Vesicle Size Research Articles

Impact of exercise on uncoupling protein 1 and macrophage phenotype in mesenteric adipose tissue in an animal model of endometriosis

Endometriosis is a gynecological disorder characterized by growth of endometrial tissue outside the uterine cavity, with symptoms which may include chronic pelvic pain and infertility resulting in a decreased quality of life. Previous studies in our laboratory demonstrated that endometriosis animals have increased mesenteric adipose tissue (MAT), inflammation, and growth of implanted endometrial vesicles which are decreased by voluntary exercise. The levels of Uncoupling Protein 1 (UCP-1), a mitochondrial protein, are known to be increased by exercise in rodents, promoting thermogenesis and decreasing inflammation. Exercise can also increase expression of peroxisome proliferator-activated receptor γ (PPARγ), a member of the nuclear receptor superfamily. When activated PPARγ polarizes the macrophages into an anti-inflammatory M2 state, which can stimulate white adipose tissue beiging. We hypothesized that voluntary wheel running exercise can increase UCP-1 and M2 macrophages, decreasing MAT and endometrial vesicle size. Aim: To assess the impact of voluntary exercise on UCP-1 expression and macrophage phenotype in MAT of an animal model of endometriosis. Methods: Endometriosis was induced by implanting uterine tissue on the intestinal mesentery of female Sprague Dawley rats while control group only received sutures (SHAM). Exercise rats had access to a running wheel after surgery (ENDO-EX or SHAM-EX), while non-exercised animals didn’t (ENDO and SHAM; n=10-11/group). After sixty days, animals were sacrificed, and MAT was collected to determine CD68, UCP-1 and PPARγ expression by immunofluorescence staining. Results: Vesicles from ENDO-EX animals exhibited significantly decreased average weight (p<0.01) and smaller average total area (p<0.01) compared with the ENDO group. Endometriosis animals showed increased mesenteric adipose tissue when compared to sham group (p<0.01), which was reduced by voluntary exercise (p<0.001). Macrophage infiltration, UCP-1 and PPARγ expression increased with exercise. Conclusions: Increased UCP-1 and anti-inflammatory M2 macrophages following exercise may result in browning of MAT which can have a role in reducing the development of vesicles in the endometriosis model. These findings suggest that exercise may be a potential complementary therapeutic strategy for managing endometriosis symptoms. This work was supported by R15AT009915, R16GM149365, T32GM144896, PRI MD Summer Program and Porter Physiology Development Fellowship. This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

Novel loading protocol combines highly efficient encapsulation of exogenous therapeutic toxin with preservation of extracellular vesicles properties, uptake and cargo activity

Extracellular vesicles (EVs) have mostly been investigated as carriers of biological therapeutics such as proteins and RNA. Nevertheless, small-molecule drugs of natural or synthetic origin have also been loaded into EVs, resulting in an improvement of their therapeutic properties. A few methods have been employed for EV cargo loading, but poor yield and drastic modifications of vesicles remain unsolved challenges. We tested a different strategy based on temporary pH alteration through incubation of EVs with alkaline sodium carbonate, which resulted in conspicuous exogenous molecule incorporation. In-depth characterization showed that vesicle size, morphology, composition, and uptake were not affected. Our method was more efficient than gold-standard electroporation, particularly for a potential therapeutic toxin: the plant Ribosome Inactivating Protein saporin. The encapsulated saporin resulted protected from degradation, and was efficiently conveyed to receiving cancer cells and triggered cell death. EV-delivered saporin was more cytotoxic compared to the free toxin. This approach allows both the structural preservation of vesicle properties and the transfer of protected cargo in the context of drug delivery.Graphic

Enhancing the efficacy of letrozole-loaded PEGylated nanoliposomes against breast cancer cells: In vitro study

Considering its overall impact on human health, letrozole (Let) has been described as having significant efficacy that could be improved by developing drug delivery systems. Considering the side effects of Let, this study aims to encapsulate Let in liposomes and PEGylated liposome nanoparticles (Lipo-Let-PEG) and evaluate the cytotoxic effects on the MCF-7 breast cancer cell line. For this purpose, the Lipo-Let-PEG formulation was designed and characterized by SEM, DLS, and FTIR methods, and the drug release from the optimized formulation and the stability of the optimized Lipo-Let-PEG were measured. Furthermore, the cytotoxicity and apoptotic studies were performed using MTT assay and flow cytometric analysis. According to the experimental data, the vesicle size and EE% were 170.05 ± 4.15 nm and 87.21 ± 1.36 %, respectively. The cumulative release from Lipo-Let-PEG at pH 5.4 and 7.4 was also approximately 60 % and 50 %, respectively. MTT results showed that Lip-Let-PEG produced more drug cytotoxicity than Lip-Let against MCF-7 cancer cells and was more compatible with normal cells. The results of apoptosis and cell cycle arrest using flow cytometry show that Lipo-Let-PEG caused the most significant increase in apoptotic rates and cell cycle arrest in cancer cells compared to other treated groups. In conclusion, Lipo-Let-PEG can be used as an anticancer agent by arresting cell cycle progression and inducing apoptosis, which can be applied in future studies to prevent breast cancer development.

Formulation, development, and characterization of loratadine emulgel

Background: This study was to develop loratadine (LTD) emulgels to treat localized skin allergy. Method: Initially oil-in-water emulsion was prepared by 3 different types of surfactants & finally gelling agent carbopol 940 was incorporated into emulsion to produce emulgel (i.e., standard conventional method). Results: The developed formulations were characterized using various parameters including particle size (PS), zeta potential (ZP), polydispersity index (PDI), entrapment efficacy (EE), pH, extrusion efficacy, physical stability, in-vitro drug release studies, and scanning electron microscopy (SEM). PS, EE, PDI, ZP and In-vitro studies ranges between 186.25 ± 6.42 mm (LE-F4) to 395.24 ± 8.64 mm (LE-F1), 62.38 ± 0.36 % (LE-F2) to 76.48 ± 0.69 % (LE-F4), 0.276 ± 0.02 (LE-F4) to 0.652 ± 0.02 (LE-F1), 16.45 ± 2.13 mV (LE-F1) to 29.46 ± 2.78 mV (LE-F3) and 21.90 ± 0.3 % (LE-F1) to 68.30±0.9 % (LE-F4) respectively. Conclusion: Based on all physicochemical properties, LE-F4 formulation was considered to be optimized with minimum PS (186.25±6.42 nm), PDI (0.276±0.02), satisfactory positive surface charge (23.15 ± 1.89 mV) and maximum EE (76.48±0.69 %). FTIR studies were confirmed that there is no physical interaction between drug and excipients and SEM studies revealed that vesicle size was spherical with smooth texture. A significantly greater rate of drug release (i.e., 68.30 ± 0.90%) was seen in the LTD emulgels that were made with cationic surfactant (i.e., LE-F4) and found to be good spreadability and extrudability.

Characterization and Stability Test of Hydrolyzed Collagen Glyserosomes

Background: Hydrolyzed collagen is a protein obtained from enzymatic denaturation of collagen with a molecular weight of about 10 kDa, and it has been reported to produce anti-aging properties. Delivering hydrolyzed collagen into the dermis becomes a great challenge due to its large molecular weight, so glycerosome, a deformable vesicle containing glycerol as the edge activator, was developed to carry it into the dermis layer. Objective: The study aimed to determine the effect of increasing the concentration of glycerol and hydrolyzed collagen on the characteristics and stability of hydrolyzed collagen glycerosomes. Methods: Glycerosomes were composed of soy lecithin and prepared using a thin film lipid method. The lipid film was hydrated with phosphate-buffered saline pH 5 containing different glycerol concentrations (20% and 40%) and hydrolyzed collagen (2.5% and 5%). Then, characteristic tests and stability tests were carried out. Results: Hydrolyzed collagen glycerosomes had vesicle sizes of 170-180 nm, polydispersity index of 0.253-0.279, zeta potential values of -23.70 to -26.50 mV with deformability indexes of 2.25-3.49. The highest percentage of entrapment efficiency was 85.72%, achieved with a glycerol concentration of 40%. During the stability test at 25°C for 12 weeks, the hydrolyzed collagen glycerosomes did not experience pH and entrapment efficiency changes, but it increased the vesicle size. Conclusion: The use of 40% glycerol produced more deformable vesicles than 20% glycerol in hydrolyzed collagen glycerosomes; however, a formula improvement is required to improve the stability of glycerosomes.

Next-generation skin cancer treatment: A quality by design perspective on artificial neural network-optimized cationic ethosomes with bleomycin sulphate

Skin cancer, exacerbated by environmental factors like ozone layer depletion, poses a global health concern. Topical administration of chemotherapy, a promising approach, minimizes systemic toxicity. Bleomycin Sulphate (BLM), effective intravenously, faces challenges due to limited skin penetration. Cationic ethosomes, known for superior skin penetration, address this by enhancing electrostatic interactions with cancer cells. This ensures localized drug delivery, preventing systemic circulation. Employing thin film hydration, we prepared cationic ethosomes with BLM, optimizing through Artificial Neural Network (ANN) and Response Surface Methodology (RSM). Characterization included entrapment efficiency, vesicle size, morphology, and zeta potential. In-vitro drug release and various kinetic models were analyzed. Ex-vivo evaluations, like dermatokinetic studies and fluorescence microscopy, and in-vivo skin sensitivity testing were conducted. Results showed the optimized formulation with 27.1 % entrapment efficiency, 144.3 nm vesicle size, smooth surface morphology, and positive zeta potential. Ex-vivo assessments indicated favorable skin retention and in-vivo tests suggested suitability for topical administration, highlighting its potential in skin cancer treatment. This research offers a promising strategy for localized and effective skin cancer therapy, addressing challenges associated with systemic toxicity and limited drug penetration.

Berberine HCl and diacerein loaded dual delivery transferosomes: Formulation and optimization using Box-Behnken design.

Berberine is a poorly water-soluble alkaloid compound showing significant anti-inflammatory characteristics. It reduces the levels of pro-inflammatory and inflammatory cytokines, including tumour necrosis factor (TNF-α, IFN-γ) and interleukin (IL-23, IL-12, and IL-23). Diacerein significantly reduces the splenomegaly associated with psoriasis. It downregulates the production of TNF-α and IL-12. This study reported the development of transferosomes containing berberine HCl and diacerein using a film hydration method followed by optimization using a Box-Behnken design. Sodium deoxycholate was used as an edge activator. The impact of independent variables (amount of phosphatidylcholine, amount of edge activator, and sonication cycles) on dependent variables (particle size and entrapment efficiency) was examined. The optimized formulation was characterized for polydispersity index, vesicle size, entrapment efficiency, ζ potential, spectral analysis like Fourier transform infrared, thermal analysis, X-ray diffraction, deformability, transmission electron microscopy, antioxidant assay, in-vitro release, and ex-vivo skin permeation studies. The optimized formulation had a particle size of 110.90±2.8 nm with high entrapment efficiency (89.50±1.5 of berberine HCl and 91.23±1.8 of diacerein). Deformability, polydispersity index, ζ potential, and antioxidant activity of the optimized formulation were 2.44, 0.296, -13.3, and 38.36 %, respectively. Optimized transferosomes exhibited 82.093±0.81 % and 85.02±3.81 % release of berberine HCl and diacerein after 24 h of dissolution study. The transdermal flux of optimized formulation was 0.0224 μg cm-2 h-1 (2.24 cm h-1 permeation coefficient) and 0.0462 μg cm-2 h-1 (4.62 cm h-1 permeation coefficient), respectively, for berberine HCl and diacerein. Raman analysis of treated pig skin confirmed that the transferosomes can permeate the skin. No change in the skin condition or irritation was observed in BALB/c mice. Formulation stored at 4 and 25±2 °C / 60±5 % relative humidity was stable for 3 months. Thus, the results demonstrated successful optimization of the transferosomes for the efficient topical delivery of berberine HCl and diacerein in the effective management of psoriasis.

Fabrication and characterisation of nabumetone transferosomal gel for effective topical delivery

Nabumetone is a lipophilic drug with a log P value of 3.2 and belongs to BCS class II due to its poor water solubility. When administered orally, it undergoes extensive first-pass metabolism, shows low bioavailability, and has unfavourable side effects that cause discomfort. Topical route has gain importance by delivering therapeutic agents via this route due to its safety and convenience of drug administration and as of there are no any gel formulations available for it. The present investigation aimed to prepare, optimise, and characterise Nabumetone-loaded transferosomes (Nb-TF) using 32 factorial designs. Transferosomes were prepared by the thin-film hydration method. The optimised formulation was incorporated into gel form (Nb-TG) and evaluated for its mechanical characteristics, in-vitro release, ex-vivo permeation, skin irritancy test, anti-inflammatory activity, and stability studies. The Nb-TF 5 transferosomal formulation was optimised based on vesicle size and entrapment efficiency and incorporated into the gel base. Both normal (Nb-gel) and transferosome-loaded (Nb-TG) gel pH was within the range of topical skin pH. The viscosities of Nb-gel and Nb-TG were 1262 ± 54 cP and 1327 ± 12 cP, respectively. The drug content of both gels was found to be 96.31 ± 1.16 and 97.18 ± 0.94, respectively. Nb-TG showed higher diffusion and drug permeation than Nb-gel. The skin irritancy test indicates that there was no sign of any edema or erythema observed. Nb-TG showed a curative and preventive anti-inflammatory effect. Gels were found stable at both refrigerator and room temperature. Nb-TFs were proven to be potential carriers for transdermal controlled delivery of Nabumetone as it has deep penetration in the dermis layer which directly goes into the systemic circulation, enhances the bioavailability and reduced toxicities.

Magnetic resonance imaging features and classification of intraspinal echinococcosis

ObjectiveThis study aimed to investigate the magnetic resonance imaging (MRI) features and classification of echinococcosis of the spinal canal. MethodsThe clinical manifestations and MRI findings and classification of 19 patients diagnosed with intraspinal echinococcosis over 10 years (2011–2020) were retrospectively analyzed. ResultsThe mean age of the nine males and 10 females was 39 years (range 28–65 years). Among these, the number of cases with thoracic, lumbar, sacral, thoracolumbar, and lumbosacral, cervical, and lumbar segments was nine (47 %), five (26 %), one (5 %), one (5 %), two (11 %), and one (5 %) cases, respectively. Furthermore, 13 cases (69 %) involved adjacent vertebral bodies, accessories, and surrounding soft tissues. The lesion was confined to the intramedullary, extramedullary subdural, extramedullary epidural, and multiple spaces in one (Type I) (5 %), four (Type II) (21 %), one (Type III) (5 %), and thirteen (69 %) cases, respectively. Moreover, nine cases (47 %) had a history of hydatid disease in the spine or other tissues. The clinical manifestations were chest and lumbosacral pain in 18 cases (95 %) and chest and lumbosacral pain accompanied by lower limb dysfunction in four cases (21 %). The MRI revealed different sizes of T1WI low-signal and T2WI high-signal vesicles with a “grape-like” appearance, with 16 cases (84 %) showing low signals on the edge of the cyst wall. ConclusionIntraspinal echinococcosis is rare, even in endemic areas. However, intraspinal echinococcosis should be considered when there is a history of echinococcosis in other sites or when there are clear MRI characteristics for the disease.

Tailoring of Bilosomal Nanogel for Augmenting the Off-Label Use of Sildenafil Citrate in Pediatric Pulmonary Hypertension.

Pediatric pulmonary hypertension is a serious syndrome with significant morbidity and mortality. Sildenafil is widely used off-label in pediatric patients with pulmonary arterial hypertension. In this study, bile salt-stabilized nanovesicles (bilosomes) were screened for their efficacy to enhance the transdermal delivery of the phosphodiesterase type 5 inhibitor, sildenafil citrate, in an attempt to augment its therapeutic efficacy in pediatric pulmonary hypertension. A response surface methodology was implemented for fabricating and optimizing a bilosomal formulation of sildenafil (SDF-BS). The optimized SDF-BS formulation was characterized in terms of its entrapment efficiency (EE), zeta potential, vesicle size, and in vitro release profile. The optimized formula was then loaded onto hydroxypropyl methyl cellulose (HPMC) hydrogel and assessed for skin permeation, in vivo pharmacokinetics, and pharmacodynamic studies. The optimized SDF-BS showed the following characteristic features; EE of 88.7 ± 1.1%, vesicle size of 185.0 + 9.2 nm, zeta potential of -20.4 ± 1.1 mV, and efficiently sustained SDF release for 12 h. Skin permeation study revealed a remarkable improvement in SDF penetration from bilosomal gel compared to plain SDF gel. In addition, pharmacokinetic results revealed that encapsulating SDF within bilosomal vesicles significantly enhanced its systemic bioavailability (∼3 folds), compared to SDF oral suspension. In addition, pharmacodynamic investigation revealed that, compared to plain SDF gel or oral drug suspension, SDF-BS gel applied topically triggered a significant elevation (p < 0.05) in cGMP serum levels, underscoring the superior therapeutic efficacy of SDF-BS gel. Conclusively, bilosomes can be viewed as a promising nanocarrier for transdermal delivery of SDF that would grant higher therapeutic efficiency while alleviating the limitations encountered with SDF oral administration.

The Structural and Functional Integrity of Rod Photoreceptor Ribbon Synapses Depends on Redundant Actions of Dynamins 1 and 3.

Vertebrate vision begins with light absorption by rod and cone photoreceptors, which transmit signals from their synaptic terminals to second-order neurons: bipolar and horizontal cells. In mouse rods, there is a single presynaptic ribbon-type active zone at which the release of glutamate occurs tonically in the dark. This tonic glutamatergic signaling requires continuous exo- and endocytosis of synaptic vesicles. At conventional synapses, endocytosis commonly requires dynamins: GTPases encoded by three genes (Dnm1-3), which perform membrane scission. Disrupting endocytosis by dynamin deletions impairs transmission at conventional synapses, but the impact of disrupting endocytosis and the role(s) of specific dynamin isoforms at rod ribbon synapses are understood incompletely. Here, we used cell-specific knock-outs (KOs) of the neuron-specific Dnm1 and Dnm3 to investigate the functional roles of dynamin isoforms in rod photoreceptors in mice of either sex. Analysis of synaptic protein expression, synapse ultrastructure, and retinal function via electroretinograms (ERGs) showed that dynamins 1 and 3 act redundantly and are essential for supporting the structural and functional integrity of rod ribbon synapses. Single Dnm3 KO showed no phenotype, and single Dnm1 KO only modestly reduced synaptic vesicle density without affecting vesicle size and overall synapse integrity, whereas double Dnm1/Dnm3 KO impaired vesicle endocytosis profoundly, causing enlarged vesicles, reduced vesicle density, reduced ERG responses, synaptic terminal degeneration, and disassembly and degeneration of postsynaptic processes. Concurrently, cone function remained intact. These results show the fundamental redundancy of dynamins 1 and 3 in regulating the structure and function of rod ribbon synapses.

Formulation of silymarin surface modified vesicles: In vitro characterization to cell viability assessment

Silymarin (SLR) is a poorly water-soluble bioactive compound with a wide range of therapeutic activities. Nanosized silymarin vesicles (F1–F6) were prepared by the solvent evaporation rehydration method. The silymarin vesicles were evaluated for vesicle size, surface charge, entrapment efficiency, and drug release studies. The optimized SLR lipid vesicle (F3) was further modified with the addition of the cationic polymer chitosan. After that, the modified vesicle (F3C1) was assessed for permeation flux, antimicrobial activity, cell viability, and molecular docking studies. The silymarin vesicles showed nanometric size (<250 nm), low polydispersibility index (<0.05), negative surface charge, and high SLR entrapment (85–95 %). The drug release study result demonstrated a maximum drug release of 91.2 ± 2.8 %. After adding chitosan to the surface, there was a significant change in the size, polydispersibility index, surface charge (positive), and encapsulation efficiency. The drug release was found to be prolonged, and the permeation flux was also increased in comparison to free SLR. A comparative antimicrobial result was observed in comparison to the free SLR and standard drug. The cell viability assay also demonstrated a low IC50 value for F3C1 against the cell line.

Flow-driven translocation of polymer vesicle through a nanochannel

Flow-driven translocation of a polymer vesicle self-assembled from amphiphilic diblock copolymers through a nanochannel is studied using hybrid Lattice-Boltzmann Molecular Dynamics simulations. Three translocation modes are discovered: a single vesicle, small vesicles, and tiny micelles in sequence, respectively, as the width of the nanochannel is decreased, which is confirmed by the self-consistent field theory calculations. Accordingly, spherical vesicles, ellipsoidal vesicles, and worm-like micelles are formed sequentially after crossing the nanochannel. More importantly, we find that the critical flow rate monotonically decreases as the ratio of the polymer vesicle size and the width of the nanochannel decreases until it approaches 1. Our results provide a basis for characterizing the stability of vesicles or sorting vesicles with different sizes and also point to a new idea of regulating spherical vesicles to ellipsoidal vesicles or worm-like micelles.

Stabilization of Prebiotic Vesicles by Peptides Depends on Sequence and Chirality: A Mechanism for Selection of Protocell-Associated Peptides.

Cells require oligonucleotides and polypeptides with specific, homochiral sequences to perform essential functions, but it is unclear how such oligomers were selected from random sequences at the origin of life. Cells were probably preceded by simple compartments such as fatty acid vesicles, and oligomers that increased the stability, growth, or division of vesicles could have thereby increased in frequency. We therefore tested whether prebiotic peptides alter the stability or growth of vesicles composed of a prebiotic fatty acid. We find that three of 15 dipeptides tested reduce salt-induced flocculation of vesicles. All three contain leucine, and increasing their length increases the efficacy. Also, leucine-leucine but not alanine-alanine increases the size of vesicles grown by multiple additions of micelles. In a molecular simulation, leucine-leucine docks to the membrane, with the side chains inserted into the hydrophobic core of the bilayer, while alanine-alanine fails to dock. Finally, the heterochiral forms of leucine-leucine, at a high concentration, rapidly shrink the vesicles and make them leakier and less stable to high pH than the homochiral forms do. Thus, prebiotic peptide-membrane interactions influence the flocculation, growth, size, leakiness, and pH stability of prebiotic vesicles, with differential effects due to sequence, length, and chirality. These differences could lead to a population of vesicles enriched for peptides with beneficial sequence and chirality, beginning selection for the functional oligomers that underpin life.

Self-Assembled Recombinant Elastin and Globular Protein Vesicles with Tunable Properties for Diverse Applications.

Vesicles are self-assembled structures comprised of a membrane-like exterior surrounding a hollow lumen with applications in drug delivery, artificial cells, and micro-bioreactors. Lipid or polymer vesicles are the most common and are made of lipids or polymers, respectively. They are highly useful structures for many applications but it can be challenging to decorate them with proteins or encapsulate proteins in them, owing to the use of organic solvent in their formation and the large size of proteins relative to lipid or polymer molecules. By utilization of recombinant fusion proteins to make vesicles, specific protein domains can be directly incorporated while also imparting tunability and stability. Protein vesicle assembly relies on the design and use of self-assembling amphiphilic proteins. A specific protein vesicle platform made in purely aqueous conditions of a globular, functional protein fused to a glutamate-rich leucine zipper (ZE) and a thermoresponsive elastin-like polypeptide (ELP) fused to an arginine-rich leucine zipper (ZR) is discussed here. The hydrophobic conformational change of the ELP above its transition temperature drives assembly, and strong ZE/ZR binding enables incorporation of the desired functional protein. Mixing the soluble proteins on ice induces zipper binding, and then warming above the ELP transition temperature (Tt) triggers the transition to and growth of protein-rich coacervates and, finally, reorganization of proteins into vesicles. Vesicle size is tunable based on salt concentration, rate of heating, protein concentration, size of the globular protein, molar ratio of the proteins, and the ELP sequence. Increasing the salt concentration decreases vesicle size by decreasing the Tt, resulting in a shorter coacervation transition stage. Likewise, directly changing the heating rate also changes this time and increasing protein concentration increases coalescence. Increasing globular protein size decreases the size of the vesicle due to steric hindrance. By changing the ELP sequence, which consists of (VPGXG)n, through the guest residue (X) or number of repeats (n), Tt is changed, affecting size. Additionally, the chemical nature of X variation has endowed vesicles with stimuli responsiveness and stability at physiological conditions.Protein vesicles have been used for biocatalysis, biomacromolecular drug delivery, and vaccine applications. Photo-cross-linkable vesicles were used to deliver small molecule cargo to cancer cells in vitro and antigen to immune cells in vivo. pH-responsive vesicles effectively delivered functional protein cargo, including cytochrome C, to the cytosol of cancer cells in vitro, using hydrophobic ion pairing to improve cargo distribution in the vesicles and release. The globular protein used to make the vesicles can be varied to achieve different functions. For example, enzyme vesicles exhibit biocatalysis, and antigen vesicles induce antibody and cellular immune responses after vaccination in mice. Collectively, the development and engineering of the protein vesicle platform has employed amphiphilic self-assembly strategies and rational protein engineering to control physical, chemical, and biological properties for biotechnology and nanomedicine applications.

BBD assisted in-situ nanoliposomes of esculin hydrate via intranasal delivery for the amelioration of Parkinson's disease

The objective of this research was to develop and optimise esculin Hydrate loaded nanoliposomes (ESH-NLs). The nanoliposomes were prepared by solvent evaporation method and optimized using a three-factor, three-level Box-Behnken design. Phospholipid 90G (X1), cholesterol (X2), and sonication time (X3) were used as independent variables, and their effects on vesicle size (Y1), entrapment efficiency (Y2), and Polydispersity index (Y3) were observed. The opt-ESH-NLs were further characterised for in-vitro drug release, DPPH assay, confocal laser scanning microscopy, and ex-vivo permeation. Opt-ESH-NLs had a vesicle size of 88.36 nm, a PDI of 0.06, an entrapment efficiency of 94.22 ± 0.93 %, and a drug release of 76.776 ± 1.127 %. Compared to standard ascorbic acid, the antioxidant activity was found to be 78.52 ± 0.148 %. Ex-vivo permeation studies revealed that opt-ESH-NLs had significantly enhanced permeation (79.484 ± 0.754 %) compared to ESH suspension (38.326 ± 1.279 %). Pharmacokinetic study revealed that opt-ESH-NLs when compared with Oral ESH suspension had a significant bioavailability in Brain as compared to plasma. The stability study of nanoliposomes at 4 °C reveals there is no significant change in the formulation. It was concluded that opt-ESH-NLs is a promising and effective formulation for intranasal administration for direct delivery of drug in the brain for amelioration of PD.

Isolation of extracellular vesicles from corn leaves using differential ultracentrifugation and the identification of miR165a microRNA

MicroRNAs are a class of non-coding RNA molecules that act as important regulators of various biological processes in plants. MicroRNAs play a significant regulatory role in the early reaction of the cell to a stressful impact, including intercellular signalling. Extracellular vesicles of plants are essential for the transport of molecules in plants. Numerous methods for isolating extracellular vesicles have been developed over the past decade, and differential centrifugation allows the isolation of small vesicles. Using the differential centrifugation method, we managed to isolate two fractions of extracellular vesicles (P40 and P100) from corn leaves that were different in size and molecular weight. The fraction of large vesicles had a size of more than 150 nm and contained high and low molecular nucleic acids. We also determined the sizes of vesicles of the P100 fraction which contained only low molecular nucleic acids: they were from 30 to 200 nm. A specific probe was designed to identify miR165a microRNA using polymerase chain reaction. Using the stem-loop probe allowed obtaining cDNA matrices larger than 70 nucleotide pairs (bp), which was sufficient to evaluate the polymerase chain reaction (PCR). The results of RT-PCR analysis with specific primers for miR165a microRNA showed the presence of an amplification product approximately 70-80 nucleotide pairs in size, which corresponded to theoretical values. Therefore, fractions of extracellular vesicle from corn leaves contained miR165a microRNA, which may explain their physiological function as intercellular transport of small non-coding RNAs.

Quercetin-loaded transliposomal gel for effective management of skin cancer: In vitro and cell line efficacy studies

In this research work, Quercetin-loaded transliposomes (QUR-TL) were developed for dermal application against skin cancers. The utilization of nano-formulation for dermal application presents several notable advantages. Using an ethanol injection method followed by sonication, the QUR-loaded TL was developed and optimized by Box-Behnken Design (BBD). The optimized QUR-TL exhibited the vesicle size of 200.1 nm, polydispersity-index of 0.181, zeta potential of −31.89 mV and an entrapment efficiency of 82.1 %. The morphology of the optimized QUR-TL assessed by Transmission Electron Microscopy has revealed the spherical shape of the vesicles. In vitro release of QUR from QUR-TL was higher as compared to QUR-Suspn. Ex vivo skin permeation has shown ≈2-fold increased permeation of QUR from QUR-TL gel (QUR-TLG) as compared to QUR-conventional gel (QUR-CFG). This demonstrated a prolonged release pattern and an increased permeability of QUR from QUR-TLG. The QUR-TL has shown 1.3-fold higher antioxidant activity as compared to pure-QUR (QUR-SOL). The CLSM demonstrated an enhanced penetration of QUR-TLG, which was substantiated by the quantification of QUR during the dermatokinetics investigation. The QUR-TLG was evaluated for its cytotoxic potential against the B16F10 melanoma cells, which has shown lowest IC50 value (9.3 μM) as compared to QUR-CFG (16.8 μM) and QUR-SOL (23 μM). This suggested strong evidence of QUR indication for skin cancers. Conclusively, QUR-TLG has significantly (p < 0.05) targeted the deeper layers of skin and can be regarded as a promising carrier for topical application of QUR or other lipophilic drugs for the management of skin cancers.

Enhancing the hypotensive effect of latanoprost by combining synthetic phosphatidylcholine liposomes with hyaluronic acid and osmoprotective agents

The first line of glaucoma treatment focuses on reducing intraocular pressure (IOP) through the prescription of topical prostaglandin analogues, such as latanoprost (LAT). Topical ophthalmic medicines have low bioavailability due to their rapid elimination from the ocular surface. Nanotechnology offers innovative ways of enhancing the ocular bioavailability of antiglaucoma agents while reducing administration frequency. This study aims to combine LAT-loaded synthetic phosphatidylcholine liposomes with hyaluronic acid (0.2% w/v) and the osmoprotectants betaine (0.40% w/v) and leucine (0.90% w/v) (LAT-HA-LIP) to extend the hypotensive effect of LAT while protecting the ocular surface. LAT-HA-LIP was prepared as a mixture of 1,2-dioleoyl-sn-glycero-3-phosphocholine and 1,2-dimyristoyl-sn-glycero-3-phosphocholine, cholesterol and α-tocopherol acetate. LAT-HA-LIP exhibited high drug-loading capacity (104.52 ± 4.10%), unimodal vesicle sizes (195.14 ± 14.34 nm) and a zeta potential of -13.96 ± 0.78 mV. LAT-HA-LIP was isotonic (284.00 ± 1.41 mOsm L−1), had neutral pH (7.63 ± 0.01) and had suitable surface tension (44.07 ± 2.70 mN m−1) and viscosity (2.69 ± 0.15 mPa s−1) for topical ophthalmic administration. LAT-HA-LIP exhibited optimal in vitro tolerance in human corneal and conjunctival epithelial cells. No signs of ocular alteration or discomfort were observed when LAT-HA-LIP was instilled in albino male New Zealand rabbits. Hypotensive studies revealed that, after a single eye drop, the effect of LAT-HA-LIP lasted 24 h longer than that of a marketed formulation and that relative ocular bioavailability was almost three times higher (p < 0.001). These findings indicate the potential ocular protection and hypotensive effect LAT-HA-LIP offers in glaucoma treatment.Graphical abstract

Chitosan-coated double-loaded liposomes as a promising delivery system for clove essential oil

To improve the stability and bioavailability of double-loaded clove essential oil (CEO) liposomes (DLCL), chitosan was used to coat DLCL (CS-DLCL) as a promising delivery system for CEO. Considering the vesicle size, polydispersity index, zeta potential, and encapsulation efficiency, 3.5 mg/mL chitosan was selected as the optimum concentration. Electrostatic interactions between chitosan and DLCL were confirmed through Fourier transform infrared spectroscopy. Stability analysis revealed that the physical barrier formed by chitosan improved the stability of DLCL under high-temperature, high-salt, and long-term storage at 4 °C and reduced the interaction of CEO with chemicals in the digestive fluids, providing a foundation for the sustained release of CEO in the intestine. Finally, chitosan exhibited an obvious inhibitory effect on lipid oxidation secondary products and enhanced the antioxidant activity of DLCL. Thus, chitosan is a promising coating material for improving the stability of DLCL, and CS-DLCL have a good potential for application in foods.