Non-Invasive Intranasal Delivery of pApoE2: Effect of Multiple Dosing on the ApoE2 Expression in Mice Brain.

Chitosan-based polymeric micelles are promising non-viral nanocarriers for safe and targeted gene delivery. Multi-functionalized chitosan polymeric micelles were prepared by grafting fatty acid, cell-penetrating peptide, and mannose on the chitosan backbone. The polymeric micelles were subjected to surface morphology and surface topography using scanning electron microscopy and atomic force microscopy, respectively. The hemotoxic profile of the prepared polymeric micelles was established against erythrocytes and was found to be <5% hemotoxic up to the concentration of 600 µg/mL. In vitro ApoE2 expression in primary astrocytes and neurons was analyzed. Multi-functionalized polymeric micelles produced greater (p < 0.05) transfection in astrocytes and neurons in comparison to mono-functionalized micelles. Intranasal administration of polymeric micelles/pApoE2 polyplex led to significantly higher (p < 0.05) in vivo pApoE2 expression than chitosan and unfunctionalized polymeric micelles-treated mice groups. The outcomes of this study predict that the developed multi-functionalized polymeric micelles could be an effective and safe gene delivery platform to the brain through the intranasal route.

A stimuli-responsive nanocarrier for diagnosis of seizures and inhibition of glutaminase in epilepsy.

Epilepsy is marked by unpredictable and recurrent episodes of seizures. It is characterized by glutamate excitotoxicity and changes in stimuli such as pH, temperature and oxidative environment. This study aimed to formulate novel nanoparticulate theranostic nanocarrier for combined effects of diagnosis and treatment of epilepsy by: i) in-situ detection of epileptic conditions through characteristic changes in pH through the synthesis of pH-responsive polymer (CS-g-PD) and ii) 'on-demand' therapeutic alleviation of epileptic seizures through an inhibitor of glutaminase, 6-diazo-5-oxo-norleucine (DON). The formulation of DON-CS-g-PD-SLNs possessed nanodimensions (∼197.56 ± 17.87) nm and zeta potential (4.19 ± 0.29), with entrapment efficiency of (80.29 ± 0.006%). The coating pH-responsive polymer showed good sensitivity for acidic conditions by releasing the drug in pH 6.4 and resisting release in higher pH 7.2. In-vivo studies in Wistar rats showed suppression of epileptic seizures, escalation in the duration latency and reduction in duration of convulsions and recovery period. Furthermore, it was also successful in reducing the levels of glutaminase (p < 0.0001) in the brain of PTZ-kindled rats, thereby leading to a decrease in glutamate levels (p < 0.01). Hence, the nanocarriers show promising potential as 'on-demand' theranostics in epilepsy by reducing both the incidence and severity of convulsions.

Functional Reconstruction of Through and Through Cheek Defect with Free Composite Bi-pedalled Anterolateral Thigh Flap.

Extensive composite defects involving the lip and cheek present difficult reconstructive challenges in view of functional recovery like oral competence, articulation, speech and mastication. This study presents our results of reconstructing through and through cheek defect with the use of free anterolateral thigh (ALT) flaps. All the patients with carcinoma of buccal mucosa and cheek who underwent through and through cheek resection and reconstructed with free composite pedalled anterolateral thigh flap between March 2019 to March 2020 were included in the study. We have assessed the post-operative functional outcome using University of Washington Quality of Life Questionnaire at the end of 12 months duration. We have excluded lost to follow-up and flap failure patients. Fifty patients were matched our inclusion criteria. Mean age was 43 years. Male to female ratio was 4.6:1. Stage 3 disease was in 8 patients and stage 4 disease was in 42 patients. Average size of the flap required was 15 × 7.5 cm, maximum was 24 × 11 cm. Oral competence was good in 74% patients. 80% patients were able to take semisolid diet, 8% were on liquid diet and 12% were dependent on RT feed. Average physical function (70.47 ± 19.09), social-emotional sub-score (81.72 ± 16.63) and composite scores (76.09 ± 17.86) were good and within acceptable range. Chewing (53 ± 29.29) and swallowing (64 ± 21.57) scored poorly among 12 domains. For extensive through and through cheek defects bipedelled ALT provides good functional outcome. In view of advance disease status extensive resection would be responsible for bone and mucosal loss affecting chewing and swallowing. Supplementary Information: The online version contains supplementary material available at 10.1007/s12070-022-03416-3.

Experimental Models of In Vitro Blood-Brain Barrier for CNS Drug Delivery: An Evolutionary Perspective.

Central nervous system (CNS) disorders represent one of the leading causes of global health burden. Nonetheless, new therapies approved against these disorders are among the lowest compared to their counterparts. The absence of reliable and efficient in vitro blood-brain barrier (BBB) models resembling in vivo barrier properties stands out as a significant roadblock in developing successful therapy for CNS disorders. Therefore, advancement in the creation of robust and sensitive in vitro BBB models for drug screening might allow us to expedite neurological drug development. This review discusses the major in vitro BBB models developed as of now for exploring the barrier properties of the cerebral vasculature. Our main focus is describing existing in vitro models, including the 2D transwell models covering both single-layer and co-culture models, 3D organoid models, and microfluidic models with their construction, permeability measurement, applications, and limitations. Although microfluidic models are better at recapitulating the in vivo properties of BBB than other models, significant gaps still exist for their use in predicting the performance of neurotherapeutics. However, this comprehensive account of in vitro BBB models can be useful for researchers to create improved models in the future.

Integration of cyclodextrins and associated toxicities: A roadmap for high quality biomedical applications.

Cyclodextrins are extensively employed in drug delivery systems like inclusion complexes, metal-organic frameworks, functionalized or PEGylated conjugates, and other nanocarrier systems such as nanosponges or hydrogel nanoparticles for targeted effect or prolonged release action. Applications of CDs range from drug-loaded nanocarrier systems useful for disease conditions (such as cancer, diabetes, and bacterial infections, etc.) to supramolecular chemistry, diagnostics, imaging, biosensors, and medical devices. However, there is a limited data and information on the adverse effects caused by cyclodextrins and their toxicities in the medical field. Various in-vitro and ex-vivo toxic effects such as cytotoxicity, ototoxicity, etc. as well as the adverse and toxic effects depend on the role of administration of cyclodextrins. This review article focuses on the advancement of characteristics, properties and chemistry of cyclodextrins and addresses the new challenges faced in cyclodextrin-based delivery systems and the various toxicities induced by them.

An Integrative QbD Approach for the Development and Optimization of Controlled Release Compressed Coated Formulation of Water-Soluble Drugs.

Controlled release dosage forms maintain regulated pharmacokinetic profile of drug substance within its therapeutic window by ensuring constant plasma concentrations. Controlled release formulations not only increase the therapeutic efficacy of drug substances but also reduce their dose-related side effects. Present investigation was conducted to develop, optimize, and validate compressed coated controlled release tablet formulation for highly water-soluble drug substances which have no rate-controlling factor towards its release from dosage form. Drug dispersed waxy core tablet, press coated within the swellable hydrophilic polymeric barrier layer, was developed and optimized via quality by design approach (QbD) using Box-Behnken design. The optimized formulation was characterized and validated using in vitro quality control parameters. Attributes identified under SUPAC guidelines, such as drug release rates at 30 min, 6 h, and 12 h, were considered as the critical quality attributes (CQAs) that significantly affected efficiency of the compressed coated controlled release tablets. CQAs screened using risk assessment and Pareto chart analyses were used for optimizing controlled release dosage form. Findings revealed that tablets containing drug to wax ratio of 1:1, hydrophilic swellable polymer concentration of 200 mg, and prepared using compression pressure of 6.5 kg/cm2 exhibited the highest desirability indices in terms of controlling the release rate of drug substance. Optimized formulation was also evaluated for swelling rate, erosion rate, and other post-compression parameters, including release kinetics. Fickian diffusion-based zero-order controlled release of BCS class I drug substance was achieved through the developed dosage form.

Synthesis and Characterization of Fatty Acid Grafted Chitosan Polymeric Micelles for Improved Gene Delivery of VGF to the Brain through Intranasal Route.

Multifunctional fatty acid grafted polymeric micelles are an effective and promising approach for drug and gene delivery to the brain. An alternative approach to bypass the blood-brain barrier is administration through intranasal route. Multifunctional fatty acid grafted polymeric micelles were prepared and characterized for pVGF delivery to the brain. In vitro pVGF expression was analyzed in bEnd.3 cells, primary astrocytes, and neurons. Comparative in-vivo pVGF expression was analyzed to evaluate the effective route of administration between intranasal and intravenous. Biocompatible, multifunctional polymeric micelles were prepared, having an average size of 200 nm, and cationic zeta potential. Modified polymers were found to be hemo- and cyto-compatible. When transfected with the different modified chitosan formulations, significantly (p < 0.05) higher VGF expression was observed in primary astrocytes and neurons using the mannose, Tat peptide, and oleic acid grafted chitosan polymer. Compared to intravenous administration, intranasal administration of pVGF in polyplex formulation led to significantly (p < 0.05) higher pVGF expression. Developed multifunctional polymeric micelles were an effective pVGF delivery platform to the brain. Mannose and Tat ligand tagging improved the pVGF delivery to the brain.

A Review of the Common Neurodegenerative Disorders: Current Therapeutic Approaches and the Potential Role of Nanotherapeutics.

Neurodegenerative disorders are primarily characterized by neuron loss. The most common neurodegenerative disorders include Alzheimer's and Parkinson's disease. Although there are several medicines currently approved for managing neurodegenerative disorders, a large majority of them only help with associated symptoms. This lack of pathogenesis-targeting therapies is primarily due to the restrictive effects of the blood-brain barrier (BBB), which keeps close to 99% of all "foreign substances" out of the brain. Since their discovery, nanoparticles have been successfully used for targeted delivery into many organs, including the brain. This review briefly describes the pathophysiology of Alzheimer's, Parkinson's disease, and amyotrophic lateral sclerosis, and their current management approaches. We then highlight the major challenges of brain-drug delivery, followed by the role of nanotherapeutics for the diagnosis and treatment of various neurological disorders.

Nanotech-based Food: An Initiative for Alternative Pharmaceuticals.

Nanotechnology opens many avenues in the food sector and offers applications associated with food production, processing, cultivation, and packaging. Nanofood employs nanotechniques like nano-encapsulation to conjugate various phytochemicals, antioxidants, probiotics, minerals, vitamins, etc., into nanovehicles. Food fortification strategies are implemented to incorporate nano-processed substances. Nanofood is mostly used for improving health and as a supplementation in various diseases ranging from liver diseases to neurodegenerative disorders. Here, we focus on recent studies that exhibit comparable results for nanofood and conventional medicines, subsiding the limitations of traditional therapies. Nanofood holds the potential for the management of various health problems and can be used as an alternative to medicine in clinical conditions, like cancers and inflammatory bowel disease. With further advances in nanotechnology and expansion in the scope of the current nanofood industry, in addition to proper regulations set in place, nanofood may offer a wide variety of advantages in terms of safety, long-term stability, etc.

Nano-biotechnology and its Innovative Perspective in Diabetes Management.

The occurrence of the diabetes happens due to the irregular operation of glucose in the body, which is also known as glucose homeostasis, thus leading to metabolic changes in the body. The two stages whether hypoglycemia or hyperglycemia differentiates diabetes into various categories and brought new innovative management for the new routes of administration of these disease condition. Various bio-nanotechnologies which are coupled with nano particulates, polymers, Liposome, various gold plated and Solid Lipids Particulates regulate the transcellular transport, non specific cellular uptake, and paracellular transport, which leads to oral, transdermal, Pulmonary, buccal, Nasal, specific gene oriented administration to avoid the non patience compliance with the parental routes of administration. Phytochemicals have an emerging strategy for the future prospects of diabetes management.

Delivery systems for improving iron uptake in anemia.

Anemia poses a threat to a broad population globally as depleted hemoglobin leads to a plethora of conditions, and the most common cause includes iron deficiency. Iron is an essential element important for erythropoiesis, DNA synthesis, protection of the immune system, energy production, and cognitive function and hence should be maintained at appropriate levels. Various proteins are involved in transporting and absorption of iron, activation of heme synthesis, and RBC production that could be possible targets to improve iron delivery. Oral supplementation of iron either from dietary or synthetic sources has been the frontline therapy for treating iron deficiency in anemia. At the same time, intravenous administration is provided in chronic anemia, such as chronic kidney diseases (CKD). This review focuses on the strategies developed to overcome the disadvantages of available iron therapies and increase iron absorption and uptake in the body to restore iron content. Nanotechnology combined with the food fortification processes gained attention as they help develop new delivery systems to improve iron uptake by enterocytes. Furthermore, naturally obtained products such as polysaccharides, peptides, proteins, and new synthetic molecules have been used in fabrication of iron-carrier systems. The establishment of transdermal iron delivery systems such as microneedle arrays or iontophoresis, or the discovery of new molecules also proved to be an effective way for delivering iron in patients non-compliant to oral therapy.

Methanolic extract of Cucumis melo attenuates ethylene glycol-induced nephrolithiasis in Wistar rats.

Evaluation of the effects of methanolic extract of Cucumis melo in ethylene glycol-induced nephrolithiasis on Wistar rats. 0.75% solution of ethylene glycol (EG) in payable water was given to produce nephrolithiasis on Wistar rats. The action of oral intake of methanolic extract of Cucumis melo seed in nephrolithiasis is studied and is matched with the action of oral intake of Cystone (standard) on Wistar rats. EG resulted in hyperoxaluria and deposition of calcium oxalate as well as raised urinary excretion of oxalate and calcium. Supplementation with methanolic extract of Cucumis melo seed decreased the increased renal oxalate, indicating a regulatory effect on oxalate formation endogenously. The outcomes stipulate that the seed of Cucumis melo is endowed with antinephrolithiatic action.

3D Printed Bioconstructs: Regenerative Modulation for Genetic Expression.

Layer-by-layer deposition of cells, tissues and similar molecules provided by additive manufacturing techniques such as 3D bioprinting offers safe, biocompatible, effective and inert methods for the production of biological structures and biomimetic scaffolds. 3D bioprinting assisted through computer programmes and software develops mutli-modal nano- or micro-particulate systems such as biosensors, dosage forms or delivery systems and other biological scaffolds like pharmaceutical implants, prosthetics, etc. This review article focuses on the implementation of 3D bioprinting techniques in the gene expression, in gene editing or therapy and in delivery of genes. The applications of 3D printing are extensive and include gene therapy, modulation and expression in cancers, tissue engineering, osteogenesis, skin and vascular regeneration. Inclusion of nanotechnology with genomic bioprinting parameters such as gene conjugated or gene encapsulated 3D printed nanostructures may offer new avenues in the future for efficient and controlled treatment and help in overcoming the limitations faced in conventional methods. Moreover, expansion of the benefits from such techniques is advantageous in real-time delivery or in-situ production of nucleic acids into the host cells. Aspects of 3D bioprinting in gene delivery.

Biofluidic material-based carriers: Potential systems for crossing cellular barriers.

Biofluids act as a repository for disease biomarkers and are excellent diagnostic tools applied in establishing a disease profile based on clinical testing, evaluation and monitoring the progression of patients suffering from various conditions. Furthermore, biofluids and their derived components such proteins, pigments, enzymes, hormones and cells carry a potential in the development of therapeutic drug delivery systems or as cargo materials for targeting the drug to the site of action. The presence of biofluids with respect to their specific location reveals the information of disease progression and mechanism, delivery aspects such as routes of administration as well as pharmacological factors such as binding affinity, rate of kinetics, efficacy, bioavailability and patient compliance. This review focuses on the properties and functional benefits of some biofluids, namely blood, saliva, bile, urine, amniotic fluid, synovial fluid and cerebrospinal fluid. It also covers the therapeutic and targeting action of fluid-derived substances in various micro- or nano-systems like nanohybrids, nanoparticles, self-assembled micelles, microparticles, cell-based systems, etc. The formulation of such biologically-oriented systems demonstrate the advantages of natural origin, biocompatibility and biodegradability and offer new techniques for overcoming the challenges experienced in conventional therapies.

Gut microbiome a promising target for management of respiratory diseases.

The intestinal microbial flora has risen to be one of the important etiological factors in the development of diseases like colorectal cancer, obesity, diabetes, inflammatory bowel disease, anxiety and Parkinson's. The emergence of the association between bacterial flora and lungs led to the discovery of the gut-lung axis. Dysbiosis of several species of colonic bacteria such as Firmicutes and Bacteroidetes and transfer of these bacteria from gut to lungs via lymphatic and systemic circulation are associated with several respiratory diseases such as lung cancer, asthma, tuberculosis, cystic fibrosis, etc. Current therapies for dysbiosis include use of probiotics, prebiotics and synbiotics to restore the balance between various species of beneficial bacteria. Various approaches like nanotechnology and microencapsulation have been explored to increase the permeability and viability of probiotics in the body. The need of the day is comprehensive study of mechanisms behind dysbiosis, translocation of microbiota from gut to lung through various channels and new technology for evaluating treatment to correct this dysbiosis which in turn can be used to manage various respiratory diseases. Microfluidics and organ on chip model are emerging technologies that can satisfy these needs. This review gives an overview of colonic commensals in lung pathology and novel systems that help in alleviating symptoms of lung diseases. We have also hypothesized new models to help in understanding bacterial pathways involved in the gut-lung axis as well as act as a futuristic approach in finding treatment of respiratory diseases caused by dysbiosis.

Treatment of insulin resistance in obesity-associated type 2 diabetes mellitus through adiponectin gene therapy.

Global rise in obesity-associated type 2 diabetes mellitus (T2DM) has led to a major healthcare crisis. Development of efficient treatments to treat the underlying chronic inflammation in obesity-associated T2DM, is an unmet medical need. To this end, we have developed a plasmid adiponectin (pADN) based nanomedicine for the treatment of insulin resistance in type 2 diabetes mellitus. Adiponectin is a potent anti-inflammatory/anti-diabetic adipokine, which is downregulated in obesity. In this study, nanomicelles comprising chitosan conjugated to oleic acid and adipose homing peptide (AHP) were developed to deliver pADN to adipocytes. Cationic chitosan-oleic-AHP micelles were 112 nm in size, encapsulated 93% of pADN and protected gene cargo from DNase I mediated enzymatic degradation. In vitro, the nanomicellar formulation significantly increased adiponectin production compared to free plasmid as well as standard transfecting agent FuGENE®HD. Single dose subcutaneous administration of pADN-chitosan-oleic-AHP to obese-diabetic rats, resulted in improved insulin sensitivity for up to 6 weeks, which matched the glucose disposal ability of healthy rats. Serum adiponectin level in pADN-chitosan-oleic-AHP treated rats was comparable to healthy rats for up to 3 weeks post treatment. Overall, the results indicate that pADN-chitosan-oleic-AHP based therapy is a promising treatment approach for obesity-associated T2DM.