Liposomal Encapsulation Research Articles

Nano- and Micro-Encapsulation of Long-Chain-Fatty-Acid-Rich Melon Seed Oil and Its Release Attributes under In Vitro Digestion Model.

Melon seed oil (MSO) possesses plenty of long-chain fatty acids (LFCAs, oleic-linoleic acid 90%), remarkable antioxidant activity (DPPH (0.37 ± 0.40 µmol TE/g), ABTS (4.98 ± 0.18 µmol TE/g), FRAP (0.99 ± 0.02 µmol TE/g), and CUPRAC (4.94 ± 0.11 µmol TE/g)), and phenolic content (70.14 ± 0.53 mg GAE/100 g). Encapsulation is a sound technology to provide thermal stability and controlled release attributes to functional compounds such as plant seed oil. Nano-sized and micro-sized capsules harboring MSO were generated by utilizing thin film dispersion, spray drying, and lyophilization strategies. Fourier infrared transform analysis (FTIR), scanning electron microscopy (SEM), and particle size analyses were used for the authentication and morphological characterization of the samples. Spray drying and lyophilization effectuated the formation of microscale capsules (2660 ± 14 nm, 3140 ± 12 nm, respectively), while liposomal encapsulation brought about the development of nano-capsules (282.30 ± 2.35 nm). Nano-liposomal systems displayed significant thermal stability compared to microcapsules. According to in vitro release studies, microcapsules started to release MSO in simulated salivary fluid (SSF) and this continued in gastric (SGF) and intestinal (SIF) environments. There was no oil release for nano-liposomes in SSF, while limited release was observed in SGF and the highest release was observed in SIF. The results showed that nano-liposomal systems featured MSO thermal stability and controlled the release attributes in the gastrointestinal system (GIS) tract.

Antimicrobial Nanoformulations Based on Schinus areira Essential Oil

The goal of this research was to create an antibacterial formulation from Scinus areira essential oil (EO) that could spread in water. To achieve this, we developed liposomal formulations of DMPC (1,2-dimyristoyl-sn-glycero-3-phosphocholine) or DPPC (1,2-dipalmitoyl-sn-glycero-3-phosphocholine) that encapsulated the EO. In addition, we utilized the EO as a reducing and stabilizing agent to synthesize silver nanoparticles (AgNPs). The nanoformulations were characterized by determining their size and zeta potential. In the case of liposomal formulations, chemical composition, and encapsulation efficiency were also determined. Furthermore, antimicrobial activity studies against Gram-positive and Gram-negative model bacteria were carried out for both kinds of formulations. The results obtained showed the successful encapsulation of the S. areira EO in multilamellar liposomes of phosphatidylcholine with high efficiency. DPPC liposomes have proven to be a better encapsulation system, retaining more monoterpenes from the EO and therefore presenting antimicrobial activity against S. aureus with an minimal inhibitory concentration (MIC) value of 3 mg/mL of EO. On the other hand, it was also possible to obtain AgNPs by using S. areira EO, which showed antimicrobial activity against S. aureus and E. coli at low concentrations of EO, with MIC values of 6.68 µg/mL and 3.4 µg/mL of silver, respectively. The data obtained will contribute to enhancing the biotechnological value of natural products derived from native plant species in Argentina. This will be achieved through the generation of novel formulations with antibacterial activity and potential bioavailability.

PH-sensitive liposomes bearing a chemotherapeutic agent and a natural apoptosis modulator for effective intracellular delivery to the solid tumor.

The intracellular delivery of the drug to the solid tumor is a major challenge in the treatment of solid tumors. This project aims to increase cytosolic drug delivery using the endosomal escape of drugs. Topotecan (TPT) and capsaicin were used for the treatment of solid tumors. The pH-dependent conversion of active lactone form to inactive carboxylic form is a major problem of TPT that limits its therapeutic use. Liposomal encapsulation of TPT improved the stability of active lactone form and increased the therapeutic efficacy of TPT. Endosomal degradation of liposomes may reduce the content in the target cells. To solve these problems, pH-sensitive liposomes (pSLPs) were developed which improved the intracellular drug delivery by the endosomal escape of drugs. The liposomes (LPs) bearing the drug(s) were prepared using the cast film method and optimized for various formulation and process variables using the Design-Expert 7 software by employing the Box-Behnken design (BBD). The developed hyaluronic acid (HA)-conjugated pSLPs (HA-pSLPs) displayed a vesicle size of 166.5 ± 2.31nm, zeta potential - 30.53 ± 0.91, and entrapment efficiency of 44.39 ± 1.78%, and 73.48 ± 2.15% for TPT and CAP, respectively. HA-pSLPs displayed better cytotoxicity in comparison to free drugs either single or in combination on the MCF-7 cell line. The apoptosis and cellular uptake of HA-pSLPs were increased ⁓ 4.45-fold and ⁓ 6.95-fold as compared to unconjugated pSLPs, respectively. The pharmacokinetic studies in Balb/c mice demonstrated that HA-pSLPs increased the half-life, MRT, and AUC in comparison to the free drug solution. The HA-pSLPs formulation has shown remarkable tumor regression as compared to PpSLPs, pSLPs, and free drug combinations. These results demonstrated that TPT- and CAP-loaded HA-pSLPs offer a potential platform for targeted drug delivery to solid tumors.

A Local and Abscopal Effect Observed with Liposomal Encapsulation of Intratumorally Injected Oncolytic Adenoviral Therapy.

This study evaluated the in vivo therapeutic efficacy of oncolytic serotype 5 adenovirus TAV255 in CAR-deficient tumors. In vitro experiments were performed with cell lines that expressed different levels of CAR (HEK293, A549, CT26, 4T1, and MCF-7). Low CAR cells, such as CT26, were poorly transduced by Ad in vitro unless the adenovirus was encapsulated in liposomes. However, the CT26 tumor in an immune-competent mouse model responded to the unencapsulated TAV255; 33% of the tumors were induced into complete remission, and mice with complete remission rejected the rechallenge with cancer cell injection. Encapsulation of TAV255 improves its therapeutic efficacy by transducing more CT26 cells, as expected from in vitro results. In a bilateral tumor model, nonencapsulated TAV255 reduced the growth rate of the locally treated tumors but had no effect on the growth rate of the distant tumor site. Conversely, encapsulated TAV255-infected CT26 induced a delayed growth rate of both the primary injected tumor and the distant tumor, consistent with a robust immune response. In vivo, intratumorally injected unencapsulated adenoviruses infect CAR-negative cells with only limited efficiency. However, unencapsulated adenoviruses robustly inhibit the growth of CAR-deficient tumors, an effect that constitutes an 'in situ vaccination' by stimulating cytotoxic T cells.

Radiopharmaceutical Encapsulated Liposomes as a Novel Radiotracer Im - aging and Drug Delivery Protocol.

Data are presented for 5 male New Zealand white rabbits. They were injected intravenously with the 99mTc-liposomes encapsulated MIBI through a marginal ear vein, and whole-body images were acquired using a dual-head gamma camera. Cationic PEGylated liposomes were prepared using the conventional thin-film-hydration method. The liposomes were tested for particle size, zeta potential, high-performance-liquid-chromatography (HPLC), and toxicity. The liver activity was slightly greater than or equivalent to heart uptake, using 99mTcsestamibi, MIBI, without liposome as a reference. The absorbed doses in myocardium cells after injecting rabbits with 99mTc-MIBI labeled with free positive lower pH liposomes was greater than in the liver, whereas 99mTc labeled with encapsulated MIBI within positive liposomes showed a significantly higher heart-to-liver ratio. The heart-to-spleen activity uptake ratio in 99mTc-MIBI was higher than or equal to one but increased in 99mTc labeled with MIBI and free positive liposomes. Injecting rabbits with 99mTc labeled with encapsulated MIBI raised myocardium uptake to 2-4 times more than the spleen. Heart-to-bowel activity began to rise with 99m Tc-labeld-MIBI and liposomes. This study provides findings in radiopharmaceutical biodistribution using liposomal agents. Adding free liposomes using a pH gradient technique enhanced the uptake and localization of the radiotracer. However, tracer encapsulation during the formation of the liposomes showed even better specificity.

Survival outcomes with CPX-351 vs 7+3 by baseline bone marrow blast percentage in older adults with newly diagnosed high-risk or secondary acute myeloid leukemia: A 5-year follow-up study.

7027 Background: CPX-351, a dual-drug liposomal encapsulation of daunorubicin and cytarabine in a synergistic 1:5 molar ratio, is approved for newly diagnosed therapy-related acute myeloid leukemia (AML) or AML with myelodysplasia-related changes in the US and Europe following positive efficacy and safety data from the pivotal phase 3 clinical trial (NCT01696084). A previous exploratory analysis of the trial data (median follow-up 20.7 months) showed improved survival outcomes with CPX-351 vs conventional 7+3 chemotherapy in adults with newly diagnosed high-risk or secondary AML (sAML) irrespective of baseline bone marrow (BM) blast percentage. Here we report 5-year follow-up data (median follow-up 60.9 months) to provide insights into the longer-term efficacy and safety of CPX-351 in these patient subgroups. Methods: Patients (60–75 years) with a pathologic diagnosis of AML (high-risk/sAML) per the WHO 2008 criteria (≥20% blasts in peripheral blood or BM) and an ECOG PS of 0–2 were randomized to receive CPX-351 or 7+3 chemotherapy. In these post hoc subgroup analyses at 5 years follow-up, overall survival (OS), event-free survival (EFS) and complete remission (CR) were assessed by baseline BM blast percentage. The analysis included patients with low ( < 20%) BM blasts to account for changes in blast count thresholds in the updated WHO AML classification. Results: Overall, median OS and EFS were longer with CPX-351 vs 7+3 in each subgroup; median OS was greatest in low blast subgroups for both treatments (Table). Higher CR rates were observed with CPX-351 vs 7+3 irrespective of baseline BM blasts. Median Kaplan-Meier (KM)-estimated survival rates increased with CPX-351 vs 7+3 across BM blast subgroups. The CPX-351 safety profile was comparable across BM blast subgroups and consistent with the known safety profile of 7+3. Conclusions: These 5-year follow-up data suggest that CPX-351 results in improved outcomes irrespective of baseline BM blast percentage vs conventional 7+3 chemotherapy in older adults with AML. Clinical trial information: NCT01696084 . [Table: see text]

Cardiotoxicity of CPX-351 vs 7+3 in patients with untreated high-risk acute myeloid leukemia.

7029 Background: CPX-351, a dual-drug liposomal encapsulation of daunorubicin and cytarabine in a synergistic 1:5 molar ratio, is approved for newly diagnosed, therapy-related acute myeloid leukemia (AML) or AML with myelodysplasia-related changes in the United States and Europe. While CPX-351 demonstrated improved overall survival and a similar safety profile vs 7+3 in the phase 3 clinical trial (NCT01696084), the safety of CPX-351 related to cardiac function has not been fully described, an important consideration, due to the known cardiotoxicity risk from anthracyclines. This post hoc analysis assessed the impact of CPX-351 vs 7+3 chemotherapy on cardiac impairment in a subset of adults with newly diagnosed high-risk or secondary AML (sAML) from the phase 3 trial. Methods: In the phase 3 trial, 309 patients (60–75 years old; AML according to WHO 2008 criteria) were randomized 1:1 to receive CPX-351 (100 units/m2 on days 1, 3 and 5; days 1 and 3 for second induction) or 7+3 (cytarabine 100 mg/m2/day for 7 days [5 days for second induction] and daunorubicin 60 mg/m2 on days 1–3 [days 1–2 for second induction]). Cardiac impairment was determined by cardiac adverse events (AEs) as well as echocardiogram (ECHO) measures including left ventricular ejection fraction (LVEF) and global longitudinal strain (GLS), which were assessed at baseline and two follow-up visits. Results: The study population included 102 patients (CPX-351, n = 57; 7+3, n = 45) with normal baseline LVEF (LVEF > 53%) and at least one post-baseline ECHO measure (LVEF or GLS). Median (min, max) age was 68 (60, 75) years for CPX-351 vs 66 (60, 75) years for 7+3. The mean cumulative dose of daunorubicin while on study was lower for patients receiving CPX-351 vs 7+3 (378 vs 570 mg, respectively). Lower proportions of CPX-351 vs 7+3-treated patients had LVEF < 53% (12% vs 31%) or GLS ≤18% (25% vs 38%) at follow-up 1 or 2. A clinically significant change in LVEF ( > 10% decrease from baseline and LVEF < 53%) or GLS ( > 12% relative decrease from baseline and GLS < 18%) was less common in patients treated with CPX-351 vs 7+3 (9% vs 20% and 21% vs 44%, respectively) at follow-up 1 or 2. The frequency of any cardiac AE by treatment group was similar for both CPX-351 and 7+3 (40% vs 42%). Tachycardia was the most common cardiac AE in CPX-351-treated patients (21% vs 9% in 7+3-treated patients), while atrial fibrillation was the most common cardiac AE in 7+3-treated patients (11% vs 4% in CPX-351-treated patients). Conclusions: The results of this post hoc analysis of data from the phase 3 trial show that CPX-351 is associated with less cardiotoxicity in adults with newly diagnosed high-risk or sAML, when compared with 7+3. The lower mean cumulative doses of daunorubicin while on study with CPX-351 appear to result in better cardiac outcomes with CPX-351 vs 7+3 chemotherapy. Characteristics of CPX-351, such as liposomal delivery, may also contribute to this effect but require further study to confirm this. Clinical trial information: NCT01696084 .

Fortification of chocolates with high-value-added plant-based substances: Recent trends, current challenges, and future prospects.

High consumption of delicious foods, such as chocolates, is considered excellent snacks, capable of converting from health-threatening to great functional foods. The fortification of chocolates with high-value-added plant-based substances might improve their healthful effects, nutritional properties, and shelf life. Chocolate could be an effective carrier for plant-based substances delivery, and it could be an effective vehicle to treat and reduce the indications of disease, such as obesity, overweight, hypertension, stress, cardiovascular failure, congestive heart failure, and diabetes. Referring to the recent studies in chocolate fortification with high-value-added plant-based substances, it seems that the recent trends are toward its therapeutic effects against noncommunicable diseases. Despite the undeniable functional effects of fortified chocolates, there are some challenges in the fortification way of chocolates. In other words, their functional characteristics, such as rheological and sensory attributes, may undesirably change. It seems that encapsulation techniques, such as spray drying, antisolvent precipitation, nanoemulsification, and liposomal encapsulation, could almost overcome these challenges. Thus, several studies focused on designing and fabricating nanoscale delivery systems with the aim of chocolate fortification, which is discussed.

A systemically administered detoxified TLR4 agonist displays potent antitumor activity and an acceptable tolerance profile in preclinical models.

Bacterial lipopolysaccharides (LPS) are potent innate immunostimulants targeting the Toll-like receptor 4 (TLR4), an attractive and validated target for immunostimulation in cancer therapy. Although LPS possess anti-tumor activity, toxicity issues prevent their systemic administration at effective doses in humans. We first demonstrated that LPS formulated in liposomes preserved a potent antitumor activity per se upon systemic administration in syngeneic models, and significantly enhance the antitumor activity of the anti-CD20 antibody rituximab in mice xenografted with the human RL lymphoma model. Liposomal encapsulation also allowed a 2-fold reduction in the induction of pro-inflammatory cytokines by LPS. Mice receiving an intravenous administration demonstrated a significant increase of neutrophils, monocytes and macrophages at the tumor site as well as an increase of macrophages in spleen. Further, we chemically detoxified LPS to obtain MP-LPS that was associated with a 200-fold decrease in the induction of proinflammatory cytokines. When encapsulated in a clinically approved liposomal formulation, toxicity, notably pyrogenicity (10-fold), was limited while the antitumor activity and immunoadjuvant effect were maintained. This improved tolerance profile of liposomal MP-LPS was associated with the preferential activation of the TLR4-TRIF pathway. Finally, in vitro studies demonstrated that stimulation with encapsulated MP-LPS reversed the polarization of M2 macrophages towards an M1 phenotype, and a phase 1 trial in healthy dogs validated its tolerance upon systemic administration up to very high doses (10µg/kg). Altogether, our results demonstrate the strong therapeutic potential of MPLPS formulated in liposomes as a systemically active anticancer agent, supporting its evaluation in patients with cancer.

Unraveling effects of anti-aging drugs on C. elegans using liposomes

Liposome-mediated delivery is a possible means to overcome several shortcomings with C. elegans as a model for identifying and testing drugs that retard aging. These include confounding interactions between drugs and the nematodes’ bacterial food source and failure of drugs to be taken up into nematode tissues. To explore this, we have tested liposome-mediated delivery of a range of fluorescent dyes and drugs in C. elegans. Liposome encapsulation led to enhanced effects on lifespan, requiring smaller quantities of compounds, and enhanced uptake of several dyes into the gut lumen. However, one dye (Texas red) did not cross into nematode tissues, showing that liposomes cannot ensure the uptake of all compounds. Of six compounds previously reported to extend lifespan (vitamin C, N-acetylcysteine, glutathione (GSH), trimethadione, thioflavin T (ThT), and rapamycin), this effect was reproduced for the latter four in a condition-dependent manner. For GSH and ThT, antibiotics abrogated life extension, implying a bacterially mediated effect. With GSH, this was attributable to reduced early death from pharyngeal infection and associated with alterations of mitochondrial morphology in a manner suggesting a possible innate immune training effect. By contrast, ThT itself exhibited antibiotic effects. For rapamycin, significant increases in lifespan were only seen when bacterial proliferation was prevented. These results document the utility and limitations of liposome-mediated drug delivery for C. elegans. They also illustrate how nematode-bacteria interactions can determine the effects of compounds on C. elegans lifespan in a variety of ways.

Enhanced Single-Particle Collision Electrochemistry at Polysulfide-Functionalized Microelectrodes for SARS-CoV-2 Detection.

Single-particle collision electrochemistry (SPCE) has shown great promise in biosensing applications due to its high sensitivity, high flux, and fast response. However, a low effective collision frequency and a large number of interfering substances in complex matrices limit its broad application in clinical samples. Herein, a novel and universal SPCE biosensor was proposed to realize sensitive detection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) based on the collision and oxidation of single silver nanoparticles (Ag NPs) on polysulfide-functionalized gold ultramicroelectrodes (Ps-Au UMEs). Taking advantage of the strong interaction of the Ag-S bond, collision and oxidation of Ag NPs on the Ps-Au UME surface could be greatly promoted to generate enhanced Faraday currents. Compared with bare Au UMEs, the collision frequency of Ps-Au UMEs was increased by 15-fold, which vastly improved the detection sensitivity and practicability of SPCE in biosensing. By combining magnetic separation, liposome encapsulation release, and DNAzyme-assisted signal amplification, the SPCE biosensor provided a dynamic range of 5 orders of magnitude for spike proteins with a detection limit of 6.78 fg/mL and a detection limit of 21 TCID50/mL for SARS-CoV-2. Furthermore, SARS-CoV-2 detection in nasopharyngeal swab samples of infected patients was successfully conducted, indicating the potential of the SPCE biosensor for use in clinically relevant diagnosis.

Liposomal Lactoferrin Exerts Antiviral Activity against HCoV-229E and SARS-CoV-2 Pseudoviruses In Vitro.

A limited number of effective therapies are currently available to treat human coronavirus SARS-CoV-2 and other human coronaviruses, which are responsible for nearly a third of global cases of the common cold. The possibility of new emerging coronaviruses demands powerful new antiviral strategies. Lactoferrin is a well-known protein that possesses anti-inflammatory and immunomodulatory activities, and it has previously shown antiviral activity against several viruses, including SARS-CoV-2. To increase this antiviral activity, here we present bovine liposomal lactoferrin. Liposomal encapsulation of the compound was proven to increase permeability, bioavailability, and time release. In the present work, we compare the antiviral activity of free and liposomal bovine lactoferrin against HCoV229E and SARS-CoV-2 in vitro and in human primary bronchial epithelial cells, and we demonstrated that the liposomal form exerts a more potent antiviral activity than its free form at non-cytotoxic doses.

Transforming a Toxic Non-Ionizable Drug into an Efficacious Liposome via Ionizable Prodrug and Remote Loading Strategies against Malignant Breast Tumors.

Liposomes (lipos), one of the most successful nanotherapeutics in the clinic, have made a rapid advance over the past few years. However, still, several challenges exist for lipos for clinical practice, such as low drug loading and premature drug leakage during in vivo circulation. Paclitaxel (PTX), a commonly used first-line drug for cancer chemotherapy, was chosen as the model drug. Due to its non-ionizable and water-insoluble characteristics, the drug-loading efficiency of the marketable PTX lipos, Lipusu, is only 6.76%. Herein, we designed an ionizable PTX prodrug (PTXP) by modifying phenylboronic acid on the C2' hydroxyl group of PTX for the remote loading of liposomal formulations through the pH gradient method. Compared with Lipusu, PTXP lipos displayed a 34% higher loading efficiency and an encapsulation efficiency of approximately 95%. A series of in vitro/vivo experiments indicated that PTXP lipos possess colloidal stability, prolonged blood circulation, high tumor site accumulation, potent anti-tumor effects, and safety. A combination of ionizable prodrugs and remote loading has proved to be an effective and simple strategy to achieve high liposomal encapsulation efficiency of poorly soluble non-ionizable drugs for clinical application.

Abstract 1999: Repurposing an antibiotic for breast cancer treatment using liposomes as a delivery system

Abstract Purpose: Scientific community is always searching for alternatives for cancer treatment and promoting drug repurposing is one of the ways. Rifamycin is a well-known antibiotic, that inhibits RNA synthesis and probably can be explored as cytotoxic agent. Thus, our goal was to repurpose rifamycin for breast cancer treatment using liposomes as a delivery system. Methods: Liposomes were produced by thin-film evaporation method using egg phosphatidylcholine, cholesterol and DOPE-PEG5k (70:25:5, molar ratio). Non-encapsulated drug was removed using dialysis bags and encapsulated drug was directly determined by UV spectrophotometer. Liposomes were characterized regarding size and surface charge by dynamic light scattering. Liposomes were stored at 4°C and 25°C for stability evaluation, monitoring nanoparticles parameters and drug encapsulation. Formulation biocompatibility was evaluated by hemolysis assay. Finally, cytotoxicity of rifamycin SV and liposomes was evaluated in MCF-7 and MDA-MB-231 cell lines. Results: Blank liposomes (without drug) and rifamycin-loaded liposomes showed size smaller than 100 nm and polydispersity around 0.2, with slightly negative zeta potential. Rifamycin encapsulation in liposomes was 3mg of drug/10mg of lipid films. Both formulations were stable for at least 21 days when stored at 4°C, keeping above 60% of drug encapsulated. In the concentrations tested (0.5mg/mL of drug, 2mg/mL of lipids), liposomes formulation (blank and loaded) and free rifamycin were biocompatible and non-hemolytic, since hemolysis was below 6%. Blank liposomes were not cytotoxic in tested cell lines. Free rifamycin and rifamycin-loaded liposomes demonstrated the IC50 in the same range between 41 and 46 µg/mL in 24h and 48h for both MCF-7 and MDA-MB-231. Conclusions: Rifamycin was successfully encapsulated in liposomes, leading to a stable, biocompatible, and non-hemolytic formulation. The liposomal rifamycin demonstrated strong cytotoxicity towards cancer cells similar to that of the free drug. Citation Format: Janaina Artem Ataide, Nina Filipczak, Satya Siva Kishan Yalamarty, Priscila Gava Mazzola, Vladimir P. Torchilin. Repurposing an antibiotic for breast cancer treatment using liposomes as a delivery system [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 1999.

KARATERISTIK ENKAPSULASI LIPOSOM EKSTRAK SUPEROKSIDA DISMUTASE (SOD) Bacillus altitudinis

Photoaging causes the dermis and collagen fibers to decrease which results in skin aging. Genus Bacillus has SOD activity as an anti-photoaging agent that can neutralize ROS (Reactive Oxygen Species) due to exposure to UV-A rays. This study aimed to examine the activity of crude extract and the encapsulation characteristics of liposomes from B. altitudinis. The study began with the isolation of the SOD enzyme, determination of the total protein content of the crude extract of the SOD enzyme and liposome encapsulation, then characterization liposome tes including particle size analysis, sorption efficiency and zeta potential. The results of the crude extract activity test were 85.09% ± 1.24%. The characteristic test was produced with a size of 112.96 ± 17.3 ; the adsorption efficiency was 81.1% and the zeta potential was 7.0 ±5.1 mV. The results showed that the SOD B. altitudinis liposomes were in the nanoparticle range.

Liposomal encapsulated oregano, cinnamon, and clove oils enhanced the performance, bacterial metabolites antioxidant potential, and intestinal microbiota of broiler chickens

Encapsulated phytochemicals with augmented therapeutic and nutritional characteristics have become promising alternatives to antimicrobials in the poultry industry. Hence, our key target was to explore the efficacy of liposomal encapsulation, as a novel carrier, for essential oils (LEOs) on growth, digestibility, intestinal microbiota, and bacterial metabolites of broiler chickens. Moreover, the impact of encapsulated EOs on transcription mechanisms targeting the genes encoding digestive enzymes, gut barrier functions and antioxidant potential of broiler chickens was evidenced. Four equal broiler groups were fed 4 basal diets fortified with LEOs (oregano, cinnamon, and clove) at the levels of 0, 200, 300, and 400 mg/kg diet. Our findings revealed significant improvement in body weight gain and feed conversion ratio of birds fed higher levels of LEOs. These results came concurrently with increasing the activities of digestive enzymes at both serum and molecular levels and consequently nutrient digestibility (dry matter, ether extract, crude protein, and crude fiber) in these groups. Remarkably, the abundance of beneficial bacteria as well as the bacterial metabolites (valeric acid, butyric acid, propionic acid, acetic acid, and total short-chain fatty acids) was increased, while that of pathogenic ones was reduced following dietary inclusion of LEOs. Of note, the mRNA expression of genes encoding antioxidant stability [catalase (CAT), superoxide dismutase 1 (SOD-1), glutathione peroxidase 1 (GPX-1), nuclear factor erythroid 2-related factor 2 (NRF2), NAD(P)H dehydrogenase quinone 1 (NQO1), and heme oxygenase-1 (HO-1)] as well as barrier functions [mucin-2 (MUC-2)] and tight junction proteins, TJP [junctional adhesion molecule-2 (JAM-2) and occludin] were noticeably upregulated in broilers fortified with 400 mg/kg diet of LEOs. Overall, the present work recommended dietary inclusion of LEOs as beneficial additives for attaining targeted performance, gut health and antioxidant stability in poultry farming.

Liposomes loaded with quercetin for resolution of lung inflammation in a lipopolysaccharide-induced mouse model of sepsis

Quercetin (QU) has been widely used as a dietary supplement and proved useful to treat lung diseases. However, the therapeutic potential of QU may be restricted because of its low bioavailability and poor water solubility. In this study, we investigated the effects of developed QU-loaded liposomes on macrophage-mediated lung inflammation. In vivo, a mouse model of sepsis induced by lipopolysaccharide challenge was used to detect the anti-inflammatory effects of liposomal QU. Hematoxylin/eosin staining and immunostaining were utilized to reveal pathological damage and leukocyte infiltration into the lung tissues. Quantitative reverse transcription-polymerase chain reaction and immunoblotting were used to determine cytokine production in the mouse lungs. In vitro, mouse RAW 264.7 macrophages were treated with free QU and liposomal QU. Cell viability assay and immunostaining were utilized to detect cytotoxicity and distribution of QU in the cells. The in vivo results showed that liposomal encapsulation promoted the inhibitory effects of QU on lung inflammation. Liposomal QU decreased mortality in septic mice with no obvious toxicity on vital organs. Mechanistically, the anti-inflammatory effects of liposomal QU were associated with inhibition of nuclear factor-kappa B-dependent cytokine production and inflammasome activation in macrophages. Collectively, the results showed that QU liposomes mitigated lung inflammation in septic mice through inhibition of macrophage inflammatory signaling.

DOPE/CHEMS-Based EGFR-Targeted Immunoliposomes for Docetaxel Delivery: Formulation Development, Physicochemical Characterization and Biological Evaluation on Prostate Cancer Cells

Docetaxel (DTX) is a non-selective antineoplastic agent with low solubility and a series of side effects. The technology of pH-sensitive and anti-epidermal growth factor receptor (anti-EGFR) immunoliposomes aims to increase the selective delivery of the drug in the acidic tumor environment to cells with EFGR overexpression. Thus, the study aimed to develop pH-sensitive liposomes based on DOPE (dioleoylphosphatidylethanolamine) and CHEMS (cholesteryl hemisuccinate), using a Box-Behnken factorial design. Furthermore, we aimed to conjugate the monoclonal antibody cetuximab onto liposomal surface, as well as to thoroughly characterize the nanosystems and evaluate them on prostate cancer cells. The liposomes prepared by hydration of the lipid film and optimized by the Box-Behnken factorial design showed a particle size of 107.2 ± 2.9 nm, a PDI of 0.213 ± 0.005, zeta potential of -21.9 ± 1.8 mV and an encapsulation efficiency of 88.65 ± 20.3%. Together, FTIR, DSC and DRX characterization demonstrated that the drug was properly encapsulated, with reduced drug crystallinity. Drug release was higher in acidic pH. The liposome conjugation with the anti-EGFR antibody cetuximab preserved the physicochemical characteristics and was successful. The liposome containing DTX reached an IC50 at a concentration of 65.74 nM in the PC3 cell line and 28.28 nM in the DU145 cell line. Immunoliposome, in turn, for PC3 cells reached an IC50 of 152.1 nM, and for the DU145 cell line, 12.60 nM, a considerable enhancement of cytotoxicity for the EGFR-positive cell line. Finally, the immunoliposome internalization was faster and greater than that of liposome in the DU145 cell line, with a higher EGFR overexpression. Thus, based on these results, it was possible to obtain a formulation with adequate characteristics of nanometric size, a high encapsulation of DTX and liposomes and particularly immunoliposomes containing DTX, which caused, as expected, a reduction in the viability of prostate cells, with high cellular internalization in EGFR overexpressing cells.

Preparation of Baicalein Long-Circulating Liposome and Its Effect on Bone Implantation in Diabetic Rats

Gradual increase in blood glucose is the major characteristics of a metabolic disease like diabetes. Inflammatory microenvironment and diabetes induced osteoporosis seriously affect prognosis of patients that undergo bone implantation. Baicalein is a natural flavonoid compound with hypoglycemic, anti-inflammatory and other pharmacological functions. Clinically, the application of baicalein is hampered by inadequate solubility, low bioavailability and other shortcomings. In this report, we designed a long-circulating liposome for potential delivery of baicalein. The best formula for long-circulating liposome preparation was obtained by screening excipients, viz., Phospholipids:Cholesterol= 10:1, Phospholipids:DSPEPEG2000 = 6:1 and Phospholipids:Baicalein= 5:1. Respectively, average size of liposomal particles and encapsulation rate were approximately 190 nm and 91%. The drug release percentage within 24 h was 60%, amid being significantly better than that of the free drug. During in vivo pharmacokinetics experiment, we observed a significantly higher plasma baicalein concentration in mice of liposome group comparable to those in free drug group amid consistency with the drug release observation. It was confirmed through in vivo studies that baicalein long-circulating liposomes could reduce blood glucose and pro-inflammatory factors in rats, thus improving the state of liver, kidney and pancreas in diabetic model rats and significantly reversing diabetes induced osteoporosis.

Inclusion complexation and liposomal encapsulation of an isoniazid hydrazone derivative in cyclodextrin for pH-dependent controlled release

Inclusion complexation and liposomal encapsulation of an isoniazid hydrazone derivative in cyclodextrin for pH-dependent controlled release