Average Zeta Potential Research Articles

Loratadine oral bioavailability enhancement via solid dispersion loaded oro-dispersible films: Formulation, characterization and pharmacokinetics.

Loratadine (LRD) belongs to second-generation tricyclic H1 antihistamine class, known for its non-sedating properties in allergic reactions. H1 antihistamines avoid and block the responses to allergens or histamine in nose and conjunctivae, thereby abolishing itching, congestion and sneezing. LRD is a Biopharmaceutical Class System (BCS) class II drug with dissolution or solubility limited absorption which limited the oral bioavailability and therapeutic efficacy of LRD. To improve the oral bioavailability of LRD for allergic disease (urticaria) treatment, LRD solid dispersions (LRD-SDs) were integrating into oro-dispersible films (ODFs). LRD-SDs were prepared through hot-melt extrusion method (HME) using d-alpha-tocopherol polyethylene glycol 1000 succinate (TPGS-1000), and polyvinyl caprolactam-polyvinyl acetate-polyethylene glycol graft copolymer (SP). Subsequently, LRD-SDs were incorporated in ODFs by solvent casting method. The physicochemical and mechanical properties of LRD solid dispersions-loaded oro-dispersible films (LRD-SDs-ODFs), were evaluated. The in-vitro dissolution, ex-vivo permeation, oral bioavailability, and pharmacodynamics studies were conducted to evaluate LRD-SDs-ODFs efficiency. LRD-SDs-ODFs showed superior solubility and in-vitro dissolution results compared to that of pure LRD (p<0.05). The solubility of the LRD-SD coded as LTS-4 was 190 times higher than the pure drug in aqueous media. The average hydrodynamic particle size (PS), polydispersity index (PDI), and zeta potential (ZP) of SD particles were 76±2.1nm, 0.20±0.08 and -19.16±1.4mV, respectively. Moreover, differential scanning calorimetry (DSC) and X-ray diffraction (XRD) results confirmed the amorphousness of LRD in LRD-SDs-ODFs. The permeability flux of LRD was 44.6±3.1μg/cm2/h from DPF-5 formulation. Likewise, in vivo oral bioavailability of DPF-5 in Sprague-Dawley rats was significantly increased (p<0.05) compared to free LRD. Further, wheal area was reduced 20 % higher than LRD in 8h (p<0.05). Overall, LRD-SDs-ODFs considerably enhanced LRD solubility, dissolution rate, bioavailability, and antihistaminic efficacy. Our findings show that SDs-ODFs is an effective carrier system for delivering poorly soluble LRD.

Effects of clove essential oil (Caryophyllus aromaticus L.) nanoemulsion incorporated edible coating on shelf-life of fresh cut apple pieces

Fresh cut fruit is ready to eat and can be taken on the go. Freshly cut apple pieces are perishable and prone to contamination that can damage its quality. In the current investigation, clove essential oil was used to formulate nanoemulsion (NE) in three distinct concentrations, NE-1 (0.1%), NE-2 (0.5%), and NE-3 (1%), which were then incorporated with Sodium Alginate (SA) based coating solution to impart their qualities to the outer surface of apple pieces. Sodium alginate layer helps to extend the shelf life of fruits by reducing moisture loss, inhibiting microbial growth, and protecting against physical damage. Physico-chemical properties of this solution were investigated using Box–Behnken design after it was employed to coat the freshly cut apple pieces and stored at 4 °C for 14 days. The clove essential oil nanoemulsion's average droplet size and Zeta Potential were in the nanometric range. The addition of increasing concentrations of clove nanoemulsion to the alginate coating solution decreased the zeta potential of the colloidal dispersion from −51.0 to −3.02 mV. The average droplet diameter of the edible coatings containing different clove nanoemulsion concentrations (81.04–3342 nm), whereas sodium alginate edible coating solution (SA) was in micrometric range. The Total phenol content and Antioxidant content of CEO based samples were found to be high as 38.9 mg/ml and 79.1% respectively because of bioactive chemicals and essential oil properties. NE-3 coated apple samples were found to more affective compared with the control and SA coated samples in terms of sensory and shelf-life metrics among all the treatments.

Dual-silane coupling agents co-grafted on black particles for fast response and high contrast ratio electrophoretic display

As the Internet of Things (IoT) significantly improves people's lifestyles, flexible and low-power displays will become essential to real-time human-computer interaction. Electrophoretic display (EPD) has many advantages, such as low power consumption, flexibility, and high sunlight visibility, which is necessary for IoT terminal display. However, preparing electrophoretic particles with high charge and spatial resistance is difficult. Herein, dual-silane coupling agents, (3-Aminopropyl) trimethoxysilane (APS) and vinyltrimethoxysilane (VTS), were co-grafted on the black particles to optimize the electrophoretic particles' dispersity and charge. To evaluate the charge content and dispersity of particles, we measured the particles with zeta potential, dynamic light scattering, and long-term standing treatment. Fe3O4-APS0.5VTS0.5-PLMA, selected as the black particles for EPD, had an average particle size and zeta potential of 300 nm and 25 mV, respectively. Besides, the concentration of the black and white particle would significantly affect the EPD's performance. When optimizing the white and black particle concentration to 4:1, the contrast ratio of EPD was up to 13.2. In addition, the EPD has a response time of 200 ms and exhibits an invertible black and white state under ±15 V. Finally, to improve the stability of electrophoretic particles, the particles were encapsulated in microcups and driven to the black and white state under 30 V. This work demonstrated that dual-silane coupling agent particles have the potential to achieve the fast response and high contrast ratio EPD.

ε-Polylysine and soybean protein isolate form nanoscale to microscale electrostatic complexes in solution: properties, interactions and as antimicrobial edible coatings on citrus

A feasible approach to enhance the antimicrobial efficacy of ε-polylysine (PL) in applications is to form delivery complexes with delicate structures and good dispersion properties. This work aims to study the multiscale structures, properties and interactions, and edible coating applications of the electrostatic complex formed by PL and soy protein isolate (SPI). When the mass ratio of SPI to PL (SE) was between 5 and 15, especially 11, microscale solid-liquid phase separation occurred in the system due to the small absolute zeta potential. When the SE was in the range of 15–20, the system formed a stable nanoscale suspension, the average particle size and zeta potential were 191 nm and −20 mV, respectively. The physicochemical properties of the complexes were investigated including the colloidal properties, spectroscopy and interactions analysis, viscosity, contact angle, and antimicrobial activities against Escherichia coli, Staphylococcus aureus, and Penicillium expansum. Finally, the in vivo application on citrus demonstrated that the nanoscale PL/SPI electrostatic complex (SE = 20) as functional coatings has both barrier and antimicrobial activities. The study provides a novel application strategy for PL and nanoscale electrostatic complexes as postharvest coatings.

Polysaccharides-based adsorbents for removal of Congo red dyes from wastewater

Gums Arabic (GA) and Xanthan (GX) are natural polysaccharides that can be extracted by natural means for applications, such adsorption of pollutants. Structural modifications such as carboxymethylation on bio-adsorbent materials, can be performed to improve its adsorptive properties. Development of polymeric nanoparticles is an economical and favorable option for the adsorption of Congo red dye, which has high toxicity and is resistant to traditional removal processes. In this work, it was developed nanoparticles (NPs) of natural GA (NPGA) and GX (NPGX) and their carboxymethylated forms (NPCMGA and NPCMGX) aiming to evaluate the adsorption of Congo red (CR) dye. NPs are sized from 133 to 1099 nm at an average zeta potential of -13 mV, suggesting stability to absorb dyes. NPGX and NPCMGX absorbed a substantially higher amount of dye than the other NPs. The kinetic studies showed that adsorption process follow pseudo-second order model, suggesting that chemisorption control the process, and the isotherm test revealed that samples fit Langmuir model, with a homogeneous adsorption profile for the two carboxymethylated samples with a maximum adsorption capacity of 182,82 mg/g for NPCMGX and 757,58 mg/g for NPGX. These findings indicate that NPs from Xanthan Gum can be used for removal CR in contaminated water and wastewater.

Conjunctive and concentration dependent effects of nanoscale zinc and boron on the physiological, biochemical, nutrient uptake, and translocation processes in peanut (Arachis hypogaea L.)

In the present investigation, different concentrations (150, 200, 300, 400, and 500 mg/L) of nanoscale zinc oxide (ZnO) and boron (B) were applied through foliar spray (pot culture) to evaluate their effects on the growth and biochemical parameters of peanut. Nanoscale zinc oxide and nano boron were prepared using modified oxalate decomposition and encapsulation methods, respectively, and were characterized using techniques such as, X-ray diffraction (XRD), dynamic light scattering (DLS), ultraviolet–visible (UV–vis) spectroscopy, and transmission electron microscopy (TEM). The average size (37.2 nm, 53.6 nm) and zeta potential (−37.7 mV, −28.3 mV) of nano ZnO and nano boron were measured, respectively. Fourier transform infrared spectroscopic (FT-IR) studies revealed the functional groups that are present in the hydrosols. Highest germination percentage (98.7% and 96%), seedling vigor index (2359 and 1325) and total chlorophyll content (3.28 and 2.24 mg g−1) were recorded at 500 mg/L of nano zinc oxide and nano boron. Significantly higher peroxidase, catalase, superoxide dismutase (SOD) enzymatic activities, pod yield (39.76 g pot−1 and 38.14 g pot−1) and haulm yield (49.56 g pot−1 and 47.05 g pot−1) were also recorded with the application of nano ZnO and boron @ 500 mg/L. Translocation of nanoscale nutrients through stomata was clearly observed. Significant nutrient uptake with the foliar application of nanoscale nutrients has been recorded. The performance of groundnut (in terms of yield, quality and nutrient uptake) was significantly enhanced with the application of nanoscale nutrients and this point to achieving the nutrient bio-fortification with the desired levels for the betterment of human health.

Nanoliposomal oxaliplatin ameliorates chemotherapy-induced neuropathy

Chemotherapy-induced peripheral neuropathy (CIPN) is an important adverse effect of treatment with oxaliplatin (OXA). We have developed PEGylated nanoliposomal oxaliplatin (OXA-LIP) and tested its activity in an animal model of CIPN. OXA-LIPs were prepared using a combination of egg yolk lecithin, cholesterol, and DSPE-mPEG2000 (at ratios 400, 80, and 27 mg). These liposomes were characterized using several different methods (e.g., polydispersity index (PDI), and zeta potential, FESEM). The in vivo study was performed in 15 male rats comprising three groups: a negative control (normal saline) OXA, and OXA-LIP. These were injected intraperitoneally at a concentration of 4 mg/kg on two consecutive days every week, for 4 weeks. After that, CIPN was assessed using the hotplate and acetonedropmethods. Oxidative stress biomarkers such as SOD, catalase, MDA, and TTG were measured in the serum samples. The functional disturbances of the liver and kidney were assessed by measuring the serum levels of ALT, AST, creatinine, urea, and bilirubin. Furthermore, hematological parameters were determined in the three groups. The OXA-LIP had an average particle size, PDI, and zeta potential of 111.2 ± 1.35 nm, 0.15 ± 0.045, and −52.4 ± 17 mV, respectively. The encapsulation efficiency of OXA-LIP was 52% with low leakage rates at 25 °C.Thermal hyperalgesia changes showed OXA has significant effects in the induction of neuropathy on days 7, 14, and 21 compared to the control group. OXA had a significantly greater sensitivity than the OXA-LIP and control groups in the thermal allodynia test (P < 0.001). OXA-LIP administration did not show significant effects on the changes of oxidative stress, biochemical factors, and cell count. Our findings provide a proof of concept on the potential application of oxaliplatin encapsulated with PEGylated nanoliposome to ameliorate the severity of neuropathy, supporting further studies in clinical phases to explore the value of this agent for Chemotherapy-induced peripheral neuropathy.

The internal aqueous phase gelation improves the viability of probiotic cells in a double water/oil/water emulsion system.

This research studied the viability of probiotic bacterium Lactobacillus plantarum (L. plantarum) encapsulated in the internal aqueous phase (W 1) of a water-in-oil-in-water (W 1/O/W 2) emulsion system, with the help of gelation and different gelling agents. Additionally, the physicochemical, rheological, and microstructural properties of the fabricated emulsion systems were assessed over time under the effect of W 1 gelation. The average droplet size and zeta potential of the control system and the systems fabricated using gelatin, alginate, tragacanth gum, and carrageenan were 14.7, 12.0, 5.1, 6.4, and 7.3 μm and - 21.1, -34.1, -46.2, -38.3, and -34.7 mV, respectively. The results showed a significant increase in the physical stability of the system and encapsulation efficiency of L. plantarum after the W 1 gelation. The internal phase gelation significantly increased the viability of bacteria against heat and acidic pH, with tragacanth gum being the best gelling agent for increasing the viability of L. plantarum (28.05% and 16.74%, respectively). Apparent viscosity and rheological properties of emulsions were significantly increased after the W 1 gelation, particularly in those jellified with alginate. Overall, L. plantarum encapsulation in W 1/O/W 2 emulsion, followed by the W 1 gelation using tragacanth gum as the gelling agent, could increase both stability and viability of this probiotic bacteria.

Effect and Mechanism of CircPTPN4 Expression in Carboxylated Single-Walled Carbon Nanotubes on Renal Fibrosis

This research was aimed to investigate the effect of circPTPN4 overexpression in carboxylated single-walled carbon nanotubes (SWCNT-COOH) on renal fibrosis (RF) in mice. SWCNT was oxidized to SWCNT-COOH by concentrated sulfuric acid and nitric acid (3:1, v/v). After characterization and analysis, the effect of SWCNT-COOH on erythrocyte status was detected. The RF mice (RFM) model was induced by SWCNT-COOH, normal mice as normal control (NC), and 1×108 TU/mL lenti-NC was injected into the tail vein as negative control, and 1×108 TU/mL lenti-circPTPN4 was injected for circPTPN4 overexpression. The expression of circPTPN4 was tested by qRT-PCR. Serum creatinine (Cr), blood urea nitrogen (BUN), interleukin 1β (IL-1β), IL-6, transforming growth factor β1 (TGF-β1), kidney injury molecule 1 (KIM-1), and neutrophil gelatinase-associated lipocalin (NGAL) were tested by ELISA. HE, Masson, and immunohistochemistry were used to detect the pathological changes of kidney tissue. The protein expressions of collagen I (COL1), fibronectin (Fn), α-SMA, TGF-β1, Smad3, p-Smad3, and Smad7 were tested by Western blot (WB). It presented SWCNT-COOH was successfully prepared by chemical oxidation method, with an average particle size of 98.1 nm and an average Zeta potential of −38.91 mV. SWCNT-COOH had good hydrophilicity, and did not cause aggregation and morphology changes of red blood cells. As against NC, the expression of circPTPN4 in RFM and lenti-NC groups was decreased; the kidney coefficient was increased, and the normal structure of the kidney was abnormal, accompanied by inflammatory cell infiltration and collagen microformation; serum Cr, BUN, IL-1β, IL-6, and TGF-β1 were increased, and KIM-1 and NGAL in renal cortex were decreased; the protein expressions of COL1, Fn, α-SMA, TGF-β1, and p-Smad3 in the kidney were increased, and the protein expression of Smad7 was decreased (P &lt;0.05). As against RFM, the expression of circPTPN4 in lenti-circPTPN4 group was increased; the kidney coefficient decreased and the kidney structure returned to normal; COL1, Fn, α-SMA, TGF-β1, and p-Smad3 in the kidney were decreased, and that of Smad7 was increased (P &lt;0.05). SWCNT-COOH has good blood compatibility characteristics and can induce RF in mice. The expression of circPTPN4 was down-regulated in the RFM. Overexpression of circPTPN4 can restore the normal renal function of RFM, inhibiting the inflammatory response and RF process.

Designing Vitamin D3 Formulations: An In Vitro Investigation Using a Novel Micellar Delivery System

Vitamin D is an essential nutrient with important immunomodulatory properties. As a fat-soluble compound, Vitamin D (and its D3 form) is immiscible with water, which presents challenges to absorption. In an in vitro setting, the current study characterizes novel micellar formulations of Vitamin D3 designed to improve absorption. Techniques used to evaluate and compare the micellar formulations against a non-micellar formula include the following: cryo-SEM to determine morphology; laser diffraction to determine particle size and distribution; zeta potential to determine stability of the particles; solubility assays to determine solubility in water and gastrointestinal media; and Caco-2 cell monolayers to determine intestinal permeability. Results show advantageous features (particle size range in the low micrometres with an average zeta potential of −51.56 ± 2.76 mV), as well as significant improvements in intestinal permeability, in one optimized micellar formula (LipoMicel®). When introduced to Caco-2 cells, LipoMicel’s permeability was significantly better than the control (p &lt; 0.01; ANOVA). Findings of this study suggest that the novel micellar form of Vitamin D3 (LipoMicel) has the potential to promote absorption of Vitamin D3. Thus, it can serve as a promising candidate for follow-up in vivo studies in humans.

Hyaluronidase inhibitor surface-modified niosomes: An efficient strategy for antibacterial and anti-biofilm drug delivery

Bacterial infections caused by resistant strains have become one of the most significant life-threatening challenges. Therefore, the exploration of competent approaches is instantly required. We report multifunctional niosomes loaded with vancomycin (VCM) antibiotic and coated with antibacterial polyhexanide layer (PH) followed by a novel antivirulence polymer hyaluronic acid-N,N-Dibutylethylenediamine (HD), which exhibited efficient in vitro and in vivo anti-methicillin-resistant Staphylococcus aureus (MRSA) activity. HD-PH-coated niosomes had an average diameter, polydispersity index, zeta potential, and drug encapsulation efficiency of 153.7 ± 2.1 nm, 0.29 ± 0.04, − 9.46 ± 1.46 mV and 84.15 ± 1.11 %, respectively. The in vitro release showed a diffusion-based controlled VCM release profile. The binding affinity studies using Microscale thermophoresis and in silico techniques showed that HD competitively inhibited hyaluronate lyase enzyme even in the presence of its natural substrate, hyaluronic acid, resulting in better inhibition of colonization and biofilm formation of hyaluronate lyase producing bacteria. HD-PH-coated niosomes exhibited higher antibacterial and antibiofilm activity than the bare VCM and non-coated niosomes. Furthermore, the time-kill assay revealed a fast inhibition of bacterial growth with complete elimination of MRSA bacteria within the first 2 h of the study. In vivo studies against a mouse model of MRSA skin infection indicated that HD-PH-coated niosomes had a 29.6-fold reduction in MRSA load compared to bare VCM (p = 0.0006). Therefore, HD-PH coating was confirmed as a promising strategy for enhancing antibacterial and anti-biofilm drug delivery systems.

Development of positively charged amphiphile containing self-nanoemulsifying drug delivery system for improved therapeutic efficacy of metronidazole against Helicobacterpylori

Metronidazole (MTZ) has been extensively used to treat bacterial infections for over five decades, but MTZ bacterial resistance has been on the rise, making it less effective against Bacteroides species. This study aimed to improve the antibacterial activity of MTZ-loaded self-emulsifying drug delivery systems (SNEDDS) against H. pylori by the incorporation of alkylated morpholine serving as a positively charged amphiphile. The studies of drug solubility and emulsification led to the development of a ternary phase diagram for selecting formulation ingredients. The key ingredients of the formulation were selected as cinnamon oil (lipid), cremophor EL (surfactant), and polyethylene Glycol (PEG) 200 (co-surfactant). The optimized formulations were characterized for various physicochemical properties. The average droplet size and zeta potential of MTZ-SNEDDS were determined to be 146.03 ± 2.1 nm and −15.3 ± 0.50 mV while for alkylated morpholine incorporated MTZ-SNEDDS, these were 183.36 ± 21.42 nm and −16.2 ± 1.30 mV, respectively. The thermodynamically robust alkylated morpholine incorporated SNEDDS was successful in maintaining the chemical integrity of the MTZ. The antibacterial potential of MTZ-SNEDDS was significantly enhanced against H. pylori upon the incorporation of alkylated morpholine. It can be concluded that incorporating amphiphilic positively charged morpholine into SNEDDS can lead to more intake of them by H. pylori, and prolonging the contact duration of the SNEDDS with the outer membrane of H. pylori increased the transmembrane penetration of SNEDDS.

Preparation and in vitro property evaluation of β-cyclodextrin-daidzein/PEG_(20000)/Carbomer_(940) nanocrystals

This study aims to improve the solubility and bioavailability of daidzein by preparing the β-cyclodextrin-daidzein/PEG_(20000)/Carbomer_(940) nanocrystals. Specifically, the nanocrystals were prepared with daidzein as a model drug, PEG_(20000), Carbomer_(940), and NaOH as a plasticizer, a gelling agent, and a crosslinking agent, respectively. A two-step method was employed to prepare the β-cyclodextrin-daidzein/PEG_(20000)/Carbomer_(940) nanocystals. First, the insoluble drug daidzein was embedded in β-cyclodextrin to form inclusion complexes, which were then encapsulated in the PEG_(20000)/Carbomer_(940) nanocrystals. The optimal mass fraction of NaOH was determined as 0.8% by the drug release rate, redispersability, SEM morphology, encapsulation rate, and drug loading. The inclusion status of daidzein nanocrystals was determined by Fourier transform infrared spectroscopy(FTIR), thermogravimetric analysis(TGA), and X-ray diffraction(XRD) analysis to verify the feasibility of the preparation. The prepared nanocrystals showed the average Zeta potential of(-30.77±0.15)mV and(-37.47±0.64)mV and the particle sizes of(333.60±3.81)nm and(544.60±7.66)nm before and after daidzein loading, respectively. The irregular distribution of nanocrystals before and after daidzein loading was observed under SEM. The redispersability experiment showed high dispersion efficiency of the nanocrystals. The in vitro dissolution rate of nanocrystals in intestinal fluid was significantly faster than that of daidzein, and followed the first-order drug release kinetic model. XRD, FTIR, and TGA were employed to determine the polycrystalline properties, drug loading, and thermal stability of the nanocrystals before and after drug loading. The nanocrystals loaded with daidzein demonstrated obvious antibacterial effect. The nanocrystals had more significant inhibitory effects on Staphylococcus aureus, Escherichia coli, and Pseudomonas aeruginosa than daidzein because of the improved solubility of daidzein. The prepared nanocrystals can significantly increase the dissolution rate and oral bioavailability of the insoluble drug daidzein.

In Situ Encapsulation of Camptothecin by Self-Assembly of Poly(acrylic acid)-b-Poly(N-Isopropylacrylamide) and Chitosan for Controlled Drug Delivery.

Camptothecin (CPT) has been shown to exhibit anticancer activity against several cancers. Nevertheless, CPT is very hydrophobic with poor stability, and thus its medical application is limited. Therefore, various drug carriers have been exploited for effectively delivering CPT to the targeted cancer site. In this study, a dual pH/thermo-responsive block copolymer of poly(acrylic acid-b-N-isopropylacrylamide) (PAA-b-PNP) was synthesized and applied to encapsulate CPT. At temperatures above its cloud point, the block copolymer self-assembled to form nanoparticles (NPs) and in situ encapsulate CPT, owing to their hydrophobic interaction as evidenced by fluorescence spectrometry. Chitosan (CS) was further applied on the surface through the formation of a polyelectrolyte complex with PAA for improving biocompatibility. The average particle size and zeta potential of the developed PAA-b-PNP/CPT/CS NPs in a buffer solution were 168 nm and -30.6 mV, respectively. These NPs were still stable at least for 1 month. The PAA-b-PNP/CS NPs exhibited good biocompatibility toward NIH 3T3 cells. Moreover, they could protect the CPT at pH 2.0 with a very slow-release rate. At pH 6.0, these NPs could be internalized by Caco-2 cells, followed by intracellular release of the CPT. They became highly swollen at pH 7.4, and the released CPT was able to diffuse into the cells at higher intensity. Among several cancer cell lines, the highest cytotoxicity was observed for H460 cells. As a result, these environmentally-responsive NPs have the potential to be applied in oral administration.

Lipid-based nanoformulation optimization for achieving cutaneous targeting: Niosomes as the potential candidates to fulfill this aim

The present study screened the utility of topically-applied nanoformulations to target the drugs/actives into the skin reservoir with the reduction of possible systemic absorption. The lipid-based nanoformulations selected in this study included solid lipid nanoparticles (SLNs), nanostructured lipid carriers (NLCs), nanoemulsions (NEs), liposomes, and niosomes. We loaded flavanone and retinoic acid (RA) as the penetrants. The prepared nanoformulations were assessed for their average diameter, polydispersity index (PDI), and zeta potential. An in vitro permeation test (IVPT) was utilized to determine the skin delivery into/across pig skin, atopic dermatitis (AD)-like mouse skin, and photoaged mouse skin. We found an increased skin absorption of lipid nanoparticles following the increase of solid lipid percentage in the formulations (SLNs > NLCs > NEs). The use of liposomes even reduced the dermal/transdermal selectivity (S value) to lessen the cutaneous targeting. The niosomes resulted in significantly greater RA deposition and reduced permeation in the Franz cell receptor compared to the other nanoformulations. The S value of the RA delivery via stripped skin was increased by 26-fold in the niosomes compared to the free RA. The dye-labeled niosomes displayed a strong fluorescence in the epidermis and upper dermis through the visualization of fluorescence and confocal microscopies. The cyanoacrylate skin biopsy manifested greater hair follicle uptake of the niosomes compared to the free penetrants by 1.5 to three-fold. The 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) assay indicated an increase in antioxidant ability from 55% to 75% after flavanone entrapment in the niosomes. In the activated keratinocytes, the niosomal flavanone could suppress the overexpressed CCL5 to the baseline control because of the facile cell internalization. After the formulation optimization, the niosomes with higher phospholipid amount had a superior effect in delivering penetrants into the skin reservoir, with limited permeation to the receptors.

Biosynthesis of Silver Nanoparticles Using the Biofilm Supernatant of Pseudomonas aeruginosa PA75 and Evaluation of Their Antibacterial, Antibiofilm, and Antitumor Activities.

As an under-explored biomaterial, bacterial biofilms have a wide range of applications in the green synthesis of nanomaterials. The biofilm supernatant of Pseudomonas aeruginosa PA75 was used to synthesize novel silver nanoparticles (AgNPs). BF75-AgNPs were found to possess several biological properties. In this study, we biosynthesized BF75-AgNPs using biofilm supernatant as the reducing agent, stabilizer, and dispersant and investigated their biopotential in terms of antibacterial, antibiofilm, and antitumor activities. The synthesized BF75-AgNPs demonstrated a typical face-centered cubic crystal structure; they were well dispersed; and they were spherical with a size of 13.899 ± 4.036 nm. The average zeta potential of the BF75-AgNPs was -31.0 ± 8.1 mV. The BF75-AgNPs exhibited strong antibacterial activities against the methicillin-resistant Staphylococcus aureus (MRSA), extended-spectrum beta-lactamase Escherichia coli (ESBL-EC), extensively drug-resistant Klebsiella pneumoniae (XDR-KP), and carbapenem-resistant Pseudomonas aeruginosa (CR-PA). Moreover, the BF75-AgNPs had a strong bactericidal effect on XDR-KP at 1/2 × MIC, and the expression level of reactive oxygen species (ROS) in bacteria was significantly increased. A synergistic effect was observed when the BF75-AgNPs and colistin were used for the co-treatment of two colistin-resistant XDR-KP strains, with fractional inhibitory concentration index (FICI) values of 0.281 and 0.187, respectively. Furthermore, the BF75-AgNPs demonstrated a strong biofilm inhibition activity and mature biofilm bactericidal activity against XDR-KP. The BF75-AgNPs also exhibited a strong antitumor activity against melanoma cells and low cytotoxicity against normal epidermal cells. In addition, the BF75-AgNPs increased the proportion of apoptotic cells in two melanoma cell lines, and the proportion of late apoptotic cells increased with BF75-AgNP concentration. This study suggests that BF75-AgNPs synthesized from biofilm supernatant have broad prospects for antibacterial, antibiofilm, and antitumor applications.

Formulation and evaluation of a two-stage targeted liposome coated with hyaluronic acid for improving lung cancer chemotherapy and overcoming multidrug resistance

Multidrug resistance (MDR) has emerged as a prominent challenge contributing to the ineffectiveness of chemotherapy in treating non-small cell lung cancer (NSCLC) patients. Currently, mitochondria of cancer cells are identified as a promising target for overcoming MDR due to their crucial role in intrinsic apoptosis pathway and energy supply centers. Here, a two-stage targeted liposome (HA/TT LP/PTX) was successfully developed via a two-step process: PTX-loaded cationic liposome (TT LP/PTX) were formulated by lipid film hydration & ultrasound technique, followed by further coating with natural anionic polysaccharide hyaluronic acid (HA). TT, an amphipathic polymer conjugate of triphenylphosphine (TPP)-tocopheryl polyethylene glycol succinate (TPGS), was used to modify the liposomes for mitochondrial targeting. The average particle size, zeta potential and encapsulation efficiency (EE%) of HA/TT LP/PTX were found to be 153 nm, −30.3 mV and 92.1% based on the optimal prescription of HA/TT LP/PTX. Compared to cationic liposome, HA-coated liposomes showed improved stability and safety, including biological stability in serum, cytocompatibility, and lower hemolysis percentage. In drug-resistant A549/T cells, HA was shown to improve the cellular uptake of PTX through CD44 receptor-mediated endocytosis and subsequent degradation by hyaluronidase (HAase) in endosomes. Following this, the exposure of TT polymer facilitated the accumulation of PTX within the mitochondria. As a result, the function of mitochondria in A549/T cells was disturbed, leading to an increased ROS level, decreased ATP level, dissipated MMP, and increased G2/M phase arrest. This resulted in a higher apoptotic rate and stronger anticancer efficacy.

Abstract 2004: Selective cytotoxicity of multiple myeloma using novel cell membrane lipid-extracted nanoliposomes

Abstract Background: Multiple Myeloma (MM) is a highly malignant hematologic disease that originates in the bone marrow from tumorigenic plasma cells that rapidly proliferate and often metastasize before detection or symptom manifestation. Due to the enhanced anaerobic metabolism of cancer vis-a-vis normal cells, elevated media levels of LDH following cell death can serve as a reliable benchmark for in-vitro targeted MM treatment efficacy. Cell membrane Lipid Extracted Nanoliposomes (CLENs) are novel drug delivery vehicles that demonstrate preferential uptake by neoplastic cells. By varying the composition of CLENs, they can be tailored to optimize liposomal uptake by MM cells. This study evaluates the degree to which optimized in-vitro cellular uptake of CLENs translates to cytotoxicity by comparing the attrition rate of doxorubicin hydrochloride (DOX)-loaded MM CLENs compared to relevant liposomal and cellular controls. Methods: The RPMI 8226 cell line was this study’s cellular model for MM. RPMI-8226 cells were seeded at 10,000 cells/mL in 48 well plates. The Y79 (retinoblastoma) cell line served as the negative control for all in vitro experiments. Both cell lines were exposed to two different liposome preparations loaded with 3 mol% doxorubicin hydrochloride (DOX), a MM targeted CLENs preparation of 95% DOPC and 5 mol% RPMI-8226 cell lipid extract (LE) and a non-specific, 100 % DOPC liposome. Liposome preparations were separated from free drug by centrifugation and overnight dialysis. Following overnight liposome exposure, the CyQUANTTM LDH Cytotoxicity Assay Kit (Invitrogen) was utilized to assess extracellular LDH activity and evaluate cytotoxicity of DOX-loaded RPMI-8226 LE in vitro. Results: The mean diameter for the DOX-loaded DOPC (100%) and DOPC/LE (95/5) nanoliposomes was 203 ± 24.3 nm (n=4) and 235 ± 24.6 nm (n=4), respectively. The average zeta potential was -15.79 ± 0.33 mV (n=2) and 0.26 ± 0.19 mV (n=2) for DOPC (100%) and DOPC/LE (95/5), respectively. The mean percent of doxorubicin incorporated into DOPC (100%) and DOPC/LE (95/5) was 72% (n=4) and 84% (n=4), respectively. Following overnight exposure of RPMI 8226 cells to DOX-loaded DOPC (100%) and DOX-loaded DOPC/LE (95/5) at 50 nM and 150 nM, preliminary results showed that the inclusion of LE significantly improved cytotoxicity when compared to the DOX-loaded DOPC control. Additionally, DOX-loaded DOPC/LE (95/5) displayed enhanced extracellular LDH activity compared to the DOX-loaded DOPC control, indicated by higher red formazan fluorescence intensity. There was no difference in LDH activity or cytotoxicity between the two DOX-loaded preparations against Y79 (negative control) cells. Conclusions: Preliminary results suggest that RPMI 8226-CLENs represent an improvement over the use of more conventional nanoliposome preparations. Future studies will evaluate additional drug-loaded CLENs in vitro. Citation Format: Matthew Amorin, Peter Daley, William Smullen, Robert Campbell. Selective cytotoxicity of multiple myeloma using novel cell membrane lipid-extracted nanoliposomes [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 2004.

Abstract 831: Selective targeting of pediatric rhabdoid tumor using G401-CLENs: Cytotoxicity studies

Abstract Background: Rhabdoid tumor is an aggressive soft tissue sarcoma that arises in the kidney, liver, peripheral nerves, and miscellaneous soft parts throughout the body. Rhabdoid tumors usually occur in infancy or during early childhood and the most common sites for metastasis are lungs, abdominal lymph nodes, liver, and brain. Current treatment options include surgery, radiation therapy, high-dose chemotherapy, and stem cell transplant but the overall survival rate is approximately 23%. Nano-sized liposomes are drug-delivery vehicles used to enhance the delivery of drugs to target cells. The composition, size, and overall charge of nanoliposomes determine targeting and influence the degree of drug uptake. The objective of this study was to develop G401-CLENs prepared from lipid extracts derived from rhabdoid tumor cells and conventional lipid components, and to evaluate the optimized drug loaded preparations of rhabdoid-CLENs in vitro. Methods: Rhabdoid tumor cell line, G401, was the chosen model for this study. G401 cells were cultured and expanded in vitro for cellular extraction purposes. The G401 cell membrane lipid extract (LE) was included in the preparation of G401-CLENs. Additional conventional lipid ingredients include DOPC, DOTAP, and DPPE-PEG5000. DPPE-rhodamine lipids were employed for fluorescence studies only. Chemo agents' doxorubicin and vinblastine were used in cell viability and cytotoxicity assays to evaluate formulation efficacy. Phase I of evaluation included physiochemical characterization and cellular uptake studies of preparations of CLENs, including 0 to 20 mol% of G401 LE content. Phase II of evaluation included the use of Hoechst33342/propidium iodide cell viability and cytotoxicity assays with varied amounts of doxorubicin and vinblastine loaded CLENs added to the G401 cells. The control cell line used in the study represents another solid tumor, SK-N-DZ (neuroblastoma). Results: The optimal composition for G401-CLENs was DOPC/DPPE-PEG5000/LE (93/2/5). The average size and zeta potential was 198 ± 16 nm (n=5) and -12 ± 1.2 mV (n=5), respectively. Cell selectivity studies revealed an increase in the uptake of the optimized G401-CLENs by G401 cells when compared to SK-N-DZ control cells. In general, the inclusion of DOTAP increased overall cellular uptake, but diminished selective uptake of G401-CLENs. Results support a significant cellular uptake of preparations consisting of 5 mol% LE compared to 0, 10, and 20 mol% G401 LE content. Doxorubicin loaded in G401-CLENs decreased rhabdoid tumor cell viability in a dose-dependent manner compared to doxorubicin loaded in conventional liposome preparations. Additional cytotoxicity studies are currently underway. Conclusion: To this date, G401-CLENs represents a promising selective drug-targeting vehicle for the treatment of rhabdoid tumors. Citation Format: Varun Kudchadker, Dongxiao Chen, Yeji Seo, Robert Campbell. Selective targeting of pediatric rhabdoid tumor using G401-CLENs: Cytotoxicity studies [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 831.

Mechanism of Triptolide Liposome Nanoparticles on Apoptosis of Multiple Myeloma Cells Induced by Bortezomib

The aim of this research was to investigated the effects of triptolide (TP) and its liposome nanoparticles (TP-LNP) on the apoptosis of multiple myeloma (MM) cells induced by Bortezomi. The encapsulation rate (ER), drug loading (DL), particle size (PS), and Zeta potential characteristics of the TP-LNP were evaluated. MMT assay was employed to detect the inhibitory effect of TP single agent and TP-LNP on proliferation of MM cells. MM cells were enrolled into a control group (Ctrl group, no intervention), a Bortezomi group (75 nM Bortezomi intervention 12 h), a Bortezomi +TP group (Bortezomi combined with TP monotherapy), and a Bortezomi+TP-LNP group (Bortezomi combined with TP-LNP). CCK-8, flow cytometry, and Western blot were utilized to detect the cell survival rate (SR), apoptosis, and MAPK/ERK pathway related protein, respectively. The results revealed that the shape of TP-LNP was similar to circular and uniformly distributed. Its average PS was (181.2±6.3) nm, the average Zeta potential was −29.15 mV, the average ER was 82.1%, and the average DL was 1.09%. When the drug concentration was 100 nmol/L, the inhibition rates of cell proliferation of TP and TP-LNP were (66.8±0.5)% and (81.4±0.6)%, respectively; and their median inhibitory concentrations (MICs) after 72 h were (89.5±1.8) nmol/L and (55.3±2.2) nmol/L, respectively. Based on the Ctrl group, the SRs in the Bortezomi, Bortezomi+TP, and Bortezomi+TP-LNP groups were decreased obviously, while the apoptosis rates were increased, and the comparisons herein exhibited great differences with P &lt;0.05. The SR in the Bortezomi+TP-LNP group was the lowest and the apoptosis rate was the highest, showing great differences among the three groups (P &lt; 0.05). Based on the Ctrl group, the expressions of P-P38 and P-ErK in the Bortezomi, Bortezomi+TP, and Bortezomi+TP-LNP groups were decreased (P &lt;0.05). The levels of P-P38 and P-Erk in the Bortezomi+TP-LNP group were the lowest, and the differences in those in this group and the Bortezomi and the Bortezomi+TP groups statistically great (P &lt;0.05). TP-LNP could inhibit the activation of MAPK/ERK pathway, enhance the apoptosis of MM cells induced by Bortezomi, and inhibit the cell proliferation.