Nanoscale Liposomes Research Articles

A Novel Acoustic Modulation of Oscillating Thin Elastic Membrane for Enhanced Streaming in Microfluidics and Nanoscale Liposome Production.

Liposomes are widely utilized in therapeutic nanosystems as promising drug carriers for cancer treatment, which requires a meticulous synthesis approach to control the nanoprecipitation process. Acoustofluidic platforms offer a favorable synthesis environment by providing robust agitation and rapid mixing. Here, a novel high-throughput acoustofluidic micromixer is presented for a solvent and solvent-free synthesis of ultra-small and size-tunable liposomes. The size-tunability is achieved by incorporating glycerol as a new technique into the synthesis reagents, serving as a size regulator. The proposed device utilizes the synergistic effects of vibrating trapped microbubbles and an oscillating thin elastic membrane to generate vigorous acoustic microstreaming. The working principle and mixing mechanism of the device are explored numerically and experimentally. The platform exhibits remarkable mixing efficacy for aqueous and viscous solutions at flow rates up to 8000 µL/h, which makes it unique for high-throughput liposome formation and preventing aggregation. As a proof of concept, this study investigates the impact of phospholipid type and concentration, flow rate, and glycerol on the size and size distribution of liposomes. The results reveal a significant size reduction, from ≈900 nm to 40 nm, achieved by merely introducing 75% glycerol into the synthesis reagents, highlighting an innovative approach toward size-tunable liposomes.

Three Diterpene Lactones from Andrographis paniculata (Burm. f) Nees In vitro, In silico Assessment of the Anticancer and Novel Liposomal Encapsulation Efficiency

: Three compounds from Andrographis paniculata (Burm. f) Nees leaf were isolated and identified using 1H, 13C, 2D-NMR, and HR-MS techniques for the first time. Compound 3,19-Di-O-acetylandrographolide (3,19-DAA) or (4) is produced by acetylating compound (2). Compounds (2) and (4) have been investigated for their cytotoxic effects on three human cancer cell lines (SK-LU-1, Hela, and HepG2) using the MTT method. Compound (4) demonstrated significant cytotoxicity against all three cancer cell lines, with IC50 values ranging from 8.38 to 10.15 μM. This represents an increase in cytotoxicity of 2.67 to 3.12-fold compared to compound (2). One way to deal with the problem of low water solubility is by encapsulating (4) into liposomes using a thin-film hydration technique. The optimal conditions for maximizing encapsulation efficiency involve molar ratios of phosphatidylcholine, 3,19-DAA, and cholesterol at 4:1:1. Encapsulating compound (4) within nanoscale liposomes increases its water solubility compared to the free form of compound (4). Pose 324 of compound (4) demonstrated the best conformation among 500 docking conformations when docked to enzyme 1T8I in a in silico docking study. The free Gibbs energy and inhibition constant were determined to be -7.09 Kcal/mol and 6.32 μM, respectively. These values help elucidate the strong interaction between compound (4) and the enzyme in the ligand interaction model. The molecular dynamics simulation using Desmond software in the Linux environment was conducted for a duration of 0 to 100 nanoseconds on the complex formed by pose 324 and 1T8I. The results showed effective interactions within the complex, with stability observed from 0 to 60 nanoseconds. Throughout the simulation, specific amino acids such as Ala 499 (involved in 90% of the simulation time with hydrogen bonding via a water bridge) and Thr 501 (involved in 50% of the simulation time with one hydrogen bond via a water bridge) were found to play significant roles. The majority of torsion bondings are C-O bondings in the acetyl group of compound (4), with torsion energy values of 13.47 Kcal/mol. Carbon atom C-29 at position 324 exhibits the highest fluctuation.

Integration of acoustic micromixing with cyclic olefin copolymer microfluidics for enhanced lab-on-a-chip applications in nanoscale liposome synthesis

The integration of acoustic wave micromixing with microfluidic systems holds great potential for applications in biomedicine and lab-on-a-chip technologies. Polymers such as cyclic olefin copolymer (COC) are increasingly utilized in microfluidic applications due to its unique properties, low cost, and versatile fabrication methods, and incorporating them into acoustofluidics significantly expands their potential applications. In this work, for the first time, we demonstrated the integration of polymer microfluidics with acoustic micromixing utilizing oscillating sharp edge structures to homogenize flowing fluids. The sharp edge mixing platform was entirely composed of COC fabricated in a COC-hydrocarbon solvent swelling based microfabrication process. As an electrical signal is applied to a piezoelectric transducer bonded to the micromixer, the sharp edges start to oscillate generating vortices at its tip, mixing the fluids. A 2D numerical model was implemented to determine the optimum microchannel dimensions for experimental mixing assessment. The system was shown to successfully mix fluids at flow rates up to 150 µl h−1 and has a modest effect even at the highest tested flow rate of 600 µl h−1. The utility of the fabricated sharp edge micromixer was demonstrated by the synthesis of nanoscale liposomes.

Stabilization of cationic liposomes using hyaluronic acid-modified gold nanoparticles by post-microfluidics conjugation approach for drug delivery applications

lipid-based nanoparticles (LNPs), particularly liposomes, have gained prominence throughout the pharmaceutical sector as a hopeful means for transporting a range of therapeutic agents. Improving LNPs stability is crucial in pharmaceuticals, and drug delivery systems to ensure their effectiveness and reliability. The current research was undertaken to design a pH-responsive hyaluronic acid-modified gold nanoparticle-stabilized cationic liposome (SPC/DOPE/DOTAP) system (HA-SH/AuNPs@liposomes) to improve the stability of liposomes. This was achieved through a post-microfluidics conjugation technique to encapsulate the 5-fluorouracil (5-FU) anticancer drug, usually employed in nonmelanoma skin cancer (NMSC) treatment. Microfluidics is an emerging technology that can be employed as a powerful tool for designing nanoscale liposomes with highly uniform size distributions and acceptable colloidal stability. Utilizing a microfluidics toroidal mixer, affixed HA-SH/AuNPs (size: 12.56 ± 1.65 nm) to positively charged liposomes (size: 75 ± 0.68 nm and PDI: 0.032 ± 0.0021). These liposomes exhibited strong stability, limited fusion propensity, and minimal cargo release at neutral pH. The gold stabilizers detached in an acidic environment, releasing encapsulated therapeutic agents. The dynamic light scattering (DLS) analysis results indicate that HA-SH/AuNPs@Liposomes (size: 102.2 ± 1.3 nm and PDI: 0.11 ± 0.01) exhibit greater stability than bare liposomes over 4 weeks at 4 °C. This enhanced stability can be attributed to the presence of HA-SH/AuNPs in the liposomal structure. This research outlines a systematic method to optimize the size rapidly and PDI of liposomes by employing the Design of Experiments (DoE). The results highlighted that utilizing both the microfluidic technique and gold nanoparticle-stabilized liposomes offers benefits for creating controlled drug release formulations. This approach leads to enhanced biopharmaceutical characteristics and improved scalability of liposomal formulations.

Retention of E-selectin functionalized liposome fanny packs on Jurkat cells following invasion through collagen

Circulating immune cells are an appealing candidate to serve as carriers of therapeutic cargo via nanoparticles conjugated to their surface, for several reasons: these cells are highly migratory and can squeeze through small pores of diameter smaller than their resting size; they are easily accessible in the peripheral blood via minimally invasive IV injection of particles, or can be harvested, processed ex vivo, and reintroduced to the body; they are adept at traveling through the circulation with minimal destruction and thus have access to various tissue beds of the body; and immune cells have built-in signal transduction machinery which allows them to actively engage in chemotaxis and home to regions of the tissue containing tumors, invading microorganisms, or injuries in need of wound healing. In this study, we sought to examine and quantify the degree to which nanoscale liposomes, functionalized with E-selectin adhesion receptor, could bind to a model T cell line and remain on the surface of the cells as they migrate through collagen gels of varying density in a transwell cell migration chamber. It is demonstrated that physiological levels of fluid shear stress are necessary to achieve optimal binding of the E-selectin liposomes to the cell surface as expected, and that CD3/CD28 antibody activation of the T cells was not necessary for effective liposome binding. Nanoscale liposomes were successfully conveyed by the migrating cells across a layer of rat tail type 1 collagen gel ranging in composition from 1 to 3 mg/mL. The relative fraction of liposomes carried through the collagen decreased at higher collagen density, likely due to the expected decrease in average pore size, and increased fiber content in the gels. Taken together, these results support the idea that T cells could be an effective cellular carrier of therapeutic molecules either attached to the surface of nanoscale liposomes or encapsulated within their interior.

Improving Cancer Therapy: Design, Synthesis, and Evaluation of Carboplatin-Based Nanoliposomes against Breast Cancer Cell Lines

Objective: Cancer poses a significant challenge to modern medicine, with breast carcinoma being one of the most prevalent forms of the disease. Carboplatin is a commonly used chemotherapy drug for treating breast cancer, but its efficacy can be limited due to its poor water solubility and associated side effects. This study intended to enhance the therapeutic potency of carboplatin by encapsulating it within liposomal nanocarriers. Methods: We fabricated nanoscale liposomes loaded with carboplatin via a technique called reverse phase evaporation, and examined their physical attributes. We also studied the toxicity of these nanoliposomes on MDA-MB 231 breast cancer cells. Results: Our findings indicated that the liposomal nanoparticles possessed a negative zeta potential of -18.2 mV and an average size of 278 nm. The drug loading level was 2.2%, while the efficiency of drug encapsulation reached 58.5%. Of note, the cytotoxicity of carboplatin in its nanoliposomal form was markedly more potent against the MDA-MB 231 breast cancer cell line than the free drug (p-value less than 0.05). Conclusion: Our findings suggest that carboplatin-loaded liposomal nanocarriers could potentially serve as an advanced chemotherapeutic approach for the treatment of breast cancer, offering enhanced efficacy and reduced side effects compared to conventional carboplatin therapy. Further research is warranted to explore this novel delivery system’s benefits fully.

Spatial distribution of tumor-associated macrophages in an orthotopic prostate cancer mouse model.

Mounting evidence suggests that the immune landscape within prostate tumors influences progression, metastasis, treatment response, and patient outcomes. In this study, we investigated the spatial density of innate immune cell populations within NOD.SCID orthotopic prostate cancer xenografts following microinjection of human DU145 prostate cancer cells. Our laboratory has previously developed nanoscale liposomes that attach to leukocytes via conjugated E-selectin (ES) and kill cancer cells via TNF-related apoptosis inducing ligand (TRAIL). Immunohistochemistry (IHC) staining was performed on tumor samples to identify and quantify leukocyte infiltration for different periods of tumor growth and E-selectin/TRAIL (EST) liposome treatments. We examined the spatial-temporal dynamics of three different immune cell types infiltrating tumors using QuPath image analysis software. IHC staining revealed that F4/80+ tumor-associated macrophages (TAMs) were the most abundant immune cells in all groups, irrespective of time or treatment. The density of TAMs decreased over the course of tumor growth and decreased in response to EST liposome treatments. Intratumoral versus marginal analysis showed a greater presence of TAMs in the marginal regions at 3 weeks of tumor growth which became more evenly distributed over time and in tumors treated with EST liposomes. TUNEL staining indicated that EST liposomes significantly increased cell apoptosis in treated tumors. Additionally, confocal microscopy identified liposome-coated TAMs in both the core and periphery of tumors, highlighting the ability of liposomes to infiltrate tumors by "piggybacking" on macrophages. The results of this study indicate that TAMs represent the majority of innate immune cells within NOD.SCID orthotopic prostate tumors, and spatial density varies widely as a function of tumor size, duration of tumor growth, and treatment of EST liposomes.

Magnetic–Plasmonic Nanoscale Liposomes with Tunable Optical and Magnetic Properties for Combined Multimodal Imaging and Drug Delivery

Magnetic–Plasmonic Nanoscale Liposomes with Tunable Optical and Magnetic Properties for Combined Multimodal Imaging and Drug Delivery

Tuning of TRAIL clustering on the surface of nanoscale liposomes by phase separation.

Phase-separated liposomes were used to formulate tumor necrosis factor-related apoptosis-inducing ligand (TRAIL), a protein that selectively kills cancer cells while sparing most healthy ones. By controlling the average number of TRAIL molecules per liposome, we demonstrate the ability to tune the formation of TRAIL clusters and their resulting apoptotic activity.

Bromelain-Decorated Nanoscale Liposomes for Mucus Permeation and Intestinal Absorption in Oral Drug Delivery

Bromelain-Decorated Nanoscale Liposomes for Mucus Permeation and Intestinal Absorption in Oral Drug Delivery

Influence of Lipid Composition on Physicochemical and Antibacterial Properties of Vancomycin-Loaded Nanoscale Liposomes

Influence of Lipid Composition on Physicochemical and Antibacterial Properties of Vancomycin-Loaded Nanoscale Liposomes

Phase 2 Study of CPX-351 in Combination with Venetoclax in Patients with Newly Diagnosed, High Risk Acute Myeloid Leukemia

Background CPX-351 is a nanoscale liposome of a fixed 5:1 molar ratio of cytarabine and daunorubicin which has improved survival of secondary acute myeloid leukemia (AML) 1. Venetoclax (ven) is a BCL2 inhibitor which, in combination with azacitidine or cytarabine, has become the standard of care for elderly AML. Ven has shown synergism with chemotherapy, with high complete remission (CR) rates in newly diagnosed (ND) and relapsed / refractory (RR) AML 2. In this study, we aim to assess the efficacy and safety of CPX-351 and ven (CPX-ven) in RR and ND AML. Methods This was a phase 2 study with a lead-in safety phase. Enrolment was allowed if the blasts were ≥10%. Patients (pts) age ≥18 with RR AML were eligible for the safety lead-in phase (LIP) or arm A. Arm B enrolled pts age 18 - 69 with ND AML. Other inclusion criteria included adequate liver, renal and cardiac function, and ECOG performance status ≤2. Pts with CNS involvement or prior CPX-351 were excluded. The recommend phase 2 dose was CPX-351 (daunorubicin 44 mg/m 2 and cytarabine 100 mg/m 2 IV on D1, 3 and 5 of induction, and daunorubicin 22 mg/m2 IV on D1 and 3 during consolidation) combined with ven on D2-8 of each cycle. Ven was dosed at 300mg daily and reduced to 150mg or 50mg daily for pts on a moderate or strong CYP3A inhibitors, respectively. The primary outcome measure was complete remission (CR) or CR with incomplete count recovery (CRi). Here, we report the outcomes of arm B including pts with ND AML. Results 14 pts were enrolled on arm B. One pt withdrew and one was too early in treatment, therefore 12 were evaluable. Baseline characteristics are shown in Table 1. The median age was 59.5 years (range, 44 - 69); Four (33.3%) were female. All 12 (100%) pts were classified as adverse risk per the 2017 European Leukemia Net Guidelines. Two (16.7%) pts had treatment for a prior non-myeloid malignancy (therapy-related AML). Eight (66.7%) had an antecedent haematological disorder (secondary AML), of whom seven had treatment (median 1 line, range 1 - 2). Four (33.3%) pts had a complex karyotype, of whom three harbored a TP53 mutation. Nine (75.0%) pts achieved a response, including four CR (33.3%), five CRi (41.6%), and one induction mortality (8.3%) prior to disease assessment. 3/4 (75.0%) pts with complex cytogenetics (including 2/3 (66.7%) with a TP53 mutation) achieved a CR/CRi. Of the secondary AML pts with an evaluable response, 5/7 (71.4%) achieved a CR/CRi. Of the nine pts achieving CR/CRi, five (55.5%) were negative for minimal residual disease (MRD) by flow cytometry. The median number of cycles to CR/CRi was one (range 1 - 2). The median overall survival (OS) was 12.9 months (95% CI, 3.1 - not reached, NR, Figure 1). The median relapse-free survival (RFS) was 8.7 months (95% CI, 3.1 -NR). 7/9 (77.8%) of responding pts underwent an allogeneic stem cell transplant (SCT) after attaining CR/CRi. There were 90 adverse events (AE) in 10 (83.3%) pts. 80 (88.9%) were grade 1 or 2, 10 (11.1%) were grade ≥3. The most common AE was gastrointestinal with seven (7.8%) occurrences. Four (33.3%) pts developed grade ≥3 infections; of these, one (8.3%) died. The cause of death was neutropenic septic shock due to pneumonia. Conclusion CPX-ven is a feasible option for remission induction in pts with high-risk, newly diagnosed AML. The combination produced high rates of response and MRD negativity in a cohort of pts with adverse risk features, including pts with adverse karyotype and AML following prior HMA-based treatment, allowing them to proceed with SCT. Toxicities are as expected for an intensive chemotherapy regimen. Further studies are needed to confirm the efficacy of this combination.

Nanoscale Liposome Synthesis for Drug Delivery Applications via Ultrafast Acoustofluidic Micromixing

Nowadays, lipid nanoparticles have gained profound interest in chemical and biomedical engineering. The rapid development of therapeutic nanosystems has led to a need to design suitable approaches to synthesize bio-carriers for efficient drug delivery. Microfluidic methods provide an excellent opportunity to acquire desirable nanoparticle properties, including stability, size, shape, and size distribution, which are often challenging to obtain using conventional bulk synthesis methods. Rapid mixing is a crucial factor in the nanoprecipitation process as it influences the size and size distribution of the nanoparticles. Within this regard, in this work, we report an ultrafast acoustofluidic micromixer to synthesize liposome nanoparticles, which have been widely investigated in the literature as drug carriers due to their biocompatibility and biodegradability. This research has also investigated the influence of glycerol addition to the solvent to control the size of the liposomes. Our findings indicate that utilizing the acoustofluidic platform resulted in the production of nanoscale liposomes with small mean sizes compared to the hydrodynamic flow-focusing (HFF) method. Furthermore, the inclusion of glycerol led to a significant reduction in liposome size. These results emphasize the potential of the proposed approach for the efficient and precise synthesis of liposome nanoparticles with improved characteristics, which can be utilized in various biomedical and drug delivery applications.

Rapid Production of Nanoscale Liposomes Using a 3D-Printed Reactor-In-A-Centrifuge: Formulation, Characterisation, and Super-Resolution Imaging.

Nanoscale liposomes have been extensively researched and employed clinically for the delivery of biologically active compounds, including chemotherapy drugs and vaccines, offering improved pharmacokinetic behaviour and therapeutic outcomes. Traditional laboratory-scale production methods often suffer from limited control over liposome properties (e.g., size and lamellarity) and rely on laborious multistep procedures, which may limit pre-clinical research developments and innovation in this area. The widespread adoption of alternative, more controllable microfluidic-based methods is often hindered by complexities and costs associated with device manufacturing and operation, as well as the short device lifetime and the relatively low liposome production rates in some cases. In this study, we demonstrated the production of liposomes comprising therapeutically relevant lipid formulations, using a cost-effective 3D-printed reactor-in-a-centrifuge (RIAC) device. By adjusting formulation- and production-related parameters, including the concentration of polyethylene glycol (PEG), temperature, centrifugation time and speed, and lipid concentration, the mean size of the produced liposomes could be tuned in the range of 140 to 200 nm. By combining selected experimental parameters, the method was capable of producing liposomes with a therapeutically relevant mean size of ~174 nm with narrow size distribution (polydispersity index, PDI ~0.1) at a production rate of >8 mg/min. The flow-through method proposed in this study has potential to become an effective and versatile laboratory-scale approach to simplify the synthesis of therapeutic liposomal formulations.

Microbubbles-assisted ultrasonication to promote tumor accumulation of therapeutics and modulation of tumor microenvironment for enhanced cancer treatments

Abnormal tumor vasculature is reported to severely hinder the therapeutic potency of diverse cancer therapeutics by restricting their intratumoral accumulation and/or causing therapeutic resistance. Herein, a microbubble-assisted ultrasonication technology (MAUT) of systemic administration of octafluoropropane-filled microbubbles together with tumor localized ultrasound (US) exposure is developed to generally promote intratumoral accumulation efficacy of three kinds of anti-tumor drugs with varying sizes through the cavitation effect-induced disruption of tumor blood vessels. MAUT was further shown to enable selective tumor hypoxia attenuation by filling microbubbles with high-purity oxygen and thus reducing the production of immunosuppressive lactic acids by suppressing glycolysis in cancer cells. Resultantly, MAUT markedly enhanced the therapeutic outcome of systemically administered anti-programmed death-1 (anti-PD-1) and chemotherapeutic doxorubicin (DOX) with and without using nanoscale liposomes as delivery vehicles. This work highlights that MAUT is a biocompatible yet versatile strategy to effectively reinforce the therapeutic potency of a broad range of cancer therapeutics, promising for future clinical usage.

Neutralization of the new coronavirus by extracting their spikes using engineered liposomes

The devastating COVID-19 pandemic motivates the development of safe and effective antivirals to reduce morbidity and mortality associated with infection. We developed nanoscale liposomes that are coated with the cell receptor of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the virus that causes COVID-19. Lentiviral particles pseudotyped with the spike protein of SARS-CoV-2 were constructed and used to test the virus neutralization potential of the engineered liposomes. Under TEM, we observed for the first time a dissociation of spike proteins from the pseudovirus surface when the pseudovirus was purified. The liposomes potently inhibit viral entry into host cells by extracting the spike proteins from the pseudovirus surface. As the receptor on the liposome surface can be readily changed to target other viruses, the receptor-coated liposome represents a promising strategy for broad spectrum antiviral development.

'Garlic-lipo'4Plants: Liposome-Encapsulated Garlic Extract Stimulates ABA Pathway and PR Genes in Wheat (Triticum aestivum).

Recently, environmentally friendly crop improvements using next-generation plant biostimulants (PBs) come to the forefront in agriculture, regardless of whether they are used by scientists, farmers, or industries. Various organic and inorganic solutions have been investigated by researchers and producers, focusing on tolerance to abiotic and biotic stresses, crop quality, or nutritional deficiency. Garlic has been considered a universal remedy ever since antiquity. A supercritical carbon dioxide garlic extract encapsulated in nanoscale liposomes composed of plant-derived lipids was examined as a possible PB agent. The present study focused on the characterization of the genes associated with the pathways involved in defense response triggered by the liposome nanoparticles that were loaded with supercritical garlic extracts. This material was applied to Triticum aestivum in greenhouse experiments using foliar spraying. The effects were examined in a large-scale genome-wide transcriptional profiling experiment by collecting the samples four times (0 min, used as a control, and 15 min, 24 h, and 48 h after spraying). Based on a time-course expression analysis, the dynamics of the cellular response were determined by examining differentially expressed genes and applying a cluster analysis. The results suggested an enhanced expression of abscisic acid (ABA) pathway and pathogenesis-related (PR) genes, of which positive regulation was found for the AP2-, C2H2-, HD-ZIP-, and MYB-related transcription factor families.

Mixed Nanoscale Liposomes Co-Encapsulating Curcumin and Berberine Inhibit Activated Hepatic Stellate Cell-Induced Drug Resistance and Tumor Metastasis

The combination therapy based on nanoscale materials has been proven an effective approach for cancer treatment. In this study, berberine (BBR) and curcumin (CUR) were loaded into mixed liposomes (CUR-GL/BBR-HL). To simulate a tumor microenvironment (TME), the dual-cell research model (SMMC7721 + LX-2) and orthotopic tumor-bearing mice model were established for antiproliferation and antimetastasis evaluation. The results showed that the mixed liposomes exhibited strong cytotoxicity and antimigration effect against the dual-cell research model. Notably, CUR-GL/BBR-HL effectively inhibited the activation of hepatic stellate cells (HSCs), extracellular matrix deposition, and tumor angiogenesis in vivo, resulting in an enhanced anticancer effect. The anti-HCC mechanism revealed that CUR-GL/BBR-HL could inhibit drug resistance and the metastasis of tumor cells by downregulating the expression of P-gp and vimentin, respectively. Hence, the combined therapeutic strategy could effectively inhibit cancer growth, and CUR-GL/BBR-HL might be a potential formulation for anticancer therapy.

Microfluidic vortex focusing for high throughput synthesis of size-tunable liposomes

Control over vesicle size during nanoscale liposome synthesis is critical for defining the pharmaceutical properties of liposomal nanomedicines. Microfluidic technologies capable of size-tunable liposome generation have been widely explored, but scaling these microfluidic platforms for high production throughput without sacrificing size control has proven challenging. Here we describe a microfluidic-enabled process in which highly vortical flow is established around an axisymmetric stream of solvated lipids, simultaneously focusing the lipids while inducing rapid convective and diffusive mixing through application of the vortical flow field. By adjusting the individual buffer and lipid flow rates within the system, the microfluidic vortex focusing technique is capable of generating liposomes with precisely controlled size and low size variance, and may be operated up to the laminar flow limit for high throughput vesicle production. The reliable formation of liposomes as small as 27 nm and mass production rates over 20 g/h is demonstrated, offering a path toward production-scale liposome synthesis using a single continuous-flow vortex focusing device.

Liposomal Formulation of Botanical Extracts may Enhance Yield Triggering PR Genes and Phenylpropanoid Pathway in Barley (Hordeum vulgare).

This work aimed to study the plant conditioning effect and mode of action of a plant-based biostimulant used in organic farming. This new generation plant biostimulant, named ELICE16INDURES®, is rich in plant bio-active ingredients containing eleven supercritical botanical extracts encapsulated in nano-scale liposomes. The dose-response (10 to 240 g ha-1) relationship was tested in a field population of autumn barley (Hordeum vulgare) test crop, and underlying molecular mechanisms were studied. Applying nanotechnology, cell-identical nanoparticles may help the better uptake and delivery of active ingredients increasing resilience, vitality, and crop yield. The amount of harvested crops showed a significant increase of 27.5% and 39.9% interconnected to higher normalized difference vegetation index (NDVI) of 20% and 25% after the treatment of low and high dosages (20 and 240 g ha-1), respectively. Illumina NextSeq 550 sequencing, gene expression profiling, and KEGG-pathway analysis of outstanding dosages indicated the upregulation of pathogenesis-related (PR) and other genes-associated with induced resistance-which showed dose dependency as well.