Objective: Pseudomonas aeruginosa and Staphylococcus aureus dual-species biofilm infections are notoriously difficult to manage. This study aimed at investigating the influence of four different culture media on the planktonic growth, adhesion, and biofilm formation of P. aeruginosa and S. aureus. Materials and Methods: We monitored four different culture media including Nutrient Broth, Brain Heart Infusion (BHI) broth, Luria-Bertani broth, and RPMI 1640 medium on the planktonic growth, adhesion, and biofilm formation of P. aeruginosa (ATCC 27853) and S. aureus (ATCC 25923) using MTT assay and scanning electron microscopy (SEM). Results: The most robust growth of the mono- and dual-species cultures was noted in BHI broth. On the contrary, RPMI 1640 medium promoted maximal initial adhesion of both the mono- and dual-species, but BHI broth fostered the maximal biofilm growth. SEM images showed profuse extracellular polysaccharide production in biofilms, particularly in coculture, in BHI medium. Conclusion: Our data demonstrate that BHI broth, relative to the other tested media, is the most conducive for in vitro evaluation of biofilm and planktonic growth kinetics of these two pathogens, both in mono- and coculture.
The study aimed to determine the antibacterial/antibiofilm effect and mechanism of interaction of curcuminoids-intercalated Mg/Al layered double hydroxide (curcuminoids-LDH) against three different bacteria. Antimicrobial effect of curcuminoids-LDH nanohybrid was investigated against P. aeruginosa, S. aureus, and E. faecalis (for both standard strains and clinical isolates), using agar well diffusion method. Minimum inhibitory concentrations (MIC) of planktonic bacteria were determined using the broth microdilution method. MIC of biofilms (MBIC ) and killing time for 48 hr matured biofilms were determined by MTT (3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay. Scanning electron microscopy (SEM) was used to determine pre- and postexposure architecture of biofilms. The mechanism of the antibiofilm activity of curcuminoids-LDH was determined using UV-visible spectroscopy. All tested bacteria had given a zone of inhibition in the presence of curcuminoids-LDH. The MIC values were 0.200 g/ml for P. aeruginosa, 0.025 g/ml for S. aureus, and 0.100 g/ml for E. faecalis. The 48 hr matured biofilms were reduced by curcuminoids-LDH with an MBIC of 0.100 g/ml. The minimum time to achieve MBIC was 3 hr, and the reduction was constant until 48 hr. SEM images showed a significant reduction of biofilm cell density and exopolymer matrics for all biofilms in the presence of curcuminoids-LDH. UV-visible studies revealed the antibiofilm activity of curcuminoids-LDH as due to the auto-oxidation of curcuminoids. The oxidation products are more limited in both product concentration per unit time and the variety of products, compared to pure curcuminoids, resulting in sharper UV-visible peaks than in the case of the latter. Curcuminoids-LDH has a potential antibacterial activity against P. aeruginosa, S. aureus, and E. faecalis. An antibiofilm activity has been achieved within 3 hr of the treatment. Curcuminoids released from the LDH showed the antibacterial activity due to oxidation products interfering with bacterial cell functions, and also encapsulation in the LDH causes curcuminoids to exhibit the activity in a persistent manner compared to pure curcuminoids.
Crystalline biofilm formation in indwelling urinary catheters is a serious health problem as it creates a barrier for antibacterial coatings. This emphasizes the failure of antibacterial coatings that do not have a mechanism to reduce crystal deposition on catheter surfaces. In this study, trifluoropropyl spray-coated polydimethylsiloxane (TFP–PDMS) has been employed as an antibiofilm forming surface without any antibacterial agent. Here, TFP was coated on half-cured PDMS using the spray coating technique to obtain a durable superhydrophobic coating for a minimum five cycles of different sterilization methods. The crystalline biofilm-forming ability of Proteus mirabilis in artificial urine, under static and flow conditions, was assessed on a TFP-PDMS surface. In comparison to the commercially available silver-coated latex and silicone catheter surfaces, TFP–PDMS displayed reduced bacterial attachment over 14 days. Moreover, the elemental analysis determined by atomic absorption spectroscopy and energy-dispersive X-ray analysis revealed that the enhanced antibiofilm forming ability of TFP-PDMS was due to the self-cleaning activity of the surface. We believe that this modified surface will significantly reduce biofilm formation in indwelling urinary catheters and further warrant future clinical studies.
Improving the superhydrophobicity of polydimethylsiloxane (PDMS) is a current interest in a wide range of applications from biomedical to aerospace. Although many fabrication techniques are available to improve the superhydrophobicity of PDMS, a significant problem occurs when the fabrication technique applies as a scalable but simple one. Here, we have described simple methods to achieve superhydrophobicity of PDMS using short-chained fluorinated polyhedral oligomeric silsesquioxanes (FPOSS). Two species of FPOSS were incorporated into PDMS using four different methods; non-solvent blending, solvent blending, spraying FPOSS/PDMS solution onto a partially cured PDMS matrix and spraying only FPOSS solution onto partially cured PDMS surfaces. Among two FPOSS species, spraying FPOSS onto partially cured PDMS produced a superhydrophobic surface with a static water contact angle of 167° ± 1°. Trifluoropropylisobutyl POSS (TFP) resulted in a higher hydrophobicity than trifluoropropyl POSS cages (CM). The multi-scale structured surface morphology, compatibility of functional groups attached to FPOSS and the fluorine content have shown a significant contribution on the superhydrophobicity in FPOSS/PDMS systems. Amorphous nature of the PDMS has improved upon incorporating FPOSS. Hence, this work presents a detailed study on the effect of the preparation method of FPOSS/PDMS composite on its superhydrophobicity.
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