Liposome Method Research Articles

Comparison of free vs. liposomal naringenin in white adipose tissue browning in C57BL/6j mice

Naringenin may play a role in browning by increasing thermogenic gene expression. In this study, we encapsulated naringenin using a liposomal formulation and examined the effects of both free and liposomal naringenin on white adipose tissue browning in C57BL6/J mice. In the first phase of the study, naringenin was encapsulated by the liposome method, which is biocompatible and biodegradable. The physical and chemical properties of liposomal naringenin were tested. In the second phase, a total of 48 six-week-old mice were divided into two main groups: prevention and recovery. Each main group was divided into four subgroups: nano-naringenin, void, free-naringenin, and control. The prevention group received a high-fat diet for 10 weeks along with weekly intravenous injections of 20 µM naringenin. On the other hand, the recovery group was first subjected to a high-fat diet for 10 weeks, followed by an additional 10 weeks of the same diet, along with weekly intravenous injections of 20 µM naringenin. Body weight was measured once per week, and brown adipose tissue, inguinal white adipose tissue, and serum samples were collected from each mouse. The mean particle size, polydispersity index and zeta potential values of liposomal naringenin were ∼207 nm, 0.35, and −27 mV, respectively. The encapsulation and loading efficiencies of liposomal naringenin were 94.6 and 19.2%, respectively. Liposomal naringenin exhibited sustained-release behavior, while free naringenin showed a burst-release profile. Liposomal naringenin showed the best physical stability in light and at 4 °C, while free naringenin was more chemically stable in light and at 4 and 22 °C. Free and liposomal naringenin did not significantly reduce weight gain. In the prevention group, liposomal naringenin increased PRDM16 gene expression in inguinal white adipose tissue 4.29 times more than free naringenin (p = 0.010). However, neither formulation significantly altered the expression levels of other browning or adipogenesis markers in the tissues. The results suggest that free naringenin can be efficiently encapsulated in biocompatible and biodegradable nanoparticles. Further research is needed to better understand the physiological effects of liposomal naringenin.

Liposomal Drug Delivery for Neurological Disorders: Advances and Challenges

Abstract: Liposomal drug delivery methods are becoming increasingly viable options for improving treatment outcomes for neurological illnesses. These systems provide a flexible framework for the formulation of medications intended for delivery to the brain, protecting the medication from enzymatic breakdown and enhancing its bioavailability. To maximize liposome-drug interactions and improve brain-targeted delivery efficiency, a variety of formulation strategies are used, such as surface modification and remote loading. By utilizing various pathways to cross the blood- -brain barrier (BBB), such as passive diffusion and receptor-mediated transcytosis, liposomes facilitate the effective transport of therapeutic drugs to the brain parenchyma. Liposomal formulations show potential for targeted drug delivery, reducing off-target effects, and improving treatment efficacy in neurological conditions like Parkinson's disease, Alzheimer's disease, stroke, multiple sclerosis, and brain cancers. For instance, in Parkinson's disease, liposomal delivery of neuroprotective agents can help maintain dopamine levels and protect dopaminergic neurons. In Alzheimer's disease, liposomes can be engineered to deliver drugs that reduce amyloid-beta plaques or tau tangles. For brain cancer, liposomal chemotherapy can target tumor cells more precisely while minimizing damage to surrounding healthy tissue. In stroke, liposomal delivery of neuroprotective agents can reduce the extent of brain damage, while in multiple sclerosis, liposomes can be used to deliver drugs that modulate the immune response. However, the clinical translation of liposomal drug delivery systems for brain diseases faces challenges related to scalability, stability, and immunogenicity, in addition to regulatory barriers. Scalability issues arise from the complex manufacturing processes required to produce liposomes consistently on a large scale. Stability concerns involve maintaining the integrity of liposomes during storage and after administration. Immunogenicity can be a problem if the liposomes trigger an unwanted immune response, potentially reducing their effectiveness or causing adverse effects. To overcome these obstacles, multidisciplinary cooperation is essential. Collaboration among materials scientists, pharmacologists, neurologists, and regulatory experts can drive the development of more robust liposomal formulations. Continuous research is needed to refine liposome designs, such as by optimizing lipid composition, surface charge, and size to improve stability and targeting capabilities. Advanced techniques like PEGylation (coating liposomes with polyethylene glycol) can help reduce immunogenicity and extend circulation time in the bloodstream. Despite these challenges, liposomal methods present intriguing prospects for transforming medication administration to the brain and offering effective treatments for neurological illnesses. The development of more sophisticated liposomal technologies, combined with a deeper understanding of their mechanisms of action, could lead to significant breakthroughs in the treatment of neurological disorders. For example, research into ligand- targeted liposomes, which use specific molecules to bind to receptors on the BBB, holds promise for enhancing delivery specificity and efficiency. To fully realize the therapeutic promise of these novel drug delivery systems, further advancements in liposomal technologies and a deeper understanding of their mechanisms are necessary. This includes not only technical improvements but also comprehensive preclinical and clinical studies to evaluate safety, efficacy, and long-term effects. As our knowledge expands and technology progresses, liposomal drug delivery could become a cornerstone of neurological disease treatment, providing new hope for patients with previously intractable conditions.

Propolis-loaded liposomes: characterization and evaluation of the in vitro bioaccessibility of phenolic compounds.

Propolis has low water solubility, poor stability, and limited bioaccessibility of phenolic constituents when subjected to in vitro digestion. To overcome these drawbacks, the liposomal encapsulation method can be employed. Soybean phosphatidylcholine lecithin mixed with Tween 80 (T80) and ammonium phosphatides (AMP) was used to produce propolis extract (PE)-loaded liposomes. The mean particle size, zeta potential, encapsulation efficiency values, and transmission electron microscopy analysis were used to characterize liposomes. Individual phenolics were determined for digested and nondigested propolis-loaded liposomes and propolis extract. Tween 80 incorporation reduced the size of unloaded liposomes, whereas AMP inclusion yielded larger liposomes. In both formulations, PE loading significantly increased the size and reduced the zeta potential values and homogeneity of the size distribution. In free PE, the most bioaccessible polyphenols were phenolic acids (3.20 to 5.63 %), and flavonoids such as caffeic acid phenethyl ester, galangin, pinobanksin, and pinocembrin (0.03 to 2.12 %) were the least bioaccessible. Both liposomal propolis provided significantly higher bioaccessibility of phenolic compounds. The liposomes with T80 and AMP in their compositions recovered 52.43 and 185.90 % of the total amount of phenolic compounds in the nondigested samples, respectively. The liposomes containing AMP not only exhibited high solubility for PE but also provided protection to the phenolic compounds during in vitro digestion. Liposomal encapsulation could be a promising approach to improving the solubility and stability of PE in digestive fluids, making it suitable for the delivery of propolis in oral formulations.

Exploring the Anti-Photoaging Potential of Bacillus cereus Superoxide Dismutase in New Zealand Rabbit Skin

Bacillus cereus is a heat-resistant bacteria that produces the enzyme superoxide dismutase (SOD). SOD is a metalloenzyme that can eliminate free radicals as the primary cause of photoaging. The point of this study is to find out how well B. cereus works as an anti-photoaging agent on the skin of rabbits’ backs. The SOD enzyme was isolated from B. cereus using the colorimetric method and characterized for its activity. It was encapsulated by the liposome method, while the activity was measured on the rabbit’s back skin exposed to ultraviolet-A (UV-A) radiation with the parameters of moisture, elasticity, and collagen levels using a skin analyzer. At the end of the treatment, a histological test for the animal skin was performed. The particle size analysis (PSA) results on the SOD in liposomes showed nanoparticles with an adsorption efficiency of 85.0% and a zeta potential of 28.0±11.7 mV. SOD encapsulated in the liposomes also indicated anti-photoaging activity potential by increasing collagen, elasticity, and moisture levels. In addition, histological tests revealed that the collagen density in the SOD liposome treatment was higher than the negatively of control and normally of control but lower than the liposome collagen.

ApoE-modified liposomes encapsulating resveratrol and salidroside alleviate manifestations of Alzheimer’s disease in APP/PS-1 mice

Objective Alzheimer’s disease (AD) is a neurodegenerative disease that is associated with aging and is influenced by both genetic and environmental factors. Several studies and clinical trials have demonstrated that resveratrol (Res) and salidroside (Sal) are not only biologically safe but also influence AD biomarker trajectories. However, their clinical applications have been quite limited due to poor specificity, low solubility, and insufficient blood–brain barrier (BBB) penetration. Therefore, we developed a nano-drug delivery system in which Res and Sal were encapsulated in liposomes, which were surface-modified with ApoE (ApoE-Res/Sal-Lips) to compensate for these deficiencies. Method In this study, ApoE-Res/Sal-Lips were prepared using a standard thin-film hydration method for liposomes. Then, cellular uptake of the loaded liposomes was assessed in vitro using fluorescent staining assays. A BBB model was constructed to investigate the capacity of the liposomes to cross the BBB in vitro, and the ability of liposomes to target the brain was observed by in vivo imaging. In addition, the neuroprotective effects of the different liposome formulations in APP/PS-1 mice were evaluated by measuring the changes in levels of oxidative, anti-inflammatory, and anti-apoptotic factors in the mice brains. Results In vitro, ApoE-Res/Sal-Lips increased the uptake of Res and Sal by bEnd.3 and N2a cells, enhanced BBB penetration, and improved transport efficiency. In vivo, the ApoE-Res/Sal-Lips were found to alleviate AD pathological symptoms, reduce learning and memory impairments, and improve brain function. Conclusion ApoE-Res/Sal-Lips provide a new method for the treatment of AD.

Characterization of hutbak based papers activated with limonene for food packaging

Limonene has a great potential to create an active paper for citrus products. The aim of this study is first to encapsulate limonone by liposome method and cyclodextrin inclusion methods and second to characterize and measure antifungal activity of hutbak papers that have active properties. Active hutbak papers are produced by dipping method with free limonene solution (H-L), encapsulated limonene containing liposome solution (H-Lp), hydroxypropyl-beta-cyclodextrin/limonene inclusion complex solutions (H-HPβCD) and beta-cyclodextrin/limonene (H-βCD) inclusion complex solutions. The effects of the production methods on the physical properties of the papers were analyzed by tensile test and color analysis. The produced active papers were stored for 30 days at 4⁰C. The amount of limonene remaining in the papers and antifungal activity analyzes against P. digitatum and P. expansium were performed on intervals of storage. The highest limonene content values were determined in H-βCD samples. Only the H-HPβCD samples showed zone diameters according to the disk diffusion method.

Short Review of Liposome Composition and Methods of Preparation

Liposomes are amongst the various nano-carriers that are used to target drugs to a specific tissue. They are colloidal, vesicular structures of phospholipid molecules that are surrounded by equal number of aqueous compartments. The lipid bilayer encapsulates an aqueous interior which can be loaded with cargo such as peptides and proteins, hormones, enzymes, antibiotics, antifungal and anticancer agents. This structural property of liposomes makes it an important nano-carrier for drug delivery

Liposomal Entrapment or Chemical Modification of Relaxin2 for Prolongation of Its Stability and Biological Activity.

Relaxin (RLX) is a protein that is structurally similar to insulin and has interesting biological activities. As with all proteins, preservation of RLX’s structural integrity/biological functionality is problematic. Herein, we investigated two methods for increasing the duration of relaxin-2’s (RLX2) biological activity: synthesis of a palmitoyl RLX2 conjugate (P-RLX2) with the use of a Palmitoyl-l-Glu-OtBu peptide modifier, and encapsulation into liposomes of P-RLX2, RLX2, and its oxidized form (O-RLX2). For liposomal encapsulation thin-film hydration and DRV methods were applied, and different lipid compositions were tested for optimized protein loading. RLX2 and O-RLX2 were quantified by HPLC. The capability of the peptides/conjugate to stimulate transfected cells to produce cyclic adenosine monophosphate (cAMP) was used as a measure of their biological activity. The stability and bioactivity of free and liposomal RLX2 types were monitored for a 30 d period, in buffer (in some cases) and bovine serum (80%) at 37 °C. The results showed that liposome encapsulation substantially increased the RLX2 integrity in buffer; PEGylated liposomes demonstrated a higher protection. Liposome encapsulation also increased the stability of RLX2 and O-RLX2 in serum. Considering the peptide’s biological activity, cAMP production of RLX2 was higher than that of the oxidized form and the P-RLX2 conjugate (which demonstrated a similar activity to O-RLX2 when measured in buffer, but lower when measured in the presence of serum proteins), while liposome encapsulation resulted in a slight decrease of bioactivity initially, but prolonged the peptide bioactivity during incubation in serum. It was concluded that liposome encapsulation of RLX2 and synthetic modification to P-RLX2 can both prolong RLX2 peptide in vitro stability; however, the applied chemical conjugation results in a significant loss of bioactivity (cAMP production), whereas the effect of liposome entrapment on RLX2 activity was significantly lower.

Impact of critical process parameters and critical material attributes on the critical quality attributes of liposomal formulations prepared using continuous processing

Liposomes were one of the earliest drug delivery vehicles used for anti-cancer therapeutics and similarly, lipid-based nanoparticles have been used for abundance of applications as gene therapies. The methods to produce these particles have remained relatively unchanged until the recent emergence of continuous manufacturing. Continuous processing enables accelerated development of nanoparticle formulations while providing a scalable manufacturing solution. For this work a continuous processing platform for the production of lipid and polymeric-based nanoparticle formulations has been developed at the University of Connecticut. This research focuses on the formation of liposomes encompassing multiple design of experiments (DoEs) to identify functional relationships between critical process parameters (CPPs), critical material attributes (CMAs), and critical quality attributes (CQAs) for liposomal formulations produced using this continuous processing platform. Liposomes of various sizes and of low polydispersity index (PDI) were produced with different material attributes under various processing conditions. In general, lower mole percentages of cholesterol produced larger particles whereas the mole percent of phosphatidylglycerol did not seem to have a s impact on the size of the liposomes that were produced. The results showed that similarly sized liposomes could be produced with different processing conditions allowing for the flexibility to operate in regions most suitable for formulation components that may be sensitive to certain processing conditions. For example, if the target size of a formulation is 100 nm but the active pharmaceutical ingredient is sensitive to temperature, then the formulation can be manufactured at high (55 °C) or low (30 °C) depending on its characteristics. Additionally, the relationships between CMAs and CPPs were different from conventional liposomal manufacturing methods, allowing for more flexibility when using a continuous processing system. Models that can effectively predict the hydrodynamic diameter of monodispersed liposomes produced using continuous processing were developed. The models developed from the DoEs in this study may be useful for accelerated development of new lipid formulations as well as facilitate the translation from traditional manufacturing methods to continuous manufacturing for products already on the market.

Influence of Linker Orientation and Regulative Factor(s) in Liposomal Gene Delivery: A Molecular Level Investigation.

Molecular level understanding of liposome-gene interaction is immensely important for the research progress and technological advancement of gene delivery, which is highly significant due to a wide range of applications of gene therapy. The liposomal gene delivery method is one of the most promising techniques due to its efficacy to easily fuse with the cell membrane and its lower toxicity. In vivo gene delivery using liposomes is reported to be extremely successful. However, the success of gene delivery depends on various factors including the chemical nature of the structural unit of the liposome. To explore the regulative factor(s) for liposomal gene delivery, we systematically analyze the linker orientation effect on the gene delivery efficiency of liposomes through a density functional theory (DFT) study. Interestingly, it is observed that the liposome-gene interaction is not the regulating factor for successful gene delivery. The success depends on the gel to liquid melting temperature of the liposome.

Regulated bioanalysis of liposomal amphotericin B to support pharmacokinetic studies of liposomal drugs.

Background: Because of the delicate nature of liposomes, bioanalysis of free and liposomal-encapsulated drugs is among the most challenging assays to perform. Current regulatory guidance for bioanalysis is not sufficient to address the complexity of this particular formulation. Method & results: Three individual LC-MS/MS methods to quantify free amphotericin B (10-3000ng/ml) and encapsulated amphotericin B (100-50,000ng/ml) in pretreated human plasma and total amphotericin B (100-50,000ng/ml) in human plasma were fully validated and applied to a bioequivalence study. The acceptance criteria and experimental design of additional validation tests using cross quality control were carefully deliberated a priori and included in the sample analysis as well. Discussion: Additional validation tests are necessary to demonstrate that the measured concentration of the intended component is accurate and free of interference from other coexisting components in the sample. These practices can be used as guidance for future liposomal drug method validation.

Method for loading liposomes with soybean protein isolate hydrolysate influences the antioxidant efficiency of liposomal systems: Adding after liposomes formation or before lipid film hydration

This research investigated the method for loading liposomes with soybean protein isolate hydrolysate (SPIH) to create stable and antioxidative protein hydrolysate-containing liposomes. SPIH (SPIH-AF) prepared via a two-step hydrolysis by alcalase and flavourzyme was added at different concentrations to liposomes by two methods. More effective adsorption of SPIH-AF onto liposomal membrane occurred when it was added at 0.5 mg/mL and distributed in both the inner and outer water phases (addition before lipid film hydration), or added at lower concentrations (< 0.192 mg/mL) and distributed in the outer water phase (addition after liposomes formation). SPIH-AF concentration and distribution directly influenced its early rate of inhibition on lipid oxidation and membrane properties. SPIH-AF in the outer water phase functioned more effectively to inhibit lipid oxidation products accumulation. The interactions between SPIH-AF and liposomes altered SPIH-AF's secondary structures. Compared with Trp-liposomes interactions, Tyr-liposomes interactions were more sensitive to SPIH-AF concentration and distribution. SPIH-AF distribution affected strongly the electrostatic interactions (but weakly the hydrophobic interactions). Tyr/Trp-liposomes interactions were more powerful when SPIH-AF was located in both the inner and outer water phases at higher concentrations.

Investigation of the Role of Induced Overexpression of the Isolated Secreted Klotho on the A-172 Human Glioblastoma Cells.

Klotho gene, identified in 1997 as an anti-aging gene, can manufacture two protein products: transmembrane and secreted forms. The later research revealed the involvement of klotho in carcinogenesis. However, little is known about the action of different Klotho forms on antitumor effects is still. The purpose of this article is to evaluate the effect of isolated secreted Klotho overexpression on the growth features of human glioblastoma cell line A-172.A-172 was transfected by a plasmid vector incorporating secreted Klotho sequence by the liposomal method. Overexpression assay was carried out quantitatively on both mRNA and protein using RT-qPCR and ELISA, correspondingly. It was shown that the relative expression of secreted Klotho in the experimental group was significantly higher than in the untransfected group by both methods (p < 0.001). At the same time, the growth curves and MTT proliferation assay demonstrated significantly decreased values under induced overexpression (p < 0.01). The increased amount of cells with activated caspases and annexin V (p < 0.001) corresponded with the expression of secreted Klotho. This mechanism, as suggested, maybe causative of the observed effects.

Novel and efficient method for loading aptamer-conjugated liposomes with arsenic trioxide for targeting cancer cells

Although the therapeutic effect of liposomal arsenic trioxide arsenic trioxide (ATO) in the treatment of solid tumours has been confirmed, its dose-limiting loading is a challenging issue. To solve the problems in the preparation of liposomal ATO, different loading strategies were evaluated and compared. In addition, liposomes decorated with anti-nucleolin aptamers were developed as a novel formulation for targeted delivery with high loading efficiency and sustained releasing property in order to treat solid tumours. The liposomes were prepared by a thin-film method exploiting the passive loading strategy of Co(II) hydrogen arsenite (CHA). The structural characteristics of the liposomes were also investigated by Fourier transform infra-red spectroscopy (FT-IR), dynamic light scattering (DLS), zeta potentiometry, field emission scanning electron microscopy (FESEM), and Energy Dispersive X-ray Diffraction (EDX) techniques. To evaluate the potential cytotoxicity of this liposomal drug vehicle in vitro, MTT assay was performed on HT-29 cancer cell line. The results showed that the synthesised liposomes loaded with CHA exhibited high entrapment efficiency (77%). MTT assays showed a significant difference between the percentage of viable cells when HT −29 cells were treated with free ATO and liposomal formulation which can be corresponded to the sustained release of the drug from the liposomes. The results of this study may lead to a promising strategy for the effective treatment of solid tumours.

Molecular mechanism of miR-369-3p regulating hepatocellular carcinoma cell proliferation and apoptosis by targeting ACTN4

Objective: To investigate the effect of miR-369-3p targeting ACTN4 expression on proliferation and apoptosis of hepatocellular carcinoma cells. Methods: Real-time quantitative polymerase chain reaction (RT-qPCR) and western blot were used to detect the expression levels of miR-369-3p and ACTN4 in hepatocarcinoma tissues and adjacent tissues. MiR-369-3p mimics, miR-negative control (NC), si-ACTN4, and si-NC were transfected into hepatocellular carcinoma MHCC97H cells by liposome method. Cell proliferation was detected by 3-(4, 5-Dimethylthiazol-2-yl)-2, 5-dipheny-ltetrazolium bromide (MTT) assay. Flow cytometry was used to detect cell cycle and apoptotic rates. The dual luciferase reporter assay was used to verify the targeted regulation of ACTN4 by miR-369-3p. Western blot was used to detect the expressions of cyclin D1, p21, Bcl-2 and Bax. Results: The expression level of miR-369-3p in liver cancer tissue was lower than that in adjacent tissues [(0.46±0.04) vs (1.00±0.08), P<0.001)], while the expression level of ACTN4 was higher than that in adjacent tissues [mRNA (3.12±0.29) vs (1.01±0.09); protein (0.61±0.06) vs (0.25±0.03), P<0.001]. Overexpression of miR-369-3p significantly decreased the cell viability[(0.71±0.06) vs (1.26±0.11), P<0.001)], increased cell apoptosis rate [(20.16±2.11)% vs (6.25±0.64)%, P<0.001], increased the proportion of cells in G(1) phase [(31.14±3.36)% vs (51.56±5.23)%, P<0.001], decreased the proportion of cells in S phase [(32.44±3.56)% vs (14.33) ±1.45)%, P<0.001], increased the levels of p21 and Bax protein (P<0.001), and decreased the levels of cyclin D1 and Bcl-2 protein (P<0.001). Inhibition of the expression of ACTN4 significantly reduced the cell viability [(0.78±0.07) vs (1.24±0.12), P<0.001], increased the apoptosis rate [(6.58±0.66)% vs (18.32±1.82)%, P<0.001], increased the proportion of cells in G(1) phase [(48.69±4.21)% vs (30.33±3.01)%, P<0.001], decreased the proportion of cells in S phase [(36.21±3.42)% vs (18.54±1.61)%, P<0.001], increased the protein levels of p21 and Bax (P<0.001), and decreased the levels of cyclin D1 and Bcl-2 protein (P<0.001). Compared with the miR-369-3p+ pcDNA group, overexpression of ACTN4 increased the proliferation ability of hepatocellular carcinoma MHCC97H cells at 72 hours of culture[(1.12±0.11) vs (0.68±0.06), P<0.001], significantly reduced the proportion of cells in G(1) stage [(38.81±3.24)% vs (51.80±4.57)%, P<0.001], significantly increased the proportion of S-phase cells [(31.65±3.11)% vs (15.69±1.44)%, P<0.001], decreased cell apoptosis rate [(13.86±1.37)% vs (22.69±2.24)%, P<0.001], increased protein expressions of cyclin D1 and Bcl-2 (P<0.001), decreased the protein expressions of p21 and Bax (P<0.001). Conclusion: MiR-369-3p can induce cell cycle arrest in G(1) phase, inhibit the proliferation and promote apoptosis of liver cancer cells by regulating the expression of ACTN4.

Effect of miRNA-223-3p Targeting Stromal Interaction Molecule 1 on Proliferation and Apoptosis of Hypoxia/Reoxygenation-Applied H9c2 Cardiomyocytes

This study was established to determine the effect of miRNA-223-3p on the proliferation and apoptosis of hypoxia/reoxygenation-applied H9c2 cardiomyocytes and the associated mechanisms. A hypoxia/reoxygenation (H/R) model was established, with normal cells also used as a control. miRNA-NC, miRNA-223-3p, anti-miRNA-NC, and anti-miRNA-223-3p plasmids were transfected into normally cultured cardiomyocytes, defined as the miRNA-NC, miRNA-223-3p, anti-miRNA-NC, and anti-miRNA-223-3p groups. In addition, miRNA-223-3p was co-transfected into normally cultured cardiomyocytes with pcDNA3.1 and pcDNA3.1-STIM1 plasmids, followed by treatment with H/R for cells in the miR-NC and miR-223-3p groups, defined as the H/R+miRNA-NC, H/R+miRNA-223-3p, H/R+miRNA-223-3p+pcDNA3.1, and H/R+miRNA-223-3p+pcDNA3.1-STIM1 groups. A liposome method was adopted for assessing transfection. qRT-PCR was used to detect miRNA-223-3p expression, while western blotting was used to detect protein expression. MTT assay was used to detect cell viability, flow cytometry to detect apoptosis, and dual luciferase reporter gene assay to detect fluorescence activity. After H/R treatment, miR-223-3p, cyclin D1, and Bcl-2 expression of cardiomyocytes decreased, p21 and Bax expression significantly increased, cell activity decreased, and the apoptosis rate increased. miRNA-223-3p achieved the targeted regulation of STIM1 expression. miRNA-223-3p overexpression promoted the H/R-induced cardiomyocyte proliferation and inhibited cardiomyocyte apoptosis. STIM1 overexpression reversed the proliferation-promoting and apoptosis-inhibiting effects of miRNA-223-3p on cardiomyocytes treated with H/R. The findings show that miRNA-223-3p overexpression promotes H/R-induced cell proliferation, inhibits apoptosis, and protects H/R-induced cardiomyocytes from injury, via a mechanism probably associated with STIM1 expression. miRNA-223-3p thus provides a new target for treating cardiomyocyte injury.

Differential Expression of MicroRNA-3148 in Patients with Osteoporosis and Its Impacts on the Osteogenic Differentiation of Rat Bone Marrow Mesenchymal Stem Cells

We aimed to explore the effects of rat bone marrow mesenchymal stem cells (BMSCs) on osteogenic differentiation via analyzing miR-3148 expression in patients with osteoporosis. Realtime quantitative PCR was conducted for assessing microRNA-3148 expression. BMSCs from SD rats were transfected with microRNA-3148 mimics and microRNA-3148 inhibitor via liposomal trans-fection method utilizing Lipo2000, followed by analysis of microRNA-3148 level. After 10-days of osteogenic differentiation induction, alkaline phosphatase (ALP) staining and alizarin red (ARS) staining were done to investigate the osteogenic differentiation potential. Simultaneously, qRT-PCR measured the expression of osteogenesis marker genes (BMP and Runx2) in each group. qRT-PCR analysis revealed a high expression of miR-3148 in the bone tissue and the serum samples from patients with osteoporosis in comparison with healthy individuals. In addition, miRNA-3148 mimics could retard the osteogenic differentiation of BMSCs, while microRNA-3148 inhibitor could prompt the procedure. MicroRNA-3148 was highly expressed in the skeletal tissues and the serum samples from patients with osteoporosis and it could restrain the differentiation of BMSCs into osteoblasts, suggesting that it might be a novel therapeutic target for treating osteoporosis.

Niosome as a Novel Pharmaceutical Drug Delivery: A Brief Review Highlighting Formulation, Types, Composition and Application

Abstract: Aim: Formation of niosome by using non-ionic surfactant. The particle size of niosome must be required in the range in between 10 nm -100 nm. This is just due to avoid the aggregation of niosome and show proper result. Materials and Methods: There are many types of niosome, their types and size depend on which method used for preparation. In this article we covered method of preparation of pro-niosome and niosome. Many factors are affecting formation of niosome such as drugs, its chemical and physical property, amount and type of surfactant, cholesterol content and its charge, resistance to osmatic stress as well as membrane composition. Various method is used for separation of unentrapped drug from final product such as dialysis, gel filtration and centrifugation. There are several routes used for administration of niosome such as oral, parenteral, transdermal and ocular etc. Results: The evaluation and characterization of niosome done by many methods such as entrapment efficacy, vesicle diameter, in-vitro release and loading efficiency. Niosome have many pharmaceutical and non-pharmaceutical applications; which having marketed product of niosome are available in market are explain in this article. Conclusion: Niosome formation methods are completely based on liposome methods of preparation. Niosome are having more storage capacity than liposome i.e. niosome are more advance than liposome. The cost of preparation of niosome is also less than liposome. Many pharmaceutical formulations of niosome are now available in market. Key words: Niosome, Pro-niosome, Non-ionic surfactant, Preparation, Application.

Monitoring Ravulizumab effect on complement assays

Ravulizumab is a new C5 inhibitor therapeutic monoclonal antibody with a longer half-life than eculizumab. Monitoring complete complement blockade by eculizumab has allowed personalized therapy in specific settings. Similar action is expected with ravulizumab. Ravulizumab has 4 different amino acids from eculizumab, which allow greater affinity for the FcRn immunoglobulin receptor and change the affinity of the molecule for C5. Here we investigate if clinical lab tests traditionally used to monitor complement blockade for eculizumab are appropriate for monitoring complement blockade caused by ravulizumab.De-identified serum samples with known normal complement activity were spiked with increasing amounts of ravulizumab, from zero to 1000 μg/mL. Measurement of classical pathway function (CH50) and C5 function using a liposome method (Wako Diagnostics) showed >50% complement inhibition starting with 50 μg/mL of ravulizumab, but inhibition >95% of complement activity was not achieved, with residual measurements of 11% at 700 μg/mL. In contrast, measurement of alternative pathway function using an ELISA (AH50, Wieslab) showed alternative pathway function inhibition of 80% at 50 μg/mL of ravulizumab and > 95% at 200 μg/mL, which is consistent with expected therapeutic concentrations of ravulizumab >175 μg/mL. If replicated in patient sera, AH50 could be a suitable therapeutic monitoring tool.

Scalable solvent-free production of liposomes.

A major challenge faced with the manufacture of liposomes is the high volumes of organic solvents used during manufacturing. Therefore, we have implemented an organic solvent-free production method for drug-loaded liposomes and demonstrated its applicability with both aqueous core-loaded and bilayer-loaded drugs. Liposomes were produced by high shear mixing dry powder lipids with an aqueous buffer, followed by down-sizing using a Microfluidizer processor. Liposomes were purified via tangential flow filtration and characterised in terms of size, polydispersity index, zeta potential and drug loading. Doxorubicin-loaded PEGylated liposomes can be manufactured using this solvent-free method with particle sizes of 100-110nm, low polydispersity index (PDI) (<0.2) and high drug loading (97-98%). If required, liposomes can be further down-sized via microfluidic processing without impacting drug loading. Similar results were achieved with non-PEGylated liposomes. With bilayer-loaded amphotericin B liposomes, again liposomes can be prepared within a clinically appropriate size range (100-110nm in size, low PDI) with high drug loading (98-100%). We apply a simple and scalable solvent-free method for the production of both aqueous core or bilayer drug-loaded liposomes.