NP Formulation Research Articles

Preparation of nanoparticle and nanoemulsion formulations containing repaglinide and determination of pharmacokinetic parameters in rats

Repaglinide (RPG) belongs to the class of drugs known as meglitinides and is used for improving and maintaining glycemic control in the treatment of patients with Type 2 diabetes. RPG is a Class II drug (BCS) because of its high permeability and low water solubility. It also undergoes hepatic first-pass metabolism. The oral bioavailability of RPG is low (about 56 %) due to these drawbacks. Our aim in this study is to prepare two different nano-sized drug carrier systems containing RPG (nanoparticle: RPG-PLGA-Zein-NPs or nanoemulsion: RPG-NE) and to carry out a pharmacokinetic study for these formulations. We prepared NPs using PLGA and Zein. In addition, a single NE formulation was developed using Tween 80 and Pluronic F68 as surfactants and Labrasol as co-surfactant. The droplet size values of the blank-NE and RPG-NE formulations were found to be less than 120 nm. The mean particle sizes of blank-Zein-PLGA-NPs and RPG-Zein-PLGA-NPs were less than 260 nm. The Cmax and tmax values of RPG-Zein-PLGA-NPs and RPG-NE (523 ± 65 ng/mL and 770 ± 91 ng/mL; 1.41 ± 0.46 h and 1.61 ± 0.37 h, respectively) were meaningfully higher than those of free RPG (280 ± 33 ng/mL; 0.72 ± 0.28 h) (p < 0.05). The AUC0-∞ values calculated for RPG-Zein-PLGA-NPs and RPG-NE were approximately 4.04 and 5.05 times higher than that calculated for free RPG. These nanosized drug delivery systems were useful in increasing the oral bioavailability of RPG. Moreover, the NE formulation was more effective than the NP formulation in improving the oral bioavailability of RPG (p < 0.05).

Receptor Targeting Using Copolymer-Modified Gold Nanoparticles for pCMV-Luc Gene Delivery to Liver Cancer Cells In Vitro.

The formulation of novel delivery protocols for the targeted delivery of genes into hepatocytes by receptor mediation is important for the treatment of liver-specific disorders, including cancer. Non-viral delivery methods have been extensively studied for gene therapy. Gold nanoparticles (AuNPs) have gained attention in nanomedicine due to their biocompatibility. In this study, AuNPs were synthesized and coated with polymers: chitosan (CS), and polyethylene glycol (PEG). The targeting moiety, lactobionic acid (LA), was added for hepatocyte-specific delivery. Physicochemical characterization revealed that all nano-formulations were spherical and monodispersed, with hydrodynamic sizes between 70 and 250 nm. Nanocomplexes with pCMV-Luc DNA (pDNA) confirmed that the NPs could bind, compact, and protect the pDNA from nuclease degradation. Cytotoxicity studies revealed that the AuNPs were well tolerated (cell viabilities > 70%) in human hepatocellular carcinoma (HepG2), embryonic kidney (HEK293), and colorectal adenocarcinoma (Caco-2) cells, with enhanced transgene activity in all cells. The inclusion of LA in the NP formulation was notable in the HepG2 cells, which overexpress the asialoglycoprotein receptor on their cell surface. A five-fold increase in luciferase gene expression was evident for the LA-targeted AuNPs compared to the non-targeted AuNPs. These AuNPs have shown potential as safe and suitable targeted delivery vehicles for liver-directed gene therapy.

RETRACTED ARTICLE: Bio synthesis, comprehensive characterization, and multifaceted therapeutic applications of BSA-Resveratrol coated platinum nanoparticles

This study examines the manufacturing, characterization, and biological evaluation of platinum nanoparticles, which were synthesized by Enterobacter cloacae and coated with Bovine Serum Albumin (BSA) and Resveratrol (RSV). The formation of PtNPs was confirmed with the change of color from dark yellow to black, which was due to the bioreduction of platinum chloride by E. cloacae. BSA and RSV functionalization enhanced these nanoparticles' biocompatibility and therapeutic potential. TGA, SEM, XRD, and FTIR were employed for characterization, where PtNPs and drug conjugation-related functional groups were studied by FTIR. XRD confirmed the crystalline nature of PtNPs and Pt-BSA-RSV NPs, while TGA and SEM showed thermal stability and post-drug coating morphological changes. Designed composite was also found to be biocompatible in nature in hemolytic testing, indicating their potential in Biomedical applications. After confirmation of PtNPs based nanocaompsite synthesis, they were examined for anti-bacterial, anti-oxidant, anti-inflammatory, and anti-cancer properties. Pt-BSA-RSV NPs showed higher concentration-dependent DPPH scavenging activity, which measured antioxidant capability. Enzyme inhibition tests demonstrated considerable anti-inflammatory activity against COX-2 and 15-LOX enzymes. In in vitro anticancer studies, Pt-BSA-RSV NPs effectively killed human ovarian cancer cells. This phenomenon was demonstrated to be facilitated by the acidic environment of cancer, as the drug release assay confirmed the release of RSV from the NP formulation in the acidic environment. Finally, Molecular docking also demonstrated that RSV has strong potential as an anti-oxidant, antibacterial, anti-inflammatory, and anticancer agent. Overall, in silico and in vitro investigations in the current study showed good medicinal applications for designed nanocomposites, however, further in-vivo experiments must be conducted to validate our findings.

Administration sequence- and formation-dependent vaccination using acid-degradable polymeric nanoparticles with high antigen encapsulation capability.

Vaccines aim to efficiently and specifically activate the immune system via a cascade of antigen uptake, processing, and presentation by antigen-presenting cells (APCs) to CD4 and CD8 T cells, which in turn drive humoral and cellular immune responses. The specific formulation of vaccine carriers can not only shield the antigens from premature sequestering before reaching APCs but also favorably promote intracellular antigen presentation and processing. This study compares two different acid-degradable polymeric nanoparticles that are capable of encapsulating a moderately immunogenic antigen, GFP, at nearly full efficacy via electrostatic interactions or molecular affinity between His tag and Ni-NTA-conjugated monomners. This resulted in GFP-encapsulating NPs composed of ketal monomers and crosslinkers (KMX/GFP NPs) and NTA-conjugated ketal monomers and crosslinkers (NKMX/GFP NPs), respectively. Encapsulated GFP was found to be released more rapidly from NKMX/GFP NPs (electrostatic encapsulation) than from KMX/GFP NPs (affinity-driven encapsulation). In vivo vaccination studies demonstrated that while repeated injections of either NP formulation resulted in poorer generation of anti-GFP antibodies than injections of the GFP antigen itself, sequential injections of NPs and GFP as prime and booster vaccines, respectively, restored the humoral response. We proposed that NPs primarily assist APCs in antigen presentation by T cells, and B cells need to be further stimulated by free protein antigens to produce antibodies. The findings of this study suggest that the immune response can be modulated by varying the chemistry of vaccine carriers and the sequences of vaccination with free antigens and antigen-encapsulating NPs.

A new approach on essential oil production of Origanum onites L.: Microbial fertilization and microwave extraction

In many studies plant and microbe interaction has been observed due to its stimulating effect on plant oil content. Bacillus spp. isolates used as microbial fertilizer/biostimulant have shown a higher potential for enhancing plant production and quality. However, there is still a need for investigating the essential oil content of plants, particularly for the pharmaceutical industry, to increase the targeted essential oil quantity and quality. Thyme is a crucial plant in medicinal aromatic herbal production and export, and its economic value is steadily increasing due to its high essential oil content. This study explores a novel approach to increase essential oil content through microbial fertilization (biostimulation) and microwave extraction methods. The microwave extraction method offers comparable yields to other methods, requiring less time, energy, and only a small amount of water to hydrate the dried plants. Microbial fertilizer formulations were applied to İzmir thyme (Origanum onites L.) to assess their effects on yield and quality. Three distinct microbial fertilizer formulations (N, P, NP) containing bacterial isolates of Bacillus subtilis and Bacillus megaterium, as well as a consortium of these two bacteria, were employed in this study. Among the applications on İzmir thyme, the NP formulation (a combination of two bacteria at a dosage of 6 L/ha−1) exhibited the highest efficacy in terms of yield and quality criteria. Moreover, the essential oil ratios (1–2%) obtained from the same fertilizer application, using microwave extraction to enhance quality and accelerate the extraction process, yielded higher than other methods.This study provides preliminary data and a new approach to regulate and enhance the targeted yield and essential oil content of thyme plants by utilizing microbial fertilizer applications and the microwave extraction method.

Preparation and optimization of hyaluronic acid decorated irinotecan-loaded poly(lactic-co-glycolic acid) nanoparticles by microfluidics for cancer therapy applications

Polymer-based nanoparticles have been largely investigated in the cancer therapy field for their features to generate colloidal delivery systems of hydrophobic drugs to improve drug stability, specific delivery and reduce toxicity. In particular, nanoparticles surface functionalized with hyaluronic acid have been widely employed for their active tumor-targeting efficiency. Furthermore, the control of the physicochemical properties as well as high drug encapsulation efficiency, yield and batch-to-batch reproducibility of nanoparticles obtained by means of microfluidics, pave the way to overcome the drawbacks related to the fabrication of NPs obtained via traditional bulk methods and consequently increase cancer cell killing effect. For these reasons, we here investigated the effects of process parameters on the preparation and optimization of poly(lactic-co-glycolic acid)-poloxamers nanoparticle formulations (PLGA/PF68+PF127 NPs) decorated with HA (HA-based NPs) by using microfluidics for intravenous delivery of a model chemotherapeutic agent, irinotecan (IRIN). The study on the aggregation behavior and morphology of NPs were investigated by Dynamic Light Scattering (DLS), Electrophoretic Light Scattering (ELS), Transmission Electron Microscopy (TEM) and Differential Scanning Calorimetry (DSC). Our results revealed that by increasing the polymer weight fractions of organic and water phases the size, the NP yield, and the drug encapsulation efficiency of NPs are increased. Furthermore, by increasing the flow rate ratios from 0.05 to 0.3, a raise of dH from ∼85.6 to ∼176.4, for PLGA/PP NPs, and from ∼101.6 to ∼283.3 for HA/PLGA/PP is detected. The optimized HA-based NP formulation showed a spherical morphology, with a core-shell structure, small hydrodynamic diameter (dH∼120 nm), uniform size distribution (PDI<0.06) and high: i) drug loading (DL∼22.7%), ii) drug encapsulation efficiency (DEE∼87.8%) and iii) yield (92.4%). Moreover, the in vitro IRIN kinetic release and stability of IRIN-loaded HA-based NPs were evaluated showing the complete release in 15 days. Thus, this study highlighted a microfluidic method to fabricate IRIN-loaded HA-based NPs without any chemical modifications to provide novel understanding of the effect of process parameters, by varying the polymer weight fractions of both organic and water phases and flow rate ratio, on the physicochemical properties and consequently on nanoparticle formulation qualities for cancer therapy applications.

Protein-polymer Bioconjugate Nanoparticles To Investigate Innate and Antigen-Specific CD4 T Cell Responses

Abstract Nanoparticles (NP) comprised of poly(lactic-co-glycolic acid) (PLGA) and poly(lactic acid) (PLA) have been shown to be effective delivery systems for peptide and protein antigens (Ags) capable of modulating Ag-specific T cell responses. Encapsulation and surface conjugation are common Ag association methods using NPs. However, these methods are associated with caveats such as imprecise Ag loading, high Ag burst release, and immune recognition. To improve the efficiency of Ag delivery and address these limitations, we prepared ovalbumin (OVA)-PLGA conjugates that were systematically combined with unmodified PLGA to produce NPs (acNP-OVA) with three precise Ag loadings, negligible immune recognition, and a reduced burst release. acNP-OVA was used to elucidate how NP formulation parameters, such as Ag content, influence innate and Ag-specific CD4 +T cell responses. We observed Ag loading-dependent innate immune cell activation (MHCII, CD80/86) and Ag-specific CD4 +T cell proliferation, cytokine secretions, and regulatory T cell induction in vitro. Ag-encapsulated NPs were employed as a control. CD25 expression and T cell expansion were achieved using both Ag delivery methods and observed at low Ag loading. However, high Ag loading was required to induce IFNg and IL-2 cytokine secretions. Notably, nanostring analysis identified inflammation-associated genes, such as S100a9, was significantly downregulated in dendritic cells due to NP treatment. We anticipate this study will aid in designing NP-based immunotherapies for Ag-specific immunomodulation.

PEGylated Erlotinib HCl Injectable Nanoformulation for Improved Bioavailability.

The present study was undertaken to synthesize PEGylated monomethoxy poly (ethylene glycol)-poly (ε-Caprolactone) (mPEG-PCL) block copolymer and formulate Erlotinib HCl-loaded mPEG-PCL nanoparticles for enhancing the bioavailability of the drug. Using the ring-opening polymerization technique, PEGylated mPEG-PCL block copolymer was synthesized, and the structure of the copolymer was characterized using FTIR, 1H-NMR, and DSC techniques. The solvent evaporation approach was used to effectively encapsulate Erlotinib HCl within block copolymeric nanoparticles. Erlotinib HCl-loaded mPEG-PCL nanoparticles had a mean particle size of 146.5 ± 2.37 nm and a zeta potential of -27.8 ± 2.77mV. The nanoparticles had a percent entrapment efficiency of 80.78 ± 0.09%. The in vitro drug release of Erlotinib HCl-loaded copolymeric nanoparticles showed a slow and sustained release behavior which could be maintained for up to 72 h. The Korsmeyer-Peppas fitting findings indicated that the drug release process followed a non-Fickian diffusion mechanism. The pharmacokinetic (PK) behavior of the developed nanoformulation was studied in albino Wistar rats, and the relative bioavailability of the optimized NP formulation given by intravenous route was found to be 187.33%. The PK data suggested that Erlotinib HCl-loaded mPEG-PCL copolymeric nanoparticles can dramatically alter the PK behavior of Erlotinib HCl and greatly improve the drug's bioavailability by as much as three times when compared to the oral formulation. As a result, it was established that the block copolymeric nanoparticles have promise for the effective encapsulation of Erlotinib HCL for an injectable formulation with increased bioavailability.

Hydroponic Ginseng ROOT Mediated with CMC Polymer-Coated Zinc Oxide Nanoparticles for Cellular Apoptosis via Downregulation of BCL-2 Gene Expression in A549 Lung Cancer Cell Line

The unique and tailorable physicochemical features of zinc oxide nanoparticles (ZnO-NPs) synthesized from green sources make them attractive for use in cancer treatment. Hydroponic-cultured ginseng-root-synthesized ZnO-NPs (HGRCm-ZnO NPs) were coated with O-carboxymethyl chitosan (CMC) polymer, which stabilized and enhanced the biological efficacy of the nanoparticles. Nanoparticles were characterized by X-ray diffraction (XRD), UV-Vis spectroscopy, transmission electron microscopy (TEM), Fourier-transform infrared spectroscopy (FT-IR), and energy-dispersive X-ray spectroscopy (EDS). The flower-shaped nanoparticles were crystalline in nature with a particle size of 28 nm. To evaluate if these NPs had anti-lung cancer activity, analysis was performed on a human lung carcinoma cell line (A549). HGRCm-ZnO nanoparticles showed less toxicity to normal keratinocytes (HaCaTs), at concentrations up to 20 µg/mL, than A549 cancer cells. Additionally, these NPs showed dose-dependent colony formation and cell migration inhibition ability, which makes them more promising for lung cancer treatment. Additionally, Hoechst and propidium iodide dye staining also confirmed that the NP formulation had apoptotic activity in cancer cells. Further, to evaluate the mechanism of cancer cell death via checking the gene expression, HGRCm ZnO NPs upregulated the BAX and Caspase 3 and 9 expression levels but downregulated Bcl-2 expression, indicating that the nanoformulation induced mitochondrial-mediated apoptosis. Moreover, these preliminary results suggest that HGRCm ZnO NPs can be a potential candidate for future lung cancer treatment.

Synthesis and characterization of silver nanoparticles developed using a novel lipopeptide(s) biosurfactant and evaluating its antimicrobial and cytotoxic efficacy.

Novel bacterial lipopeptide(s) (LPs) biosurfactant obtained from Pseudomonas sp. OXDC12 were used to synthesizeLPs silver nanoparticles (LPs-Ag-NPs). LPs provided excellent colloidal stability by acting as a stabilizing/capping agent. The LPs-Ag-NPswere characterized using Field-Emission Scanning Electron Microscopy, Ultraviolet-visible spectroscopy,Fourier-transform infrared, Atomic force microscopy andX-Ray Diffraction. The formulation exhibited the production of monodisperse NPs with spherical morphology withan average diameter of 42.43 ± 5.2 nm.The metallic NP (LPs-Ag-NPs) formulation displayed remarkable antimicrobial/antifungal activity against Candida albicans (22.0 ± 1.3 mm), Fusarium oxysporum (28.0 ± 1.4 mm),Mucor sp. (26.0 ± 1.6 mm),Staphylococcus aureus MTCC96 (05.0 ± 0.6 mm), Salmonella typhimurium NCTC 74 (11.0 ± 0.2 mm), Klebsiellapneumnoniae (14.0 ± 0.3 mm) and Escherichiacoli MTCC1687 (12.6 ± 0.7 mm). The minimum inhibitory concentration (MIC) value was very less for LPs-Ag-NPs formulation when compared to purified LPs biosurfactant. LPs-Ag-NPs formulations showed cytotoxic activity against cancerous cells (L20B and RD; 80–94% at 80 µg/ mL) and were significantly less toxic against normal cells (PMC) showing only 22% cell reduction at 80 µg/ mL.In all, itwas concluded that capping the LPs on Ag-NPs successfully enhanced its antimicrobial and anti-cytotoxic properties.

Cecropin-Loaded Zeolitic Imidazolate Framework Nanoparticles with High Biocompatibility and Cervical Cancer Cell Toxicity.

Cecropins (CECs) are insect venom-derived amphiphilic peptides with numerous pharmacological effects, including anti-inflammatory, antibacterial, antiviral, and anti-tumor activities. Cecropins induce tumor cell death by disrupting phospholipid membrane integrity. However, non-specific cytotoxicity and in vivo rapid degradation limit clinical application. Nanotechnologies provide novel strategies for tumor eradication, including nanocarriers that can precisely target drugs to tumor tissue. We report the fabrication of CEC-encapsulated zeolitic imidazolate framework 8 (ZIF-8) nanoparticles (CEC@ZIF-8 NPs) via the preparation of CEC@ZIF-8 NPs in pure water by one-pot stirring. This method yielded morphologically uniform NPs with 20 wt% drug loading capacity and 9% loading efficiency. The NP formulation protected CECs from proteasome degradation, enhanced peptide bioavailability, promoted HeLa tumor cell uptake, and increased antitumor efficacy compared to free CECs. In conclusion, this ZIF-8 encapsulation strategy may enhance the clinical applicability of CECs and other antitumor peptides.

Chitosan-Hyaluronan Nanoparticles for Vinblastine Sulfate Delivery: Characterization and Internalization Studies on K-562 Cells.

In the present study, we developed chitosan/hyaluronan nanoparticles (CS/HY NPs) for tumor targeting with vinblastine sulfate (VBL), that can be directed to the CD44 transmembrane receptor, over-expressed in cancer cells. NPs were prepared by coating with HY-preformed chitosan/tripolyphosphate (CS/TPP) NPs, or by polyelectrolyte complexation of CS with HY. NPs with a mean hydrodynamic radius (RH) of 110 nm, 12% polydispersity index and negative zeta potential values were obtained by a direct complexation process. Transmission Electron Microscopy (TEM) images showed spherical NPs with a non-homogeneous matrix, probably due to a random localization of CS and HY interacting chains. The intermolecular interactions occurring between CS and HY upon NPs formation were experimentally evidenced by micro-Raman (µ-Raman) spectroscopy, through the analysis of the spectral changes of characteristic vibrational bands of HY during NP formation, in order to reveal the involvement of specific chemical groups in the process. Optimized NP formulation efficiently encapsulated VBL, producing a drug sustained release for 20 h. In vitro studies demonstrated a fast internalization of labeled CS/HY NPs (within 6 h) on K-562 human myeloid leukemia cells. Pre-saturation of CD44 by free HY produced a slowing-down of NP uptake over 24 h, demonstrating the need of CD44 for the internalization of HY-based NPs.

Evaluation of novel conjugated resveratrol polymeric nanoparticles in reduction of plasma degradation, hepatic metabolism and its augmentation of anticancer activity in vitro and in vivo

Resveratrol (RSV) is a natural product with multiple biological benefits including anticancer properties. Unfortunately, its biological benefits are limited by its low bioavailability and rapid hepatic metabolism and degradation in the body. The aim of this study was to develop an effective delivery system for RSV that would enhance the plasmatic stability and decrease the metabolism rate of RSV through a dual strategy of chemical modification and nanoparticle formulation. The effectiveness of this strategy was tested for the application of RSV anticancer treatment in a mouse cancer model. Chemical modification of RSV was achieved by conjugating RSV to a low molecular weight co-polymer mPEG-PLA. This conjugated RSV together with free RSV were formulated into mPEG-PLA nanoparticles (conjugated RSV NPs). These NPs showed a stable plasma stability profile and decreased liver metabolism rate compared to nanoparticles encapsulating free RSV in mPEG-PLA (encapsulated RSV NPs) and free RSV alone. However, in vitro cell studies using B16-F10 cancer cells showed that conjugated RSV NPs were less effective compared to encapsulated RSV NPs, possibly due to the lack of biotransformation of conjugated RSV to the active form RSV in the simple cell studies. To study the actual effect of our strategy, an in vivo C57BL/6J mouse model with subcutaneous B16-F10 melanoma using intraperitoneal administration was used to reveal the relationship between the improved plasma stability and reduced liver metabolism rate of RSV in conjugated RSV NPs, and suppression of the tumour growth in mice. In vivo, a better tumour suppression trend with conjugated RSV NPs was noted. Our study suggests that the use of chemical conjugation with NP formulation is an effective strategy to reduce the degradation and metabolism rate of RSV and consequently increase the antitumour activity of RSV in vivo. This strategy has potential to be further developed for the suppression of early growth of tumours with no side effects.

Antiparasitic properties of miltefosine-based nanoformulations against protozoan pathogen, Acanthamoeba castellanii

Background: Acanthamoeba castellanii genotype T4 is the causative agent of the progressively increasing sight-threatening Acanthamoeba keratitis and central nervous system infections. Because of the increased prevalence and the ineffectiveness of the current antiamoebic drugs, we synthesized miltefosine poly(lactic-co-glycolic acid) nanoparticles (miltefosine PLGA NP) as a potential potent and biocompatible antiamoebic drug. The advantage to use PLGA NP is to preserve the cells from the toxic effect of miltefosine drug. In particular, miltefosine PLGA nanoformulation offers a better cellular uptake and a sustained drug release compared with the free drug that presents potent cytotoxicity at high concentrations against human colon cancer cell lines. Methods: The miltefosine NP were synthesized using a double emulsion-solvent evaporation method, characterized, and then assessed for their antiamoebic activity against A. castellanii belonging to the T4 genotype. Blank PLGA NP and miltefosine were used as controls. Results: Amoebicidal assays revealed that at 25 and 50 µM, unmodified miltefosine eradicated 83% and 93% of amoebae, respectively. At these same concentrations of 25 and 50 µM, the amount of miltefosine released form PLGA NP formulation was limited to 22.6%. However, it killed 36% and 56% of the protozoa, respectively. Thus, the efficacy of PLGA NP formulation was similar to that of the unmodified miltefosine. Both miltefosine and its PLGA NP significantly inhibited the pretreated amoebae (minimum inhibitory concentration 50% = 37.23 and 55.26 µM, respectively, compared with 147.2 µM of the blank NP; P < 0.05) and reduced amoebae-mediated host cell death. The blank NP and miltefosine NP exhibited minimal cytotoxicity against colon epithelial cell lines. In contrast, the unmodified miltefosine caused 37%, 71%, and 88% of cytotoxicity at 10, 25, and 50 µM, respectively. Conclusion: Overall, these findings suggest that controlling the release of miltefosine from PLGA NP for a short time was almost as effective as miltefosine alone against A. castellanii genotype T4 while reducing host cell toxicity. Hence, this study demonstrates the feasibility of using PLGA NP for the treatment of Acanthamoebic infections.

Development of Oral Chewable Tablets Containing Montelukast Nanoparticles for the Treatment of Childhood Asthma: Preclinical Study in Animal Model

The aim of the present study was to formulate oral chewable tablets of Montelukast (MTL) in the form of nanoparticles (NP’s). The MTL loaded NP’s were formulated by ionotropic external gelation method using tripolyphosphate (TPP) as crosslinking agent and Tween 60 as surfactant. NP’s were characterized for drug loading, encapsulation efficiency, surface morphology, saturation solubility, particle size, zeta potential and polydispersity index. The optimized NP formulation was used for development of chewable tablets using direct compression method. The prepared tablets were characterized for disintegration test, dissolution, thickness, hardness, friability and assay. The optimized formulation was evaluated in asthamatic animals to demonstrate the efficiency in asthama. The encapsulation efficiency of NP’s was found between 91.24 to 98.21% while drug loading was in the range of 10.09–14.25%. All formulations were found of nanosized in nature (110 to 200 nm) with excellent zeta potential (20.12 to 22.27 mV). PDI of all NP formulations were found within acceptable limit (less than 0.3). The nanoparticles were found spherical in shape with smooth surface. The saturation solubility of MTL was enhanced nearly 10 times (92 mg/ml) as compared to pure MTL saturation solubility. All physical parameters of the tablets were found within range. The optimized tablets showed disintegration time of 20 sec while other formulations showed DT in the rage of 35–57 sec. Tab1 (Optimized formulation) showed almost 100% MTL release from chewable tablets within the period of 30 min. Reduction in lung resistance (RI) was found in animals treated with Tab1. This reduction in RI was found nearly two fold and three fold as compare to MTL treated and control group animals. These observations clearly support the efficacy of chewable tablets containing nanoparticulate MTL in asthmatic animals.

Intraocular Pressure Reduction Effect of 0.005% Latanoprost Eye Drops in a Hyaluronic Acid-Chitosan Nanoparticle Drug Delivery System in Albino Rabbits.

PurposeThe purpose of this study was to evaluate the intraocular pressure (IOP) reduction efficiency of hyaluronic acid-chitosan-latanoprost link nanoparticle (HA-CS-latanoprost link NP) formulated eye drops.MethodsThe IOP reduction study was performed in 24 normotensive albino rabbits. The test animals were randomized and grouped accordingly to treatment namely, HA-CS-latanoprost link NP, plain latanoprost, and the commercially available Xalatan eye drop, all were formulated with 0.005% latanoprost. The 9 days of the experiment were divided into baseline period (days 1–2), treatment period (days 3–6), and recovery period (days 7–9). A wireless noncontact tonometer was used to measure IOP at a time interval of 2 hours for 12 hours per day with 5 readings each.ResultsThe highest mean daily IOP reduction during the treatment period was 24% for plain latanoprost, 23% for Xalatan, and 29% for HA-CS-latanoprost link NP. The maximum reduction in IOP for plain latanoprost and Xalatan all occurred at the sixth hour with the peak effects of 4.85 mm Hg (37%) and 4.8 mm Hg (36%), respectively. Although HA-CS-latanoprost link NP had peak effects of 5.75 mm Hg (43%) at the sixth hour and 5.22 mm Hg (39%) at the eighth hour. Daily mean IOP measurements of each treatment group showed that HA-CS-latanoprost link NP has a greater IOP reduction effect compared with the other two treatments (P < 0.001).ConclusionsThe results showed that the formulation of latanoprost with CS and HA is more effective in reducing the IOP than by drug alone.Translational RelevanceThe results provide evidence from animal experiment that HA-CS-latanoprost link NP formulation could improve and sustain drug concentration in the anterior segment of the eye. The improved reduction in IOP with that HA-CS-latanoprost link NP formulation can serve as a basis that latanoprost eye drops can be formulated with decreased concentration of benzalkonium HCl, an irritant preservative and penetration enhancer.

Dual Encapsulated Dacarbazine and Zinc Phthalocyanine Polymeric Nanoparticle for Photodynamic Therapy of Melanoma.

Melanoma is an invasive and very aggressive skin cancer due to its multi-drug resistance that results in poor patient survival. There is a need to test new treatment approaches to improve therapeutic efficacy and reduce side effects of conventional treatments. PLA/PVA nanoparticles carrying both Dacarbazine and zinc phthalocyanine was produced by double emulsion technique. The characterization was performed by dynamic light scattering and atomic force microscopy. In vitro photodynamic therapy test assay using MV3 melanoma cells as a model has been performed. In vitro cell viability (MTT) was performed to measure cell toxicity of of nanoparticles with and without drugs using human endothelial cells as a model. The in vivo assay (biodistribution/tissue deposition) has been performed using radiolabeled PLA/PVA NPs. The nanoparticles produced showed a mean diameter of about 259nm with a spherical shape. The in-vitro photodynamic therapy tests demonstrated that the combination is critical to enhance the therapeutic efficacy and it is dose dependent. The in vitro cell toxicity assay using endothelial cells demonstrated that the drug encapsulated into nanoparticles had no significant toxicity compared to control samples. In-vivo results demonstrated that the drug loading affects the biodistribution of the nanoparticle formulations (NPs). Low accumulation of the NPs into the stomach, heart, brain, and kidneys suggested that common side effects of Dacarbazine could be reduced. This work reports a robust nanoparticle formulation with the objective to leveraging the synergistic effects of chemo and photodynamic therapies to potentially suppressing the drug resistance and reducing side effects associated with Dacarbazine. The data corroborates that the dual encapsulated NPs showed better in-vitro efficacy when compared with the both compounds alone. The results support the need to have a dual modality NP formulation for melanoma therapy by combining chemotherapy and photodynamic therapy.

Encapsulation of Apoptotic Proteins in Lipid Nanoparticles to Induce Death of Cancer Cells

According to previous reports, cationic nanoparticles (cNPs) consisting of 1,2-dioleoyl-3-trimethylammonium-propane (DOTAP), a cationic lipid, without any cargos, induce generation of reactive oxygen species (ROS) and toxicity in cells. We investigated the effect of DOTAP-based cNPs containing encapsulated human pro-apoptotic proteins (P-cNPs), tBid, Bax, or cytochrome c (Cyt c) on the death of HepG2, cancer cells. Upon the treatment to cells, cNPs containing Cyt c-cargo were the most effective in inducing cell death, followed by tBid- and Bax-cNPs. Among three P-cNPs, Cyt c-cNPs also induced the highest levels of ROS production and caspase-3 activity in the cells under the treatment with the same DOTAP concentration. The incorporation of dioleoylglycerol (DOG), a neutral phospholipid, in cNPs at the expense of DOTAP resulted in an increased amount of the encapsulated protein and consequently enhanced cell death in a DOG concentration-dependent manner. Thus, the present cNP formulation with protein cargo may elicit apoptotic effects in cancer cells and serve as a rational background for the preparation of cationic lipid-based NP formulation for cancer treatment.

High‐fat diet induced alteration in lipid enzymes and inflammation in cardiac and brain tissues: Assessment of the effects of Atorvastatin‐loaded nanoparticles

Treatment with Lipitor is associated with several adverse impacts. Here we investigated the effects of low Lipitor nanoparticles (atorvastatin calcium nanopartilcle [AC-NP]), with size less than 100 , on enzymes of lipid metabolism and inflammation in cardiac, hepatic, and brain tissues of hypercholestremic adult male rats. Adult male rats were divided into five experimental groups. In group 1, the intact control (normal pellet diet), animals were fed a normal control diet; the other four groups were fed a high-fat diet (HFD) for 6 weeks. After 6 weeks, groups from 2 to 5 were assigned as a positive control (HFD), HFD + Lipitor, HFD + AC-NP-R1, or HFD + AC-NP-R2. Different treatments were administrated orally for two regimen periods (R1 daily and R2 once every 3 days). The treatment was conducted for two consecutive weeks. The HFD group faced a significant elevation in 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA), associated with a significant reduction in low-density lipoprotein receptor (LDL-R) along with cholesterol 7 α-hydroxylase enzyme in hepatic tissues, compared with the control group. Also, the HFD group induced hepatic, cardiac, and brain inflammation, evidenced by increased hepatic oxidative stress markers and cardiac homocysteine, together with elevated proinflammatory cytokines interleukin-1β (IL-1β) and IL-6 levels in brain tissue, compared with the control group. Different AC-NP treatments significantly augmented both mRNA LDL-R and mRNA 7α-hydroxylase expression in hepatic tissues, associated with significant depletion in mRNA HMG-CoA expression, compared with HFD + Lipitor. The inflammation symptoms were ameliorated by the AC-NP treatments, compared to HFD + Lipitor. Lipitor encapsulation in NP formulation results in increased efficiency and reduced dose-related adverse effects known to be associated with the Lipitor chronic administration.

Enhanced anti-angiogenic effects of bevacizumab in glioblastoma treatment upon intranasal administration in polymeric nanoparticles

Glioblastoma multiforme (GBM) is one of the most aggressive cancers, where the aggressiveness of tumor has been associated to its high vascularization rate. Bevacizumab (Avastin®), an anti-angiogenic monoclonal antibody, has been used to decrease the angiogenic profile. To circumvent the blood-brain barrier (BBB) and decrease off-target organ toxicity, bevacizumab-loaded poly(D,L-lactic-co-glycolic acid) nanoparticles (PLGA NP) were developed and intranasally administrated in CD-1 mice to study their pharmacokinetic and pharmacodynamic profile. After 7 days of administration, PLGA NP showed a higher brain bioavailability of bevacizumab when compared to intranasally administrated free bevacizumab. On the other hand, bevacizumab-loaded PLGA NP were able to increase the penetration (higher Cmáx) and the residence time of bevacizumab into the brain (higher Clast). Furthermore, PLGA NP formulation totally prevented bevacizumab systemic exposure. The efficacy of this nanosystem was next evaluated in a validated orthotopic GBM nude mice model, studying the tumor growth over time by bioluminescence and the anti-angiogenic effects. After 14 days, bevacizumab-loaded PLGA NP demonstrated a reduction in the tumor growth accompanied by a higher anti-angiogenic effect compared to the free bevacizumab. These results can be explained by the fact that bevacizumab was found in the brain just for bevacizumab-loaded PLGA NP group, after 14 days of formulation administration. Therefore, we believe that our strategy would be an efficient alternative to improve GBM treatment with high impact for patient life quality and survival.