In Vivo Cancer Microenvironment Responsive Glycan Receptor-Targeted Nanoparticles for Gemcitabine Delivery to Benzo[a]pyrene-Induced Lung Cancer Model.

Targeted gemcitabine (GEB) loaded 5-N-acetyl-neuraminic acid (Neu5Ac) assembled chitosan nanoparticles (CA-NPs) were formulated by ionotropic gelation process and evaluated for physicochemical and morphological characterization, in vitro and in vivo studies in A-549 cells and lung cancer mice model, respectively. The mean diameter of GEB-CA-Neu5Ac-NPs determined by dynamic light scattering was 161.16 ± 7.70 nm with a polydispersity index (PDI) value of 0.303 ± 0.011 and its zeta potential and entrapment efficiency (%EE) were 40.3 ± 3.45 mv and 66.11 ± 1.94%, respectively. The in vitro cellular uptake studies showed that glycan receptor-targeted nanoparticles deliver significantly more amount (p < 0.001) of GEB into the A-549 lung cancerous cells than non-targeted nanoparticles. The cytotoxicity study using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay clearly demonstrated that GEB-CA-Neu5Ac-NPs have lower IC50 value (6.39 ± 3.78 µg/ml) than others groups that showed that the greater lung cancerous cells inhibition potential of targeted nanoparticles. The in vivo biodistribution of the GEB-loaded 5-N-acetyl-neuraminic acid conjugated chitosan nanoparticles was revealed that targeted nanoparticles showed higher accumulation and retention for an extended period of time due to the active targeting ability of Neu5Ac to glycan receptors. Histopathological examination showed significant recovery in the physiological architecture upon administration of targeted nanoparticles. The glycan receptor-targeted nanoparticles treated groups showed a significant decline in the number of metastatic lung epithelial cells, as compared to the untreated positive control group (p < 0.001) confirming higher anticancer efficacy of the GEB-CA-Neu5Ac-NPs.

pH-influenced self-assembled stealth nanoscaffolds encapsulating memantine for treatment of Alzheimer's disease.

Alzheimer's disease (AD) is the most common neurological disorder characterized by the accumulation of ß-amyloid peptides. The only medication used to treat moderate-to-severe AD progression is memantine. In this study, polyethylene glycol (PEG)-coated poly D, L-lactic-co-glycolic acid (PLGA) nanostructures were prepared, as their self-assembling ability helps to penetrate the drug at disease sites with altered pH range (5.7-6.8) due to AD. Drug and polymer interaction studies by FTIR showed no interaction among them, and the thermal properties of drugs, polymers, and nanostructures tell us about their melting behaviour, thermal degradation, and glass transition temperatures. Characterization of prepared self-assembled nanoscaffolds signifies that the acquired properties such as size, structure, surface charges, zeta-potential, stability, thermal properties, biodegradability, biocompatibility, swelling ability, encapsulation, and drug loading provide an efficient therapeutic activity to the nanostructures for the treatment of AD. In addition, parallel artificial membrane permeability assay (PAMPA) has revealed the paracellular transport mechanism of nanoscaffolds across the blood-brain barrier. In vitro release kinetics showed a sustained release pattern exhibiting the Korsmeyer-Peppes drug release kinetic model with a correlation coefficient (R2) value of 0.9905, which describes the drug release pattern from a polymeric system. In vitro enzymatic studies demonstrated the inhibition activity of nanostructures on acetylcholinesterase (AChE), butyrylcholinestrase (BUChE), and ß-secretase enzyme which prevents the breakdown of acetylcholine, butyrylcholine, and amyloid precursor protein, and retention of these neurotransmitters constituted the primary therapeutic strategy for AD. Also, behavioural studies have shown a significant (p<0.05) improvement in cognition behaviour among nanostructures- administered animal groups in a scopolamine-induced amnesia model. The designed nanocarrier can also accelerate the treatment strategies for AD by incorporating stem cells and self-assembled nanoscaffolds that could provide a 3D extracellular matrix to facilitate neuron regeneration, hence improving cognition behaviour effectively.

Evaluation of methotrexate encapsulated polymeric nanocarrier for breast cancer treatment.

Herein, Methotrexate-loaded chitosan nanoparticles (Meth-Cs-NPs) was formulated through single-step self-assembly by incorporating the ionic-gelation method. Chitosan was cross-linked with Methotrexate via a sodium tripolyphosphate (STPP) where 49 % Methotrexate was loaded in the nanoparticles (∼143 nm) and zeta potential of 34 ± 3 mV with an entrapment efficiency of 87 %. The efficacy of nanoparticles was assessed for chemically induced breast cancer treatment in the Sprague Dawley rats model. These Meth-Cs-NPs followed the Korsmeyer-Peppas model in-vitro release kinetics. Nanoparticles were further evaluated for in-vitro efficacy on triple-negative breast cancer (MDA-MB-231) cell lines. The MTT assay studies revealed that even slight exposure to Meth-Cs-NPs (IC50 = 15 µg/mL) caused 50 % cell death in 24 h. Further, hemocompatibility studies of Meth-Cs-NPs were performed, deciphered that Meth-Cs-NPs were biocompatible (hemolysis < 2 %). Additional cellular uptake was evaluated by confocal imaging. Moreover, an in-vivo pharmacokinetic study of nanoparticles in rats displayed increased plasma concentration of the drug and retention time, whereas a decrease in cellular clearance compared to free Methotrexate. Further, anti-tumor efficacy studies revealed that nanoparticles could reduce tumor volume from 1414 mm3→385 mm3 compared to free Methotrexate (1414 mm3→855 mm3). The current study presents encouraging prospects of Meth-Cs-NPs to be used as a viable breast cancer treatment modality.

In-vivo anticancer efficacy of self-targeted methotrexate-loaded polymeric nanoparticles in solid tumor-bearing rat.

Here, cytotoxicity and antitumor efficacy against a chemically (N-methyl-N-nitrosourea) generated mammary tumor in rats were assessed using methotrexate-loaded chitosan nanoparticles (Meth-Cs-NPs). Meth-Cs-NPs intravenous administrated resulted in noticeably decreased tumor incidence, multiplicity, and weight. Further, kidney function tests for the treated groups resulted in noticeably decreased ALP (Meth-Cs-NPs; 244 ± 15, diseases control; 403 ± 14 U/L), Creatinine (Meth-Cs-NPs; 0.81 ± 0.05, diseases control; 2 ± 0.05 mg/dl), and Urea (Meth-Cs-NPs; 56.62 ± 5, diseases control; 113 ± 6 mg/dl) levels, close to a normal control group. Similarly, liver function tests showed significantly decreased serum biomarkers, SGPT (Meth-Cs-NPs; 40 ± 1.8, diseases control; 84 ± 1.9 U/L) and SGOT (Meth-Cs-NPs; 15 ± 2, diseases control; 55 ± 4 U/L) levels in treated groups as compared to the untreated group (diseases control). From the results, pro-inflammatory cytokines were also markedly reduced in the treated group such as, TNF-α (Meth-Cs-NPs; 17.31 ± 1.15, diseases control; 36.9 ± 5 pg/mL), IL-1ß (Meth-Cs-NPs; 433.3 ± 66.5, diseases control; 1540 ± 131.1 pg/mL), and IL-6 (Meth-Cs-NPs; 1515 ± 53, diseases control; 2200.6 ± 69 pg/mL) levels. Whereas Meth-Cs-NPs not only helped in lowering tumor multiplicity rates but also decrease inflammation. The studies could be successfully performed in chemically induced mammary tumors due to their easy, quick tumor growth and low mortality rates in rat models. According to the current study, Meth-Cs-NPs have high treatment potency and represent a possible therapeutic alternative for breast cancer treatment.

Role of pro-inflammatory cytokines in Alzheimer's disease and neuroprotective effects of pegylated self-assembled nanoscaffolds.

Neurodegeneration and synaptic loss in Alzheimer's disease (AD) lead to impairment in memory functions. Neuroinflammation causes activation of microglia and astrocytes cells that locally and systemically produces inflammatory cytokines which can serve as early diagnostic markers or therapeutic targets in AD. Pro-inflammatory cytokines (Interleukins (IL-1ß, IL-6 and IL-10) and tumor necrosis factor (TNF α)) levels were estimated in serum, cerebral tissue, hepatic tissue, and renal tissue in treatment groups of scopolamine-induced amnesia mice model using ELISA protocol. The results showed that cerebral tissue of AD mice exhibited elevated levels of IL1ß, IL6, IL10 and TNFα which indicate contribution of pro-inflammatory cytokines in the progression of AD. A significant reduction in the concentration of IL1ß, IL-10 and TNF-α were noticed in serum, cerebral tissue and hepatic tissue of animal group treated with marketed memantine tablet (Admenta), pure memantine drug (MEMp), memantine-poly (lactic-co-glycolic acid) self-assembled nanoscaffolds (MEM-PLGA) SANs, Polyethylene Glycol coated memantine-poly (lactic-co-glycolic acid) self-assembled nanoscaffolds [(PEG-MEM-PLGA) SANs] and Polyethylene Glycol coated memantine-poly [(lactic-co-glycolic acid)] self-assembled nanoscaffolds grafted with Bone Marrow Derived Stem Cell ((PEG-MEM-PLGA) SANs-BMSc), whereas a high level of IL-6 was observed in hepatic tissue, cerebral tissue and renal tissues of normal and AD induced mice which showed the emerging potential of IL-6 cytokines that can trigger either neurons survival after injury or causing neurodegeneration and cell apoptosis. The Neuroregenerative potential of stem cells helps in the proliferation of neuronal cell and thus improves cognition in AD animal model.