Enhancing Osseointegration and Mitigating Bacterial Biofilms on Medical-Grade Titanium with Chitosan-Conjugated Liquid-Infused Coatings

Titanium alloys, in particular, medical-grade Ti-6Al-4V is heavily used in orthopaedic applications due to its high moduli, strength, and biocompatibility. Implant infection can result in biofilm formation and failure of prosthetics. The formation of a biofilm on implants protect bacteria from antibiotics and the immune response, resulting in the propagation of the infection and ultimately result in device failure. Recently, slippery liquid-infused surfaces (LIS) have been investigated for their stable liquid interface, which provide excellent repellent properties to suppress biofilm formation. One of the current limitations of LIS coatings lies in the indistinctive repellency of bone cells in orthopaedic applications, therefore causing poor integration between tissue and implant. Here, we report a chitosan impregnated LIS coating that facilitates cell adhesion and osseointegration while preventing biofilm formation. Our results indicate that chitosan-conjugated LIS increased cell adhesion of osteoblast-like SaOS-2 cells and significantly promoted proliferation compared to conventional titanium liquid-infused surfaces. Furthermore, the chitosan conjugated LIS significantly reduced biofilm formation of methicillin-resistant Staphylococcus aureus (MRSA) when compared to untreated and chitosan-coated titanium. Our engineered coating can be easily modified with other biopolymers or capture molecules to be applied to other biomaterials where both tissue integration and biofilm prevention is needed.

When Beneficial Biofilm on Materials Is Needed: Electrostatic Attachment of Living Bacterial Cells Induces Biofilm Formation

Bacterial attachment is crucial in many biotechnological applications, but many important bacterial strains cannot form biofilms. Biofilms can damage materials, and current strategies to manage biofilms are focused on inhibition and removal of biofilm. Biofilm formation is inevitable when materials are exposed to microbes and instead of biofilm prevention, we propose management of microbial composition by formation of biofilms with beneficial microbes. Since bacteria need to overcome a high repulsive force to attach to the surface and later to grow and multiply on it, electrostatic modification of the surfaces of cells or the material by polyelectrolytes (PE) was used in our approach, enabling efficient attachment of viable bacterial cells. Since highly positively charged PEs are known to be bactericidal, they were acetylated to reduce their toxicity, while preserving their net positive charge and ensuring cell viability. In our study bacterial strains were selected according to their intrinsic capability of biofilm formation, their shape variety and cell wall structure. These strains were tested to compare how the artificially prepared vs. natural biofilms can be used to populate the surface with beneficial bacteria. Using an artificial biofilm constructed of the potentially probiotic isolate Bacillus sp. strain 25.2. M, reduced the attachment and induced complete inhibition of E. coli growth over the biofilm. This study also revealed that the modification of the surfaces of cells or material by polyelectrolytes allows the deposition of bacterial cells, biofilm formation and attachment of biofilm non-forming cells onto surfaces. In this way, artificial biofilms with extended stability can be constructed, leading to selective pressure on further colonization of environmental bacteria.

Glutaraldehyde Enhancement by D-limonene for Mitigating Biocorrosion of Carbon Steel by An Oilfield Biofilm Consortium

Microbiologically influenced corrosion (MIC) is one of the corrosion threats in the oil and gas industry. It is caused by environmental biofilms. Glutaraldehyde is a popular green biocide for mitigating biofilms and MIC. This work investigated the enhancement of glutaraldehyde efficacy by food-grade green chemical D-limonene in the biofilm prevention and MIC mitigation using an oilfield biofilm consortium. After 7–d incubation at 37oC with enriched artificial seawater (EASW) in 125 mL anaerobic vials, the 100 ppm glutaraldehyde + 200 ppm D-limonene combination treatment reduced the sessile cell counts on C1018 carbon steel coupons by 2.1 − log, 1.7 − log, and 2.3 − log for SRB (sulfate reducing bacteria), APB (acid producing bacteria), and GHB (general heterotrophic bacteria), respectively in comparison with the untreated control. The treatment achieved 68% weight loss reduction and 78% pit depth reduction. The 100 ppm glutaraldehyde + 200 ppm D-limonene combination treatment was found more effective in biofilm prevention and MIC mitigation than glutaraldehyde and D-limonene used individually. Electrochemical tests corroborated weight loss and pit depth data trends.

2-(2-Methyl-2-nitrovinyl)furan but Not Furvina Interfere with Staphylococcus aureus Agr Quorum-Sensing System and Potentiate the Action of Fusidic Acid against Biofilms

Quorum sensing (QS) plays an essential role in the production of virulence factors, in biofilm formation and antimicrobial resistance. Consequently, inhibiting QS is being considered a promising target for antipathogenic/anti-virulence therapies. This study aims to screen 2-nitrovinylfuran derivatives structurally related to Furvina (a broad-spectrum antibiotic already used for therapeutic purposes) for their effects on QS and in biofilm prevention/control. Furvina and four 2-nitrovinylfuran derivatives (compounds 1–4) were tested to assess the ability to interfere with QS of Staphylococcus aureus using bioreporter strains (S. aureus ALC1742 and ALC1743). The activity of Furvina and the most promising quorum-sensing inhibitor (QSI) was evaluated in biofilm prevention and in biofilm control (combined with fusidic acid). The biofilms were further characterized in terms of biofilm mass, viability and membrane integrity. Compound 2 caused the most significant QS inhibition with reductions between 60% and 80%. Molecular docking simulations indicate that this compound interacts preferentially with the protein hydrophobic cleft in the LytTR domain of AgrA pocket. Metabolic inactivations of 40% for S. aureus ALC1742 and 20% for S. aureus ALC1743 were reached. A 24 h-old biofilm formed in the presence of the QSI increased the metabolic inactivation by fusidic acid to 80%, for both strains. The overall results highlight the effects of compound 2 as well as the potential of combining QSI with in-use antibiotics for the management of skin and soft tissues infections.

Efficient Biofilms Eradication by Enzymatic-Cocktail of Pancreatic Protease Type-I and Bacterial α-Amylase

Removal of biofilms is extremely pivotal in environmental and medicinal fields. Therefore, reporting the new-enzymes and their combinations for dispersal of infectious biofilms can be extremely critical. Herein, for the first time, we accessed the enzyme “protease from bovine pancreas type-I (PtI)” for anti-biofilm properties. We further investigated the anti-biofilm potential of PtI in combination with α-amylase from Bacillus sp. (αA). PtI showed a very significant biofilm inhibition effect (86.5%, 88.4%, and 67%) and biofilm prevention effect (66%, 64%, and 70%), against the E. coli, S. aureus, and MRSA, respectively. However, the new enzyme combination (Ec-PtI+αA) exhibited biofilm inhibition effect (78%, 90%, and 93%) and a biofilm prevention effect (44%, 51%, and 77%) against E. coli, S. aureus, and MRSA, respectively. The studied enzymes were found not to be anti-bacterial against the E. coli, S. aureus, and MRSA. In summary, the PtI exhibited significant anti-biofilm effects against S. aureus, MRSA, and E. coli. Ec-PtI+αA exhibited enhancement of the anti-biofilm effects against S. aureus and MRSA biofilms. Therefore, this study revealed that this Ec-PtI+αA enzymatic system can be extremely vital for the treatment of biofilm complications resulting from E. coli, S. aureus, and MRSA.

Recent Advances in Surface Nanoengineering for Biofilm Prevention and Control. Part II: Active, Combined Active and Passive, and Smart Bacteria-Responsive Antibiofilm Nanocoatings

The second part of our review describing new achievements in the field of biofilm prevention and control, begins with a discussion of the active antibiofilm nanocoatings. We present the antibiofilm strategies based on antimicrobial agents that kill pathogens, inhibit their growth, or disrupt the molecular mechanisms of biofilm-associated increase in resistance and tolerance. These agents of various chemical structures act through a plethora of mechanisms targeting vital bacterial metabolic pathways or cellular structures like cell walls and cell membranes or interfering with the processes that underlie different stages of the biofilm life cycle. We illustrate the latter action mechanisms through inhibitors of the quorum sensing signaling pathway, inhibitors of cyclic-di-GMP signaling system, inhibitors of (p)ppGpp regulated stringent response, and disruptors of the biofilm extracellular polymeric substances matrix (EPS). Both main types of active antibiofilm surfaces, namely non-leaching or contact killing systems, which rely on the covalent immobilization of the antimicrobial agent on the surface of the coatings and drug-releasing systems in which the antimicrobial agent is physically entrapped in the bulk of the coatings, are presented, highlighting the advantages of each coating type in terms of antibacterial efficacy, biocompatibility, selective toxicity, as well as drawbacks and limitations. Developments regarding combined strategies that join in a unique platform, both passive and active elements are not omitted. In such platforms with dual functionality, passive and active strategies can be applied either simultaneously or sequentially. We especially emphasize those systems that can be reversely and repeatedly switched between the non-fouling status and the bacterial killing status, thereby allowing several bacteria-killing/surface regeneration cycles to be performed without significant loss of the initial bactericidal activity. Eventually, smart antibiofilm coatings that release their antimicrobial payload on demand, being activated by various triggers such as changes in local pH, temperature, or enzymatic triggers, are presented. Special emphasis is given to the most recent trend in the field of anti-infective surfaces, specifically smart self-defensive surfaces for which activation and switch to the bactericidal status are triggered by the pathogens themselves.

Investigation on the effect of vitamin C on growth & biofilm-forming potential of Streptococcus mutans isolated from patients with dental caries

Background: Streptococcus mutans is a major cause of dental caries. Its capacity to produce biofilm is fundamental in the pathogenesis of this ubiquitous condition. As maintaining a healthy dentition is a genuine goal given the contemporary advance in caries control, researchers are striving to achieve a breakthrough in caries therapy. We are taking the anti-cariogenic properties of vitamin C a step-further, considering the well-known evidence of the inversely proportionate relationship between salivary levels of vitamin C and dental caries. The aim of this study was to determine MIC, MBC, biofilm prevention concentration (BPC), and derivative measures of vitamin C against fresh clinical isolates of S. mutans to evaluate its efficacy as an anti-cariogenic agent.Results: Based on the data of four independent experiments done in quadruplicates, we found a concentration-dependent inhibitory effect of vitamin C on all S. mutans strains tested. The average MBC, MIC, and BPC of vitamin C were found to be 10.16, 9.38, and 5.61 mg/ml, respectively. Spectrophotometric quantitation of crystal violet showed diminished biofilm formation in the presence of vitamin C (p < 0.05). When compared with gentamicin, vitamin C produced a zone of inhibition that was three times as large against the clinical isolates.Conclusion: Our results show that vitamin C has a negative effect on S. mutans growth and biofilm formation. Being the first to meticulously utilize BPC to explore a well-known effect of vitamin C, this report aims to help in the instigation of trials of higher evidence that will ultimately culminate in repurposing vitamin C as a novel anti-cariogenic agent, albeit further studies are required to provide auxiliary evidence in this context.

A review of iodine-based compounds, with a focus on biofilms: results of an expert panel

Biofilms play a central role in the chronicity of non-healing lesions such as venous leg ulcers and diabetic foot ulcers. Therefore, biofilm management and treatment is now considered an essential part of wound care. Many antimicrobial treatments, whether topical or systemic, have been shown to have limited efficacy in the treatment of biofilm phenotypes. The antimicrobial properties of iodine compounds rely on multiple and diverse interactions to exert their effects on microorganisms. An expert panel, held in Las Vegas during the autumn Symposium on Advanced Wound Care meeting in 2018, discussed these properties, with the focus on iodine and iodophors and their effects on biofilm prevention and treatment.