Double Emulsion Solvent Evaporation Technique Research Articles

Optimisation of microencapsulation efficiency of propolis phenolic compounds by double emulsion method using response surface methodology

AbstractThe aim of this study was to optimise the microencapsulation efficiency of propolis phenolic compounds by double emulsion solvent evaporation technique (W1/O/W2). The solvent/sample ratio and the polymer and surfactant concentration parameters were optimised using response surface methodology (RSM) through Box–Behnken Design (BBD). For each parameter studied, total phenolic content encapsulation efficiency (TPCEE), free radical scavenging activity (DPPH), and ferric reducing antioxidant power (FRAP) were evaluated. The results showed that the optimal parameters were: 31.60 mg mL−1 for sample/solvent ratio, 606.28 mg mL−1 for poly(ε-caprolactone) (PCL) concentrations, and 2.05 g mL−1 for poly(vinyl alcohol) (PVA) concentration. The optimum values obtained were: 84.62% for encapsulation efficiency of phenolic content, 51.89% for DPPH, and 48,733 mg Trolox Equivalent/100 g dry weight for FRAP. The experimental checking of results revealed the validity of elaborated models and their suitability for the prediction of both responses. The developed mathematical models have expressed a high level of significance through RSM optimisation processes for phenolic antioxidants of propolis.

Synthesis and characterization of tenofovir disoproxil fumarate loaded nanoparticles for HIV‐1 treatment

AbstractThe remarkable ability of the human immunodeficiency virus (HIV) to evade the host's immune system and conventional antiretroviral therapy, has posed significant challenges in achieving complete eradication of the virus in people living with HIV (PLWHIV). However, nanotechnology has emerged as promising avenue for addressing some of the obstacles associated with the use of antiretroviral drugs by modifying drug molecules in nanoscale dimensions. Hence, the present study explores the utilization of poly(epsilon‐caprolactone) (PCL) as a carrier for encapsulating tenofovir disoproxil fumarate (TDF), offering an alternative treatment approach for HIV infection. TDF‐loaded polymeric nanoparticles were successfully prepared using double emulsion solvent evaporation technique and characterized. The characterization of TDF‐loaded polymeric nanoparticles at varied drug to polymer ratios showed that TDF was loaded in PCL with an encapsulation efficiency and drug loading capacity in the range of 23–46% and 4.8–19.9%, respectively. Of note, the neutralization efficacy of TDF‐loaded polymeric nanoparticles was more improved compared to free TDF. Encapsulation of TDF with PCL did not hinder the antiviral activity of TDF against HIV‐1 infection but rather enhanced its potency.

Aerosolizable Pyrazinamide-Loaded Biodegradable Nanoparticles for the Management of Pulmonary Tuberculosis.

Background: Pyrazinamide is a Biopharmaceutical Classification System class III antibiotic indicated for active tuberculosis. Methods: In the present work, pyrazinamide-loaded biodegradable polymeric nanoparticles (PNPs) based dry powder inhaler were developed using the double emulsion solvent evaporation technique and optimized using design of experiments to provide direct pulmonary administration with minimal side effects. Batches were characterized for various physicochemical and aerosol performance properties. Results: Optimized batch exhibited particle size of 284.5 nm, % entrapment efficiency of 71.82%, polydispersibility index of 0.487, zeta potential of -17.23 mV, and in vitro drug release at 4 hours of 79.01%. Spray-dried PNPs were evaluated for drug content, in vitro drug release, and kinetics. The particle mass median aerodynamic diameter was within the alveolar region's range (2.910 μm). In the trachea and lung, there was a 2.5- and 1.2-fold increase in in vivo deposition with respect to pure drug deposition, respectively. In vitro drug uptake findings showed that alveolar macrophages with pyrazinamide PNPs had a considerably higher drug concentration. Furthermore, accelerated stability studies were carried out for the optimized batch. Results indicated no significant change in the evaluation parameters, which showed stability of the formulation for at least a 6-month period. Conclusion: PNPs prepared using biodegradable polymers exhibited efficient pulmonary drug delivery with decent stability.

PLGA-Encapsulated Haemonchus contortus Antigen ES-15 Augments Immune Responses in a Murine Model.

Haemonchus contortus is a gastrointestinal parasite that adversely impacts small ruminants, resulting in a notable reduction in animal productivity. In the current investigation, we developed a nanovaccine by encapsulating the recombinant protein rHcES-15, sourced from the excretory/secretory products of H. contortus, within biodegradable poly (D, L-lactide-co-glycolide) (PLGA) nanoparticles (NPs). The development of this nanovaccine involved the formulation of PLGA NPs using a modified double emulsion solvent evaporation technique. Scanning electron microscopy (SEM)verified the successful encapsulation of rHcES-15 within PLGA NPs, exhibiting a size range of 350-400 nm. The encapsulation efficiency (EE) of the antigen in the nanovaccine was determined to be 72%. A total of forty experimental mice were allocated into five groups, with the nanovaccine administered on day 0 and the mice euthanized at the end of the 14-day trial. The stimulation index (SI) from the mice subjected to the nanovaccine indicated heightened lymphocyte proliferation (*** p < 0.001) and a noteworthy increase in anti-inflammatory cytokines (IL-4, IL-10, and IL-17). Additionally, the percentages of T-cells (CD4+, CD8+) and dendritic cell phenotypes (CD83+, CD86+) were significantly elevated (** p < 0.01, *** p < 0.001) in mice inoculated with the nanovaccine compared to control groups and the rHcES-15 group. Correspondingly, higher levels of antigen-specific serum immunoglobulins (IgG1, IgG2a, IgM) were observed in response to the nanovaccine in comparison to both the antigenic (rHcES-15) and control groups (* p < 0.05, ** p < 0.01). In conclusion, the data strongly supports the proposal that the encapsulation of rHcES-15 within PLGA NPs effectively triggers immune cells in vivo, ultimately enhancing the antigen-specific adaptive immune responses against H. contortus. This finding underscores the promising potential of the nanovaccine, justifying further investigations to definitively ascertain its efficacy.

Microfluidic-assisted preparation of PLGA nanoparticles loaded with insulin: a comparison with double emulsion solvent evaporation method

Poly lactic-co-glycolic acid (PLGA) is an ideal polymer for the delivery of small and macromolecule drugs. Conventional preparation methods of PLGA nanoparticles (NPs) result in poor control over NPs properties. In this research, a microfluidic mixer was designed to produce insulin-loaded PLGA NPs with tuned properties. Importantly; aggregation of the NPs through the mixer was diminished due to the coaxial mixing of the precursors. The micromixer allowed for the production of NPs with small size and narrow size distribution compared to the double emulsion solvent evaporation (DESE) method. Furthermore, encapsulation efficiency and loading capacity indicated a significant increase in optimized NPs produced through the microfluidic method in comparison to DESE method. NPs prepared by the microfluidic method were able to achieve a more reduction of trans-epithelial electrical resistance values in the Caco-2 cells compared to those developed by the DESE technique that leads to greater paracellular permeation. Compatibility and interaction between components were evaluated by differential scanning calorimetry and fourier transform infrared analysis. Also, the effect of NPs on cell toxicity was investigated using MTT test. Numerical simulations were conducted to analyze the effect of mixing patterns on the properties of the NPs. It was revealed that by decreasing flow rate ratio, i.e. flow rate of the organic phase to the flow rate of the aqueous phase, mixing of the two streams increases. As an alternative to the DESE method, high flexibility in modulating hydrodynamic conditions of the microfluidic mixer allowed for nanoassembly of NPs with superior insulin encapsulation at smaller particle sizes.

Treatment of methicillin resistant Staphylococcus aureus wound infection usingvancomycin-loaded nanoparticles: Aninvitro and in vivo study.

Due to the rise of bacteria that are resistant to multiple drugs, treating infections in burn wounds has become a worldwide problem. It is important to find new ways to treat these infections. Lipid-based drug delivery systems have emerged as a promising type of carrier for antibiotics as well as antibacterial agents, such as antimicrobial peptides and nucleic acids, for wound infections. To this end, the current study assessed the antibacterial effects and wound healing properties of vancomycin-loaded solid lipid nanoparticles (SLN) against methicillin-resistant Staphylococcus aureus (MRSA) surgical wound infections. vancomycin-SLN was synthesized by double emulsion solvent evaporation techniques, and then in vitro antibacterial activity was evaluated by well diffusion and broth microdilution methods. A surgical wound was inflicted on 36 male rats; the infection was caused by MRSA, and free vancomycin and vancomycin-SLN were topically applied. Particle size, PDI, zeta potential, drug loading, and encapsulation efficiency of the optimum vancomycin-SLN were 350 ± 24 nm, 0.402 ± 0.014, -16.3 ± 2.5 mV, 13.9 ± 1.4%, and 92.14 ± 3.1%, respectively. In vitro antibacterial assays showed lower MICs for free vancomycin and killed bacteria more efficiently than vancomycin-SLN. However, the synthesized nanoparticles indicated long-term antibacterial activity against MRSA. Animals that were treated with vancomycin-SLN showed the best wound healing procedure. Treatment of rats with vancomycin-SLN increased re-epithelialization and decreased granulation tissue development, as seen by histological analysis. Antibiotic-loaded SLN could not only manage wound infection but also enhance wound healing. Therefore, this kind of treatment should be considered by scientists as an applicable treatment for wounds.

In vitro performance of composition-equivalent PLGA microspheres encapsulating exenatide acetate by solvent evaporation

The once-weekly Bydureon® (Bdn) PLGA microsphere formulation encapsulating the GLP-1 receptor agonist, exenatide acetate, is an important complex injectable product prepared by coacervation for the treatment of type 2 diabetic patients. Encapsulation by coacervation is useful to minimize an undesirable initial burst of exenatide, but it suffers from manufacturing difficulties such as process scale-up and batch-to-batch variations. Herein we prepared exenatide acetate-PLGA formulations of similar compositions using the desirable alternative double emulsion-solvent evaporation technique. After screening several process variables, we varied the PLGA concentration, the hardening temperature, and the collected particle size range, and determined the resulting drug and sucrose loading, initial burst release, in vitro retention kinetics, and peptide degradation profiles using Bdn as a positive control. All formulations exhibited a triphasic release profile with a burst, lag, and rapid release phase, although the burst release was greatly decreased to <5% for some. Marked differences were observed in the peptide degradation profiles, particularly the oxidized and acylated fractions, when the polymer concentration was varied. For one optimal formulation, the release and peptide degradation profiles were similar to Bdn microspheres, albeit with an induction time shift of one week, likely due to the slightly higher Mw of PLGA in Bdn. These results highlight the effects of key manufacturing variables on drug release and stability in composition-equivalent microspheres encapsulating exenatide acetate and indicate the potential of manufacturing the microsphere component of Bdn by solvent evaporation.

Controlled release of inactivated influenza virus and α-galactosylceramide from PLGA microparticles results in long-term protection against influenza infection

Abstract The objective of this study was to develop biodegradable microparticles with continuous release that can enhance and extend the long-term protection of single-dose vaccines. Several formulations of Poly (lactic-co-glycolic acid) (PLGA) microparticles were synthesized using a double emulsion solvent evaporation technique to encapsulate inactivated A/PR/8/34 (PR8) H1N1 influenza virus in combination with the invariant natural killer T (NKT) cell agonist α-galactosylceramide (α-GalCer). The most efficient controlled release particle formulations were injected into C57BL/6J mice. Additional control mice were injected with PBS alone, inactivated PR8 dissolved in PBS, or inactivated PR8 and α-GalCer dissolved in PBS. Thirty weeks after vaccination, mice were challenged with a lethal dose of live homologous virus. Mice immunized with the microparticle vaccine survived the infection and had reduced clinical disease and decreased weight loss compared to unvaccinated mice. After vaccination, mice injected microparticles presented a gradual increase production of PR8-specific IgGs that was dominated by IgG1 isotype that is usually produced by T helper (Th) 2 responses. In contrast, mice that received soluble vaccine presented a rapid antibody response that was dominated by antibodies of the Th1-associated IgG2a isotype. Collectively, these results demonstrate the potential of PLGA-based single-dose microparticle vaccines for providing long-term protection against influenza infection and potentially other pathogens. Supported by grants from USAMRAA (W81XWH-19-1-0005)

Box-Behnken Design of Experiments of Polycaprolactone Nanoparticles Loaded with Irinotecan Hydrochloride.

The Box-Behnken design of experiments (BBD) is a statistical modelling technique that allows the determination of the significant factors in developing nanoparticles (NPs) using a limited number of runs. It also allows the prediction of the best levels of variables to obtain the desired characteristics (size, charge, and encapsulation efficiency) of the NPs. The aim of this study was to examine the effect of the independent variables (amount of polymer and drug, and surfactant concentration) and their interaction on the characteristics of the irinotecan hydrochloride (IRH)-loaded polycaprolactone (PCL) NPs and to determine the most optimum conditions for producing the desired NPs. The development of the NPs was carried out by a double emulsion solvent evaporation technique with yield enhancement. The NPs data were fitted in Minitab software to obtain the best fit model. By using BBD, the most optimum conditions for producing the smallest size, highest magnitude of charge, and highest EE% of PCL NPs were predicted to be achieved by using 61.02 mg PCL, 9 mg IRH, and 4.82% PVA, which would yield 203.01 nm, -15.81 mV, and 82.35% EE. The analysis by BBD highlighted that the model was a good fit to the data, confirming the suitability of the design of the experiments.

Oral PLGA-based DNA vaccines using interferons as adjuvants remarkably promote the immune protection of grass carp (Ctenopharyngodon idella) against GCRV infection

Grass carp hemorrhagic disease caused by grass carp reovirus (GCRV) results in significant economic losses to the global grass carp aquaculture industry. Oral vaccination is an ideal choice for disease precaution in aquaculture. However, oral vaccine can be degraded in the gut. Therefore, the selection of loading materials is essential. In this study, the S6 and S7 fragments (encoding the outer capsid protein VP4 and fibronectin VP56 of GCRV) and grass carp interferons (IFNs), including IFN1, IFN3, and IFNγ2 were used to create DNA vaccines and adjuvants based on pcDNA3.1, respectively. The oral DNA vaccine was encapsulated in poly(lactic-co-glycolic acid) (PLGA) and polyvinyl alcohol (PVA) with IFNs. The PLGA-PVA (PP) nano-microspheres were prepared by double emulsion-solvent evaporation technique. Using transmission electron microscopy and dynamic light scattering assays, it was determined the vaccines had a spherical structure with uniform particle size (643.5 ​± ​35.3 ​nm). The nano-microspheres possessed excellent encapsulation efficiency (81.6 ​± ​2.6%) and loading rate (0.54 ​± ​0.02%), and simultaneously exhibited negligible hemolytic activity and cell toxicity. The protection rate and tissue viral loads post-GCRV challenge in grass carp were assessed. The oral PP nano-microsphere with pVP4 ​+ ​pIFN1 (PP41) vaccine increased protection rate by 44% compared with the control group and was correlated with relatively low viral loads in the spleen, head kidney, and hindgut. Further, three crucial serum biochemical indexes, total superoxide dismutase (TSOD), complement C3 (C3), and lysozyme (LZM), were also dramatically increased. Furthermore, mRNA expressions of representative immune-related genes (IgM, IFN1, IFNγ2, MHC-Ⅰ, and CD8α) in the head kidney and spleen were significantly enhanced. In addition, mRNA expression of IgT was significantly boosted in the hindgut. The results indicate that DNA vaccine capsulated with PP is effective against GCRV infection. The present study provides insights into a prospective strategy for oral vaccine development in aquaculture.

Clindamycin phosphate and bone morphogenetic protein-7 loaded combined nanoparticle-graft and nanoparticle-film formulations for alveolar bone regeneration – An in vitro and in vivo evaluation

Commonly utilized techniques for healing alveolar bone destruction such as the use of growth factors, suffering from short half-life, application difficulties, and the ability to achieve bioactivity only in the presence of high doses of growth factor. The sustained release of growth factors through a scaffold-based delivery system offers a promising and innovative tool in dentistry. Furthermore, it is suggested to guide the host response by using antimicrobials together with growth factors to prevent recovery and achieve ideal regeneration. Herein, the aim was to prepare and an in vitro - in vivo evaluation of bone morphogenetic protein 7 (BMP-7) and clindamycin phosphate (CDP) loaded polymeric nanoparticles, and their loading into the alginate-chitosan polyelectrolyte complex film or alloplastic graft to accelerate hard tissue regeneration. PLGA nanoparticles containing CDP and BMP-7, separately or together, were prepared using the double emulsion solvent evaporation technique. Through in vitro assays, it was revealed that spherical particles were homogeneously distributed in the combination formulations, and sustained release could be achieved for >12 weeks with all formulations. Also, results from the micro-CT and histopathological analyses indicated that CDP and BMP-7 loaded nanoparticle-film formulations were more effective in treatment than the nanoparticle loaded grafts.

Harnessing Folate-Functionalized Nasal Delivery of Dox-Erlo-Loaded Biopolymeric Nanoparticles in Cancer Treatment: Development, Optimization, Characterization, and Biodistribution Analysis.

The aim of the present study is to develop Doxorubicin-Erlotinib nanoparticles (Dox-Erlo NPs) and folate-armored Dox-Erlo-NP conjugates for targeting glioma cancer. Glioma is one of the most common progressive cancerous growths originating from brain glial cells. However, the blood-brain barrier (BBB) is only semi-permeable and is highly selective as to which compounds are let through; designing compounds that overcome this constraint is therefore a major challenge in the development of pharmaceutical agents. We demonstrate that the NP conjugates studied in this paper may ameliorate the BBB penetration and enrich the drug concentration in the target bypassing the BBB. NPs were prepared using a biopolymer with a double-emulsion solvent evaporation technique and functionalized with folic acid for site-specific targeting. Dox-Erlo NPs and Dox-Erlo-NP conjugates were extensively characterized in vitro for various parameters. Dox-Erlo NPs and Dox-Erlo-NP conjugates incurred a z-average of 95.35 ± 10.25 nm and 110.12 ± 9.2 nm, respectively. The zeta potentials of the Dox-Erlo NPs and Dox-Erlo-NP conjugates were observed at -18.1 mV and -25.1 mV, respectively. A TEM image has shown that the NPs were well-dispersed, uniform, de-aggregated, and consistent. A hemolytic assay confirmed hemocompatibility with the developed formulation and that it can be safely administered. Dox-Erlo-NP conjugates significantly reduced the number of viable cells to 24.66 ± 2.08% and 32.33 ± 2.51% in U87 and C6 cells, respectively, and IC50 values of 3.064 µM and 3.350 µM in U87 and C6 cells were reported after 24 h, respectively. A biodistribution study revealed that a significant concentration of Dox and Erlo were estimated in the brain relative to drug suspension. Dox-Erlo-NP conjugates were also stable for three months. The findings suggest that the developed Dox-Erlo-NP conjugates may be a promising agent for administration in glioma therapy.

Enhanced Coating Protection of C-Steel Using Polystyrene Clay Nanocomposite Impregnated with Inhibitors.

Polymer-Clay Nanocomposite (PCN) coatings were prepared using the solution intercalation method. The raw Khulays clay was treated with NaCl to produce sodium clay (NaC). Thereafter, Cetyl Pyridinium Chloride (CPC) was used to convert NaC into the organic clay form (OC). PCN was prepared by adding polystyrene as the matrix to different weights of OC to prepare 1 wt.% and 3 wt.% PCN. To enhance the coating protection of C-steel in NaCl solution, PCN coatings were added to microcapsules loaded with some corrosion inhibitors PCN (MC). The microcapsules are prepared by the encapsulation of rare-earth metal Ce+3 ions and Isobutyl silanol into polystyrene via the Double Emulsion Solvent Evaporation (DESE) technique. Characterization techniques such as FTIR, X-Ray Diffraction (XRD), and Transmission Electron Microscopy (TEM) were employed. FTIR confirmed the success of the preparation, while XRD and TEM revealed an intercalated structure of 1 wt.% PCN while 3 wt.% PCN has a fully exfoliated structure. Electrochemical Impedance Spectroscopy (EIS), Electrochemical Frequency Modulation (EFM), and Potentiodynamic Polarization showed an enhanced protection efficiency of PCN (MC) coatings. The results demonstrated that the corrosion resistance (RCorr) of 3% PCN (MC) coating was higher than all the formulations. These PCN (MC) coatings may provide corrosion protection for C-steel pipes in many industrial applications.

Immobilization of the metagenomic lipase, LipG9, on porous pellets of poly-hydroxybutyrate produced by the double emulsion solvent evaporation technique.

This work aimed to produce porous poly-hydroxybutyrate (PHB) pellets in order to evaluate the pellets as a support for immobilization of the metagenomic lipase, LipG9. Four types of pelletized PHB particles with different morphological characteristics were obtained using the double emulsion and solvent evaporation technique (DESE). The micropores of these PHB pellets had similar average diameters (about 3nm), but the pellets had different specific surface areas: 11.7 m2 g-1 for the PHB powder, 8.4m2 g-1 for the control pellets (Ø<0.5mm, produced without the pore forming agent), 10.0m2 g-1 for the small pellets (Ø<0.5mm), 9.5m2 g-1 for the medium pellets (0.5 < Ø<0.8mm) and 8.4m2 g-1 for the large pellets (Ø>1.4mm). Purified LipG9 was immobilized by adsorption on these pellets, and the results were compared with those obtained with PHB powder. The highest immobilization yield (83%) was obtained for the medium PHB pellets, followed by large (76%) and small (55%) PHB pellets. The activity of LipG9 immobilized on the pellets, for the synthesis of ethyl oleate in n-hexane, was highest for the medium pellets (22Ug-1 ). The immobilization yield was high for PHB powder (99%) but the esterification activity was slightly lower (20Ug-1 ). These results show that pelletized PHBbeads can be used for the immobilization of lipases, with the advantage that pelletized PHB will perform better than PHB powder in large-scale enzyme bioreactors.

Preparation of polyvinyl alcohol hydrogel containing chlorogenic acid microspheres and its evaluation for use in skin wound healing.

Chlorogenic acid (CGA) is a phenolic compound widely found in plants. Several studies have shown that CGA possesses antioxidant, antibacterial, anti-inflammatory and wound healing properties. Because of their three-dimensional structure, good permeability, excellent biocompatibility and moisturizing properties, hydrogels are ideal candidates for wound dressing. The aim of the present study was to preparation and characterization of Polyvinyl alcohol (PVA) hydrogel containing CGA microspheres and evaluation its wound healing activity. The double-emulsion solvent evaporation technique was applied for preparing the CGA containing microspheres. The microspheres were characterized using scanning electron microscopy (SEM) and Fourier transformation infrared spectroscopy (FTIR) and subsequently incorporated in the structure of a PVA hydrogel. The effects of prepared hydrogel on NIH3T3 cell line viability were evaluated using MTT method and wound healing activity was investigated in full thickness wound model in rabbit. SEM images showed formation of homogenous CGA microspheres with diameters in the range of 1-2μm, embedded in the porous structure of the hydrogel. Infra-red results indicated successful incorporation of CGA microspheres into PVA hydrogel. The NIH3T3 cell viability percentage in CGA 2.5% hydrogel treated group significantly (p < .05) increased after 24 h and 48 h comparing to control group. In vivo studies showed that CGA hydrogel significantly (p < .001) stimulated the rate of wounds closures. Histological studies revealed that administration of CGA hydrogel significantly increased epithelialization and production of collagen fibers compared to the control group. It can be concluded that the CGA microsphere loaded PVA hydrogel has the potential for wound healing.

Olive Mill Pomace Extract Loaded Ethylcellulose Microparticles as a Delivery System to Improve Olive Oils Oxidative Stability

The protective effect of olive mill pomace (OMP) loaded ethylcellulose microparticles as an alternative to synthetic antioxidants against the oxidation of olive oils was assessed. OMP extract was obtained by an optimized two-step solid-liquid extraction; encapsulation was performed by double emulsion solvent evaporation technique considering a theoretical loading content in phenolic compounds of 5% (w/w). The changes in the peroxide values, the p-anisidine values, the total oxidation values, the free fatty acids content, the total antioxidant activity, and the total phenolic content were synchronized under storage at 62 °C. The results of oxidative stability were compared with plain oils, oils enriched with synthetic antioxidants, and oils fortified with OMP extract. The encapsulation efficiency of phenolic compounds was 96.0 ± 0.3%. The fortification of olive oils with microparticles retarded the appearance of peroxides, reduced the content of secondary oxidation products, and slowed down hydrolysis processes. The microparticles were efficiently designed to sustain the release of antioxidants to control the oxidative status of oil samples, retarding the free fatty acids formation rather than synthetic antioxidants. The results of this study bring new perspectives regarding the potential use of encapsulated extracts rich in antioxidants as an alternative to synthetic antioxidants to improve oil oxidative stability.

Targeting CXCR4-expressing Cancer Cells with Avidin-poly (lactic-co-glycolic acid) Nanoparticle Surface Modified with Biotinylated DV1 Peptide.

Chemokine receptor CXCR4 is frequently present in cells of various cancers. Hence, targeted therapy using CXCR4 ligands, such as DV1 peptide, on drug-loaded nanoparticles, has the potential to enhance the efficiency of cancer treatment. The present study created a CXCR4-targeting drug delivery system using avidin-poly (lactic-co-glycolic acid) (PLGA) nanoparticle surface tagged with biotinylated DV1 peptide ligand. A double-emulsion solvent evaporation technique was employed to prepare avidin-PLGA nanoparticles and characterized by transmission electron microscopy (TEM) and dynamic light scattering. Uptake was studied by confocal microscopy after incorporating fluorescein isothiocyanate (FITC)-labeled albumin inside the nanoparticles during their synthesis. Peptide-biotin-avidin-PLGA nanoparticles were tested in vitro on CXCR4-expressing U87MG cells. Photomicroscopy was done by a Nikon A1 Confocal Microscope, and pictures were analyzed by Nikon NIS-Elements BR software. Experimental results confirmed the specificity of DV1 peptide-tagged avidin-PLGA nanoparticles for cells expressing CXCR4 receptors. The avidin-PLGA nanoparticles were successfully synthesized and the same was confirmed by tagging them with FITC-labeled biotin. Avidin-PLGA nanoparticle surface tagged with biotinylated DV1 peptide ligand has potential clinical application in the treatment of various cancers as targeted therapy for CXCR4-expressing cancer cells.

Polycaprolactone based pharmaceutical nanoemulsion loaded with acriflavine: optimization and in vivo burn wound healing activity

Cutaneous burn wounds are a common and troublesome critical issue of public health. Over the last decade, many researchers have investigated the development of novel therapeutic modalities which are capable of fully regeneration and reinstatement of structure and function of the skin with no or limited scar formation. Novel pharmaceutical carriers are offering a potential platform to deliver the drug effectively and to overcome the limitation associated with conventional wound dressings. The aim of this study was to investigate a pharmaceutical acriflavine-loaded polycaprolactone nanoemulsion (ACR-PCL-NE) for burn wound healing. Nanoemulsion was prepared by using the double emulsion solvent evaporation technique and it was subjected to thermodynamic stability testing, droplet size, polydispersity, zeta potential, pH, and surface morphology analysis. The in vivo study was performed to evaluate the efficacy of nanoemulsion using Sprague-Dawley rats as an animal model. The results of this study revealed that the optimized nanoemulsion was stable and had desirable physicochemical properties. The pH was about 4.02 at 25 °C and the particle size was found to be in the range of 302 ± 4.62 nm while the zeta potential was −7.8 ± 1.22 mV and the polydispersity index of 0.221 ± 0.017. The wound regeneration process was evaluated in vivo by different techniques, the formulation group (FG) showed high wound healing potential as compared to the standard group (SD) and control group (CG). These findings reveal that this nanoemulsion formulation can be used effectively for wound healing.

Biodegradable porous micro/nanoparticles with thermoresponsive gatekeepers for effective loading and precise delivery of active compounds at the body temperature

Stimuli-responsive controlled delivery systems are of interest for preventing premature leakages and ensuring precise releases of active compounds at target sites. In this study, porous biodegradable micro/nanoparticles embedded with thermoresponsive gatekeepers are designed and developed based on Eudragit RS100 (PNIPAM@RS100) and poly(N-isopropylacrylamide) via a double emulsion solvent evaporation technique. The effect of initiator types on the polymerization of NIPAM monomer/methylene-bis-acrylamide (MBA) crosslinker was investigated at 60 °C for thermal initiators and ambient temperature for redox initiators. The crosslinked PNIPAM plays a key role as thermal-triggered gatekeepers with high loading efficiency and precise release of a model active compound, Nile Blue A (NB). Below the volume phase transition temperature (TVPT), the gatekeepers possess a swollen conformation to block the pores and store NB within the cavities. Above its TVPT, the chains rearrange, allowing gate opening and a rapid and constant release rate of the compound until completion. A precise “on–off” switchable release efficiency of PNIPAM@RS100 was demonstrated by changing the temperatures to 4 and 40 °C. The materials are a promising candidate for controlled drug delivery systems with a precise and easy triggering mechanism at the body temperature for effective treatments.

Preparation and characterization of PLA-PEG/Chitosan-FA/DNA for gene transfer to MCF-7 cells

The ability of Chitosan-Folic acid and PLA-PEG polymers for gene delivery into MCF-7 cells was evaluated. The PLA-PEG copolymer is one of the most appealing nanoparticles for gene transfer to human cells because of its biocompatibility and high blood circulation. However, one of the barriers to the use of these polymers in gene therapy has always been the low effectiveness of these nanoparticles. In this study, Chitosan and folic acid were utilized to improve gene transfer efficiency and encapsulation of PLA-PEG/Chitosan-FA/DNA nanoparticles. PLA-PEG/Chitosan-FA/DNA nanoparticles with different concentrations of Chitosan-FA were prepared using double emulsion-solvent evaporation technique. The biological properties (i.e., biocompatibility, DNA release, and gene transfer ability to MCF-7 cells in vitro) as well as the physical and chemical properties (i.e., particle size, zeta potential, and the morphology of the resultant nanoparticles) of PLA-PEG/Chitosan-FA/DNA nanoparticles were investigated. The results showed that increasing the Chitosan-FA concentration in the PLA-PEG/Chitosan-FA/DNA nanoparticles resulted in an increase in zeta potential, DNA release, encapsulation, and gene delivery efficiency. The MTT assay showed PLA-PEG/Chitosan-FA/DNA nanoparticles exhibit low cytotoxicity and good compatibility. On the other hand, fluorescence microscopy and flow cytometry were used to test the ability of PLA-PEG/Chitosan-FA/DNA nanoparticles with varied concentrations of chitosan-folic acid to transfer the gene to MCF-7 cells. Flow cytometry and fluorescence microscopy analysis showed that increasing the concentration of chitosan-folic acid in PLA-PEG/Chitosan-FA/DNA nanoparticles improved gene transfer efficiency to MCF-7 cells.