Quaternary Lipophilic Chitosan and Gelatin Cross-Linked Antibacterial Hydrogel Effectively Kills Multidrug-Resistant Bacteria with Minimal Toxicity toward Mammalian Cells.

Wounds or tissue openings in the skin are susceptible to bacterial attack, which can deteriorate and slow down the healing process. In this regard, antimicrobial gels are valuable as they mitigate the infection spread and assist in the healing. Despite the success, commercially available release-active antimicrobial gels suffer from narrow-spectrum activity, resistance induction, reservoir exhaustion, and in some cases may be associated with toxicity. To circumvent these limitations, herein, we have developed new quaternary lipophilic chitosan derivatives (QuaChi) synthesized by modifying the primary alcohol of the sugar moieties without altering the free amino groups of glucosamines. Compared to protonated chitosan, the synthesized derivatives exhibited improved water solubility and enhanced antibacterial activity against multidrug-resistant Gram-positive and Gram-negative bacteria including clinical isolates. The enhanced antibacterial activity was evident from the bacterial membrane depolarization leading to rapid inactivation of ∼105-106 bacterial cells within 2 h. The applicability of the chitosan derivatives was further demonstrated by developing antibacterial hydrogels by cross-linking the free amino groups of QuaChi with biocompatible gelatin through amide linkages. The hydrogel showed ∼5-7 log reduction of various multidrug-resistant bacteria including the stationary-phase cells within 6 h. Scanning electron microscopy revealed the loss of integrity of the bacterial structure when treated with the hydrogel, whereas mammalian cells (human embryonic kidney-293 (HEK-293)), when exposed to the hydrogel, appeared to be healthy with retained morphology. Collectively, these findings suggest that the developed hydrogel formulation can find potential applications to combat notorious drug-resistant bacterial infections in the healthcare settings.

Dual Function Injectable Hydrogel for Controlled Release of Antibiotic and Local Antibacterial Therapy.

We present vancomycin-loaded dual-function injectable hydrogel that delivers antibiotic locally suitable for treatment of infections in avascular or necrotic tissues. The syringe-deliverable gels were developed using polydextran aldehyde and an inherently antibacterial polymer N-(2-hydroxypropyl)-3-trimethylammonium chitosan chloride along with vancomycin. The antibiotic was primarily encapsulated via reversible imine bonds formed between vancomycin and polydextran aldehyde in the hydrogel which allowed sustained release of vancomycin over an extended period of time in a pH-dependent manner. Being inherently antibacterial, the gels displayed excellent efficacy against bacteria due to dual mode of action (killing bacteria upon contact as well as by releasing antibiotics into surroundings). Upon subcutaneous implantation, the gel was shown to kill methicillin-resistant Staphylococcus aureus (>99.999%) when bacteria were introduced directly into the gel as well as at distal site from the gel in a mice model. These materials thus represent as novel noninvasive drug-delivery device suitable for local antibiotic therapy.

Recent Progress in Polymer Research to Tackle Infections and Antimicrobial Resistance.

Global health is increasingly being threatened by the rapid emergence of drug-resistant microbes. The ability of these microbes to form biofilms has further exacerbated the scenario leading to notorious infections that are almost impossible to treat. For addressing this clinical threat, various antimicrobial polymers, polymer-based antimicrobial hydrogels and polymer-coated antimicrobial surfaces have been developed in the recent past. This review aims to discuss such polymer-based antimicrobial strategies with a focus on their current advancement in the field. Antimicrobial polymers, whose designs are inspired from antimicrobial peptides (AMPs), are described with an emphasis on structure-activity analysis. Additionally, antibiofilm activity and in vivo efficacy are delineated to elucidate the real potential of these antimicrobial polymers as possible therapeutics. Antimicrobial hydrogels, prepared from either inherently antimicrobial polymers or biocide-loaded into polymer-derived hydrogel matrix, are elaborated followed by various strategies to engineer polymer-coated antimicrobial surfaces. In the end, the current challenges are accentuated along with future directions for further expansion of the field toward tackling infections and antimicrobial resistance.

Functionalized chitosan based antibacterial hydrogel sealant for simultaneous infection eradication and tissue closure in ocular injuries.

Management of infections at ocular injury often requires prolonged and high dose of antibiotic, which is associated with challenges of antibiotic resistance and bacterial biofilm formation. Tissue glues are commonly used for repairing ocular tissue defects and tissue regeneration, but they are ineffective in curing infection. There is a critical need for antibacterial ocular bio-adhesives capable of both curing infection and aiding wound closure. Herein, we present the development of an imine crosslinked N-(2-hydroxypropyl)-3-trimethylammonium chitosan chloride (HTCC)­silver chloride nanocomposites (QAm1-Agx) and poly-dextran aldehyde (PDA) based bactericidal sealant (BacSeal). BacSeal exhibited potent bactericidal activity against a broad spectrum of bacteria including their planktonic and stationary phase within a short duration of 4 h. BacSeal effectively reduced biofilm-embedded MRSA and Pseudomonas aeruginosa by ∼99.99 %. In ex-vivo human cornea infection model, BacSeal displayed ∼99 % reduction of ocular infection. Furthermore, the hydrogel exhibited excellent sealing properties by maintaining ocular pressure up to 75 mm-Hg when applied to human corneal trauma. Cytotoxicity assessment and hydrogel-treated human cornea with a retained tissue structure, indicate its non-toxic nature. Collectively, BacSeal represents a promising candidate for the development of an ocular sealant that can effectively mitigate infections and may assist in tissue regeneration by sealing ocular wounds.

Versatile and User-Friendly Anti-infective Hydrogel for Effective Wound Healing.

Wound dressings play a crucial role in facilitating optimal wound healing and protecting against microbial infections. However, existing commercial options often fall short in addressing chronic infections due to antibiotic resistance and the limited spectrum of activity against both Gram-positive and Gram-negative bacteria frequently encountered at wound sites. Additionally, complex fabrication processes and cumbersome administration strategies pose challenges for cost-effective wound dressing development. Consequently, there is a pressing need to explore easily engineered biocompatible biomaterials as alternative solutions to combat these challenging wound infections. In this study, we present the development of an anti-infective hydrogel, P-BAC (polymeric bactericidal hydrogel), which exhibits simple administration and promotes efficient wound healing. P-BAC is synthesized via a one-step fabrication method that involves the noncovalent cross-linking of poly(vinyl alcohol), N-(2-hydroxypropyl)-3-trimethylammonium chitosan chloride-AgCl nanocomposite, and proline. Remarkably, P-BAC demonstrates broad-spectrum antibacterial activity against both planktonic and stationary cells of clinically isolated Gram-positive and Gram-negative bacteria, resulting in a significant reduction of bacterial load (5-7 log reduction). Moreover, P-BAC exhibits excellent efficacy in eradicating bacterial cells within biofilm matrices (>95% reduction). In vivo experiments reveal that P-BAC accelerates wound healing by stimulating rapid collagen deposition at the wound site and effectively inactivates ∼95% of Pseudomonas aeruginosa cells. Importantly, the shear-thinning property of P-BAC simplifies the administration process, enhancing its practicality and usability. Taken together, our findings demonstrate the potential of this easily administrable hydrogel as a versatile solution for effective wound healing with potent anti-infective properties. The developed hydrogel holds promise for applications in diverse healthcare settings, addressing the critical need for improved wound dressing materials.

Biocide loaded shear-thinning hydrogel with anti-biofilm efficacy cures topical infection.

The continuous intervention of multidrug-resistant (MDR) bacterial infections worsens and slows the dynamicity of natural wound healing processes. Fortunately, antibiotics, metal ions, or metal nanoparticle-loaded antimicrobial hydrogels have been developed to tackle infections at injury sites and speed up the healing process. Despite their success, these marketed released based hydrogels are still limited owing to their lack of broad-spectrum activity, inability to tackle biofilm-associated infections, susceptibility towards resistance development, fast release kinetics, and mild to moderate toxicity. To address these shortcomings, we report the development of a biocompatible, shear-thinning, injectable gellan-gelatin hydrogel loaded with a peptidomimetic potent biocide (ASAM-10). The hydrogel upon sustained biocide release (60% within 72 h), displays a broad-spectrum antibacterial activity with negligible in vitro (hemolysis < 20%) and in vivo toxicity (no adverse effects on dermal layer of mice). Besides tackling bacterial dormant subpopulation (1-6 Log reduction), the optimized hydrogel is able to disrupt the preformed bacterial biofilm and even kill the biofilm-trapped pathogens with enhanced pathogenicity. Above all, the lead hydrogel was proficient in tackling methicillin-resistant Staphylococcus aureus (MRSA) wound infections in a mouse model through its safe topical administration. Overall, the biocide-loaded hydrogel can be considered as a promising candidate to combat MDR chronic infections at the wound site.

Isoamphipathic antibacterial molecules regulating activity and toxicity through positional isomerism.

Peptidomimetic antimicrobials exhibit a selective interaction with bacterial cells over mammalian cells once they have achieved an optimum amphiphilic balance (hydrophobicity/hydrophilicity) in the molecular architecture. To date, hydrophobicity and cationic charge have been considered the crucial parameters to attain such amphiphilic balance. However, optimization of these properties is not enough to circumvent unwanted toxicity towards mammalian cells. Hence, herein, we report new isoamphipathic antibacterial molecules (IAMs: 1-3) where positional isomerism was introduced as one of the guiding factors for molecular design. This class of molecules displayed good (MIC = 1-8 µg mL-1 or µM) to moderate [MIC = 32-64 µg mL-1 (32.2-64.4 µM)] antibacterial activity against multiple Gram-positive and Gram-negative bacteria. Positional isomerism showed a strong influence on regulating antibacterial activity and toxicity for ortho [IAM-1: MIC = 1-32 µg mL-1 (1-32.2 µM), HC50 = 650 µg mL-1 (654.6 µM)], meta [IAM-2: MIC = 1-16 µg mL-1 (1-16.1 µM), HC50 = 98 µg mL-1 (98.7 µM)] and para [IAM-3: MIC = 1-16 µg mL-1 (1-16.1 µM), HC50 = 160 µg mL-1 (161.1 µM)] isomers. Co-culture studies and investigation of membrane dynamics indicated that ortho isomer, IAM-1 exerted more selective activity towards bacterial over mammalian membranes, compared to meta and para isomers. Furthermore, the mechanism of action of the lead molecule (IAM-1) has been characterized through detailed molecular dynamics simulations. In addition, the lead molecule displayed substantial efficacy against dormant bacteria and mature biofilms, unlike conventional antibiotics. Importantly, IAM-1 exhibited moderate in vivo activity against MRSA wound infection in a murine model with no detectable dermal toxicity. Altogether, the report explored the design and development of isoamphipathic antibacterial molecules to establish the role of positional isomerism in achieving selective and potential antibacterial agents.

Biocompatible Hemostatic Sponge Exhibiting Broad-Spectrum Antibacterial Activity.

Hemorrhage during accidents or surgery is a significant challenge that can contribute to mortality. This is further aggravated due to bacterial infections at the injured site. Therefore, rapid application of a hemostatic and antibacterial material is highly necessary as a pretreatment for patients' survival. Herein, we have developed a hemostatic sponge (Hemobac) through amide crosslinking of gelatin and an N-(2-hydroxy) propyl-3-trimethylammonium chitosan (HTCC)-silver chloride nanocomposite (QAm1-Ag0.1) to mitigate bacterial infections, while aiding hemostasis. This Hemobac sponge completely eradicated (∼4-5 log) a wide range of Gram-positive and Gram-negative bacteria encompassing various clinical isolates within 6 h. The antihemorrhagic ability of Hemobac was ascertained through SEM images, which exhibited the presence of agglomerated blood cells onto the sponge with a significantly low blood-clotting index value (∼23 ± 1). Notably, Hemobac reduced the blood loss by ∼70-80% in the liver puncture model and femoral vein injury model in mice, displaying its improved hemostatic ability over a marketed gelatin-based sponge. Negligible hemolytic activity (∼6%) and retained healthy morphology of mammalian cells were observed upon exposure to the Hemobac sponge. Minimal immune response was noticed at the Hemobac-treated wound in mice through histopathology analysis. Collectively, these findings indicate that this biocompatible Hemobac sponge can stop the bleeding instantaneously and combat bacterial infections.

An easy-to-use antimicrobial hydrogel effectively kills bacteria, fungi, and influenza virus.

Various drug resistant pathogens such as bacteria, fungi and viruses enter a host through different routes, which can lead to health-related problems and even fatalities. Propagation of these infectious microbes majorly occurs through the mucosal openings or upon topical contact. To curb their transmission or to cure infections associated with these pathogens, herein we describe the development of an antimicrobial hydrogel, based on a water soluble quaternary lipophilic polyethyleneimine derivative (QPEINH-C6). The cationic polymer QPEINH-C6 exhibited antibacterial activity against drug-resistant Gram-positive bacteria (MIC = 10-62 µg mL-1) and Gram-negative bacteria (MIC = 117-123 µg mL-1). The derivative showed killing of human pathogenic fungi (MIC = 58-67 µg mL-1), including their clinical isolates. The rapid bactericidal and fungicidal nature were confirmed from the fast inactivation kinetics of bacterial cells (methicillin resistant S. aureus and vancomycin resistant S. aureus) within 3-6 hours and C. albicans within 1 h with ∼5-6 log reduction in the microbial burden. This antibacterial and antifungal cationic polymer was then used to construct an antimicrobial shear-thinning hydrogel (Bacfuvir), through non-covalent crosslinking with biocompatible gellan and polyvinyl alcohol (PVA). This hydrogel displayed ∼5-7 log reduction of numerous multidrug-resistant bacteria and their stationary phase cells which are insusceptible to conventional antibiotics. In addition, >99.9 % viable bacterial burden was reduced from preformed biofilm matrices of drug-resistant bacteria. Alongside, fluconazole-resistant C. albicans strains were killed completely within 15-60 min upon exposure to Bacfuvir gel. Most importantly, MRSA and C. albicans cells were reduced (3-4 log) in polymicrobial biofilms after hydrogel treatment. The hydrogel exhibited 99.9 % reduction of influenza viruses in a rapid manner. Due to the biocompatibility of Bacfuvir gel on topical application in a murine model and easy administration owing to its shear-thinning behaviour, this hydrogel can markedly contribute to mitigating drug-resistant bacterial, fungal and viral infections in healthcare settings.

Advancements in release-active antimicrobial biomaterials: A journey from release to relief.

Escalating medical expenses due to infectious diseases are causing huge socioeconomic pressure on mankind globally. The emergence of antibiotic resistance has further aggravated this problem. Drug-resistant pathogens are also capable of forming thick biofilms on biotic and abiotic surfaces to thrive in a harsh environment. To address these clinical problems, various strategies including antibacterial agent delivering matrices and bactericidal coatings strategies have been developed. In this review, we have discussed various types of polymeric vehicles such as hydrogels, sponges/cryogels, microgels, nanogels, and meshes, which are commonly used to deliver antibiotics, metal nanoparticles, and biocides. Compositions of these polymeric matrices have been elaborately depicted by elucidating their chemical interactions and potential activity have been discussed. On the other hand, various implant/device-surface coating strategies which exploit the release-active mechanism of bacterial killing are discussed in elaboration. This article is categorized under: Therapeutic Approaches and Drug Discovery > Nanomedicine for Cardiovascular Disease Implantable Materials and Surgical Technologies > Nanomaterials and Implants Therapeutic Approaches and Drug Discovery > Nanomedicine for Infectious Disease.

Easy Fabrication of a Polymeric Transparent Sheet to Combat Microbial Infection.

Surges in infectious diseases and their transmission in households and commercial and healthcare settings have increased the use of polymeric materials as protective covers. Despite ongoing efforts, conventional polymeric materials still pose the threat of surface-associated transmission of pathogens due to the fact that they lack antimicrobial properties. Here, we have developed an easy-to-fabricate polymeric sheet [quaternary polymeric transparent sheet (QPTS)] that shows an excellent antimicrobial property and is also transparent in nature, increasing its practical applications in a wide range of surfaces. The sheet was fabricated by combining cationic amphiphilic water-soluble polyethylenimine derivative (QPEINH-C6) and poly(vinyl alcohol) (PVA). The optimum composition (QPTS-3) exhibited a complete reduction of bacterial and fungal infection (∼3-4 log reduction) within 15 min. QPTS-3 also exhibited activity against antibiotic-insusceptible metabolically inactive bacterial cells. The sheet prevented the growth of MRSA biofilm even after 72 h of incubation, which was confirmed through electron microscopy on the QPTS sheet. Most importantly, ∼99.9% of the influenza viral load was reduced completely within 30 min of exposure of the sheet. Apart from the antimicrobial property, the sheet successfully retained its transparency (∼88%) and maintained a significant mechanical strength (∼15 N), highlighting its potential applications in commercial and healthcare settings.