Ethanol Injection Method Research Articles

Does the technical methodology influence the quality attributes and the potential of skin permeation of Luliconazole loaded transethosomes?

Luliconazole (LCZ) is a novel topical imidazole medication with broad-spectrum antifungal activity. Unfortunately, the high potential of LCZ for topical antifungal activity is hampered by its low solubility and low skin permeability that required long-term therapy with frequent administration for a full recovery. So, this study aimed to enhance the skin permeation of LCZ through its loading in ultradeformable nanovesicles, transethosomes (TE), using either ethanol injection sonication or thin-film hydration methods. The prepared nanovesicles were characterized for vesicles size, zeta potential, entrapment efficiency, and in vitro drug release. Moreover, the ex vivo skin permeation and deposition via rat skin were estimated. In vitro characterization of the LCZ-TE displayed that the average vesicles size prepared by the ethanol injection method was 246.3 ± 0.56 versus 62.75 ± 0.16 nm with a thin-film method. Also, the zeta potential and encapsulation efficiency were 36.3 ± 0.81 mV and 82.35 ± 3.68% versus 49.4 ± 0.67 mV and 93.85 ± 1.78% in the ethanol injection sonication and thin-film hydration, respectively. Furthermore, the deposited percentage of LCZ in the skin by TE formulation prepared by thin film hydration (54.79 ± 5.23%) is more than 2-fold the percentage of LCZ solution in propylene glycol (PG) (25.26 ± 2.84%) and 1.5 fold more than TE prepared by ethanol injection (35.65 ± 4.354) due to the enhanced skin permeation and deposition of LCZ-TE. These results revealed the superiority of the thin film hydration technique in the enhancement of the quality attributes of the prepared nanovesicles and pointed out their potential in the treatment of fungal infections of the skin.

Drug Release, Stability And Efficiency Of Vitamin E Loaded In Liposomes For Bovine Sperm Protection In Cryopreservation Medium

BACKGROUND:It is known that a considerable number of drugs in clinical use or under development are water-insoluble drugs with poor bioavailability. The liposomal delivery system has drawn attention as one of the noteworthy approaches to increase both dissolution and absorption because of its biocompatibility and ability to encapsulate hydrophobic molecules in the lipid domain. However, several drawbacks have been reported, the most common is liposome structural instability .OBJECTIVE:To encapsulate alpha tocopherol into liposomes, to determine the new formulation stability and to study the drug-release of alpha tocopherol into the sperm cryopreservation medium.MATERIALS AND METHODS:The liposomes prepared by an ethanol injection method were characterized for size stability, alpha tocopherol release and sperm motility tests.RESULTS:The prepared unilamellar vesicles had both narrow size distribution (around 99 nm) and a good physical and chemical stability at 4°C during 12 months. The liposomes did not release the vitamin E immediately, but retained the protectant for 24 hours, probably due to the rigidity of the liposomal fence which was reinforced by adding cholesterol. Then, all vitamin E molecules were released by 48 hours. Release was potentially by Fickian diffusion probably by the creation of mini-ducts due to both agitation and fence hydration. Moreover, semen motility treated with vitamin E liposome preparations was significantly improved compared to all other treatments (including commonly used sperm conservation media).CONCLUSION :The stable vitamin E liposomes formulated in this work are a promising alternative for semen cryopreservation protection.

Novel nano spanlastic carrier system for buccal delivery of lacidipine

Lacidipine, a calcium channel antagonist with its highly lipophilic and poorly water-soluble nature (BCS Class II Drug), when administered via the conventional oral route undergoes extensive hepatic metabolism by Cytochrome P450 3A4 (CYP3A4), results in low bioavailability (∼10%) and summons a challenge to anti-hypertensive therapy. This present research study aimed to override this drug delivery obstacle by formulating lacidipine loaded spanlastic orally dissolving films. A 23 factorial design model was employed in the spanlastics development process to examine the influence of the formulation variables over the response variables. Spanlastics composed of Span 60 and Tween 80 were prepared by modified ethanol injection method and developed into an orally dissolving film using hypromellose E5, polyvinyl alcohol and polyethylene glycol 400 by the solvent casting technique. The characterization results of the formulation showed that the spanlastics possessed nano-sized vesicles and good entrapment efficiency (86.84%). Optical microscopy and high resolution transmission electron microscopy displayed lamellar, circularly shaped, and uniformly dispersed nanovesicles. Evaluation studies revealed that the oral film possessed considerable flexibility, good mechanical strength, disintegrated within 35 s and revealed a drug release of 92.96% within 5 min. The scanning electron microscopy displayed a smooth, uniform surface of the film with pores. Stability studies of the films at 25 ± 2 °C and 60 ± 5% RH for 3 months showed no alteration in physical appearance or mechanical properties. The developed spanlastic vesicles, owing to their nano size have an enhanced barrier permeation profile while the oral films offer better bioavailability of the drug by absorption through the oro-mucosal tissue. In combination with the increased permeability, the circumvention of hepatic metabolism, improved dissolution and fast onset of action, this new drug delivery approach makes lacidipine efficiently available for antihypertensive therapy.

Cell-penetrating peptide-surface modified liposomes to enhance the intestinal absorption of enoxaparin

Enoxaparin is low-molecular-weight heparin administered by subcutaneous/intravenous injection. The oral bioavailability of enoxaparin is restricted by its low absorption through the intestine. In this study cell-penetrating peptide-surface functionalized liposomes (CPPs-L) were prepared to improve the intestinal absorption of enoxaparin. Liposomal formulations were prepared by the ethanol injection method and the intestinal absorption of the formulation was evaluated using single-pass intestinal perfusion (SPIP) technique in rats. Meanwhile, the human fraction dose absorbed value (Fa (human)) of the formulations were predicted based on the calculated effective intestinal permeability (P effect (rat)) values obtained from the SPIP study. Liposomal enoxaparin revealed an increased intestinal absorption by ten-time compared with the free drug solution. Meanwhile, CPPs-L formulation revealed an enhanced intestinal absorption compared with the un-modified liposomal formulation. Regarding Fa (human), it is predicted that liposomal formulations could have the potential to improve the fraction dose absorbed of enoxaparin from low to intermediate level. Overall, the liposomal formulation can be considered as a mighty drug carrier for the oral delivery of enoxaparin.

Liposomes loaded with betel leaf (Piper betle L.) ethanolic extract prepared by thin film hydration and ethanol injection methods: Characteristics and antioxidant activities.

Betel leaf ethanolic extract (BLEE), which was dechlorophyllized by sedimentation process was loaded in liposomes at 1 and 2% (w/v) concentrations using two different methods, namely thin film hydration (TF) and ethanol injection (EI) methods. Liposomes loaded with 1% BLEE and prepared by TF method (BLEE/L-T1) had the smallest particle size and paler color than BLEE/L-E1, BLEE/L-E2, and BLEE/L-T2 (p<.05). BLEE/L-T1 also showed strong stability as judged by its lowest zeta potential and polydispersity index. The highest encapsulation efficiency (EE) and lowest releasing efficiency (RE) were also found with BLEE/L-T1. No significant difference (p>.05) in the antioxidant activities was detected between the BLEE-loaded liposomes and BLEE solutions, indicating that encapsulation had no adverse effect on BLEE antioxidant potency. BLEE/L-T1 showed higher antioxidant stability than unencapsulated BLEE at the equivalent amount based on EE (BLEE/U-T1) during in vitro gastrointestinal tract digestion system. Therefore, BLEE/L-T1 could be an efficient delivery system for improving stability of antioxidant activities of BLEE. PRACTICAL APPLICATIONS: Despite the many benefits of betel leaf ethanolic extract, it still has some distinctive odor and slightly greenish color as well as instability induced by environment factors, which can limit applications in foods. Encapsulation of the betel extract in liposomes can be a good approach to mask its undesirable color and odor and to augment its antioxidant stability. Liposomal technology can be used to load betel leaf extract. However, different methods have been implemented to prepare liposomes that exhibit varying encapsulation efficacy as well as bioactivities. Thin film hydration method was shown to yield the liposome with better physical characteristics, higher encapsulation efficiency, slower release, and higher antioxidant stability than the ethanol injection method. Therefore, the thin film hydration method could be adopted to prepare stable liposomes loaded with betel leaf extract that possess antioxidant activity suitable for food applications.

Encapsulation of α-Pinene in Delivery Systems Based on Liposomes and Cyclodextrins.

The essential oil component α-pinene has multiple biological activities. However, its application is limited owing to its volatility, low aqueous solubility, and chemical instability. For the aim of improving its physicochemical properties, α-pinene was encapsulated in conventional liposomes (CLs) and drug-in-cyclodextrin-in-liposomes (DCLs). Hydroxypropyl-β-cyclodextrin/α-pinene (HP-β-CD/α-pinene) inclusion complexes were prepared in aqueous solution, and the optimal solubilization of α-pinene occurred at HP-β-CD:α-pinene molar ratio of 7.5:1. The ethanol-injection method was applied to produce different formulations using saturated (Phospholipon 90H) or unsaturated (Lipoid S100) phospholipids in combination with cholesterol. The size, the phospholipid and cholesterol incorporation rates, the encapsulation efficiency (EE), and the loading rate (LR) of α-pinene were determined, and the storage stability of liposomes was assessed. The results showed that α-pinene was efficiently entrapped in CLs and DCLs with high EE values. Moreover, Lipoid S100 CLs displayed the highest LR (22.9 ± 2.2%) of α-pinene compared to the other formulations. Both carrier systems HP-β-CD/α-pinene inclusion complex and Lipoid S100 CLs presented a gradual release of α-pinene. Furthermore, the DPPH radical scavenging activity of α-pinene was maintained upon encapsulation in Lipoid S100 CLs. Finally, it was found that all formulations were stable after three months of storage at 4 °C.

Anti-inflammatory activity of cyanidin-3-O-glucoside and cyanidin-3-O-glucoside liposomes in THP-1 macrophages.

Cyanidin‐3‐O‐glucoside (C3G) is a kind of water‐soluble pigment widely existing in many plants. It has strong antioxidant and anti‐inflammatory activities. However, C3G cannot exist stably for a long time because of the phenolic hydroxyl groups in its structure. Liposome technology could improve the stability and bioavailability of compounds. Based on our previous studies, C3G liposomes prepared by ethanol injection method have a certain stability in two weeks of storage. In this study, THP‐1 macrophages treated with C3G and C3G liposomes can reduce the levels of inflammatory‐related factors, such as tumor necrosis factor‐a (TNF‐a), interleukin (IL)‐1β, IL‐6, and IL‐8, stimulated by lipopolysaccharide (LPS). Further studies showed that the LPS induction could increase the level of phosphorylated nuclear transcription factor NF‐κB and phosphorylated IkBa, while C3G and C3G liposomes could inhibit the expression of phosphorylated proteins. Moreover, C3G and C3G liposomes could protect macrophages from apoptosis. In conclusion, C3G prepared by liposome technology exhibits anti‐inflammatory activity, which provides a theoretical basis for the food industry to study functional food.

Preparation, characterization, and anti-colon cancer activity of oridonin-loaded long-circulating liposomes

In this study, oridonin-loaded long-circulating liposomes (LC-lipo@ORI) were prepared with the ethanol injection method. Its physicochemical properties and the morphology were characterized, and its stability and release profiles were evaluated. Furthermore, its antitumor effects were studied using two in vitro cell models of colon cancer and two tumor-bearing models in nude mice. The prepared LC-lipo@ORI was quasi-spherical, with a mean particle size of 109.55 ± 2.30 nm. The zeta potential was −1.38 ± 0.21 mV, the encapsulation efficiency was 85.79%±3.25%, and the drug loading was 5.87%±0.21%. In vitro release results showed that the cumulative release rate of LC-lipo@ORI at 12 h was 63.83%. However, ORI dispersion was almost completely released after 12 h. In vitro cytotoxicity results showed that, the inhibiting effects of LC-lipo@ORI on the proliferation of two types of colon cancer cells were apparently higher than those of ORI dispersion, whereas those of the blank carrier were not noticeable. In vivo studies confirmed that, the encapsulation of LC-lipo enhanced the inhibitory effects of ORI on tumor growth. These results indicated that LC-lipo@ORI a promising formulations for colon cancer treatment.

Surface modification with cholesteryl acetyl carnitine, a novel cationic agent, elevates cancer cell uptake of the PEGylated liposomes

The present study aimed to synthesize cholesteryl acetyl carnitine (CAC), and surface modify the PEGylated liposomes with the intention of enhanced cancer cell uptake. For this, CAC synthesis was performed in amine-free esterification conditions and then four liposomal formulations of unmodified, CAC/PEG, and CAC + PEG-modified were prepared by ethanol injection method. Cytotoxicity of the liposomes was investigated in A549 cells, followed by cellular uptake assessments of coumarin 6 (C6)-loaded liposomes. The results of ATR-FTIR, 1HNMR, and 13CNMR demonstrated successful formation of CAC. A molecular docking study showed efficient binding affinities rather than carnitine to the active site of four carnitine transporters. Liposomal formulations possessed spherical morphology with a mean particle size range of 112–138 nm, narrow size distribution, and negative surface charge. All formulations had low cytotoxicity at 0.5 mg/ml, but high cytotoxicity at around 2.5 mg/ml. The lowest IC50 was obtained for CAC modified liposomes. CAC + PEG-modified liposomes had the highest cellular uptake.In conclusion, CAC + PEG modification of liposomes is an effective approach for increasing A549 cellular uptake, with low cytotoxicity at commonly applied liposome concentrations. The elevated uptake may be due to the involvement of the organic cation transporter, cationic structure, and the metabolic preference of CAC in cancer cells.

Development of an mRNA-LNP Vaccine against SARS-CoV-2: Evaluation of Immune Response in Mouse and Rhesus Macaque.

Among the vaccines have been developed thus far against SARS-CoV-2, the mRNA-based ones have demonstrated more promising results regarding both safety and efficacy. Two remarkable features of the mRNA vaccines introduced by the Pfizer/BioNTech and Moderna companies are the use of (N1-methyl-pseudouridine-) modified mRNA and the microfluidics-based production of lipid nanoparticles (LNPs) as the carrier. In the present study, except Anti-Reverse Cap Analog (ARCA), no other nucleoside analogs were employed to synthesize Spike-encoding mRNA using the in vitro transcription (IVT) method. Furthermore, LNPs were prepared via the ethanol injection method commonly used for liposome formation as an alternative for microfluidics-based approaches. The produced mRNA-LNP vaccine was evaluated for nanoparticles characteristics, encapsulation and transfection efficiencies, in vitro cytotoxicity as well as stability and storability. The safety of vaccine was assessed in Balb/c mice injected with mRNA-LNPs containing 10 µg of spike-encoding mRNA. Eventually, the vaccine efficacy in inducing an immune response against SARS-CoV-2 was studied in Balb/c and C57BL/6 mice (received either 1 or 10 µg of mRNA) as well as in rhesus macaque monkeys (infused with mRNA-LNPs containing 100 µg of mRNA). The ELISA and virus neutralizing test (VNT) results showed a significant augmentation in the level of neutralizing antibodies against SARS-CoV-2. Moreover, the ELISA assay showed virus-specific IFN-γ secretion in immunized mice as a marker of TH1 cell-based immune response, whereas favorably no change in the production of IL-4 was detected.

Mithramycin delivery systems to develop effective therapies in sarcomas

BackgroundSarcomas comprise a group of aggressive malignancies with very little treatment options beyond standard chemotherapy. Reposition of approved drugs represents an attractive approach to identify effective therapeutic compounds. One example is mithramycin (MTM), a natural antibiotic which has demonstrated a strong antitumour activity in several tumour types, including sarcomas. However, its widespread use in the clinic was limited by its poor toxicity profile.ResultsIn order to improve the therapeutic index of MTM, we have loaded MTM into newly developed nanocarrier formulations. First, polylactide (PLA) polymeric nanoparticles (NPs) were generated by nanoprecipitation. Also, liposomes (LIP) were prepared by ethanol injection and evaporation solvent method. Finally, MTM-loaded hydrogels (HG) were obtained by passive loading using a urea derivative non-peptidic hydrogelator. MTM-loaded NPs and LIP display optimal hydrodynamic radii between 80 and 105 nm with a very low polydispersity index (PdI) and encapsulation efficiencies (EE) of 92 and 30%, respectively. All formulations show a high stability and different release rates ranging from a fast release in HG (100% after 30 min) to more sustained release from NPs (100% after 24 h) and LIP (40% after 48 h). In vitro assays confirmed that all assayed MTM formulations retain the cytotoxic, anti-invasive and anti-stemness potential of free MTM in models of myxoid liposarcoma, undifferentiated pleomorphic sarcoma and chondrosarcoma. In addition, whole genome transcriptomic analysis evidenced the ability of MTM, both free and encapsulated, to act as a multi-repressor of several tumour-promoting pathways at once. Importantly, the treatment of mice bearing sarcoma xenografts showed that encapsulated MTM exhibited enhanced therapeutic effects and was better tolerated than free MTM.ConclusionsOverall, these novel formulations may represent an efficient and safer MTM-delivering alternative for sarcoma treatment.Graphical abstract

Implementing Spanlastics for Improving the Ocular Delivery of Clotrimazole: In vitro Characterization, Ex vivo Permeability, Microbiological Assessment and In vivo Safety Study.

PurposeThe aim of this study was to encapsulate clotrimazole (CLT), an antifungal drug with poor water solubility characteristics, into spanlastics (SPs) to provide a controlled ocular delivery of the drug.MethodsSpan 60 was used in the formulation of SPs with Tween 80, Pluronic F127, or Kolliphor RH40 as an edge activator (EA). The presence of EA offers more elasticity to the membrane of the vesicles which is expected to increase the corneal permeation of CLT. SPs were prepared using ethanol injection method applying 32 complete factorial design to study the effect of formulation variables (ratio of Span 60: EA (w/w) and type of EA) on SPs characteristics (encapsulation efficiency percent (EE%), average vesicle size (VS), polydispersity index (PDI) and zeta potential (ZP)). Design-Expert software was used to determine the optimum formulation for further investigations.ResultsThe optimum formulation determined was S1, which contains 20 mg of Tween 80 used as an EA and 80 mg of Span 60. S1 exhibited EE% = 66.54 ± 7.57%, VS = 206.20 ± 4.95 nm, PDI = 0.39 ± 0.00 and ZP = −29.60 ± 0.99 mV. S1 showed highly elastic sphere-shaped vesicles. Furthermore, S1 displayed a sustained release profile and a higher ex vivo permeation across rabbit cornea relative to CLT suspension. Also, S1 revealed superior inhibition of Candida albicans development compared to CLT suspension applying 2,3-bis(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide (XTT) reduction technique. Moreover, in vivo histopathological examination assured the safety of S1 after ophthalmic application in mature male albino rabbits.ConclusionOverall, the outcomes revealed the marked efficacy of SPs for ocular delivery of CLT.

Improved Efficacy and Stability of Silymarin Loaded Nanocochleates Over Liposomes for the Treatment of Skin Diseases

Aim: Silymarin, a complex polyphenolic component mixture with anti-oxidant, anti-inflammatory, and membrane-stabilizing property is being investigated in several dermatological conditions. Present research aims to evaluate potential of silymarin loaded nanocochleates and liposomal topical application for treating chronic skin diseases.&#x0D; Study Design: Silymarin loaded liposomes and nanocochleates were formulated and optimized using Design Expert software. Different invitro and exvivo tests were performed to compare their performance.&#x0D; Place and Duration of Study: The study was conducted in Smt. Kashibai Navale College of Pharmacy, Pune, India, between January 2019 till February 2020.&#x0D; Methodology: Liposomes were prepared using ethanol injection method and further treated with calcium chloride to form nanocochleates by trapping method. Design of experiments (32 Factorial Design) was used for optimization of nanocochleates. Cell line studies (HaCaT cell lines) and short term stability studies were performed to compare the efficacy and stability respectively.&#x0D; Results: Particle size, entrapment efficiency and drug deposition in Wistar Rat Skin was found to be statistically significant for nanocochleates over liposomes proving superiority of cochleates. Both the carriers sustained release of silymarin for 24h. Antimicrobial efficacy of nanocochleates against E.coli and S.aureus was significant. Inhibition of hyper proliferation of HaCaT cell lines (key mechanism by which most of the antipsoriatic drugs act) demonstrated the superiority of nanocochleates over liposomes.The nanocochleates also displayed better stability compared to liposomes due to decreased entrapment efficacy and leakage of drug.&#x0D; Conclusion: Silymarin loaded Nanocochleates could prove as a promising topical drug delivery system for the treatment of chronic skin diseases like psoriasis.

Down regulation of inflammatory cytokines by the bioactive resveratrol-loaded chitoniosomes in induced ocular inflammation model

Resveratrol (RSV); a natural polyphenol possess anti-inflammatory property through reduction of proinflammatory cytokines tumor-necrosis factor-alpha (TNFα) and interleukin (IL)-6 which play a key role in eye inflammation. Ocular inflammation results in irreversible tissues damage and impaired vision if left untreated. Surface coating of niosomes with chitosan can enhance their efficacy. Therefore the aim was to develop RSV-loaded chitosan-coated niosomes (Chitoniosomes) for topical ocular delivery. Chitoniosomes were formulated by ethanol-injection method. The selected formula (RSV.CsNiPolx) was composed of Span 60, Poloxamer407 and Cholesterol (3.5:3.5:3) coated with 0.6% w/v Chitosan. RSV.CsNiPolx showed particle size 363.1nm, PDI (0.21), zeta potential (+17.9), 90.3% entrapment efficiency, mucoadhesive efficiency with a sustained-release profile (56%) after 6 h. TEM showed circular shape niosomes surrounded by a relatively faded coat. The formula kept stable at 4 °C for 6 months storage. Good ocular tolerance of Chitoniosomes formulation without any inflammatory response to the rabbit eyes was confirmed by histological examination. Invivo antiinflammatory effects after 3 days treatment revealed reduced gene expressions of TNFα and IL-6, down to 49% and 55% respectively, in treated groups compared to control group. In conclusion, the results offer a promising nanoplatforms for topical application of the natural bioactive resveratrol in ocular inflammation.

Redox-sensitive irinotecan liposomes with active ultra-high loading and enhanced intracellular drug release

In this report, a novel irinotecan (IR) encapsulated redox-responsive liposome was developed. The redox-responsive liposomes were prepared based on disulfide phosphatidylcholine (SS-PC), DSPC, DSPE-PEG2000 and cholesterol by ethanol injection method. IR was actively loaded by triethylammonium sucrose octasulfate (TEA8-SOS) gradient method to generate IR/SS-PC liposomes (IR/SS-LP). The particle size of IR/SS-PC was characterized by using dynamic light scattering (DLS) and transmission electron microscopy (TEM). It was found that IR/SS-LP with 30 % content of SS-PC (IR/SS30-LP) had an average size of 125.5 ± 5.8 nm with a negative zeta potential of −19.5 ± 0.1. The encapsulation efficiency (EE) was further determined to be 98.1 ± 0.8 % and drug loading (DL) was 31.8 ± 0.1 %. The redox-responsiveness of IR/SS-LP was investigated by observing the change of particle size and morphology as well as the release behavior of IR triggered by glutathione (GSH). The data indicated GSH breaks the disulfide bonds in SS-PC and leads to the controlled release of IR. At 1 mM GSH, 60.2 % irinotecan was released from IR/SS30-LP within 24 h. Finally, the effects of IR/SS-LP in cell and animal experiments were evaluated in detail. The results showed that IR/SS30-LP had superior pharmacokinetic and antitumor efficacy compared to free irinotecan and traditional irinotecan liposome (ONIVYDE®-like). Taken together, IR/SS30-LP displayed redox-responsive release of IR, ultra-high loading and enhanced anti-tumor activity, which has great potential for clinical application as a new generation of IR liposomal formulation.

Characterization of fortified pasteurized cow milk with nanoliposome loaded with skipjack tuna eyeball oil

SummaryNanoliposomes loaded with skipjack tuna eyeballs oil (STEO‐NL) were prepared using an ethanol injection method, without and with the aid of ultrasonication. Higher amplitude (80%) and longer time (15 min) of ultrasonication reduced particle size (from 192.20 to 150.70 nm) and zeta‐potential (from −40.43 to −47.29 mV) of STEO‐NL but increased peroxide value (PV), conjugated diene (CD) and thiobarbituric acid reactive substances (TBARS) of STEO‐NL. However, encapsulation efficiency was not different among all the samples (96.82‐97.70%) (P &gt; 0.05). Cow milk was fortified by addition of STEO‐NL (without ultrasonication) at different final levels (0, 2.5 and 5.0% w/w). Subsequently, high‐temperature short‐time (72 °C, 15 s) process was employed to pasteurize the fortified milk, which were further stored up to 10 days at 4 °C. Microbiological quality and pH of pasteurized fortified cow milk were still acceptable. Addition of STEO‐NL at higher level (5%) resulted in higher PV and TBARS and lower acceptability than the control and that fortified with 2.5% STEO‐NL throughout the storage of 10 days, but sensorial acceptability was still obtained. In general, no differences in colour, and viscosity were attained among all samples. Moreover, pasteurized cow milk fortified with STEO‐NL had an increased amount of n‐3 polyunsaturated fatty acids.

Formulation of Spray-dried Proliposomes Loaded with Berberin

The aims of study was formulation and evaluation of berberin (BBR) loaded proliposomes by spray-drying method. BBR proliposomes were evaluated for appearance, spray-drying efficiency, morphology and differential scanning calorimetry (DSC). Liposomes, obtained after hydration, were evaluated for particle size, size distribution, morphology and entrapment efficiency. The results showed that BBR proliposomes were prepared by spray-drying method with molar ratio of Hydrogenated soy phosphatidyl choline (HSPC): Sodium deoxycholat (NaDC): vitamin E (vtE): BBR = 7: 1: 6: 6. Mixture of manitol and Aerosil at weight ratio of 97:3 was used as carrier. Results of DSC showed that berberin was dispersed molecularly into proliposomes powder. BBR liposomes, obtained after hydration, had average particle diameter of about 29 μm and entrapment efficiency was 22.23%.&#x0D; Keywords&#x0D; Proliposomes, liposomes, berberin, sodium deoxycholate, spray-dried.&#x0D; References&#x0D; [1] W. Kong, J. Wei, A. Parrveen et al., Berberine is A Novel Cholesterol-Lowering Drug Working Through A Unique Mechanism Distinct From Statins, Nature Medicine, Vol. 10, No. 12, 2004, pp. 1344-1351, https://doi.org/10.1038/nm1135.[2] S. K. Kulkarni, A. Dhir, on The Mechanism of Antidepressant-Like Action of Berberine Chloride, European Journal of Pharmacology, Vol. 589, No. 1-3, 2008, pp. 163-172, https://doi.org/ 10.1016/j.ejphar.2008.05.043.[3] Y. T. Ho, J. S. Yang, T. C. Li et al., Berberine Suppresses in Vitro Migration and Invasion of Human SCC-4 Tongue Squamous Cancer Cells Through the Inhibitions of FAK, IKK, NF-Κb, U-PA and MMP-2 and-9, Cancer Letters, Vol. 279, No. 2, 2009, pp. 155-162, https://doi.org/10.1016/j.canlet.2009.01.033.[4] S. Muneer, Z. Masood, S. Butt et al., Proliposomes as Pharmaceutical Drug Delivery System: A Brief Review, Journal of Nanomedicine and Nanotechnology, Vol. 8, No. 3, 2017, pp. 448-450, https://doi.org/10.4172/2157-7439.1000448.[5] H. K. Omer, N. R. Hussein, A. Ferraz et al., Spray-Dried Proliposome Microparticles for High-Performance Aerosol Delivery Using a Monodose Powder Inhaler, AAPS PharmSciTech, Vol. 19, No. 5, 2018, pp. 2434-2448, https://doi.org/10.1208/s12249-018-1058-4.[6] T. T. H. Yen, T. T. N. Quynh, D. T. Thuan, P. T. M. Hue, Preparation of Berberin Liposomes, Contained Sodium Deoxycholate by Ethanol Injection Method, Journal of Pharmaceutical Research and Drug information, Vol. 11, No. 4, 2020, pp. 11-17 (in Vietnamese). [7] T. T. H. Yen, T. T. Hue, P. T. M. Hue et al., Preparation of Berberin Proliposomes by Film Deposition on Carrier Surface Method, VNU Journal of Science: Medical and Pharmaceutical Sciences, Vol. 36, No. 2, 2020, pp. 9-15, https://doi.org/10.25073/2588-1132/vnumps.4204.[8] R. G. Ahmed, S. Sherif, Z. Zainab et al., Silymarin Spray-Dried Proliposomes: Preparation, Characterization and Cytotoxic Evaluation, Drug Delivery Letters, Vol. 10, No. 1, 2020, pp. 14-23, https://doi.org/10.2174/2210303109666190722114211.[9] A. Bangham, M. M. Standish, J. C. Watkins Diffusion of Univalent Ions Across the Lamellae of Swollen Phospholipids, Journal of Molecular Biology, Vol. 13, No. 1, 1965, pp. 238-252.&#x0D; &#x0D;

Probing influence of nanocarrier structure on red blood cell membrane perturbations

ABSTRACT In the present work, we have attempted to investigate influence of nanocarrier composition and structure on the RBC membrane perturbations which delivers nanocarrier contents inside the cell. For this purpose, blank liposomes (LIP), lipid nanoparticles (NAN) and cubosomes (CUB) were prepared using ethanol injection method. The interaction of nanocarriers with RBC membrane was investigated using confocal microscopy and small angle x-ray scattering (SAXS). ‘d’ value obtained from SAXS data was plotted against time to investigate rate of fusion of nanocarriers with RBC membrane. Amongst all the nanocarriers tested, NAN and CUB were found to be superior over conventional LIP in terms of fusion with RBC membrane. Additionally, the study showed that use of Glyceryl monooleate in the preparation of nanocarriers will help in faster drug/gene delivery inside the RBCs. Thus NAN and CUB can act as better nanocarriers for guest molecules when such molecules are to be delivered inside RBCs.

QbD-driven development of dissolving microneedle patch loaded with ultradeformable liposomes encapsulated Noopept: Exploring a patient friendly, once-daily option to manage dementia

Noopept (NPT), a potent neuroprotective agent, suffers the problem of poor oral bioavailability (~10%) and thus demands exploration of ways of bioavailability improvement. Present work focuses on confronting this issue via development of NPT loaded ultradeformable liposomes (UDL) and its further incorporation in fast dissolving microneedle patch (MNP) for transdermal route. A combination of Phospholipon 90 G and Phospholipon 90H was used as bilayer forming lipids while sodium deoxycholate was used as edge activator to formulate NPT UDL by ethanol injection method. QbD approach was adapted to optimize NPT UDL considering vesicle size and entrapment efficiency as critical quality attributes (CQA). Fractional factorial design established amount of lipids, surfactant and NPT as critical material attributes and their optimum levels were statistically derived using combined D-optimal design. These optimized NPT UDL were fabricated as fast dissolving MNP and exhaustively evaluated to establish their safety, efficacy and stability. NPT UDL MNP possessed axial needle fracture force of 0.688 N which was sufficient enough to breach stratum corneum. Physical stability evaluation revealed that NPT UDL re-dispersion obtained from MNP matrix dissolution possessed identical vesicle shape and size while retaining > 99% NPT when compared to optimized NPT UDL. In vitro viability of HaCaT cells after exposure to NPT UDL MNP matrix was found to be 89.74% supporting the formulation as safe for transdermal application. NPT UDL MNP showed a 6.5-fold increase in steady state flux across full thickness pig ear skin as compared to NPT suspension. A 3-fold increase in relative bioavailability with similar pharmacological response as compared to oral NPT suspension was also observed during PK-PD studies in Sprague Dawley rats. Results were found fairly encouraging and created a scope of reducing both dose and dosing frequency to eventually improve the associated patient compliance.

Zolmitriptan Intranasal Spanlastics for Enhanced Migraine Treatment; Formulation Parameters Optimized via Quality by Design Approach

Zolmitriptan is a potent second-generation triptan prescribed for migraine attacks. It suffers low bioavailability (40%) after oral administration due to the hepatic first-pass metabolism. Spanlastics are surfactant-based elastic vesicular drug carrier systems. This study aimed to design and optimize intranasal spanlastic formulations as an alternative approach that directly targets brain delivery, enhancing its bioavailability and avoiding the first-pass effect. The quality by design approach was applied to correlate the formulation parameters (Span 60 and Tween 80 concentrations) and critical quality attributes (entrapment efficiency (EE%) and particle size). Spanlastic formulations were designed based on response surface central composite design and prepared via an ethanol injection method. Designed formulations were characterized by EE% and particle size measurements to select the optimized formula (with a combination of small particle size and high EE%). The optimized formula was further subjected to transmission electron microscopy, zeta potential measurement and ex vivo permeation study. The optimized formulation showed a particle size of 117.5 nm and EE% of 45.65%, with a low percentage of error between the observed and predicted values. Seventy percent of zolmitriptan was permeated through the nasal membrane within 30 min, and it completely permeated within 2 h with a significantly higher steady-state flux compared to plain gel. This study introduced a successful and promising intranasal formulation suitable for further brain delivery analysis.