embedding , Stromal Cells , Statistics, Nonparametric , Endothelial Cells/metabolism , Fibroblasts/metabolism , Middle Aged
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