Nanocarrier Drug Conjugates Exhibit Potent Anti-Naegleria fowleri and Anti-Balamuthia mandrillaris Properties.

Given the opportunity and access, pathogenic protists (Balamuthia mandrillaris and Naegleria fowleri) can produce fatal infections involving the central nervous system. In the absence of effective treatments, there is a need to either develop new antimicrobials or enhance the efficacy of existing compounds. Nanocarriers as drug delivery systems are gaining increasing attention in the treatment of parasitic infections. In this study, novel nanocarriers conjugated with amphotericin B and curcumin were evaluated for anti-amoebic efficacy against B. mandrillaris and N. fowleri. The results showed that nanocarrier conjugated amphotericin B exhibited enhanced cidal properties against both amoebae tested compared with the drug alone. Similarly, nanocarrier conjugated curcumin exhibited up to 75% cidal effects versus approx. 50% cidal effects for curcumin alone. Cytopathogenicity assays revealed that the pre-treatment of both parasites with nanoformulated-drugs reduced parasite-mediated host cellular death compared with the drugs alone. Importantly, the cytotoxic effects of amphotericin B on human cells alone were reduced when conjugated with nanocarriers. These are promising findings and further suggest the need to explore nanocarriers as a means to deliver medicine against parasitic infections.

Antibacterial Properties of Ethacridine Lactate and Sulfmethoxazole Loaded Functionalized Graphene Oxide Nanocomposites.

The emergence of drug-resistant bacterial strains that reduce the effectiveness of antimicrobial agents has become a major ongoing health concern in recent years. It is therefore necessary to find new antibacterials with broad-spectrum activity against both Gram-positive and Gram-negative bacteria, and/or to use nanotechnology to boost the potency of already available medications. In this research, we examined the antibacterial efficacy of sulfamethoxazole and ethacridine lactate loaded two-dimensional glucosamine functionalized graphene-based nanocarriers against a range of bacterial isolates. Graphene oxide was first functionalized with glucosamine, which as a carbohydrate moiety can render hydrophilic and biocompatible characters to the GO surface, and subsequently loaded with ethacridine lactate and sulfamethoxazole. The resulting nanoformulations had distinct, controllable physiochemical properties. By analyzing the formulation using Fourier Transform Infrared Spectroscopy (FTIR), X-ray diffraction (PXRD), a thermogravimetric analysis (TGA), zetasizer, and a morphological analysis using Scanning Electron Microscopy and Atomic Force Microscopy, researchers were able to confirm the synthesis of nanocarriers. Both nanoformulations were tested against Gram-negative bacteria, including Escherichia coli K1, Serratia marcescens, Pseudomonas aeruginosa, Salmonella enterica, as well as Gram-positive bacteria, including Bacillus cereus, Streptococcus pyogenes, and Streptococcus pneumoniae. Importantly, ethacridine lactate and its nanoformulations exhibited significant antibacterial properties against all bacteria tested in this study. When tested for minimum inhibitory concentration (MIC), the results were remarkable and revealed that ethacridine lactate presented MIC90 at 9.7 µg/mL against S. enteric, and MIC90 at 6.2 µg/mL against B. cereus. Notably, ethacridine lactate and its nanoformulations showed limited toxicity effects against human cells using lactate dehydrogenase assays. Overall, the results revealed that ethacridine lactate and its nanoformulations possess antibacterial activities against various Gram-negative and Gram-positive bacteria and that nanotechnology can be employed for the targeted delivery of effective drugs without harming the host tissue.

Anti-Balamuthia mandrillaris and anti-Naegleria fowleri effects of drugs conjugated with various nanostructures.

Balamuthia mandrillaris and Naegleria fowleri are protist pathogens that can cause fatal infections. Despite mortality rate of > 90%, there is no effective therapy. Treatment remains problematic involving repurposed drugs, e.g., azoles, amphotericin B and miltefosine but requires early diagnosis. In addition to drug discovery, modifying existing drugs using nanotechnology offers promise in the development of therapeutic interventions against these parasitic infections. Herein, various drugs conjugated with nanoparticles were developed and evaluated for their antiprotozoal activities. Characterizations of the drugs' formulations were accomplished utilizing Fourier-transform infrared spectroscopy, efficiency of drug entrapment, polydispersity index, zeta potential, size, and surface morphology. The nanoconjugates were tested against human cells to determine their toxicity in vitro. The majority of drug nanoconjugates exhibited amoebicidal effects against B. mandrillaris and N. fowleri. Amphotericin B-, Sulfamethoxazole-, Metronidazole-based nanoconjugates are of interest since they exhibited significant amoebicidal effects against both parasites (p < 0.05). Furthermore, Sulfamethoxazole and Naproxen significantly diminished host cell death caused by B. mandrillaris by up to 70% (p < 0.05), while Amphotericin B-, Sulfamethoxazole-, Metronidazole-based drug nanoconjugates showed the highest reduction in host cell death caused by N. fowleri by up to 80%. When tested alone, all of the drug nanoconjugates tested in this study showed limited toxic effects against human cells in vitro (less than 20%). Although these are promising findings, prospective work is warranted to comprehend the mechanistic details of nanoconjugates versus amoebae as well as their in vivo testing, to develop antimicrobials against the devastating infections caused by these parasites.

Cinnamic acid and lactobionic acid based nanoformulations as a potential antiamoebic therapeutics.

Acanthamoeba castellanii causes granulomatous amoebic encephalitis, an uncommon but severe brain infection and sight-threatening Acanthamoeba keratitis. Most of the currently used anti-amoebic treatments are not always effective, due to persistence of the cyst stage, and recurrence can occur. Here in this study we synthesize cinnamic acid and lactobionic acid-based magnetic nanoparticles (MNPs) using co-precipitation technique. These nanoformulations were characterized by Fourier transform infrared spectroscopy and Atomic form microscopy. The drugs alone (Hesperidin, Curcumin and Amphotericin B), magnetic NPs alone, and drug-loaded nano-formulations were evaluated at a concentration of 100 µg/mL for antiamoebic activity against a clinical isolate of A. castellanii. Amoebicidal assays revealed that drugs and conjugation of drugs and NPs further enhanced amoebicidal effects of drug-loaded nanoformulations. Drugs and drug-loaded nanoformulations inhibited both encystation and excystation of amoebae. In addition, drugs and drug-loaded nanoformulations inhibited parasite binding capability to the host cells. Neither drugs nor drug-loaded nanoformulations showed cytotoxic effects against host cells and considerably reduced parasite-mediated host cell death. Overall, these findings imply that conjugation of medically approved drugs with MNPs produce potent anti-Acanthamoebic effects, which could eventually lead to the development of therapeutic medications.

Improving curcumin bactericidal potential against multi-drug resistant bacteria via its loading in polydopamine coated zinc-based metal-organic frameworks.

Multi-drug resistant (MDR) bactearial strains have posed serious health issues, thus leading to a significant increase in mortality, morbidity, and the expensive treatment of infections. Metal-organic frameworks (MOFs), comprising metal ions and a variety of organic ligands, have been employed as an effective drug deliveryy vehicle due to their low toxicity, biodegradability, higher structural integrity and diverse surface functionalities. Polydopamine (PDA) is a versatile biocompatible polymer with several interesting properties, including the ability to adhere to biological surfaces. As a result, modifying drug delivery vehicles with PDA has the potential to improve their antimicrobial properties. This work describes the preparation of PDA-coated Zn-MOFs for improving curcumin's antibacterial properties against S. aureus and E. coli. Powder X-ray diffraction (P-XRD), FT-IR, scanning electron microscopy (SEM), and DLS were utilized to characterize PDA-coated Zn-MOFs. The curcumin loading and in vitro release of the prepared MOFs were also examined. Finally, the MOFs were tested for bactericidal ability against E. coli and S. aureus using an anti-bacterial assay and surface morphological analysis. Smaller size MOFs were capable of loading and releasing curcumin. The findings showed that as curcumin was encapsulated into PDA-coated MOFs, its bactericidal potential was significantly enhanced, and the findings were further supported by SEM which indicated the complete morphological distortion of the bacteria after treatment with PDA-Cur-Zn-MOFs. These studies clearly indicate that the PDA-Cur-Zn-MOFs developed in this study are extremely promising for long-term release of drugs to treat a wide range of microbial infections.

Zinc Oxide Nanoconjugates against Brain-Eating Amoebae.

Naegleria fowleri and Balamuthia mandrillaris are opportunistic protists, responsible for fatal central nervous system infections such as primary amoebic meningoencephalitis (PAM) and granulomatous amoebic encephalitis (GAE) with mortality rates higher than 90%. Threatening a rise in cases is the increase in temperature due to global warming. No effective treatment is currently available. Herein, nanotechnology was used to conjugate Zinc oxide with Ampicillin, Ceftrixon, Naringin, Amphotericin B, and Quericitin, and the amoebicidal activity and host cell cytotoxicity of these resulting compounds were investigated. The compounds ZnO-CD-AMPi, ZnO-CD-CFT, ZnO-CD-Nar, ZnO-CD-AMB, and ZnO-CD-QT were found to reduce N. fowleri viability to 35.5%, 39.6%, 52.0%, 50.8%, 35.9%, and 69.9%, respectively, and B. mandrillaris viability to 40.9%, 48.2%, 51.6%, 43.8%, and 62.4%, respectively, when compared with their corresponding controls. Furthermore, the compounds reduced N. fowleri-mediated and B. mandrillaris-mediated host cell death significantly. Additionally, the compounds showed limited cytotoxicity against human cells; cell toxicity was 35.5%, 36.4%, 30.9%, 36.6%, and 35.6%, respectively, for the compounds ZnO-CD-AMPi, ZnO-CD-CFT, ZnO-CD-Nar, ZnO-CD-AMB, and ZnO-CD-QT. Furthermore, the minimum inhibitory concentrations to inhibit amoeba growth by 50% were determined for N. fowleri and B. mandrillaris. The MIC50 for N. fowleri were determined to be 69.52 µg/mL, 82.05 µg/mL, 88.16 µg/mL, 95.61 µg/mL, and 85.69 µg/mL, respectively; the MIC50 of the compounds for B. mandrillaris were determined to be 113.9 µg/mL, 102.3 µg/mL, 106.9 µg/mL, 146.4 µg/mL, and 129.6 µg/mL, respectively. Translational research to further develop therapies based on these compounds is urgently warranted, given the lack of effective therapies currently available against these devastating infections.

Enhancing efficacy of existing antibacterials against selected multiple drug resistant bacteria using cinnamic acid-coated magnetic iron oxide and mesoporous silica nanoparticles.

Developing new antibacterial drugs by using traditional ways is insufficient to meet existing challenges; hence, new strategies in the field of antibacterial discovery are necessary. An alternative strategy is to improve the efficacy of currently available antibiotics. Herein, the antibacterial efficacy of drugs (Cefixime, Sulfamethoxazole, and Moxifloxacin) and drug-loaded cinnamic acid-coated magnetic iron oxide and mesoporous silica nanoparticles (NPs) was elucidated versus Gram-negative bacteria (Pseudomonas aeruginosa, Klebsiella pneumoniae, neuropathogenic Escherichia coli K1 and Serratia marcescens) and Gram-positive bacteria (Methicillin-resistant Staphylococcus aureus (MRSA), Streptococcus pyogenes, Streptococcus pneumoniae, and Bacillus cereus). NPs were synthesized by co-precipitation and the Stöber method, and characterized by Fourier transform-infrared spectroscopy, Zetasizer, and Atomic force microscopy. Lactate dehydrogenase (LDH) assays were accomplished to determine drug cytotoxicity against human cells. Spherical NPs in the range of 118-362 nm were successfully synthesized. Antibacterial assays revealed that drugs conjugated with NPs portray enhanced bactericidal efficacies against multiple drug resistant bacteria compared to the drugs alone. Of note, Cefixime-conjugated NPs against Escherichia coli K1 and Methicillin- resistant Staphylococcus aureus, resulted in the complete eradication of all bacterial isolates tested at significantly lower concentrations compared to the antibiotics alone. Likewise, conjugation of Moxifloxacin resulted in the complete elimination of E. coli K1 and MRSA. Of note, nano-formulated drugs presented negligible cytotoxicity against human cells. These results depict potent, and enhanced efficacy of nano-formulated drugs against medically important bacteria and can be used as alternatives to current antibiotics. Future in vivo studies and clinical studies are warranted in prospective years to realize these expectations.

Electrospun Nanofiber-Based Viroblock/ZnO/PAN Hybrid Antiviral Nanocomposite for Personal Protective Applications.

Designing novel antiviral personal protective equipment (PPE) is crucial for preventing viral infections such as COVID-19 in humans. Here, we fabricate an electrospun nanofiber-based Viroblock (VB)-loaded polyacrylonitrile (PAN)/zinc oxide (ZnO) hybrid nanocomposite for PPE applications. Five different concentrations of Viroblock (0.5%, 1.5%, 2.5%, 3.5%, and 5%) were added to PAN/ZnO solution and loaded for electrospinning. The developed samples reflected antibacterial activity of 92.59% and 88.64% against Staphylococcus aureus and Pseudomonas aeruginosa bacteria, respectively, with 5% VB loading. Moreover, a significant reduction in virus titer (37%) was observed with the 5% VB/PAN/ZnO nanofiber sheet. Hence, VB-loaded PAN/ZnO nanofibers have great potential to kill enveloped viruses such as influenzas and coronaviruses and could be the ideal candidate for the development of nanofiber-based PPE, such as facemasks and surgical gowns, which can play a key role in the protection of frontline health workers and the general public in the COVID-19 pandemic.

Synthesis, characterization and drug delivery application of Dapsone based double tailed biocompatible nonionic surfactant.

The increase in antimicrobial resistance has created a crisis that has become top priority for global policy and public health. Antibiotics are constantly being rendered in-effective due to the emergence of bacterial resistance; therefore, novel strategies for improving therapeutic efficacies of existing drugs must be focused. Advancements in nanotechnology have opened up new avenues for enhancing therapeutic efficacy of existing drugs via construction of intelligent and efficient delivery systems. This study reports the synthesis of Dapsone based nonionic surfactant and its utilization as delivery system for Ceftriaxone sodium. The synthesized nonionic surfactant was characterized via mass spectrometry and 1H NMR and IR spectroscopic techniques. The drug loaded vesicles of newly synthesized sulfur based nonionic were formed through thin film hydration method and characterized for drug entrapment efficiency, vesicles size, zeta potential, morphology using UV-vis spectrometry, dynamic light scattering (DLS) and atomic force microscopic (AFM) techniques. The biocompatibility of newly synthesized surfactant was assessed using blood hemolysis and in-vitro cells cytotoxicity. Antibacterial potential of drug loaded vesicles was assessed in gram positive and gram negative bacterial cultures. The spectroscopic results confirm successful synthesis of novel sulfur based nonionic surfactant that formed spherical shaped drug loaded vesicles with an average size of 97.95 ± 3.45 nm and 56.3 ± 3.15 % entrapment of the model drug (Ceftriaxone sodium). The vesicles displayed negative surface charge of -16.8 ± 3.72 mV and released the entrapped drug in a controlled way in-vitro drug release. The drug loaded vesicular formulation showed enhanced cellular uptake and greater antibacterial potentials when compared with control. Results of this study show that the Dapsone based surfactant is safe, biocompatible, non-toxic and can be used as promising vesicular carrier for enhancing therapeutic efficacy of antibacterial drug, Ceftriaxone sodium.

Galactosylated iron oxide nanoparticles for enhancing oral bioavailability of ceftriaxone.

The current study focuses on the development, characterization, biocompatibility investigation and oral bioavailability evaluation of ceftriaxone (CFT)-loaded lactobionic acid (LBA)-functionalized iron oxide magnetic nanoparticles (MNP-LBA). Atomic force microscopy and dynamic light scattering showed that the developed CFT-loaded MNP-LBA is spherical, with a measured hydrodynamic size of 147 ± 15.9 nm and negative zeta potential values (-35 ± 0.58 mV). Fourier transformed infrared analysis revealed interactions between the nanocarrier and the drug. Nanoparticles showed high drug entrapment efficiencies of 91.5 ± 2.2%, and the drug was released gradually in vitro and shows prolonged in vitro stability using simulated gastrointestinal (GI) fluids. The formulations were found to be highly biocompatible (up to 100 µg/mL) and hemocompatible (up to 1.0 mg/mL). Using an albino rabbit model, the formulation showed a significant enhancement in drug plasma concentration up to 14.46 ± 2.5 µg/mL in comparison with its control (1.96 ± 0.58 µg/mL). Overall, the developed MNP-LBA formulation was found promising for provision of high-drug entrapment, gradual drug release and was appropriate for enhancing the oral delivery of CFT.

Enhancement in Oral Absorption of Ceftriaxone by Highly Functionalized Magnetic Iron Oxide Nanoparticles.

The present study aims at the development, characterization, biocompatibility investigation and oral bioavailability evaluation of ceftriaxone (CFT)-loaded N'-methacryloylisonicotinohydrazide (MIH)-functionalized magnetic nanoparticles (CFT-MIH-MNPs). Atomic force microscopy (AFM) and dynamic light scattering (DLS) showed that the developed CFT loaded MIH-MNPs are spherical, with a measured hydrodynamic size of 184.0 ± 2.7 nm and negative zeta potential values (-20.2 ± 0.4 mV). Fourier transformed infrared spectroscopic (FTIR) analysis revealed interactions between the nanocarrier and the drug. Nanoparticles showed high drug entrapment efficiency (EE) of 79.4% ±1.5%, and the drug was released gradually in vitro and showed prolonged in vitro stability using simulated gastrointestinal tract (GIT) fluids. The formulations were found to be highly biocompatible (up to 100 µg/mL) and hemocompatible (up to 1.0 mg/mL). Using an albino rabbit model, the formulation showed a significant enhancement in drug plasma concentration up to 14.4 ± 1.8 µg/mL in comparison with its control (2.0 ± 0.6 µg/mL). Overall, the developed CFT-MIH-MNPs formulation was promising for provision of high drug entrapment, gradual drug release and suitability for enhancing the oral delivery of CFT.

Design and synthesis of mixed micellar system for enhanced anticancer efficacy of Paclitaxel through its co-delivery with Naringin.

Emergence of multidrug resistance (MDR) has limited the success of chemotherapeutic agents. Reversal of drugs efflux systems through combination therapy has got wider attention for increasing anticancer drugs efficacy. This study aims at co-encapsulation of Paclitaxel with Naringin in mixed polymeric micelles for enhanced anticancer activity of the drug. Drug-loaded micelles were prepared using two different amphiphilic block co-polymers and were characterized for morphology, size, zeta potential, drug encapsulation, in vitro release and stability using atomic force microscope (AFM), zetasizer, UV spectrophotometer, and FT-IR. MTT assay and fluorescence microscopy were used for in vitro cytotoxicity and cellular uptake studies. Nano-size micelles with spherical morphology and negative charge encapsulated 76.52 ± 0.94% and 32.87 0.61% Paclitaxel and Naringin, respectively. The micelles were thermally stable and retained 87.05 ± 0.69% and 92.88 ± 2.17% Paclitaxel and Naringin upon one-month storage. Maximum drug release was achieved at fourth hour of the study for both the loaded drugs. Paclitaxel co-encapsulation with Naringin synergistically improved its intracellular uptake and 65% in vitro cytotoxicity against breast cancer cells was achieved at its lower dose of 15 µg/mL. Results suggest that co-encapsulation of Paclitaxel with Naringin in mixed micelles is an effective strategy for achieving its higher anticancer activity.

Fabrication of lecithin-gum tragacanth muco-adhesive hybrid nano-carrier system for in-vivo performance of Amphotericin B.

Nano-carriers are excellent systems for improving bioavailability of poor aqueous soluble drugs. This study focuses fabrication of lecithin-gum tragacanth muco-adhesive hybrid NPs for enhancing Amphotericin B (AmpB) oral bioavailability. AmpB loaded lecithin NPs were synthesized through solvent diffusion method. Green synthesis of stable muco-adhesive gum tragacanth (GT) gold NPs was confirmed through UV-vis spectrophotometer and FT-IR. AmpB loaded lecithin NPs hybrid with GT gold NPs were characterized for shape, size, polydispersity index (PDI), zeta potential, drug entrapment efficiency and drug-excepients interactions using atomic force microscope (AFM), zetasizer, UV-vis spectrophotometer and FT-IR respectively. In-vivo bioavailability of AmpB loaded in NPs was investigated in rabbits. AmpB loaded muco-adhesive NPs were found polydispersed with 358.3 ±â€¯1.78 nm mean size and -19.9 ±â€¯0.51 mV zeta potential. They entrapped 78.91 ±â€¯2.44% AmpB and enhanced its oral bioavailability in animals. Results reveal the hybrid NPs as efficient carriers for enhancing AmpB oral bioavailability in controlled manner.

Gum acacia stabilized silver nanoparticles based nano-cargo for enhanced anti-arthritic potentials of hesperidin in adjuvant induced arthritic rats.

Nanomedicines anticipate drug delivery to inflamed tissues in rheumatoid arthritis (RA) with greater efficacy and lesser side effects. This study investigates the anti-arthritic potentials of Hesperidin (HP) loaded in gum acacia (GA) stabilized green silver nanoparticles (AgNPs). Synthesized GA-AgNPs were characterized through UV-vis spectrophotometer, zetasizer and atomic force microscope (AFM). The HP and its loaded NPs were tested for RA in Complete Freund's adjuvant (CFA) induced arthritis model. GA-AgNPs were found in nano-range size with negative charge, spherical shape and loaded increased HP amount. HP loaded GA-AgNPs showed minimal arthritic score exhibiting mild to moderate tissue swelling, reduced degenerative changes along with mild articular changes. Histopathological analysis revealed comparatively lesser influx of inflammatory cells and diminished granulamatous inflammation in ankle joints tissues in the presence of HP loaded GA-AgNPs. RT-PCR revealed that HP loaded GA-AgNPs significantly reduced the TLRs mRNA expression. Results validate GA stabilized green AgNPs as stable nano-cargos for targeted delivery of HP for restoring the progression of RA.

Bergenin loaded gum xanthan stabilized silver nanoparticles suppress synovial inflammation through modulation of the immune response and oxidative stress in adjuvant induced arthritic rats.

Bergenin (BG) is a naturally occurring C-glycoside with demonstrated anti-arthritic potential. Its therapeutic efficacy is compromised due to its lower absorption and instability at neutral-basic pH. The present study reports fabrication of gum xanthan (GX) stabilized silver nanoparticles (AgNPs) with BG for anti-arthritic activity in a CFA-induced arthritis model targeting ROS, cytokines and TLR expression. NPs were characterized through UV-vis, zetasizer, FT-IR and AFM. Oral administration of BG loaded NPs (1 mg kg-1) exhibited potent anti-arthritic activity with a minimal arthritic score, mild to moderate paw tissue swelling, reduced degenerative changes along with mild articular changes and less influx of inflammatory cells in macroscopic X-ray and histological examination. Administration of BG and its NPs suppressed the levels of reactive oxygen species (ROS) significantly as compared to the arthritic control group. Moreover, increased production of O2˙- in human neutrophils, stimulated by opsonized zymosan (OZ) and phorbol-12-myristate-13-acetate (PMA) was also suppressed. BG and its loaded NPs were revealed to antagonize the oxidative stress via interference with the NADPH oxidase metabolic pathway. Their anti-oxidant activity was further assessed by their inhibitory effect against TLR (TRL-2 & -4) and cytokine (IL-1ß, IL-6 and TNF-α) production. The current investigation validates GX stabilized AgNPs as stable and promising multi-targeted therapeutic nano-cargo for BG delivery with efficient treatment of RA.

Gum tragacanth stabilized green gold nanoparticles as cargos for Naringin loading: A morphological investigation through AFM.

Gold nanoparticles (AuNPs) have attracted greater scientific interests for the construction of drugs loading cargos due to their biocompatibility, safety and facile surface modifications. This study deals with the fabrication of gum tragacanth (GT) green AuNPs as carrier for Naringin, a less water soluble therapeutic molecule. The optimized AuNPs were characterized through UV-vis spectroscopy, FT-IR and atomic force microscope (AFM). Naringin loaded nanoparticles were investigated for their bactericidal potentials using Tetrazolium Microplate assay. Morphological studies conducted via AFM revealed spherical shape for AuNPs with nano-range size and stabilized by GT multi-functional groups. The AuNPs acted as carrier for increased amount of Naringin. Upon loading in AuNPs, Naringin An increased in the bactericidal potentials of Naringin was observed after loading on AuNPs against various tested bacterial strains. This was further authenticated by the surface morphological analysis, showing enhanced membrane destabilizing effects of loaded Naringin. The results suggest that GT stabilized green AuNPs can act as effective delivery vehicles for enhancing bactericidal potentials of Naringin.