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In the present study, the effect of vitamin E supplementation 450 mg/kg diet was appraised in the process of induced wound healing in Nile tilapias Oreochromis niloticus. Fish were distributed into 18 tanks (10 fish each). Nine tanks were fed the non-supplemented diet and the other 9 tanks were fed 450 mg vitamin E for 60 days. Subsequently, the fish were anesthetized and the epidermis and dermis were surgically removed. The rate of cicatricial retraction and appearance of the wounds, and the histomorphometry of mucous cells, chromatophores, revascularization, inflammatory cells, presence of fibroblasts, collagen fibers, and scales were checked after 3-, 7-, 14-, 21-, and 28-days post-wounding. The retraction rate of the wound was significantly higher in the supplemented fish. The higher concentrations of inflammatory cells, mucous cells, and chromatophores, as well as the production and organization of collagen fibers, resulted in a higher retraction rate. We concluded that a dietary supplementation diet improves specific aspects of the cutaneous healing process in Nile tilapia fish.(AU)
No presente estudo, o efeito da suplementação com vitamina E de 450 mg / kg de dieta foi avaliado no processo de cicatrização induzida de feridas em tilápias do Nilo, Oreochromis niloticus. Os peixes foram distribuídos em 18 tanques (N=10), sendo 9 tanques com dieta não suplementada e os outros 9 tanques suplementados com 450 mg de vitamina E por 60 dias. Posteriormente, os peixes foram anestesiados e a epiderme e derme foram removidas cirurgicamente. Nos tempos pré-determinado de 3, 7, 14, 21 e 28 dias após a ferida foi analisado a taxa de retração cicatricial, a aparência das feridas e a histomorfometria das células mucosas, dos cromatóforos, das células inflamatórias, a revascularização, a presença de fibroblastos, de fibras de colágeno e escamas. A taxa de retração da ferida foi significativamente maior nos peixes suplementados. As maiores concentrações de células inflamatórias, mucosas e cromatóforos, bem como a produção e organização das fibras de colágeno, resultaram em uma maior taxa de retração. Concluímos que a dieta de suplementação melhora aspectos específicos do processo de cicatrização cutânea em peixes de tilápia do Nilo.(AU)
Subject(s)
Animals , Vitamin E , Wound Healing , Cichlids/physiology , Cichlids/injuries , alpha-Tocopherol , InflammationABSTRACT
ABSTRACT Seaweeds are related to anti-inflammatory, anti-bacterial and anti-noceptive effects. This work aimed to verify the potential of seaweed Padina gymnospora (Kützing) Sonder 1871 to improve wound healing in vitro. P. gymnospora was collected at a bethonic area in Espirito Santo. Methanolic extract of P. gymnospora was obtained by percolation. To determine cytotoxicity, colorimetric MTT tests were performed against normal fibroblasts (L929), macrophages (RAW 264.7) and human ovarian carcinoma (OVCAR-3) cell lines using concentration range of 12–110 µg ml-1. To evaluate in vitro wound healing, monolayer of fibroblasts L929 was seeded and artificial wounded. Cell proliferation was blocked by 5 µg ml-1 Mytomycin C. Nitric oxide inhibition was quantified with Raw 264.7 by Griess reaction. Minimal inhibitory concentration (MIC) against Staphylococcus aureus was determined. Eletrospray ionization with Fourier transform ion cyclotron resonance mass spectrometry (ESI-FT-ICR MS) was applied to detail composition of P. gymnospora methanolic extract. No cytotoxic effect in all cell lines was detected until the maximum concentration of 110 µg ml-1. P. gymnospora promoted significantly migration at the concentration of 25 µg ml-1 (p < 0.05). A prominent inhibition of nitric oxide formation was achieved in a concentration of 20 µg ml-1 of methanolic extract of P. gymnospora (62.06 ± 1.20%). Antibacterial activity against S. aureus could be demonstrated with MIC of 500 µg ml-1. ESI-FT-ICR MS analysis indicated eleven molecules between then, linolenic, oleic and linoleic acid. P. gymnospora favored wound repair in vitro what could be related to its fatty acid composition. In addition, its antimicrobial effect, and NO inhibition activity contribute for a new approach of P. gymnospora as a promise natural product for treatment of cutaneous wound.
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Objective:To explore the wound-healing effect and the anti-inflammatory activity of the aqueous extracts from the fresh roots and rhizomes of A. calamus. Methods: The image analysis techniques and the histological analysis were used to determine the wound-healing effect in the excised wound test, and the real-time RT-PCR techniques was used to evaluate the anti-inflammatory activi-ty in the lipopolysaccharide-induced RAW 264. 7 cells test. Results:The aqueous extracts were given 3 times a day since the model was established. The skin wound area was reduced significantly in the aqueous extracts group when compared with that in the control group since the 3rd day, the wound area in the aqueous extracts group was only 15% of that in the control group on the 13th day, and the wound-healing rate was enhanced significantly by the extracts. Moreover, the mRNA expressions of the inflammatory mediators such as TNF-α, IL-1β and IL-6 in the inflammatory RAW 264. 7 cells induced by lipopolysaccharide were inhibited effectively by the ex-tracts in a dose-dependant manner. Conclusion:The results indicate that the aqueous extracts from the fresh roots and rhizomes of A. calamus have significant wound-healing activity in animal excised wound model and anti-inflammatory activity in vitro.
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Aims: To investigate the scientific basis for the wound-healing properties of Funtumia elastica (Apocynaceae) leaf extracts using relevant in vitro fibroblast growth stimulation, antimicrobial and DPPH-antioxidant assays. Place and Duration of Study: School of Health, Sports and Bioscience (Bioscience Laboratories), University of East London in the United Kingdom, between July 2007 and May 2010. Methodology: Methanolic extract of the leaves, and petroleum ether, ethyl acetate, nbutanol and aqueous fractions partitioned thereof were tested for antimicrobial activities against common wound pathogens (such as Staphylococcus spp, Pseudomonas aeroginosa and Escherichia coli). The Broth dilution method was used to determine the minimum inhibitory concentrations (MIC) of the extracts and fractions. The antioxidant activities were also determined using a 2, 2-diphenyl-1-picrylhydrazyl (DPPH) free radical assay; whilst the ability to stimulate fibroblast growth was investigated using the MTT (3- [4, 5-dimethylthiazol-2-yl]-2, 5-diphenyltetrazolium bromide) assay. Results: The n-butanol fraction exhibited the greatest overall activities. It stimulated the growth of fibroblast cells by 28%, and showed MIC range of 0.13 - 1.0 mg/mL against the Staphylococci species, P. aeruginosa, Bacillus subtilis and E. coli. The non-polar petroleum ether fraction exhibited MICs greater than 2.0 mg/mL against all the organisms. All the fractions exhibited antioxidant activities greater than or comparable to that of ascorbic acid. Conclusion: Collectively, the antioxidant activity, fibroblast growth stimulation and the antimicrobial activities demonstrated by F. elastica leaf extracts provide some evidence to support the use of the plant to manage wounds in African folklore medicine.
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[ABSTRACT]AIM:Toinvestigatecellapoptosisindiabeticfootulcersandtheeffectofadvancedglycosylation end products (AGEs) on apoptosis in human fibroblast cells.METHODS: Diabetic foot patients (n=18) and 18 age-matched non-diabetic controls were recruited .The clinical and biochemical features were compared by statistics methods . Skin biopsies were obtained from foot .Cleaved caspase-3 was measured by immunohistochemistry using the technique of streptavidin-biotin complex ( SABC ) staining.Terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling ( TUNEL) technique was used to detect apoptosis of the skin tissues .Human primary foreskin fibroblasts were isolated and cultured in the presence of 5.6 mmo/L glucose, 25 mmo/L glucose, fluctuant glucose ( changing the glucose from 5.6 mmo/L to 25 mmo/L every 8 h) or AGEs (150 mg/L, containing 5.6 mmo/L glucose).After 72 h treatment, Western blotting was used to determine the levels of the apoptotic protein cleaved-caspase-3.Other cells were trypsinized , washed with cold PBS and incubated with PI and Annexin V-FITC, then analyzed by flow cytometry to detect cell apoptosis .RE-SULTS:Diabetic patients had higher levels of fasting blood glucose (FBG), 2-hour postprandial blood glucose (2 h PBG) and glycosylated hemoglobin A1c (HbA1c), and longer wound duration.The protein level of cleaved caspase-3 was signifi-cantly higher in diabetic group , suggesting that apoptosis was increased in diabetic skin tissues .TUNEL analysis showed that apoptotic index was higher in diabetic group compared with that in non-diabetic group (8.4%±1.5% vs 3.8%± 0.8%) , which further confirmed that cell apoptosis was increased in diabetic foot tissues .In human fibroblasts , the levels of cleaved caspase-3 in normal group , sustained high glucose group , fluctuant high glucose group and AGEs group were 0.80 ±0.13, 1.22 ±0.18, 1.46 ±0.32 and 1.83 ±0.25, respectively.The apoptotic rates detected by flow cytometry were 2.43%±0.19%, 2.89%±0.51%, 3.99% ±0.24% and 6.83% ±0.36%, respectively.Both the level of cleaved caspase-3 and the apoptotic rate in AGEs group were higher than those in normal glucose group and sustained high glucose group .CONCLUSION:Increased apoptosis in diabetic foot ulcers is one of the most important reasons for im-paired wound healing .As compared to sustained high glucose and glucose fluctuations , AGEs induce greater apoptosis in human fibroblast cells .
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Hedgehog pathway is an osteogenesis-related signaling pathway . During embryonic development , it regulates the growth and proliferation of progenitor cells and tissue formation. This pathway can be activated during liver injury. Activated Hedge-hog signaling pathway is involved in many aspects of liver wound-healing responses, including hepatic progenitor cell pro-liferation, myofibroblast transdifferentiation, apoptosis of various types of liver cells, inflammatory reactions, and vascular remod-eling. This article reviews the research progress in the role of Hedgehog signaling pathway in liver injury and the underlying mechanisms. The potential drug targets are also discussed. This review is to provide novel insights into antifibrotic research and therapeutic targets.
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Objective To evaluate the effects of Annexin A1 on migratory and invasive ability of colon cancer cells. Methods Eukaryotic expression vector pcDNA3. 1-Annexin A1 was constructed and analyzed by restriction analysis and sequencing. The recombinant plasmid pcDNA3. 1-Annexin A1 was transfected into SW480 cells by liposome-mediated gene transferred method. The stable transfectants were obtained after screening with G418. Real-time PCR and Western blotting analysis were used to examine the expression of Annexin A1 mRNA and protein in SW480 cells before and after transfection. Wound-healing experiment and Transwell invasion assays were used to study the effects of Annexin A1 on the migratory and invasive ability of SW480 cells. Results The recombinant plasmid pcDNA3. 1-Annexin A1 was successfully constructed. The results of real-time PCR and Western blotting analysis showed that the Annexin A1 expression was significantly higher in cells transfected with pcDNA3. 1-Annexin A1 than in un-transfected cells or those transfected with empty vectors (P0.05). The migratory rate of SW480 cells in pcDNA3. 1- Annexin A1 group was significantly higher than those in un-transfected cells or transfected with empty vectors ([0. 415 ± 0. 002) vs [0. 267± 0. 003] and [0. 271 ±0. 002], P<0.05), and there was no significant difference between the latter two groups. The migrating SW480 cells in pcDNA3. 1-Annexin A1 group was significantly more than those in the other two groups ([ 221. 75± 12. 07] vs [162. 80± 12. 07] and [164. 25±9. 50], P<0. 05). Conclusion Overexpression of Annexin A1 can increase the migratory and invasive ability of SW480 cell line.
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Objective:To observe the biological behaviors of colorectal cancer LS174 cells before and after pcDNA3.0-hugl-1 transfection,so as to investigate the association of hugl-1 with colorectal cancer.Methods:The eukaryotic expression vector pcDNA3.0-hugl-1 was constructed and transfected into LS174 cells.RT-PCR and Western blotting methods were used to analyze the expression of hugl-1 mRNA and protein in LS174 cells before and after transfection.Soft agar colony formation assay,wound-healing experiment,adhesion assay and Matrigel invasion assays were used to study the effects of hugl-1 expression on the proliferation,adhesion,movement and invasion in LS174 cells.Results:The recombinant plasmid pcDNA3.0-hugl-1 was successfully constructed.RT-PCR and Western blotting showed that the hugl-1 expression was higher in cells transfected with pcDNA3.0-hugl-1 than in those un-transfected or empty vector-transfected cells (P