RESUMEN
PURPOSE: Alumina substrates are one of the commonly used scaffolds applied in cell culture, but in order to prevent formation of biofilm on the alumina substrate, these substrates are modified with carbon nanotube. METHODS: The alumina substrate was made by a two-step anodization method and was then modified with carbon nanotubes by simple chemical reaction. The substrates were characterized with FTIR, SEM, EDX, 3D laser scanning digital microscope, contact angle (CA) and surface free energy (SFE). To determine how this modification influences the reduction of biofilm, biofilm of two various bacteria, Escherichia coli (E.coli) and Staphylococcus aureus (S. aureus), were investigated. RESULTS: The biofilm on the modified substrate decreased due to the presence of carbon nanotubes and increased antibacterial properties. Dental pulp stem cells (DPSCs) were cultured onto flat alumina (FA) and nanoporous alumina-multiwalled carbon nanotubes (NAMC) substrates to examine how the chemical modification and surface topography affects growth of DPSCs. CONCLUSION: Cell attachment and proliferation were investigated with SEM and Presto Blue assay, and the findings show that the NAMC substrates are suitable for cell culture.
Asunto(s)
Técnicas de Cultivo de Célula/instrumentación , Técnicas de Cultivo de Célula/métodos , Pulpa Dental/citología , Nanotubos de Carbono , Células Madre/citología , Óxido de Aluminio , Adhesión Bacteriana , Biopelículas , Proliferación Celular , Células Cultivadas , Escherichia coli/fisiología , Humanos , Microscopía Confocal , Microscopía Electrónica de Rastreo , Nanoporos , Espectroscopía Infrarroja por Transformada de Fourier , Staphylococcus aureus/fisiologíaRESUMEN
The cytosine embedded copper based metal-organic framework (Bio-MOF) was synthesized by facile one-step sonochemical method by simply mixing of 4-4, biphenyldicarboxylic, cytosine and copper nitrate (Bio-Cu-H2bpdc-Cy). The prepared bio-MOF was characterized by XRD, FTIR and FE-SEM techniques. The effect of Cu-H2bpdc-Cy on the expression of the rsbA gene was evaluated in the clinical and standard Proteus mirabilis and study of MIC of Cu-H2bpdc-Cy by microdilution against them that have the rsbA gene. According to different concentrations of MIC, MBC concentrations was cultured on blood agar culture medium. Regarding to the concentration of MIC, gene expression changes were obtained by real-time PCR. MIC for standard and clinical strains of Proteus mirabilis was 1.6 and 1.8â¯mg/ml, and also MBC was obtained to be 1.8 and 2.0â¯mg/ml, respectively. Finally, in the real time PCR method, expression of the rsbA gene in presences of bio-Cu-H2bpdc-Cy was reduced, but has no effect on the gene expression of the Housekeeping DNA Gyrase-B gene. Considering the effect of Cu-H2bpdc-Cy on the rsbA gene in Proteus mirabilis bacteria, it is possible to use of Cu-H2bpdc-Cy agent as a therapeutic supplement against this bacterium.
Asunto(s)
Antibacterianos/química , Proteínas Bacterianas/genética , Cobre/química , Citosina/química , Proteus mirabilis/efectos de los fármacos , Sonicación , Infecciones Urinarias/microbiología , Antibacterianos/farmacología , Humanos , Difracción de Polvo , Proteus mirabilis/genética , Reacción en Cadena en Tiempo Real de la PolimerasaRESUMEN
The aim of this work was comparison study of dilution and plating method for evaluation of the synergism effect of metal-organic framework nanocubes (MOF-5-NCs) and broccoli extract (Brassica oleracea) on antibacterial activity of standard and clinical Pseudomonas aeruginosa strains. For this purpose, sonochemical synthesis of MOF-5-NCs was performed and it was characterized using XRD, FT-IR, FESEM and EDS techniques. Maceration extraction (ME) and ultrasound assisted extraction (UAE) methods in three different solvents were prepared and applicability of their extracts were compared in some cases such as radical scavenging and antioxidant activity. The HPLC/UV analysis was applied for separation, identification and evaluation of phenolic acids in prepared broccoli extracts. Then, antimicrobial activity of MOF-5NCs and broccoli extract against gram-negative bacteria, Pseudomonas aeruginosa was evaluated by detection of minimal inhibition concentration (MIC), minimal bactericidal concentration (MBC) and zone of inhibition (ZOI). The results of in vitro assays showed that dilution method due to flase estimation of 4% viability percentage which is not logic by consideration of MBC well could not be able to estimate MBC. Therefore, plate count method was performed for precise calculation of MBC. MIC of broccoli extract and MOF-5-NCs on Pseudomonas aeruginosa strains were 7.81mgmL-1 and 3.13mgmL-1, respectively.