Nanoemulsion Formulation and Evaluation of Terminalia chebula Fruit to Improve Bioavailability

Tretinoin has shown its effectiveness in the treatment of several skin diseases, such as acne vulgaris, photoaging, psoriasis, and epithelial skin cancer. Nevertheless, formulation of effective delivery systems of tretinoin is challenging due to its poor solubility as well as high chemical- and photo-instability. Furthermore, skin irritation cause by tretinoin upon topical application is also a concern. In addition, sustained and prolonged drug release is required at the disease site for effective treatment. Keeping all these in mind, we hypothesized that encapsulation of tretinoin in solid lipid nanoparticles (SLNs) will avoid the above mentioned drawbacks associated with tretinoin. Additionally, SLNs are expected to penetrate into skin due to its nano size and provide sustained and prolonged drug release due to solid matrix of SLNs. Thus, SLNs containing tretinoin were developed by varying different formulation parameters, such as types of lipids, types of surfactants, lipid concentration, surfactant concentration, drug concentration, homogenization time, and sonication time. Emulsification-ultrasonification method was adopted to prepare SLNs. Particle size, polydispersity index (PI), and zeta potential (ZP) were measured by photon correlation spectroscopy. Surface morphology and shape of SLNs were checked by cryogenic scanning electron microscopy (cryo-SEM). Encapsulation efficiency (EE) of the SLNs was determined by HPLC. Drug release study was performed in 10 mM phosphate buffer containing 2% Tween® 80 (pH 7.4). Further, short-term stability study was also performed to check the stability of SLNs. Formulation parameters exhibited significant impact on the size, PI, ZP, and EE. The developed SLNs demonstrated 113.50±0.89 nm particle size, 0.182±0.007 PI, -38.70±0.56 mV ZP, and 78.18±4.26% EE. Cryo-SEM image revealed smooth and spherical SLNs. SLNs were stable during storage at 4°C. In vitro drug release study showed sustained and prolonged drug release from SLNs. Hence, the developed tretinoin-loaded SLNs can be incorporated into a gel formulation for topical application.

Zanthoxylum acanthopodium DC., known as andaliman, contains bioactive compounds with potential anticancer properties. This study investigated the cytotoxic activity of the ethanolic extract of Z. acanthopodium and its phytosomal formulation (Phytosome-EEA) against the MCF-7 breast cancer cell line. Phytosomes were prepared using antisolvent precipitation and optimized based on phospholipid ratios, reflux temperature, and n-hexane droplet rate. The phytosomes were characterized by particle size, polydispersity index, zeta potential, entrapment efficiency, stability tests, and morphology study by Transmittion Electron Microscopy (TEM). WST-1 assay at 450 nm was used to assess cytotoxicity against HEK 293A and MCF-7 cells, while apoptosis was evaluated using Annexin V-PI staining. The optimized phytosome formulation yielded nanoparticles with an average size of 192.4 nm, polydispersity index of 0.783, a zeta potential of −36.7 V, and an entrapment efficiency of 90.26%. When stored at 4, 25 and 40 °C for 14 days, there were no significant changes in particle size, zeta potential, or cytotoxicity, indicating high physical stability. Both the ethanolic extract and phytosomes exhibited cytotoxic activity against MCF-7 and HEK 293A cells, with IC₅₀ values of 100.2 and 109.2 µg/mL, respectively. The cell death induced by EEA occurred primarily via apoptosis, while Phytosome-EEA promoted increased late apoptosis. Although the selectivity index remained < 1, the phytosomal formulation maintained consistent cytotoxic activity and improved physicochemical characteristics compared to the crude extract. HIGHLIGHTS The ethanolic extract of acanthopodium fruit contains bioactive compounds such as alkaloids, flavonoids, and tannins that contribute to the anticancer effect. The phytosome formulation produced stable particles with a particle size of 192.4 nm, zeta potential of -33.2 mV, and high entrapment efficiency of 90.26%. The andaliman extract showed cytotoxic activity against breast cancer cells (MCF-7), but less selective (SI < 1). IC50 value against MCF-7: 100.2 µg/mL (moderately active category) Phytosome formulation did not improve selectivity, but provided good physical stability and was stable during storage. Phytosomes have potential as an herbal-based anticancer delivery system, but further optimization is needed. Further considerations such as targeting modifications are needed to be more selective towards cancer cells GRAPHICAL ABSTRACT

Cancer remains a global health challenge, with conventional treatments limited by toxicity and drug resistance. Propolis, a natural resin with promising anticancer properties but restricted in clinical applications due to low bioavailability and poor solubility. Nanotechnology, offers a potential approach to enhance propolis' therapeutic efficacy through more efficient delivery and improved pharmacokinetics. Propolis-loaded niosomes (PLNs) were prepared using the ethanol injection method, optimized using response surface methodology (RSM) for surfactant type (Tween 80), cholesterol-to-surfactant ratio, and propolis content. Physicochemical properties, including particle size, polydispersity index (PDI), and zeta potential were characterized. Stability was assessed under various storage conditions, and total polyphenol content (TPC) and entrapment efficiency (EE%) were determined. Anticancer activity wasin vitroassessed against MCF7 breast cancer and L929 fibroblast cell lines. The optimized PLN formulation (at a mass ratio 4:1:8 of propolis: cholesterol: Tween 80, respectively) achieved a particle size of 193.5 nm, PDI of 0.123, and zeta potential of -19.6 mV, with a TPC of 21.83 mg GAE g-1and EE% of 57.82%. Stability studies confirmed optimized formulation's robustness at 4 °C, with minimal changes over 42 d, though higher temperatures induced aggregation. PLNs exhibited superior cytotoxicity against MCF7 cells inhibitory concentration (IC50equivalent to 106.85 µg ml-1) compared to L929 cells (IC50equivalent to 127.14 µg ml-1). The formulation's uniformity and moderate stability support its potential for targeted drug delivery. PLNs effectively enhance propolis' anticancer efficacy and bioavailability, offering a promising delivery system for cancer therapy. Future studies should focus on improving zeta potential,in vivovalidation, and encapsulation efficiency to advance clinical translation.

Fisetin (3,3,4,7-tetrahydroxyflavone) is a natural antioxidant that has shown to posses anticancer, antioxidant and anti-inflammatory properties. However, the poor solubility leads to poor bioavailability and limits its development.The aim of the research is to investigate the effect of fisetin nanosuspension using a nanoprecipitation technique and additional stabilizers polysorbat 80, SLS, PVA and Eudragit on particle size average, polydispersity index and zeta potential.The suspensions of microcrystalline FIS were prepared by a nanoprecipitation technique with different proportion of stabilizers fixed. The nanosuspension produced was then characterized using Photon Correlation Spectroscopy (PCS) in term of particle size distribution, polydispersity index, zeta potential and morphology nanosuspensiom (TEM). Result showed fisetin nanosuspension were successfully prepared by anti-solvent precipitation with additional stabilizer SLS and polysorbat 80. The nanosuspension containing polysorbat 80 showed smaller average particle size of 225.7 nm ± 1.31, a polydispersity index of 0.272 ±0.02 and zeta potential -39.3 ± 0.26 was obtained. Conclusion, FIS nanosuspension successfully prepared by nanoprecipitation tecnique with the polysorbate 80 as stabilizer and ethanol as solvent were spherical in shape..

The main objective of this study was to optimize squalene-loaded nanostructured lipid carriers using a lipid matrix of palm kernel stearin, rice bran oil, and Tween 80 as surfactants. Nanostructured lipid carrier (NLC) was prepared using the ultrasonication method. Optimization process was performed using Box-Behnken Design (BBD) response surface methodology (RSM). Three ultrasonication parameters were employed including temperature, time, and amplitude. The optimization was evaluated for its response on particle size (X1), polydispersity index (X2), zeta potential (X3), and encapsulation efficiency (X4). Furthermore, transmission electron microscopy (TEM) photograph, peroxide value (PV), free fatty acid value, anisidine value, stability, particle size (PS), polydispersity index (PDI), zeta potential (ZP), and encapsulation efficiency (EE) were tested on nanostructured lipid carrier squalene-loaded (NLC-s) and control samples within a storage time of 28 days. These NLC-s were prepared by ultrasonication time of 24.8 minutes, amplitude of 80, and temperature of 43.7°C. The characteristics of the NLC-s were evaluated before and after storage for 28 days at room temperature resulted in particle size (nm) of 38.6 and 40.3, PDI of 0.282 and 0.213, zeta potential (mV) of -34.1 and -34.8, and EE of (%) 85.69 and 84.04, respectively. The empty NLC had particle size (nm) of 37.4 and 38.5, PDI of 0.240 and 0.276, and zeta potential (mV) of -41.5 and -32.9.

Abstract: Background: Azilsartan medoxomil (AZL) is an orally active nonpeptide angiotensin II receptor antagonist with less water solubility and oral bioavailability. Objectives: Increase the solubility and dissolving rate of AZL. Materials and Methods: For formulation we used a probe sonication approach to create nanocrystals. The impacts of independent factors such as % polymer concentration (X1) and sonication duration (X2 min) on dependent variables such as particle size (Y1 nm) and % drug release (DR) were optimised using a 32 response surface methodology (Y2). Results: The prepared batches were examined for size, polydispersity index (PDI), zeta potential, solubility study and dissolution study. AZL nanocrystal (PS2 batch) particle size and zeta potential was found to be 168 ± 10 nm, 0.314 ± 0.02 and - 22.72 ± 2.6 mV respectively. The batch (PS2) with the best results was chosen and subjected to additional testing. In vitro dissolution of all 13 batches and pure drug was in ranges of 51.98-81.99% and 11.23 %, respectively. Conclusion: The FTIR analysis indicated that AZL and soluplus have no physical interaction. DSC, XRD, and SEM investigations revealed that the crystalline form of the medication was converted to an amorphous form, resulting in an improve water solubility and dissolution rate. Thus the studies exhibited nanocrystals prepared by probe sonication method showed significant enhancement in solubility and dissolution rate. Key words: Azilsartan medoxomil (AZL), Nanocrystal, Sonication, Solubility, Drug release.

This work aims to develop nanostructured lipid carriers (NLCs) to improve the oral bioavailability of febuxostat (FEB). High shear homogenization, a well-known technique, followed by bath sonication with slight modifications was used to prepare FEB-loaded NLCs using oleic acid as liquid lipid and stearic acid as solid lipid. A total of 3² full factorial design was utilized to examine the effect of 2 independent variables, namely, X1 (liquid to solid lipid ratio) and X2 (surfactant concentration) on the Y1 (particle size) and Y2 (% entrapment efficiency) of the drug. The prepared NLCs were evaluated for particle size, polydispersity index, zeta potential, and (%) entrapment efficiency. The drug's highest solubility was found in the stearic (solid lipid) and oleic acid (liquid lipid), which were further chosen for NLC preparation. Result of the present study showed an increase in entrapment efficiency and a decrease in particle size with the increase in liquid lipid to solid lipid ratio. With increased surfactant concentration, a small particle size is observed. The optimized formulation's particle size and (%) entrapment efficiency was found to be 99 nm and 80%, respectively. The formulations' zeta potential and polydispersity index were found within the range. Compared to plain drug suspension, the optimized formulation showed higher drug release, which may be due to the presence of the higher amount of liquid lipid. The particles shown in the transmission electron microscopy were round in shape and have smooth surface. Stability studies showed that the NLC formulation can be stored for a longer time period under room condition. FEB-loaded NLC were successfully prepared using full 3² factorial design, and can be further used for oral delivery of FEB for gout treatment.

IntroductionThe application of biomimetic and stimuli-responsive nanocarriers displays considerable promise in improving the management of bacterial sepsis and overcoming antimicrobial resistance. Therefore, the study aimed to synthesize a novel hyaluronic acid-lysine conjugate (HA-Lys) and to utilize the attributes of the synthesized HA-Lys with Tocopherol succinate (TS) and Oleylamine (OLA) in the formulation of multifunctional biomimetic pH-responsive HNLCs loaded with vancomycin (VCM-HNLCs), to combat bacterial sepsis.MethodsA novel hyaluronic acid-lysine conjugate (HA-Lys) was synthesized and characterized using FTIR and 1H NMR spectroscopy. Vancomycin-loaded hybrid nanosystems (VCM-HNLCs) were prepared through hot homogenization ultrasonication and evaluated for particle size, polydispersity index (PDI), zeta potential (ZP), and encapsulation efficiency (EE%). In vitro biocompatibility was assessed via MTT assay and red blood cell hemolysis test. The binding affinity to TLR2 and TLR4 was measured using microscale thermophoresis (MST). Drug release was evaluated using the dialysis bag method. Antimicrobial activity against MRSA and efflux pump inhibition were also determined. Efficacy was demonstrated in an MRSA-induced sepsis mice model.ResultsThe VCM-HNLCs, produced via hot homogenization ultrasonication, exhibited particle size (PS), polydispersity index (PDI), zeta potential (ZP), and encapsulation efficiency (EE%) of 110.77 ± 1.692 nm, 0.113 ± 0.022, − 2.92 ± 0.210 mV, and 76.27 ± 1.200%, respectively. In vitro, biocompatibility was proven by hemolysis and cytotoxicity studies. The VCM-HNLCs demonstrated targetability to human Toll-like receptors (TLR 2 and 4) as validated by microscale thermophoresis (MST). VCM-HNLCs showed a twofold reduction in MIC values at physiological pH compared to the bare VCM against S. aureus and MRSA for 48 h. While at pH 6.0, MIC values were reduced by fourfold in the first 24 h and by eightfold in the subsequent 48 and 72 h against tested strains. Furthermore, VCM-HNLCs showed inhibitory effects against MRSA efflux pumps, reactive oxygen species (ROS), and lipopolysaccharide (LPS)-induced hyperinflammation. In an MRSA-induced sepsis mice model, VCM-HNLCs demonstrated superior efficacy compared to free VCM, significantly eliminated bacteria and improved survival rates.ConclusionsOverall, these results highlight the potential of VCM-HNLCs as novel multifunctional nanocarriers to combat antimicrobial resistance (AMR) and enhance sepsis outcomes.Graphical

Context: Arthritis is inflammation of one or more joints. Terminalia chebula Retz. (Combretaceae) fruit is mentioned in Ayurveda as useful in treating arthritic disorders.Objective: This work was undertaken to evaluate the anti-inflammatory, antioxidant, anti-lipid peroxidative and membrane-stabilizing effects of hydroalcoholic extract of Terminalia chebula fruits and also to establish a possible association between them.Materials and methods: In vivo anti-inflammatory activity of T. chebula fruit extract at different doses ranged from 50 to 500 mg/kg, p.o. was evaluated against carrageenin-induced inflammation in rats. Human erythrocyte hemolytic assay was used for in vitro anti-inflammatory activity testing with 50 to 500 µg/ml fruit extract. Antioxidant potential of test fruit extract (10 to 100 µg/ml) was evaluated using TBARS and DPPH methods. The fruit extract was standardized for total phenolic content using Folin-Ciocalteu method.Results: The standardized extract at 250 mg/kg, p.o. dose caused 69.96% reduction in carrageenin-induced rat paw edema and demonstrated 96.72% protective effect on human RBC membrane stability. Besides, T. chebula fruit extract significantly reduced the in vivo formation of TBARS in carrageenin-induced rat liver with IC50 94.96 mg/kg, p.o. and also in vitro radical scavenging activities in DPPH assay method with IC50 42.14 µg/ml. The standardized extract contains phenolics 118.5 mg gallic acid equivalent/g of extract.Discussion and conclusion: These promising findings support the traditional use of T. chebula fruits in the treatment of arthritic disorders and suggest that radical quenching may be one of the mechanisms for its anti-inflammatory activity.

Purpose: To prepare, characterize, and determine the purity of 2,4,6-trihydroxygeranylacetophenone (tHGA), and also to develop and characterize a liposomal formulation of tHGA to overcome its poor water solubility. Methods: The tHGA was synthesized and then purified in two steps using two types of column chromatography separation techniques. The compound was characterized using different analytical techniques, while the purity of tHGA was determined by quantitative-nuclear magnetic resonance (qNMR). Proliposomes method was developed to produce liposome-encapsulated tHGA which was characterized based on particle size, polydispersity, stability, and encapsulation efficiency. A selective and rapid high-performance liquid chromatography (HPLC) method was developed and validated to quantify tHGA in a liposomal formulation in order to evaluate the encapsulation efficiency. Results: The tHGA was successfully prepared and characterized with 98.4 % purity. A simple and reproducible proliposomes method was successfully developed to produce liposome-encapsulated tHGA. The liposomal formulation exhibited excellent encapsulation efficiency (90.4 %). This formulation also yielded a homogenous liposome population (polydispersity index = 0.39) with a small particle size (250.8 nm). The prepared liposome-encapsulated tHGA was stable at refrigerated temperature (4 ºC) for at least four weeks. The developed HPLC method showed good linearity over the range of 10 to 500 μg/mL with high precision and accuracy. Conclusion: The compound produced has a high purity which can be used as an analytical reference standard. The developed formulation is effective for dissolving and entrapping a high amount of tHGA which helps to overcome its poor solubility. Keywords: Liposomes, Proliposomes, Trihydroxygeranylacetophenone, tHGA, Spectroscopic characterization, Poor solubility

Optimisation and biological evaluation of palm glyceryl monocaprylate antimicrobial nanoemulsion for combating S. aureus wound infection

Red palm oil (RPO) is a potential carotenoid source mostly containing β-carotene. RPO instability requires a delivery system such as nanostructured lipid carriers (NLC). This research aimed to develop an NLC delivery system and study the characteristics of NLCRPO. The melting-emulsification-ultrasonication method was employed to produce the NLC-RPO based on 6% (w/w) lipids, with solid lipid to RPO ratio (SRR) varied 6:4, 7:3, and 8:2. Tween 80 (24% w/w) to lipids ratio of 4:1, and distilled water 70% (w/w). The solid lipids employed in this investigation were palm stearin (PS), and palm kernel stearin (PKS), while the liquid lipid was RPO. The stability of NLC-RPO was evaluated using βcarotene entrapment efficiency (EE), centrifugation, cooling, heating test, color, and pH for 90 days of storage. The NLC-RPO was characterized by particle size, polydispersity index (PDI), zeta potential, and viscosity before and after 90 days. Regression analysis was performed to evaluate the relationships between the storage and stability parameters. The highest encapsulation efficiency of β-carotene in NLC-RPO as a carrier β-carotene from the RPO was achieved when solid lipid to RPO ratio (SRR) of 6:4 and there was no significant difference in the type of solid lipid (PS and PKS) used. Generally, NLC-RPO stored for 90 days at room temperature showed good stability after centrifugation, cooling and heating tests with greenish-yellow color (-a*;+b*), and pH of 6.38-6.54. The particle size (38-87 nm), PDI (0.01-0.54), and zeta potential (-10.17 to -22.67 mV) did not significantly change over 90 days of storage, while the viscosity (8.36-9.11cP) was significantly different. The NLC-RPO with SRR of 6:4 had the highest β-carotene entrapment efficiency and remained stable after 90 days of storage at room temperature.

Optimization of linalool-loaded solid lipid nanoparticles using experimental factorial design and long-term stability studies with a new centrifugal sedimentation method

This study aimed to investigate the use of Sucrose acetate isobutyrate (SAIB) and Glyceryl monooleate (GMO) as co-formers for creating Cubosomes and SAIB-based nanodispersions of Rivaroxaban (RXB). The process utilized a modified melt dispersion technique with varying polymer: drug ratios (0.5:1, 0.75:1, and 1:1) and a fixed polymer: poloxamer 407 ratio (0.1:1). Particle size (PS), polydispersity index (PDI), zeta potential (ZP), and entrapment efficiency (EE) were measured to determine the optimal formulas. The best-lyophilized formulas were then analyzed using Fourier transform infrared spectroscopy (FT-IR), powder X-ray diffraction (PXRD), differential scanning calorimetry (DSC), dissolution testing, and Pharmacokinetic (PK) studies. The results revealed significant correlations between polymer concentrations and various variables in cubosomal and SAIB-based nanodispersions. An increase in GMO concentration led to a decrease in PS, PDI, and ZP but an increase in EE and yield. Maintaining optimal GMO concentration is crucial for consistent nanoparticle formulations. In contrast, increasing SAIB concentration led to a decrease in PS and PDI but an increase in EE and yield. The drug release rates of different preparations were measured during the dissolution test. The best-lyophilized cubosome (L4) and the best-lyophilized SAIB-based nanodispersions (L8) showed significantly improved drug release compared to XARELTO®. L4 displayed the best dissolution rate, and L8 also had a reasonable rate. A PK study demonstrated that L4 and L8 had significantly better bioavailability than XARELTO®, possibly due to their improved solubility. This study suggests that SAIB and GMO can significantly enhance the solubility and bioavailability of RXB in nano preparations, leading to more efficient drug delivery. This new approach can also reduce the required dosage for the desired therapeutic effect. However, further research is needed to fully understand these polymers' potential benefits and limitations.

Objectives Breast cancer remains the most frequently diagnosed cancer worldwide and the second leading cause of cancer-related mortality among women. Drug resistance and significant adverse effects limit anticancer drugs efficacy despite their availability. Oxaliplatin (OXA), a platinum-based chemotherapeutic agent, has shown potential in metastatic breast cancer, however, its toxicity limit its use. Melatonin (MEL) demonstrates anticancer activity and improves chemotherapy efficacy, but its therapeutic use is limited by poor stability and short half-life. Nanocarriers, particularly lipid-polymer-hybrid nanoparticles (LPNs), offer an innovative approach to improve drug delivery, enhance bioavailability, and limited systemic toxicity. Methods OXA and MEL co-encapsulated LPNs were prepared. Design of Experiments (DoE)-based optimization was employed using the Box-Behnken design (BBD) to systematically evaluate critical formulation parameters. Particle size, polydispersity index, zeta potential, encapsulation efficiency, release, FTIR, DSC, and stability studies were carried out. The effects of OXA-MEL-loaded-LPNs were assessed on human breast cancer cell line (MCF-7). Results Optimized LPNs exhibited a particle size of ∼240 nm, PDI of 0.17, zeta potential of −30 mV, and encapsulation efficiencies of 99.1% for MEL and 96.1% for OXA. In vitro release studies showed sustained release, with ∼26% OXA and ∼18% MEL released over 8 h. FTIR and DSC analysis showed all substances were loaded into particles. Stability data indicate that particle size remains within the acceptable range for up to one week. Lyophilization resulted in a slight increase in particle size and PDI. Co-loaded LPNs exhibited significant cellular cytotoxicity. Conclusion The findings suggest that LPNs offer a promising platform for combination therapy, potentially improving treatment outcomes in breast cancer.