Targeted Macrophage Re-Programming: Synergistic Therapy With Methotrexate and RELA siRNA Folate-Liposome in RAW264.7 Cells and Arthritic Rats.

Rheumatoid arthritis (RA) is a chronic autoimmune disorder characterized by joint inflammation and destruction. Current treatments, such as Methotrexate (MTX), though effective, often face limitations such as high plasma Cmax and lack of sustained release. This study explores a synergistic approach to RA therapy using folate-liposomal co-delivery of MTX and RELA siRNA (short interfering RNA), targeting RAW264.7 macrophage repolarization via nuclear factor kappa B (NF-κB) pathway inhibition. Extensive in vitro characterizations demonstrate the stability and biocompatibility of this therapy via folate-liposomes. In the collagen-induced arthritis (CIA) rat model, treatment leads to reduced synovial inflammation and improved mobility. The combined MTX and RELA siRNA approach indirectly inhibits inflammatory cytokines, rheumatoid factor (RF), and C-reactive protein (CRP). Targeted macrophage delivery shows marked therapeutic effects in RAW264.7 murine macrophages, potentially modulating M1 to M2 polarization. This research presents a promising avenue for innovative RA therapies by inhibiting the inflammatory cascade and preventing joint damage.

Bortezomib-loaded immunoliposomes against CD44 expressing macrophages: an interplay for inflammation resolution.

Macrophage-driven inflammation is the central player in a range of pathological conditions, comprising autoimmune disorders, various cancers, as well as chronic inflammatory states like rheumatoid arthritis. Therapeutic strategies tailored to specifically target macrophage behavior have acquired substantial interest for their potential to alleviate chronic inflammation effectively. In this study, we introduce a pioneering therapeutic approach utilizing specialized CD44-targeted immunoliposomes carrying bortezomib to address inflammation at the cellular level and the significance of this strategy lies in its precision nature. Bortezomib's inhibition of the proteasome interferes with the finely-tuned mechanism that controls NFκB activation, ultimately leading to a downregulation of the inflammatory response. After performing computational docking demonstrating its strong binding affinity to the proteasome molecule, the resulting nano-construct displayed a hydrodynamic size of 144.26 ± 74.4 nm and a quasi-spherical morphology. Moreover, the nano-construct ensured a minimum shelf-life of 30 days, aiming for targeted delivery with practical longevity. Upon internalization of immunoliposomes, the interaction with CD44 receptors exhibited downstream signaling events. This included the activation of Jun amino-terminal kinases 1/2 (JNK1/2) and the extracellular-signal-regulated kinases (ERK) pathway. JNK1/2 activation may lead to the release of mitochondrial pro-apoptotic factors, triggering the intrinsic apoptotic pathway and activation of caspases, which was confirmed from the level of apoptotic gene and protein expression. The precise targeting and anti-inflammatory action of this therapy against macrophages hold promise for therapeutic interventions in a wide range of inflammatory conditions, offering a novel avenue for precision medicine in the battle against excessive inflammation.

Nanoemulsion-Based System as a Novel and Promising Approach for Enhancing the Antimicrobial and Antitumoral Activity of Thymus vulgaris (L.) Oil in Human Hepatocellular Carcinoma Cells.

The utilisation of medicinal plants and their essential oils is receiving more attention due to the ineffectiveness of current therapeutic methods in the treatment of various cancers and the rising incidence of bacterial antibiotic resistance. Thymol, an active ingredient of Thymus vulgaris, is known to have hepatoprotective, antibacterial, and antioxidant properties. To overcome major obstacles to their usage, such as quick oxidation and high volatility, plant essential oils must be administered through a system to improve the delivery of their active pharmaceutical ingredient. The bioavailability of active substances may be enhanced by the colloidal dispersion nanoemulsion. Therefore, this study aims to derive a comparative evaluation of the thyme oil nanoemulsion formulation and the characterisation of its antibacterial and antitumorigenic activities. A nanoemulsion (NE) with a droplet size of 122.2 ± 1.079 nm was discovered to be stable and mono-dispersed for 4 months and inhibited the growth of B. subtilis, E. coli, P. aeruginosa, and S. aureus. It also displayed antitumorigenic capabilities in HepG2 cells by arresting the cell cycle in the G2/M phase and upregulating the gene expression levels of Bcl-2-associated X protein (Bax), Caspase 3, 8, and 9, as well as a concomitant concentration-dependent decrease in B-cell leukaemia/lymphoma 2 protein (BCL2). Along with an increase in inducible nitric oxide synthase (iNOS) levels, upregulation of the expression levels of the reactive oxygen species (ROS), mitogen-activated protein kinase (MAPK), and endoplasmic reticulum (ER) stress pathways was also seen, indicating of ROS formation in the cancer cells.

Functional hydrogel-based wound dressings: A review on biocompatibility and therapeutic efficacy.

Chronic wounds, burns, and surgical incisions represent critical healthcare challenges that significantly impact patient quality of life and strain healthcare resources. In response to these pressing needs, the field of wound healing has witnessed a radical advancement with the emergence of functional hydrogel-based dressings. This review article underscores the severity and importance of this transformative study in the domain of wound healing. The hydrogel matrix offers a moist and supportive environment that facilitates cellular migration, proliferation, and tissue regeneration, vital for efficient wound closure. Their conformable nature ensures patient comfort, reducing pain and uneasiness during dressing changes, particularly in chronic wounds where frequent interventions are required. Beyond their structural merits, functional hydrogel dressings possess the capability of incorporating bioactive molecules such as growth factors and antimicrobial agents. This facilitates targeted and sustained delivery of therapeutics directly to the wound site, addressing the multifactorial nature of chronic wounds and enhancing the healing trajectory. The integration of advanced nanotechnology has propelled the design of hydrogel dressings with enhanced mechanical strength and controlled drug release profiles, amplifying their therapeutic potential. In conclusion, the significance of this study lies in its ability to revolutionize wound healing practices and positively impact the lives of countless individuals suffering from chronic wounds and burns. As this transformative technology gains momentum, it holds the promise of addressing a major healthcare burden worldwide, thus heralding a new era in wound care management.

Reprogramming M1-to-M2 Phenotype to Alleviate Inflammation: Using Liposomal Curcumin as a Tool to Redefine Macrophage Functionality.

The versatile nature of macrophages and their ability to switch between various activation states plays a pivotal role in both promoting and inhibiting inflammatory processes. In pathological inflammatory conditions, classically activated M1 macrophages are often associated with initiating and maintaining inflammation, while alternatively activated M2 macrophages are linked to the resolution of chronic inflammation. Achieving a favorable equilibrium between M1 and M2 macrophages is crucial for mitigating inflammatory environments in pathological conditions. Polyphenols are known to have strong inherent antioxidative capabilities, and curcumin has been found to reduce macrophage inflammatory reactions. However, its therapeutic efficacy is compromised due to its poor bioavailability. The present study aims to harness the properties of curcumin by loading it in nanoliposomes and enhancing the M1-to-M2 macrophage polarization. A stable liposome formulation was achieved at 122.1 ± 0.08 nm, and a sustained kinetic release of curcumin was observed within 24 h. The nanoliposomes were further characterized using TEM, FTIR, and XRD, and the morphological changes in macrophage cells, RAW264.7, were observed in SEM, indicating a distinct M2-type phenotype after the treatment with liposomal curcumin. ROS may partially control macrophage polarization and be observed to decrease after treatment with liposomal curcumin. The nanoliposomes were able to successfully internalize in the macrophage cells, and an enhanced expression of ARG-1 and CD206 with a decrease in iNOS, CD80, and CD86 levels suggested the polarization of LPS-activated macrophages toward the M2 phenotype. Also, liposomal curcumin treatment dose-dependently inhibited TNF-α, IL-2, IFN-γ, and IL-17A at secretory levels and simultaneously increased the levels of cytokines like IL-4, IL-6, and IL-10.

Development of food-grade antimicrobials of fenugreek oil nanoemulsion-bioactivity and toxicity analysis.

Antimicrobials of natural origin are proving to be an effective solution to emerging antimicrobial resistance and its physiological side effects. Recent studies have demonstrated that essential oils encapsulated in the form of nanoemulsions have better antimicrobial activity than the oil itself, possibly due to its high stability, solubility, sustained release, and increased bioavailability. In the present study, fenugreek oil-a well-known antimicrobial and antioxidant-has been used to fabricate nanoemulsion (NE), with an objective to meet potential alternative to synthesized antimicrobials. A combination of three different components, water, fenugreek oil, and Tween 80, has been used to prepare the nanoemulsions of different size and one of the most stable nanoemulsion with lowest concentration of surfactant Tween 80 was used to assess its bioactivity, antimicrobial properties, and toxicity against human hepatic cell line. Among all the formulations, nano-emulsion with 2.5% oil concentration, 30 min sonication (hydrodynamic size 135.2 nm, zeta potential 36.8 mV, PDI 0.135, and pH 5.12), was selected for all studies. The nanoemulsion showed potential antibacterial activity against all the microbial strains (Escherichia coli, Bacillus subtilis, Staphylococcus aureus, and Pseudomonas aeruginosa) used in this study. Interestingly, the nanoemulsion showed potential antibacterial activity against P. aeruginosa, known to show resistance against ampicillin. The toxicity evaluation in human hepatic cell line (WRL-68) indicated no significant toxicity of nanoemulsion up to the concentration of 800 µg/ml. The synthesized nanoemulsion thus has a translational potential as a food-grade non-toxic natural nanoantimicrobials.

Recent advances in nanoparticle-based drug delivery systems for rheumatoid arthritis treatment.

Nanotechnology has increasingly emerged as a promising tool for exploring new approaches, from treating complex conditions to early detection of the onset of multiple disease states. Tailored designer nanoparticles can now more comprehensively interact with their cellular targets and various pathogens due to a similar size range and tunable surface properties. The basic goal of drug delivery is to employ pharmaceuticals only where they are needed, with as few adverse effects and off-target consequences as possible. Rheumatoid arthritis (RA) is a chronic inflammatory illness that leads to progressive loss of bone and cartilage, resulting in acute impairment, decreased life expectancy, and increased death rates. Recent advancements in treatment have significantly slowed the progression of the disease and improved the lives of many RA sufferers. Some patients, on the other hand, attain or maintain illness remission without needing to continue immunosuppressive therapy. Furthermore, a large percentage of patients do not respond to current treatments or acquire tolerance to them. As a result, novel medication options for RA therapy are still needed. Nanocarriers, unlike standard medications, are fabricated to transport drugs directly to the location of joint inflammation, evading systemic and negative effects. As a result, researchers are reconsidering medicines that were previously thought to be too hazardous for systemic delivery. This article gives an overview of contemporary nanotechnology-based tactics for treating rheumatoid arthritis, as well as how the nanotherapeutic regimen could be enhanced in the future.

MiR-206 conjugated gold nanoparticle based targeted therapy in breast cancer cells.

MicroRNAs (miRNAs) are single-stranded, non-coding, 19-25 nucleotide RNA molecules that have been observed to be dysregulated in many diseases including cancer. miRNAs have been known to play an important role in cellular proliferation, differentiation, migration, apoptosis, survival, and morphogenesis. Breast cancer is heterogeneous in nature and contributed extensively to the increased mortality rate. miRNA can either be tumor-suppressive or oncogenic in nature. The level of expression of miRNA changes according to the subtypes of cancer and the mutation responsible for different cancers. miRNA mimicry or inhibition are emerging possible therapies to maintain the level of miRNA inside the cells. In order to have proper miRNA mimicry, the major hurdle is to deliver the miRNA mimics at the site of tumor. Metallic nanoparticles with modified surface can be used to solve the problem of miRNA delivery. MiR-206 is reported to be down-regulated in Luminal-A type of breast cancer. In the current manuscript, we aim to modify the surface of gold-nanoparticles (AuNPs) with PEG moiety and allow miRNA to attach to it. The fabricated nano-complex, not only delivered miR-206 but also caused cell death in MCF-7 by arresting cells in the G0-G1 phase and inducing apoptosis by downregulating NOTCH 3.

Fabrication of methotrexate-loaded gold nanoconjugates and its enhanced anticancer activity in breast cancer.

Methotrexate (MTX) is known antagonist of folic acid and widely used as an anti-cancer drug. The folate receptor (FR) and reduced folate carrier are mostly responsible for internalization of methotrexate in tumor cells. Mutation in reduced folate carrier (RFC) leads to resistance against MTX in various tumor cell lines including MDA-MB-231 breast cancer cells. To overcome the resistance of MTX, folate receptor targeted nanoparticles have been commonly used for targeting breast tumors. The aim of the study is to determine the ability of methotrexate gold nanoparticles (MTX-GNPs) in the induction of apoptosis and to explore the molecular changes at genomics and proteomics level. Different assays like cell viability assay, cell cycle analysis, apoptosis, real-time PCR and western blot were carried out to evaluate the anti-cancer effect of MTX-Gold NPs on MCF-7 and MDA-MB-231 cells. Our observations demonstrated the decrease in the percent viable cells after the treatment of MTX-GNPs, with an arrest in cell cycle at G0/G1 phase and a significant increase in apoptotic cell population and loss of mitochondrial membrane potential in MCF-7 and MDA-MB-231 cells. Folate receptor targeted MTX-GNPs showed significant cellular uptake in breast cancer cells along with significant down-regulation in expression of anti-apoptotic gene (Bcl-2) and up-regulation in expression of pro-apoptotic genes (Bax, Caspase-3, Caspase-9, APAF-1, p53). These results unveil the increased anti-cancer effect of MTX-GNPs in cancer cells. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s13205-021-02718-7.