RESUMO
The major resolution of the study was to develop a dynamic form of natural biopolymer material to improve the wound healing by inhibition of biofilm formation on the surface. The extraction of collagen was effectively prepared from Scomberomorus lineolatus fish skin. Lyophilized collagen sheet was liquefied in 0.5M acetic acid to form acidic solubilized collagen (ASC) for further analysis. Physicochemical characterization of ASC was performed by various techniques using a standard protocol. The yield of ASC form S.lineolatus is higher (21.5%) than the previous reported studies. The effect of collagen solubility is gradually decreases with increasing concentration of NaCl and collagen is mostly soluble in acidic pH conditions. The sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis of ASC contains α chain composition of α1 and α2 subunits and was characterized as type I collagen. Ultraviolet absorption was regulated as the appropriate wavelength to optimize the collagen. Fourier-transform infrared spectroscopy and X-ray diffraction confirmed that the isolated collagen is a triple-helical structure. The biofilm formation of Pseudomonas aeruginosa was significantly reduced by collagen incorporated with isolated 3,5,7-trihydroxyflavone (collagen-TF) sheet up to 70%. The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide assay executed on fibroblast cell lines (L929) shows that the collagen-TF sheet was 100% compatible to enrich the cell adhesion and proliferation. The current study was the first report to extract, purify, and characterize ASC from S. lineolatus fish skin and characterize as type I collagen. Based on the result, we design the natural biodegradable collagen loaded with TF compound (collagen-TF) for antibiofilm properties. Compared with different sources of polymer, fish skin collagen is more effective and can be used as a biopolymer sheet for wound healing, food, drug delivery, tissue engineering, and pharmaceutical application.
Assuntos
Colágeno Tipo I/farmacologia , Proteínas de Peixes/farmacologia , Pele/química , Cicatrização/efeitos dos fármacos , Animais , Bandagens/microbiologia , Plásticos Biodegradáveis/química , Plásticos Biodegradáveis/farmacologia , Biofilmes/efeitos dos fármacos , Linhagem Celular , Colágeno Tipo I/química , Fibroblastos/efeitos dos fármacos , Proteínas de Peixes/química , Peixes/metabolismo , Humanos , Pele/metabolismo , SolubilidadeRESUMO
The current study is to evaluate the inhibition of biofilm formation and quorum sensing activity of isolated 3, 5, 7-Trihydroxyflavone (TF) from A.scholaris leaf extract against Pseudomonas aeruginosa. The effects of isolated TF on quorum sensing-regulated virulence factors production such as swimming motility, pyocyanin production, proteolytic, EPS, metabolic assay and inhibition of biofilm formation against P.aeruginosa was evaluated by standard protocols. In addition, the interaction between the isolated TF and active sites of QS- gene (LasI/rhlI, LasR/rhlR, and AHLase) in P.aeruginosa was evaluated by molecular docking studies using AutoDock Tools version 1.5.6. Based on the structural elucidation of the isolated compound was identified as 3, 5, 7-Trihydroxyflavone. Consequently, the isolated TF shows a significant reduction of biofilm formation through the inhibition of QS-dependent phenotypes such as pyocyanin production, proteolytic, swimming motility, EPS activities against P.aeruginosa in a dose-dependent manner. Molecular docking analysis of isolated TF can interfere the signaling [N-(3-oxododecanoyl)-L-homoserine lactone (3-oxo-C12-HSL) and N-butanoyl-L-homoserine lactone (C4-HSL)] molecules in P.aeruginosa by QS genes (LasI, LasR, rhlI, and AHLase) regulation. The isolated TF compound from A.scholaris reveals a greater potential to inhibit biofilm and QS dependent virulence factor production in P.aeruginosa. Docking interaction studies of TF-LasR complex express higher binding affinity than the other QS gene in P.aeruginosa.
Assuntos
Alstonia/química , Antibacterianos/farmacologia , Biofilmes/efeitos dos fármacos , Flavonoides/farmacologia , Folhas de Planta/química , Pseudomonas aeruginosa/efeitos dos fármacos , Percepção de Quorum/efeitos dos fármacos , Antibacterianos/química , Antibacterianos/isolamento & purificação , Relação Dose-Resposta a Droga , Flavonoides/síntese química , Flavonoides/química , Testes de Sensibilidade Microbiana , Simulação de Acoplamento Molecular , Estrutura Molecular , Relação Estrutura-AtividadeRESUMO
Fibrous hemostat was produced using wet laying technique comprising of acid and water soluble chitosan with fibrous cellulose. The hemostatic efficiency and their structural properties were evaluated using blood clotting time, whole blood adhesion and analytical characterization techniques. The maximum concentration of chitosan utilised for the production of nonwoven was 1.5 w/v % upon which increased the viscosity of the solution. The absorption of phosphate buffer solution was found to be 1117% for acid soluble chitosan based nonwoven whereas for water soluble chitosan based nonwoven was 997%. Increased red blood cells and platelets were found to be higher in acid soluble chitosan compared to water soluble chitosan based cellulose nonwoven. The blood clotting time for acid soluble chitosan nonwoven was found to be 166 s and water soluble chitosan nonwoven was 170 s. The developed hemostat have potential application in reducing blood loss and also inducing the blood coagulation pathway which was confirmed by the adhesion of platelets on to the surface of nonwoven web.