Liposome Complexes Research Articles

Liposomal 5-Fluorouracil Polymer Complexes Facilitate Tumor-Specific Delivery: Pharmaco-Distribution Kinetics Using Microdialysis.

Liposomes are widely used as carriers for anticancer drugs due to their ability to prolong the retention of encapsulated drugs in blood plasma while directing their distribution increasingly into tumor tissue. We report on the development of stealth liposomal formulations for the common chemotherapy drug 5-fluorouracil, where pharmacokinetic studies were undertaken using a microdialysis probe to specifically quantify drug accumulation in tumor, which was contrasted to drug exposure to healthy tissue. Greater accumulation of the drug into the tumor than into healthy subcutaneous tissue was observed for neutral and cationic liposomal 5-fluorouracil polymer complexes in comparison to the conventional delivery by an injected solution. Increased drug accumulation in tumor also correlated to reduced tumor growth. This research has generated new mechanistic insight into liposomal-specific delivery to tumors with potential to improve the efficacy and reduce the toxicity of chemotherapy.

Easy preparation of a liposome-mediated protein delivery system by freeze-thawing a liposome-protein complex.

Homeostasis can be achieved by adding a protein supplement; however, an appropriate vector is required to deliver the protein into the cell because of the low stability of proteins in the blood and low cell membrane permeability. Here we report an easy one-step method of encapsulating proteins into liposomes for delivery. We used negatively charged superoxide dismutase (SOD) and a polycation liposome as protein and liposome models, respectively. Liposome-encapsulated SOD was prepared by freeze-thawing the SOD-liposome complex (lipoplexes). The amount of immobilized SOD within the lipoplex significantly increased on freeze-thawing. Surprisingly, subjecting the single-layered lipoplexes to freeze-thawing produced multilayered liposomes with SOD localized between the lipid layers. The amount of SOD delivered intracellularly significantly increased by freeze-thawing compared with that delivered by lipoplexes without freeze-thawing. SOD, liposomes, and endosomes were separately localized in the cells. The freeze-thawed lipoplex-encapsulated SOD samples were intravenously injected in mice. The SOD biodistribution was dramatically changed compared with the injection of free SOD or lipoplex. SOD was detached from the lipoplex in the bloodstream after the injection of non-freeze-thawed lipoplex, whereas the encapsulation of SOD in the liposomes upon freeze-thawing enabled the stable circulation of SOD with the liposomes in the bloodstream. This work paves the way for the application of the freeze-thawing technology for the easy one-step encapsulation of proteins into liposomes for protein delivery.

Reduction of plasmid vector backbone length enhances reporter gene expression

Gene therapy has a wide range of applications for various types of pathologies. Viral methods of gene delivery provide high levels of gene expression but have various safety concerns. Non-viral methods are largely known to provide lower levels of expression. We aim to address this issue by using plasmid DNA with smaller backbones to increase gene expression levels when delivered using non-viral methods. In this study we compare gene expression levels between two vectors with firefly luciferase encoding gene insert using liposome complexes and gene electrotransfer as delivery methods. A 2-fold reduction in plasmid vector backbone size, disproportionately enhanced gene expression levels more than 10-fold in rat tenocytes in vitro, and rat myocardium in vivo, while improvements in delivery to the skin were more moderate.

Interaction of multicomponent anionic liposomes with cationic pyridylphenylene dendrimer: Does the complex behavior depend on the liposome composition?

Dendrimers are individual macromolecular compounds having a great potential for biomedical application. The key step of the cell penetration by dendrimers is the interaction with lipid bilayer. Here, the interaction between cationic pyridylphenylene dendrimer of third generation (D350+) and multicomponent liquid (CL/POPC), solid (CL/DPPC) and cholesterol-containing (CL/POPC/30% Chol) anionic liposomes was investigated by dynamic light scattering, fluorescence spectroscopy, conductometry, calorimetric studies and molecular dynamic (MD) simulations. Microelectrophoresis and MD simulations revealed the interaction is electrostatic and reversible with only part of pyridinium groups of dendrimers involved in binding with liposomes. The ability of dendrimer molecules to migrate between liposomes was discovered by the labeling liposomes with Rhodamine B. The phase state of the lipid membrane and the incorporation of cholesterol into the lipid bilayer were found to not affect the mechanism of the dendrimer - liposome complex formation. Rigid dendrimer adsorption on liposomal surface does not induce the formation of significant defects in the lipid membrane pave the way for possible biological application of pyridylphenylene dendrimers.

Preparation and characterization of egg yolk immunoglobulin loaded chitosan-liposome assisted by supercritical carbon dioxide

The egg yolk immunoglobulin (IgY) loaded chitosan-liposomes (IgY-CS-LP) were prepared and assisted by supercritical carbon dioxide (SCCO2). The effects of phospholipid type and SCCO2 pressure on particle size, zeta potential, encapsulation efficiency, structural properties and stabilities were investigated. The results showed that the liposomes prepared by egg yolk phosphatidylcholine (EPC) had better homogeneity and higher encapsulation rate than those by soybean phosphatidylcholine (SPC). With the increase in critical pressure, the particle size decreased dramatically and became more uniform. Under pressure of 20 MPa, it showed a preferable stability on IgY-CS-LP and superior encapsulation efficiency of IgY (76.85%). Besides, IgY could be wrapped in the phospholipid layer which has strong interaction with chitosan. The results suggested that chitosan liposome complex could form an effective carrier for IgY with method of SCCO2, which can solve the problem of IgY inactivation in vivo, so as to enhance human immunity and other effects.

Preclinical Safety Evaluation of a Recombinant Plasmid Vector Encoding Mature Human Neutrophil Peptide-1 by Repeated Local Administrations in Nonhuman Primates.

In our previous studies, a novel gene therapy approach was developed based on a plasmid vector pSecTag2B in which recombinant HNP1 gene was regulated under a cytomegalovirus promoter to encode a mature human neutrophil peptide-1 (HNP1) form. We showed for the first time in various tumor models, including human cancer xenografts, that overexpression of HNP1 in the tumor milieu by intratumoral pSecTag-HNP1 (pHNP1) administration efficiently attenuated in vivo tumor progression, mediated host immune responses to tumors, and produced a synergistic effect when combined with chemotherapeutics. In this study, a preclinical safety investigation of HNP1 gene therapy was conducted in nonhuman primates. Eleven cynomolgus monkeys were divided into three groups of three to four animals each and received either repeated s.c. injections of pHNP1/cationic liposome complexes at a low (0.625 mg/kg) or a high (2.5 mg/kg) dose or glucose as control. Significant HNP1 in vivo accumulation was detected after consecutive administrations. All primates reached the end of the study with good body conditions. Injection site inflammation was the only obvious toxic reaction during observation period. In addition, elevation of monocyte/macrophage and neutrophil as well as decline of lymphocyte were detected in the peripheral blood of pHNP1-treated primates. These alterations were partially alleviated at the end of observation period. Besides, dose-related histopathological changes of the immune organs were observed at necropsy, including a minimal thymic lymphocyte decrease and a minimal-to-mild lymph node erythrocyte increase, but which cannot be excluded from HNP1-induced immune reactions. Together, these data support future clinical studies of pHNP1-based local gene delivery in tumor patients.

Effect of using amino acids in the freeze-drying of siRNA lipoplexes on gene knockdown in cells after reverse transfection.

Recently, small interfering RNA (siRNA)/cationic liposome complexes (siRNA lipoplexes) have become a crucial research tool for studying gene function. Easy and reliable siRNA transfection with a large set of siRNAs is required for the successful screening of gene function. Reverse (Rev)-transfection with freeze-dried siRNA lipoplexes is validated for siRNA transfection with a large set of siRNAs in a multi-well plate. In our previous study, it was shown that Rev-transfection with siRNA lipoplexes freeze-dried in disaccharides or trisaccharides resulted in long-term stability with a high level of gene-knockdown activity. In the present study, the effects of amino acids used as cryoprotectants in the freeze-drying of siRNA lipoplexes on gene knockdown via Rev-transfection were assessed. A total of 15 types of amino acids were used to prepare freeze-dried siRNA lipoplexes, and it was found that the freeze-drying of siRNA lipoplexes with amino acid concentrations >100 mM strongly suppressed targeted gene expression regardless of the amino acid type; however, some amino acids strongly upregulated or downregulated gene expression in the cells transfected with negative control siRNA. Amongst the amino acids tested, the presence of asparagine showed specific gene-knockdown activity, forming large cakes after freeze-drying and retaining a favorable siRNA lipoplex size after rehydration. These findings provide valuable information regarding amino acids as cryoprotectants for Rev-transfection using freeze-dried siRNA lipoplexes for the efficient delivery of siRNA into cells.

Effects of sterol derivatives in cationic liposomes onbiodistribution and gene-knockdown in the lungs of mice systemically injected with siRNA lipoplexes.

Cationic liposomes can be intravenously injected to deliver short interfering (si)RNAs into the lungs. The present study investigated the effects of sterol derivatives in systemically injected siRNA/cationic liposome complexes (siRNA lipoplexes) on gene-knockdown in the lungs of mice. Cationic liposomes composed of 1,2-dioleoyl-3-trimethylammonium-propane or dimethyldioctadecylammonium bromide (DDAB) were prepared as a cationic lipid, with sterol derivatives such as cholesterol (Chol), β-sitosterol, ergosterol (Ergo) or stigmasterol as a neutral helper lipid. Transfected liposomal formulations composed of DDAB/Chol or DDAB/Ergo did not suppress the expression of the luciferase gene in LLC-Luc and Colon 26-Luc cells in vitro, whereas other formulations induced moderate gene-silencing. The systemic injection of siRNA lipoplexes formulated with Chol or Ergo into mice resulted in abundant siRNA accumulation in the lungs. In comparison, systemically injected DDAB/Chol or DDAB/Ergo lipoplexes of Tie2 siRNA effectively increased the suppression of the Tie2 mRNA expression in the lungs of mice. These findings indicated that DDAB/Chol and DDAB/Ergo liposomes could function as vectors for siRNA delivery to the lungs.

Potential Of Curcumin Loaded Nanoparticles In Antimicrobial Photodyanamic Therapy

In the past decade, multi-drug resistance of pathogenic bacteria has become a serious problem to public health. Finding novel approaches to combat multi-drug resistant bacteria have therefore become increasingly important. One promising approach is antimicrobial photodynamic therapy (aPDT) which involves the use of photo-sensitizer (nontoxic dyes) that are excited by visible light and produce oxygen free radicals in the presence of oxygen. Many studies report that curcumin inhibits the growth of both gram-positive and gram-negative bacteria. Formulations of curcumin in liposome, nanoparticles, micelles and phospholipids complexes are being prospected to decline its hydrophobicity, increase solubility to amplify blood flow time, and its permeability through membrane barriers. Among these options nano-formulations of curcumin such as nano-crystals, nano-emulsions and polymeric nanoparticle encapsulated curcumin got enormous thrust and advance in the recent years in the category of natural or herbal drugs.. There are evidences that support the enhancement of antimicrobial activity and antibiofilm activities of curcumin silica nanoparticles of P.aeruginosa and S.aureus cultures using aPDT. This therapy exhibits significant antimicrobial activity against both bacteria. This review provides an updated information regarding the use of natural products to limit or prevent microbial resistance through use of photosensitizing agents like curcumin.

Carbon Nanotube–Liposome Complexes in Hydrogels for Controlled Drug Delivery via Near-Infrared Laser Stimulation

Externally controllable drug delivery systems are crucial for a variety of biological applications where the dosage and timing of drug delivery need to be adjusted based on disease diagnosis and pr...

Impact of Quercetin Encapsulation with Added Phytosterols on Bilayer Membrane and Photothermal-Alteration of Novel Mixed Soy Lecithin-Based Liposome.

This study used highly lipophilic agents with an aim to increase the oxidant inhibitory activity and enhance photothermal stability of a novel mixed soy lecithin (ML)-based liposome by changing the composition of formulation within the membrane. Specifically, the development and optimization of the liposome intended for improving Trolox equivalent antioxidant capacity (TEAC) value and %TEAC loss was carried out by incorporating a natural antioxidant, quercetin (QU). In this context, a focus was set on QU encapsulation in ML-based liposomes and the concentration-dependent solubility of QU was investigated and calculated as encapsulation efficiency (EE). To explore the combined effects of the incorporation of plant sterols on the integrity and entrapment capacity of mixed phospholipid vesicles, conjugation of two types of phytosterols (PSs), namely β-sitosterol (βS) and stigmasterol (ST), to mixed membranes at different ratios was also performed. The EE measurement revealed that QU could be efficiently encapsulated in the stable ML-based liposome using 0.15 and 0.1 g/100 mL of βS and ST, respectively. The aforementioned liposome complex exhibited a considerable TEAC (197.23%) and enhanced TEAC loss (30.81%) when exposed to ultraviolet (UV) light (280–320 nm) over a 6 h duration. It appeared that the presence and type of PSs affect the membrane-integration characteristics as well as photodamage transformation of the ML-based liposome. The association of QU with either βS or ST in the formulation was justified by their synergistic effects on the enhancement of the EE of liposomes. Parallel to this, it was demonstrated that synergistic PS effects could be in effect in the maintenance of membrane order of the ML-based liposome. The findings presented in this study provided useful information for the development and production of stable QU-loaded ML-based liposomes for food and nutraceutical applications and could serve as a potential mixed lipids-based delivery system in the disease management using antioxidant therapy.

Вірусінфікований калюс квасолі та його оздоровлення in vitro за дії ліпосомальних препаратів

The aim of the study was to develop a recovery means for beans infected by Bean yellow mosaic virus (BYMV) as well as Bean common mosaic virus (BCMV) using callus culture and liposomal glycan preparations. Methods. Cultivation of explants and callus cultures was carried out in vitro using conventional methods of plant biotechnology. The tissue culture propagation was performed during the spring or summer seasons. The presence of viral infection was tested by reverse transcription polymerase chain reaction. The virus-specific primers that allowed amplifying the conserved regions of the capsid protein gene of BCMV or BYMV were used for virus identification. Results. The culture of bean callus infected with BCMV was obtained and adapted for antiviral agents testing. It has been shown that during long-term cultivation (10–12 weeks) in the presence of liposomal preparation containing Ganoderma adspersum glucan (10–100 mg/l), plant tissue culture become free from viruses following virus eradication. This is evidenced by the absence in the callus tissue of 391 bp sequences typical for the virus coat protein gene. Conclusions. The full suppression of virus reproduction and gradual elimination of virus occurred in callus tissue obtained from BCMV-infected beans and cultured on B-5 medium supplemented with liposomal glycanglycolipid complex (10–100 mg/l). The data obtained can be useful for the development of practical control method to cure plant virus diseases using callus culture and antiviral-active glycan-glycolipid complexes.

INCREASING SAFETY OF SURGICAL INTERVENTIONS IN PATIENTS WITH VENTRAL AND INCISIONAL HERNIA UNDER COMORBIDITY CONDITIONS WHEN PERFORMING DIFFERENT TYPES OF OPEN HERNIOPLASTY

Abstract. In the period from 2001 to 2017, 1419 patients with ventral and postoperative ventral hernia were operated on. The unsatisfactory results of the surgical treatment of this pathology were due to a number of disadvantages: the choice of an inadequate method of hernioplasty in a specific clinical situation, an incomplete revision of the muscle-aponeurotic layer of the anterior abdominal wall, especially in patients with postoperative veterinary hernias, concomitant syndrome of undifferentiated connective tissue dysplasia and obesity, and weak points, the wrong choice of the type of mesh implant (“light” or “heavy” polypropylene mesh) and its size, the choice of an inappropriate type of suture material, as well as the high invasiveness of surgical intervention. Based on the given causes of complications in the operated patients with a high risk of their development, we have made a technical improvement of the existing methods of hernioplasty and development of new ones, which significantly reduced the number of postoperative complications and increased the safety of performing surgical interventions in patients with ventral and postoperative ventral hernia.
 Purpose. Based on the results of the analysis of surgical treatment of patients with comorbid conditions for primary and incisional ventral hernias to develop technical prerequisites for improving the safety of surgical interventions in this category of patients.
 Methods. Іn the period from 2001 to 2017, an in-depth comprehensive clinical-instrumental and laboratory examination of 1419 patients with primary ventral hernia (PVH) and postoperative ventral hernia (PVH) was conducted. The patients were divided into groups according to the periods of surgical treatment of patients. For this purpose, two periods were formed: from 2001 to 2009 and from 2010 to 2017. At the first period, 597 (42.07%) patients were examined and operated on. These patients formed a comparison group. The main group corresponding to patients who were treated in the period from 2010 to 2017 was 822 (57.93%) people.
 Results. Developed techniques for performing retromuscular allogernioplasty, which were based on controlled visualization of suturing when fixing polypropylene mesh to the posterior leaf of the vagina of the rectus muscle, allowed us to reliably and safely place and secure the implant in the retromuscular space, significantly reduce the trauma and duration of surgery.
 Kind of treatment using the mesh allograft with a liposomal complex with included antibacterial and immunosuppressive agents during the operation on the "onlay" method significantly reduced the number of local infectious complications. The location of the PRF membrane over the polypropylene mesh during retromuscular allogernioplasty improved neoangiogenesis at the mesh implantation site, increased fibroblast activity and the formation of collagen fibers around the mesh material, which provided maximum integration of the "light" abdominal wall with polypropylene tissue.
 Conclusion. These features of the dynamics of the used methods of surgical interventions and types of polypropylene nets with an individualized approach to each patient were reflected in a significant reduction in the number of both early local and general late postoperative complications, which improved the effectiveness of surgical treatment of patients with primary postoperative ventral hernias.

Effect of pre‑freezing and saccharide types in freeze‑drying of siRNA lipoplexes on gene‑silencing effects in the cells by reverse transfection.

Our previous study reported that reverse (Rev)-transfection with small interfering RNA (siRNA)/cationic liposome complexes (siRNA lipoplexes) freeze-dried in trehalose or sucrose solution resulted in high gene-silencing activity in cells. The current study investigated whether pre-freezing or saccharide types present during the freeze-drying of siRNA lipoplexes affected gene-silencing in cells after Rev-transfection. Three types of cationic cholesterol derivatives and three types of dialkyl or trialkyl cationic lipids were used for the preparation of cationic liposomes. Additionally, six types of siRNA lipoplexes were vacuum-dried in trehalose or sucrose solution without a pre-freezing process in multi-well plates. A strong gene-silencing activity after Rev-transfection was observed regardless of the cationic lipid types in the cationic liposomes. It was also investigated whether saccharide types in the freeze-drying of siRNA lipoplexes affected gene-silencing after Rev-transfection. siRNA lipoplexes freeze-dried in monosaccharides (glucose, fructose, galactose or mannose), disaccharides (maltose, lactose, lactulose or cellobiose) and trisaccharide solution (raffinose or melezitose) demonstrated high gene-silencing activity. However, following Rev-transfection with siRNA lipoplexes freeze-dried in monosaccharides or trisaccharides, certain saccharides induced cytotoxicity and/or off-target effects. The results of the current study indicated that disaccharides may be suitable for the preparation of vacuum-dried or freeze-dried siRNA lipoplexes for Rev-transfection.

Targeted Myoglobin Delivery as a Strategy for Enhancing the Sensitivity of Hypoxic Cancer Cells to Radiation.

SummaryThe effectiveness of cancer radiotherapy is frequently hindered by the hypoxia of the tumor microenvironment. Direct delivery of oxygen to hypoxic tumor tissues is an attractive strategy to overcome this hypoxia-associated radioresistance. Herein, we report the generation of submicron-sized particles comprising myoglobin fused to the crystal-forming domain of Cry3Aa protein for the targeted delivery of oxygen to cancer cells. We demonstrate that myoglobin-containing particles were successfully produced in Bacillus thuringiensis with the assistance of the Cry3Aa domain I. Furthermore, these particles could be genetically modified to incorporate the cell penetrating peptide TAT and cell targeting peptide A549.1, resulting in particles that exhibited improved cellular uptake and targeting toward A549 cells. Notably, these myoglobin-containing particles increased the intracellular oxygen levels of A549 cells and thereby sensitized them to radiation. These findings suggest that the targeted delivery of O2-bound myoglobin could be an effective approach to enhance the efficacy of radiotherapy.

Multifunctional carriers for controlled drug delivery

Abstract In the review we describe a method for concentration of anionic liposomes with encapsulated water-soluble substances within a small volume via electrostatic liposome adsorption on the surface of polymer particles with grafted cationic chains (spherical polycationic brushes), or cationic microgel particles. Dozens of intact liposomes can be bound to each polymer particle, the resulting polymer/liposome complex does not dissociate into the original components in a physiological solution. This allows fabrication of multi-liposomal complexes (MLCs) with a required ratio of encapsulated substances. Two approaches are discussed for the synthesis of stimuli-sensitive MLCs. The first is to incorporate the conformation switch, morpholinocyclohexanol-based lipid, into the liposomal membrane thus forming pH-sensitive liposomes capable of releasing their cargo when acidifying the surrounding solution. These liposomes complexed with the brushes release encapsulated substances much faster than the uncomplexed liposomes. The second is to adsorb liposomes on cationic thermo-responsive microgels. The resulting MLCs contracts upon heating over a volume phase transition temperature from the swollen to the collapsed state of microgel, thus causing the adsorbed liposomes to change drastically their morphology and release an encapsulated substance. Complexation of anionic liposomes with chitosan microgels and polylactide micelles gives MLCs which degrade in the presence of enzymes down to small particles, 10–15 nm in diameter. A novel promising approach suggests that immobilized liposomes can act as a capacious depot for biologically active compounds and ensure their controllable leakage to surrounding solution.

A molecular view on the escape of lipoplexed DNA from the endosome.

The use of non-viral vectors for in vivo gene therapy could drastically increase safety, whilst reducing the cost of preparing the vectors. A promising approach to non-viral vectors makes use of DNA/cationic liposome complexes (lipoplexes) to deliver the genetic material. Here we use coarse-grained molecular dynamics simulations to investigate the molecular mechanism underlying efficient DNA transfer from lipoplexes. Our computational fusion experiments of lipoplexes with endosomal membrane models show two distinct modes of transfection: parallel and perpendicular. In the parallel fusion pathway, DNA aligns with the membrane surface, showing very quick release of genetic material shortly after the initial fusion pore is formed. The perpendicular pathway also leads to transfection, but release is slower. We further show that the composition and size of the lipoplex, as well as the lipid composition of the endosomal membrane, have a significant impact on fusion efficiency in our models.

Сationic liposomes as delivery systems for nucleic acids

Objectives. Gene therapy is based on the introduction of genetic material into cells, tissues, or organs for the treatment of hereditary or acquired diseases. A key factor in the success of gene therapy is the development of delivery systems that can efficiently transfer genetic material to the place of their therapeutic action without causing any associated side effects. Over the past 10 years, significant effort has been directed toward creating more efficient and biocompatible vectors capable of transferring nucleic acids (NAs) into cells without inducing an immune response. Cationic liposomes are among the most versatile tools for delivering NAs into cells; however, the use of liposomes for gene therapy is limited by their low specificity. This is due to the presence of various biological barriers to the complex of liposomes with NA, including instability in biological fluids, interaction with serum proteins, plasma and nuclear membranes, and endosomal degradation. This review summarizes the results of research in recent years on the development of cationic liposomes that are effective in vitro and in vivo. Particular attention is paid to the individual structural elements of cationic liposomes that determine the transfection efficiency and cytotoxicity. The purpose of this review was to provide a theoretical justification of the most promising choice of cationic liposomes for the delivery of NAs into eukaryotic cells and study the effect of the composition of cationic lipids (CLs) on the transfection efficiency in vitro.Results. As a result of the analysis of the related literature, it can be argued that one of the most promising delivery systems of NAs is CL based on cholesterol and spermine with the addition of a helper lipid DOPE. In addition, it was found that varying the composition of cationic liposomes, the ratio of CL to NA, or the size and zeta potential of liposomes has a significant effect on the transfection efficiency.Conclusions. Further studies in this direction should include optimization of the conditions for obtaining cationic liposomes, taking into account the physicochemical properties and established laws. It is necessary to identify mechanisms that increase the efficiency of NA delivery in vitro by searching for optimal structures of cationic liposomes, determining the ratio of lipoplex components, and studying the delivery efficiency and properties of multicomponent liposomes.

Ultrasound-responsive neurotrophic factor-loaded microbubble- liposome complex: Preclinical investigation for Parkinson's disease treatment

Ultrasound-targeted microbubble destruction (UTMD) in conjunction with neurotrophic factors (NFs) gene delivery has the potential to facilitate the penetration of therapeutic genes into the brain for neuroprotective therapy against neurodegenerative diseases. We previously presented a gene delivery system that conjugates gene-carrying liposomes with microbubbles (MBs) to open the blood-brain barrier (BBB) for the delivery of genes into the brain. Since both glia cell line-derived neurotrophic factor (GDNF) and brain-derived neurotrophic factor (BDNF) can protect dopaminergic neurons from neurotoxicity demonstrated in Parkinson's disease (PD) animal models, the present study seeks (1) to develop a novel gene-nanocarrier MB complex carrying BDNF or GDNF gene and (2) to protect dopaminergic neurons in a mouse model of PD via the proposed UTMD system. In the experimental design, PD animals received treatment that delivered GDNF, BDNF, or combined GDNF/BDNF in conjunction with UTMD treatment, and pathological changes in dopamine neurons were histologically examined. Rotarod assay was employed to evaluate the motor behavior. Our results demonstrate that either BDNF or GDNF gene delivery via the UTMD system provides a neuroprotective effect with evidence of improvements of behavioral deficits, decreased calcium influx, GFAP and caspase 3 expression, and rescued dopaminergic neuronal loss. Simultaneously performing GDNF/BDNF gene delivery did not show additional benefits beyond individually delivering BDNF or GDNF genes, possibly due to a hampering effect of simultaneous GDNF/BDNF competing expressions, thus dampening the overall therapeutic effect. In conclusion, these results suggest that UTMD in conjunction with delivery of GDNF or BDNF gene can synergistically serve as an effective gene therapy strategy for neurodegenerative diseases.

Targeting folate receptor \u03b2 positive tumor-associated macrophages in lung cancer with a folate-modified liposomal complex

Tumor-associated macrophages (TAMs) facilitate cancer progression by promoting tumor invasion, angiogenesis, metastasis, inflammatory responses, and immunosuppression. Folate receptor β (FRβ) is overexpressed in TAMs. However, the clinical significance of FRβ-positive macrophages in lung cancer remains poorly understood. In this study, we verified that FRβ overexpression in lung cancer TAMs was associated with poor prognosis. We utilized a folate-modified lipoplex comprising a folate-modified liposome (F-PLP) delivering a BIM-S plasmid to target both lung cancer cells and FRβ-positive macrophages in the tumor microenvironment. Transfection of LL/2 cells and MH-S cells with F-PLP/pBIM induced cell apoptosis. Injection of F-PLP/pBIM into LL/2 and A549 lung cancer models significantly depleted FRβ-positive macrophages and reduced tumor growth. Treatment of tumor-bearing mice with F-PLP/pBIM significantly inhibited tumor growth in vivo by inducing tumor cell and macrophage apoptosis, reducing tumor proliferation, and inhibiting tumor angiogenesis. In addition, a preliminary safety evaluation demonstrated a good safety profile of F-PLP/pBIM as a gene therapy administered intravenously. This work describes a novel application of lipoplexes in lung cancer targeted therapy that influences the tumor microenvironment by targeting TAMs.