QbD-Based Fabrication of Biomimetic Hydroxyapatite Embedded Gelatin Nanoparticles for Localized Drug Delivery against Deteriorated Arthritic Joint Architecture.

Biomaterials such as nanohydroxyapatite and gelatin are widely explored to improve damaged joint architecture associated with rheumatoid arthritis (RA). Besides joint damage, RA is associated with inflammation of joints and cartilage, which potentiates the need for both bone nucleation and therapeutic intervention. For such purpose, a modified nanoprecipitation method is used herein to fabricate tofacitinib (Tofa)-loaded nanohydroxyapatite (nHA) embedded gelatin (GLT) nanoparticles (NPs) (Tofa-nHA-GLT NPs). The quality by design (QbD) approach is chosen to assess the key parameters that determine the efficiency of the NPs, and are further optimized via Box-Behnken design of experiment. The particle size, polydispersity, zeta potential, and encapsulation efficiency (EE) of the prepared NPs are found to be 269 nm, 0.18, -20.5 mV, and 90.7%, respectively. Furthermore, the NPs have improved stability, skin permeability, and a sustained drug release pattern at pH 6.5 (arthritic joint pH). Moreover, rhodamine-B loaded nHA-GLT NPs demonstrates considerably higher cellular uptake by the murine-derived macrophages than free rhodamine-B solution. In vitro, cell-based experiments confirm the good cell biocompatibility with insignificant toxicity. Thus, QbD-based approach has successfully led to the development of Tofa-nHA-GLT NPs with the potential to target inflamed arthritic joint.

Dual pH and microbial-sensitive galactosylated polymeric nanocargoes for multi-level targeting to combat ulcerative colitis.

Ulcerative colitis (UC) is a type of inflammatory bowel disease characterized by inflammation, ulcers and irritation of the mucosal lining. Oral drug delivery in UC encounters challenges because of multifaceted barriers. Dexamethasone-loaded galactosylated-PLGA/Eudragit S100/pullulan nanocargoes (Dexa-GP/ES/Pu NCs) have been developed with a dual stimuli-sensitive coating responsive to both colonic pH and microbiota, and an underneath galactosylated-PLGA core (GP). The galactose ligand of the GP preferentially binds to the macrophage galactose type-lectin-C (MGL-2) surface receptor. Therefore, both stimuli and ligand-mediated targeting facilitate nanocargoes to deliver Dexa specifically to the colon with enhanced macrophage uptake. Modified emulsion method coupled with a solvent evaporation coating technique was employed to prepare Dexa-GP/ES/Pu NCs. The nanocargoes were tested using in vitro, ex vivo techniques and dextran sodium sulfate (DSS) induced UC model. Prepared nanocargoes had desired physicochemical properties, drug release, cell uptake and cellular viability. Investigations using a DSS-colitis model showed high localization and mitigation of colitis with downregulation of NF-ĸB and COX-2, and restoration of clinical, histopathological, biochemical indices, antioxidant balance, microbial alterations, FTIR spectra, and epithelial junctions' integrity. Thus, Dexa-GP/ES/Pu NCs found to be biocompatible nanocargoes capable of delivering drugs to the inflamed colon with unique targeting properties for prolonged duration.

QbD-based fabrication of transferrin-anchored nanocarriers for targeted drug delivery to macrophages and colon cells for mucosal inflammation healing.

Colon mucosal inflammation attracts a plethora of immune cells with overexpressed surface receptors. Colon drug targeting can be aided by exploiting overexpressed cell surface receptors which improve drug site retention for an extended period. We developed Tofacitinib citrate (Tofa) loaded transferrin anchored PLGA nanocarriers (Tofa-P/tfr NCs) via the quality by design (QbD) approach for specific binding to the transferrin receptor (TFR-1/CD71) overexpressed on macrophages and colon epithelial cells. Nanocarriers were produced using a modified emulsion-evaporation method with a protein adsorption technique. The QbD-risk assessment method was adopted to screen the variables impacting the quality of nanocarriers, which were then optimized using the 33 Box-Behnken design of experiment (DOE). The obtained nanocarriers have the desired physicochemical properties, drug entrapment, tfr adsorption, stability, mucoadhesion, and sustained drug release pattern at pH 7.4 (colon pH). In vitro cell-based studies confirmed the cellular biocompatibility and considerable uptake of nanocarriers by colon and macrophage cells; the uptake was diminished by anti-CD71/TFR1 antibodies. Tofa-P/tfr NCs demonstrated good colon targeting potential in the dextran sulfate sodium (DSS) induced ulcerative colitis (UC) model. In vivo therapeutic efficacy against UC was established through restored morphological and histopathological scores, vascular integrity, antioxidant levels, hematological parameters, pro-inflammatory cytokine/marker levels, and microbial indices. Tofa-P/tfr NCs shut down the elevated STAT-1 and TFR-1 levels, demonstrating the enhanced efficacy of the encapsulated drug. Thus, the QbD-driven approach successfully developed Tofa-P/tfr NCs with good potential to mitigate mucosal inflammation by targeting colon and macrophage surface receptors.

Small but powerful: will nanoparticles be the future state-of-the-art therapy for IBD?

INTRODUCTION: Inflammatory bowel disease (IBD) is the inflammatory condition of the gastrointestinal tract particularly affecting the colon and the ileum. IBD patients can have a very poor quality of life because of the limited therapeutic efficacy and accompanied adverse effects. AREAS COVERED: The potential ways to employ nanoparticles to deliver drugs to a certain site of inflammation are discussed. The focus was set on the microenvironment in the gut as well as the mucosa, epithelial layer and the microbiota. Moreover, experimental animal colitis models were nanoparticles were used as a potential treatment are presented. Lastly, challenges for the potential clinical use in humans are discussed. EXPERT OPINION: Although there still remain many open questions e.g. regarding the toxicity, the metabolism or the pharmacokinetics of nanoparticles further research on this topic could overcome these challenges. For example, instead of synthetically engineered particles, biodegradable components could be used. Since there have been a lot pf promising results in the recent years, we are sure that in the future nanoparticles will be developed in a way to ensure safe and targeted delivery of drugs to the site of inflammation.

Nanomaterials in the Management of Gram-Negative Bacterial Infections.

The exploration of multiplexed bacterial virulence factors is a major problem in the early stages of Escherichia coli infection therapy. Traditional methods for detecting Escherichia coli (E. coli), such as serological experiments, immunoassays, polymerase chain reaction, and isothermal microcalorimetry have some drawbacks. As a result, detecting E. coli in a timely, cost-effective, and sensitive manner is critical for various areas of human safety and health. Intelligent devices based on nanotechnology are paving the way for fast and early detection of E. coli at the point of care. Due to their specific optical, magnetic, and electrical capabilities, nanostructures can play an important role in bacterial sensors. Another one of the applications involved use of nanomaterials in fighting microbial infections, including E. coli mediated infections. Various types of nanomaterials, either used directly as an antibacterial agent such as metallic nanoparticles (NPs) (silver, gold, zinc, etc.), or as a nanocarrier to deliver and target the antibiotic to the E. coli and its infected area. Among different types, polymeric NPs, lipidic nanocarriers, metallic nanocarriers, nanomicelles, nanoemulsion/ nanosuspension, dendrimers, graphene, etc. proved to be effective vehicles to deliver the drug in a controlled fashion at the targeted site with lower off-site drug leakage and side effects.

Simulation, In Vitro, and In Vivo Cytotoxicity Assessments of Methotrexate-Loaded pH-Responsive Nanocarriers.

In this study, pH-responsive niosomal methotrexate (MTX) modified with ergosterol was prepared for potential anticancer application. The prepared formulation had a size of 176.7 ± 3.4 nm, zeta potential of -31.5 ± 2.6 mV, EE% of 76.9 ± 2.5%, and a pH-responsive behavior in two different pHs (5.4 and 7.4). In-silico evaluations showed that MTX intended to make a strong hydrogen bond with Span 60 compartments involving N2 and O4 atoms in glutamic acid and N7 atom in pteridine ring moieties, respectively. The cytotoxic effects of free and pH-MTX/Nio were assessed against MCF7 and HUVECs. Compared with free MTX, we found significantly lower IC50s when MCF7 cells were treated with niosomal MTX (84.03 vs. 9.464 µg/mL after 48 h, respectively). Moreover, lower cell killing activity was observed for this formulation in normal cells. The pH-MTX/Nio exhibited a set of morphological changes in MCF7 cells observed during cell death. In-vivo results demonstrated that intraperitoneal administration of free MTX (2 mg/kg) after six weeks caused a significant increase in serum blood urea nitrogen (BUN), serum creatinine, and serum malondialdehyde (MDA) levels of rats compared to the normal control rats. Treatment with 2 and 4 mg/kg doses of pH-MTX/Nio significantly increased serum BUN, serum creatinine, and serum lipid peroxidation. Still, the safety profile of such formulations in healthy cells/tissues should be further investigated.

Evaluating the mucoprotective effects of glycyrrhizic acid-loaded polymeric nanoparticles in a murine model of 5-fluorouracil-induced intestinal mucositis via suppression of inflammatory mediators and oxidative stress.

OBJECTIVES: 5-Fluorouracil (5-FU), a chemotherapeutic drug, has severe deteriorating effects on the intestine, leading to mucositis. Glycyrrhizic acid is a compound derived from a common herbal plant Glycyrrhiza glabra, with mucoprotective, antioxidant and anti-inflammatory actions, however, associated with poor pharmacokinetics. Owing to the remarkable therapeutic action of glycyrrhizic acid-loaded polymeric nanocarriers in inflammatory bowel disease, we explored their activity against 5-FU-induced intestinal mucositis in mice. Polymeric nanocarriers have proven to be efficient drug delivery vehicles for the long-term treatment of inflammatory diseases, but have not yet been explored for 5-FU-induced mucositis. Therefore, this study aimed to produce glycyrrhizic acid-loaded polylactic-co-glycolic acid (GA-PLGA) nanoparticles to evaluate their protective and therapeutic effects in a 5-FU-induced mucositis model. METHODS: GA-PLGA nanoparticles were prepared using a modified double emulsion method, physicochemically characterized, and tested for in vitro drug release. Thereafter, mucositis was induced by 5-FU (50 mg/kg; IP) administration to the mice for the first 3 days (day 0, 1, 2), and mice were treated orally with GA-PLGA nanoparticles for 7 days (day 0-6). RESULTS: GA-PLGA nanoparticles significantly reduced mucositis severity measured by body weight, diarrhea score, distress, and anorexia. Further, 5-FU induced intestinal histopathological damage, altered villi-crypt length, reduced goblet cell count, elevated pro-inflammatory mediators, and suppressed antioxidant enzymes, all of which were reversed by GA-PLGA nanoparticles. CONCLUSION: Morphological, behavioral, histological, and biochemical results suggested that GA-PLGA nanoparticles were efficient, biocompatible, targeted, and sustained release drug delivery nano-vehicle for enhanced mucoprotective, anti-inflammatory, and antioxidant effects in 5-FU-induced intestinal mucositis.

DNA Based and Stimuli-Responsive Smart Nanocarrier for Diagnosis and Treatment of Cancer: Applications and Challenges.

The rapid development of multidrug co-delivery and nano-medicines has made spontaneous progress in tumor treatment and diagnosis. DNA is a unique biological molecule that can be tailored and molded into various nanostructures. The addition of ligands or stimuli-responsive elements enables DNA nanostructures to mediate highly targeted drug delivery to the cancer cells. Smart DNA nanostructures, owing to their various shapes, sizes, geometry, sequences, and characteristics, have various modes of cellular internalization and final disposition. On the other hand, functionalized DNA nanocarriers have specific receptor-mediated uptake, and most of these ligand anchored nanostructures able to escape lysosomal degradation. DNA-based and stimuli responsive nano-carrier systems are the latest advancement in cancer targeting. The data exploration from various studies demonstrated that the DNA nanostructure and stimuli responsive drug delivery systems are perfect tools to overcome the problems existing in the cancer treatment including toxicity and compromised drug efficacy. In this light, the review summarized the insights about various types of DNA nanostructures and stimuli responsive nanocarrier systems applications for diagnosis and treatment of cancer.

A holistic QBD approach to design galactose conjugated PLGA polymer and nanoparticles to catch macrophages during intestinal inflammation.

Recruited macrophages in inflammation attract various ligand-receptor drug delivery approaches. Galactose bound nanocarriers are promising to catch macrophages because of surface-expressed macrophage galactose type-lectin-C (MGL-2) receptor. The present study reported fabrication of galactose conjugated PLGA (GAL-PLGA) polymer and nanoparticles under quality by design (QBD) approach to investigate macrophages targeting potential at inflamed intestine. GAL-PLGA nanoparticles were fabricated through O/W emulsion-evaporation method under QBD approach and Box-Behnken design. Obtained GAL-PLGA nanoparticles have optimum particle size (~118 nm), drug entrapment (87%) and zeta potential (-9.5). TGA, XPRD and FTIR confirmed stability and negate drug-polymer interactions. Further, nanoparticles have considerable hemocompatibility, biocompatibility and cellular uptake; macrophage uptake was inhibited by D-galactose confirming involvement of MGL-2. Moreover, drug retention studies in the DSS-colitis model provide background for potential of nanoparticles to target and reside inflamed intestine. It is concluded that GAL-PLGA nanoparticles are suitable platform to target macrophages at the inflamed intestine through oral route.

Topical delivery of curcumin-loaded transfersomes gel ameliorated rheumatoid arthritis by inhibiting NF-κß pathway.

Aim: To fabricate and evaluate curcumin-loaded transfersomes (Cur-TF) for the targeted delivery and enhanced therapeutic efficacy of curcumin for the treatment of rheumatoid arthritis (RA). Methods: Modified thin-film hydration method was used to prepare Cur-TF which were then embedded into carbopol-934 gel. They were further evaluated through in vitro techniques and in an in vivo arthritis model. Results: Cur-TF had optimal particle size, spherical morphology, high encapsulation efficiency and sustained drug release profiles. The Cur-TF gel had better in vitro skin penetration than plain curcumin. In vivo findings demonstrated improved clinical, histological and x-ray scores and reduced pro-inflammatory cytokines through NF-κß inhibition. Conclusion: Cur-TF gel delivered curcumin to the arthritic dermal tissue through a topical route and demonstrated promising therapeutic efficacy by significantly alleviating complete Freud's adjuvant (CFA)-induced arthritis.

Aerodynamic properties and in silico deposition of isoniazid loaded chitosan/thiolated chitosan and hyaluronic acid hybrid nanoplex DPIs as a potential TB treatment.

Existing therapies yield low drug encapsulation or accumulation in the lungs, hence the site-specific drug delivery remains the challenge for tuberculosis. Lately, dry powder inhalers (DPIs) are showing promising drug deposition in the deeper lung tissues. Biocompatible polymers with the ability to naturally recognize and bind to the surface receptors of alveolar macrophages, the reservoir of the causative organism, were selected. DPIs comprised of chitosan (CS)/thiolated chitosan (TC) in conjugation with Hyaluronic acid (HA) were synthesized loaded with isoniazid (INH) by using the Design of Experiment (DoE) approach. Nanosuspensions were prepared by ionic gelation method using cross-linker, sodium-tripolyphosphate (TPP) and were optimized by using Box-Behnken 3-level screening design and later freeze-dried to obtain nanopowders. Physico-chemical compatibility of nanoplex systems was investigated using in-vitro characterization techniques. In-vitro release and permeation studies were correlated in terms of the pattern of drug content dissolved over time. In addition, the cytotoxicity studies on A549 cells demonstrated the safety profile of the nanoplexes. Moreover, in-silico studies and aerodynamic profiles verify the suitability of DPIs for further in-vivo tuberculosis therapeutics. DoE analyses affirmed the lack of linearity in the model for the certain response of studied parameters in a holistic way, which was not possible else ways.

Biomimetic hydroxyapatite as potential polymeric nanocarrier for the treatment of rheumatoid arthritis.

Rheumatoid arthritis (RA) is a chronic autoimmune disease of unknown etiology with much higher prevalence in world's population. RA is initially associated with inflammation of joints and cartilages that ultimately leads to their destruction thus requiring a therapeutic intervention. The available conventional therapeutic strategies for RA are not satisfactory because of the associated side effects such as toxicity, delayed action, and dependence. Therefore, a carrier system is required that should deliver the drug to the target site with minimal side effects. In this connection, nanocarrier systems are of prime importance because of the associated benefits such as their nano-scaled size, targeted drug delivery, and reduced toxicity that can improve the patient's compliance. Moreover, in the past few decades, nano-particulate-based drug delivery approaches that have been investigated for the treatment of RA include ceramics, polymers, and hydrogels. Among these nanocarrier systems, ceramics like hydroxyapatite have gathered striking attention due to their bioactive, biocompatible, and bio-conductive characteristics. Nano-sized hydroxyapatite (HA) permeates the bone tissues and serves as a source of calcium phosphates required for bone repairing that are damaged during disease process. The aim of this review article is to highlight the potential use of HA as nanocarrier for anti-rheumatic drugs as well its possible effect on bone remodeling.

Glycyrrhizic acid-loaded pH-sensitive poly-(lactic-co-glycolic acid) nanoparticles for the amelioration of inflammatory bowel disease.

Aim: To fabricate and evaluate the therapeutic efficacy of glycyrrhizic acid (GA)-loaded pH-sensitive nanoformulations that specifically target and combat mucosal inflammation of the colon. Methods: GA-loaded Eudragit® S100/poly-(lactic-co-glycolic acid) nanoparticles were developed through modified double-emulsion evaporation coupled with solvent evaporation coating techniques and analyzed for physicochemical characteristics, surface chemistry, release kinetics, site-retention and therapeutic effectiveness. Results: Nanoparticles have a particle size of approximately 200 nm, high encapsulation efficiency, desired surface chemistry with pH-dependent and sustained drug release behavior following the Gompertz kinetic model. In vivo retention and therapeutic effectiveness in the inflamed colon tissues were confirmed by macroscopic and microscopic indices, cytokine analysis and antioxidant assays. Conclusion: GA-loaded Eudragit S100/poly-(lactic-co-glycolic acid) nanoparticles could efficiently deliver GA to the colon and ameliorate the mucosal inflammation for a prolonged duration.

Advances in orally-delivered pH-sensitive nanocarrier systems; an optimistic approach for the treatment of inflammatory bowel disease.

Inflammatory bowel disease (IBD) is the inflammation of the gastrointestinal tract (GIT) affecting the colon and ileum in particular. Increasing IBD prevalence worldwide is alarming, and needs to be resolved. Due to the limited therapeutic efficacy, accompanying adverse effects, dependence, and pharmacokinetics issues of the available therapy, IBD patients have compromised quality of life. Meanwhile, conventional drug delivery systems (DDS) for IBD face many obstacles en-route to the colon, such as physiological and pathophysiological barriers, genetic variability, disease severity, and nutrition status. Therefore, the pH-dependent nanocarrier DDS is a recent advancement that fulfills the need for a more tolerable and effective remedy for IBD. It facilitates localized and targeted action, eliminating systemic adverse effects and unnecessary flushing of the drug from the inflamed colon tissues. The integration of a pH-sensitive polymer as a nanocarrier provides protection in drug transport to the lower region of the GIT. In this review, we will briefly explain IBD pathophysiology, the pros and cons of pH-dependent conventional DDS, and highlight a novel pH-dependent nanocarrier system for treating the disease.