Doaa A. Abdelmonsif

Intranasal Piperine‐Loaded Chitosan Nanoparticles as Brain‐Targeted Therapy in Alzheimer's Disease: Optimization, Biological Efficacy, and Potential Toxicity

Piperine (PIP) is a phytopharmaceutical with reported neuroprotective potential in Alzheimers disease (AD). Oral PIP delivery suffers from its hydrophobicity and pre-systemic metabolism. In this article, mono-disperse intranasal chitosan nanoparticles (CS-NPs) were elaborated for brain targeting of PIP. Formula optimization was based on particle size (PS), zeta potential (ZP), polydispersity index (PDI), % entrapment efficiency (% EE), release studies, and transmission electron microscopy. AD was induced in 48 male Wistar rats on which full behavioral and biochemical testing was conducted. Brain toxicity was assessed based on Caspase-3 assay for apoptosis and tumor necrosis factor for inflammation. Spherical NPs with optimum % EE (81.70), PS (248.50 nm), PDI (0.24), and ZP (+56.30 mV) were elaborated. PIP-NPs could significantly improve cognitive functions as efficient as standard drug (donpezil injection) with additional advantages of dual mechanism (Ach esterase inhibition and antioxidant effect). CS-NPs could significantly alleviate PIP nasal irritation and showed no brain toxicity. This work was the first to report additional mechanism of PIP in AD via anti-apoptosis and anti-inflammatory effects. To conclude, mucoadhesive CS-NPs were successfully tailored for effective, safe, and non-invasive PIP delivery with 20-folds decrease in oral dose, opening a gate for a future with lower AD morbidity.Piperine (PIP) is a phytopharmaceutical with reported neuroprotective potential in Alzheimers disease (AD). Oral PIP delivery suffers from its hydrophobicity and pre-systemic metabolism. In this article, mono-disperse intranasal chitosan nanoparticles (CS-NPs) were elaborated for brain targeting of PIP. Formula optimization was based on particle size (PS), zeta potential (ZP), polydispersity index (PDI), % entrapment efficiency (% EE), release studies, and transmission electron microscopy. AD was induced in 48 male Wistar rats on which full behavioral and biochemical testing was conducted. Brain toxicity was assessed based on Caspase-3 assay for apoptosis and tumor necrosis factor for inflammation. Spherical NPs with optimum % EE (81.70), PS (248.50nm), PDI (0.24), and ZP (+56.30mV) were elaborated. PIP-NPs could significantly improve cognitive functions as efficient as standard drug (donpezil injection) with additional advantages of dual mechanism (Ach esterase inhibition and antioxidant effect). CS-NPs could significantly alleviate PIP nasal irritation and showed no brain toxicity. This work was the first to report additional mechanism of PIP in AD via anti-apoptosis and anti-inflammatory effects. To conclude, mucoadhesive CS-NPs were successfully tailored for effective, safe, and non-invasive PIP delivery with 20-folds decrease in oral dose, opening a gate for a future with lower AD morbidity.

Novel piperine-loaded Tween-integrated monoolein cubosomes as brain-targeted oral nanomedicine in Alzheimer's disease: pharmaceutical, biological, and toxicological studies.

Alzheimer’s disease (AD) is one of the most patient devastating central nervous system diseases with no curative therapy. An effective oral therapy with brain-targeting potential is required that is hampered by blood–brain barrier. Piperine (PIP) is a natural alkaloid with memory enhancing potentials. Oral PIP delivery suffers from its hydrophobicity and first-pass metabolism. In this study, novel Tween-modified monoolein cubosomes (T-cubs) were elaborated as bioactive nanocarriers for brain-targeted oral delivery of PIP. Seven liquid crystalline nanoparticles (cubosomes) were prepared testing different bioactive surfactants (Tween 80, poloxamer, and Cremophor). Full in vitro characterization was carried out based on particle size, zeta potential, polydispersity index, entrapment efficiency, and in vitro release. Morphological examination and structure elucidation were performed using transmission and polarizing microscopes. Sporadic dementia of Alzheimer’s type was induced in 42 male Wistar rats on which full behavioral and biochemical testing was conducted. Brain toxicity was assessed based on Caspase-3 assay for apoptosis and tumor necrosis factor-α for inflammation. Liver and kidney toxicity studies were conducted as well. Among others, T-cubs exhibited optimum particle size (167.00±10.49 nm), polydispersity index (0.18±0.01), and zeta potential (−34.60±0.47 mv) with high entrapment efficiency (86.67%±0.62%). Cubs could significantly sustain PIP in vitro release. In vivo studies revealed T-cubs potential to significantly enhance PIP cognitive effect and even restore cognitive function to the normal level. Superiority of T-cubs over others suggested brain-targeting effect of Tween. Toxicological studies contended safety of cubs on kidney, liver, and even brain. T-cubs exhibited potential anti-inflammatory and anti-apoptotic activity of loaded PIP, indicating potential to stop AD progression that was first suggested in this article. Novel oral nanoparticles elaborated possess promising in vitro and in vivo characteristics with high safety for effective chronic treatment of AD.

Stealth, biocompatible monoolein-based lyotropic liquid crystalline nanoparticles for enhanced aloe-emodin delivery to breast cancer cells: in vitro and in vivo studies

Recently, research has progressively highlighted on clues from conventional use of herbal medicines to introduce new anticancer drugs. Aloe-emodin (AE) is a herbal drug with promising anticancer activity. Nevertheless, its clinical utility is handicapped by its low solubility. For the first time, this study aims to the fabrication of surface-functionalized polyethylene glycol liquid crystalline nanoparticles (PEG-LCNPs) of AE to enhance its water solubility and enable its anticancer use. Developed AE-PEG-LCNPs were optimized via particle size and zeta potential measurements. Phase behavior, solid state characteristics, hemocompatibility, and serum stability of LCNPs were assessed. Sterile formulations were developed using various sterilization technologies. Furthermore, the potential of the formulations was investigated using cell culture, pharmacokinetics, biodistribution, and toxicity studies. AE-PEG-LCNPs showed particle size of 190 nm and zeta potential of −49.9, and PEGylation approach reduced the monoolein hemolytic tendency to 3% and increased the serum stability of the nanoparticles. Sterilization of liquid and lyophilized AE-PEG-LCNPs via autoclaving and γ-radiations, respectively, insignificantly affected the physicochemical properties of the nanoparticles. Half maximal inhibitory concentration of AE-PEG-LCNPs was 3.6-fold lower than free AE after 48 hours and their cellular uptake was threefold higher than free AE after 24-hour incubation. AE-PEG-LCNPs presented 5.4-fold increase in t1/2 compared with free AE. Biodistribution and toxicity studies showed reduced AE-PEG-LCNP uptake by reticuloendothelial system organs and good safety profile. PEGylated LCNPs could serve as a promising nanocarrier for efficient delivery of AE to cancerous cells.

Targeting AMPK, mTOR and β-Catenin by Combined Metformin and Aspirin Therapy in HCC: An Appraisal in Egyptian HCC Patients

BackgroundHepatocellular carcinoma (HCC) is an expanding health problem with a great impact on morbidity and mortality, both in Egypt and worldwide. Recently, metformin and aspirin showed a potential anticancer effect on HCC, although the mechanism of this effect is not fully elucidated.ObjectiveThe current work aimed to investigate the possibility of targeting AMP-activated protein kinase (AMPK), mammalian target of rapamycin (mTOR), and β-catenin proteins through combined metformin/aspirin treatment in the HepG2 cell line, and to explore such molecular targets in Egyptian HCC patients.Materials and MethodsHepG2 cells were exposed to increasing concentrations of metformin, aspirin and combined treatment, and an MTT assay was performed to determine half maximal inhibitory concentration (IC50). Caspase-3 activity, cell cycle analysis, and protein expression of AMPK, phosphorylated AMPK (pAMPK) and mTOR proteins were assessed. Furthermore, the expression and localization of β-catenin protein was assessed by immunocytochemistry, and protein expression of pAMPK, mTOR and β-catenin was assessed in Egyptian HCC and cirrhotic tissue specimens.ResultsMetformin/aspirin combined treatment had a synergistic effect on cell cycle arrest at the G2/M phase and apoptosis induction in a caspase-dependent manner via downregulation of pAMPK and mTOR protein expression. Additionally, metformin/aspirin combined treatment enhanced cell–cell membrane localization of β-catenin expression in HepG2 cells, which might inhibit the metastatic potential of HepG2 cells. In Egyptian HCC specimens, pAMPK, mTOR and β-catenin proteins showed a significant increased expression compared with cirrhotic controls.ConclusionsTargeting AMPK, mTOR and β-catenin by combined metformin/aspirin treatment could be a promising therapeutic strategy for Egyptian HCC patients, and possibly other HCC patients.

Cerium oxide nanoparticles could ameliorate behavioral and neurochemical impairments in 6-hydroxydopamine induced Parkinson's disease in rats

Background: Cerium oxide nanoparticles (CeO2NPs) showed promising effects in neurodegenerative diseases including some animal models of Parkinsonism. However, the implication of CeO2NPs in 6‐hydroxydopamine (6‐OHDA) induced Parkinsonism remains to be investigated. Aim: This study was designed to assess whether CeO2NPs treatment could alleviate neurobehavioral and neurobiochemical deficits in 6‐OHDA induced neurotoxicity in rats. Material and methods: 50 rats received left intrastriatal (IS) injection of either saline (control, n = 10) or 6‐OHDA (n = 40). At the third week post‐lesion, motor dysfunction was verified using neurobehavioral tests. Then diseased rats received intraperitoneal injection of 0.1, 0.5 or 1 mg/kg of CeO2NPs or vehicle (10 rats each) for 3 weeks. Rats were subjected to behavioral assessments and then sacrificed for biochemical analyses of the striatum. Striatal dopamine levels, oxidative stress markers including total antioxidant capacity (TAC) and malondialdehyde (MDA), and caspase 3 activity as an apoptotic marker were assessed. Results: Different doses of CeO2NPs variably improved motor dysfunctions induced by 6‐OHDA injection in open field, Rota Rod and stepping tests. In addition, the neurobiochemical derangements were almost reversed by the 0.5 mg/kg dose of CeO2NPs, while 0.1 mg/kg dose was not sufficient to alter biochemical measurements in the striatum. Administration of 1 mg/kg of CeO2NPs partially ameliorated striatal dopamine and decreased apoptosis without significant effect on oxidative stress. Conclusion: The present study showed a putative therapeutic role of CeO2NPs in the treatment of 6‐OHDA‐induced Parkinsonian rats, and suggested their antioxidant and antiapoptotic effects as possible mechanisms for elevated striatal dopamine level and improved motor performance. HighlightsCeO2NPs treatment improved motor deficits in 6‐OHDA induced‐Parkinsonism in rats.CeO2NPs increased striatal dopamine and ameliorated oxidative stress and apoptosis.CeO2NPs at 0.5 mg/kg dose showed superior therapeutic effects than 0.1 and 1 mg/kg.The 1 mg/kg CeO2NPs dose seemed neurotoxic to striatal tissues.

Mucopenetrating nanoparticles for enhancement of oral bioavailability of furosemide: In vitro and in vivo evaluation/sub-acute toxicity study

The aim of this study was to formulate and evaluate chitosan (CS)/alginate (ALG) nanoparticles (NPs) loaded with furosemide (FSM) in an attempt to enhance its release, permeability and bioavailability. Non-everted gut sac method was used to evaluate the ex vivo permeation of FSM from its suspension and the selected CS/ALG NPs formulation. The pharmacokinetic parameters of FSM subsequent to oral administration of the selected formulation were assessed in rats. In vivo subacute toxicity study of the prepared blank and FSM loaded formulations was evaluated in rats. The selected optimized formulation (F3) showed optimum particle size (PS), polydispersity index (PDI), zeta potential (ZP) and acceptable percentage entrapment efficiency (%EE) of 253.8nm±4.6, 0.25±0.03, -35mV±1 and 96%±1, respectively. The release profile of FSM from the selected formulation was characterized by initial burst effect in 0.1N HCl. Scanning electron microscope (SEM) demonstrated a smooth surface and spherical shape for the lyophilized optimized NPs. Selected CS/ALG NPs (F3) presented a significant enhancement (p≤0.01) in permeation parameters of FSM as well as in Tmax, Cmax, AUC0-24 and AUC0-∞. Subacute toxicity study results revealed that the selected formulation was safe and nontoxic. The histopathological inspection of the stomach and small intestine tissues of the loaded NPs (F3) and blank groups reflected no obvious signs of cellular toxicity or inflammatory reaction. CS/ALG NPs loaded with FSM enhanced both drug release and mucus-penetrating ability leading to an overall increase in FSM bioavailability. In addition, the in vivo subacute toxicity study results indicated the safety of the prepared NPs for oral drug delivery.

Cardioprotective effect of cerium oxide nanoparticles in monocrotaline rat model of pulmonary hypertension: A possible implication of endothelin-1.

Aims: Cerium oxide nanoparticles (CeO2NPs) have been recently introduced into the medical field for their antioxidant properties. The ability of CeO2NPs alone or in combination with spironolactone (SP) to attenuate monocrotaline (MCT)‐induced pulmonary hypertension and associated right ventricular hypertrophy was studied in rats. A special emphasis was given to endothelin‐1 pathway. Materials and methods: Pulmonary hypertension was induced in albino rats by a single subcutaneous injection of MCT (60 mg/kg). Rats received either single CeO2NPs therapy or combined therapy with SP for 2 weeks. Key findings: CeO2NPs improved pulmonary function tests with concomitant decrease in serum endothelin‐1 and pulmonary expression of endothelin‐1 and its receptor ETAR. Besides, CeO2NPs diminished MCT‐induced right ventricular hypertrophy and reduced cardiac oxidative stress and apoptosis. Significance: CeO2NPs could improve pulmonary hypertension and associated right ventricular hypertrophy with no additive value for SP. Besides being an antioxidant, CeO2NPs work through endothelin‐1 pathway to improve pulmonary hypertension. Graphical abstract Figure. No caption available.

Autophagy-related Protein LC3 in Egyptian Colorectal Cancer: Impact and Possible Relation to STAT3 andmiRNA 101.

Dina Abdallah 1 , Doaa Abdelmonsif 2 , Mai Moaaz 3 and Mohamed Selimah 4 . 1. Pathology Department, Faculty of Medicine, Alexandria University, Egypt. 2. Medical Biochemistry Department, Faculty of Medicine, Alexandria University, Egypt. 3. Department of Immunology and Allergy, Medical Research Institute, Alexandria University, Egypt. 4. Department of Experimental and Clinical Surgery, Medical Research Institute, Alexandria University, Egypt.

Methyl Guanine Methyl Transferase Methylation Status and Epidermal Growth Factor Receptor expression in a cohort of Egyptian glioblastoma patients

IntroductionGlioblastoma multiforme is the most frequently occurring primary brain tumour with a dismal prognosis. Chemotherapy with an alkylating agent such as Temozolomide (TMZ) confers a significant survival benefit. Genomic data elucidating pathogenetic mechanisms underlying gliomagenesis include epidermal growth factor receptor (EGFR) amplification/overexpression and methyl guanine methyl transferase promoter hypermethylation (MPH). Study of molecular changes paves the way to optimal personalized treatment. AimIn this study we aimed to assess EGFR expression and MPH status in relation to the patients’ response to radio/TMZ-based chemotherapy. Patients and materialsOverall, 34 cases of glioblastoma multiforme were studied for immunohistochemical assessment of EGFR expression and for methylation-specific PCR assessment of MPH status. Statistical correlations assessed the relation to radiologic tumor response. ResultsThe mean±SD of the patients’ ages was 56.56±9.8 years. Of 34 cases, 20 were males and 18 showed EGFR overexpression in less than 50% of tumor cells. MPH was detected in 52.9% of cases and was significantly related to younger age (t=2.3, P=0.03) but not to EGFR overexpression (&khgr;2=0.13, P=0.72). MPH was significantly related to tumor response (&khgr;2=5.44, P=0.02). ConclusionEGFR was largely expressed but was not related to response to TMZ-based treatment. MPH was expressed in 52.9% of cases with a significant correlation to treatment response. EGFR expression was not associated with MPH.