Liposomal Carrier Research Articles

Nanoparticles use for Delivering Ursolic Acid in Cancer Therapy: A Scoping Review

Ursolic acid is a natural pentacyclic triterpenoid that exerts a potent anticancer effect. Furthermore, it is classified as a BCS class IV compound possessing low permeability and water solubility, consequently demonstrating limited bioavailability in addition to low therapeutic effectiveness. Nanoparticles are developed to modify the physical characteristics of drug and can often be produced in the range of 30–200 nm, providing highly effective cancer therapy due to the Enhanced Permeation and Retention (EPR) Effect. This study aims to provide a review of the efficacy and safety of various types of Ursolic Acid-loading nanoparticles within the setting of preclinical and clinical anticancer studies. This literature study used scoping review method, where the extracted data must comply with the journal inclusion criteria of within years of 2010–2020. The identification stage produced 237 suitable articles. Duplicate screening was then conducted followed by the initial selection of 18 articles that had been reviewed and extracted for data analysis. Based on this review, the use of nanoparticles can be seen to increase the anticancer efficacy of Ursolic Acid in terms of several parameters including pharmacokinetic data, survival rates and inhibition rates, as well as the absence of serious toxicity in preclinical and clinical trials in terms of several parameters including body weight, blood clinical chemistry, and organ histipathology. Based on this review, the use of nanoparticles has been able to increase the anticancer efficacy of Ursolic Acid, as well as show the absence of serious toxicity in preclinical and clinical trials. Evenmore, the liposome carrier provides development data that has reached the clinical trial phase I. The use of nanoparticle provides high potential for Ursolic Acid delivery in cancer therapy.

In vitro applicability of mixed soy lecithin-based liposomes with added several lipophilic agents as novel delivery systems for delivery of quercetin

The study used highly lipophilic compounds to evaluate membrane physical and constitutional changes and vesicle stabilization of mixed soy lecithin (ML)-based liposomes. The in vitro bio-accessibility of novel quercetin (QU)-loaded ML-based liposomes as a function of particle size, polydispersity index (PDI), zeta potential (ZP), different membrane stabilizers, membrane fluidity and encapsulation efficacy (EE) was examined. Three different membrane systems, empty ML-based liposomes (E-LPs), squalene (SQ) loaded LPs (SQ-LPs), and SQ and a combination of β-sitosterol (βS) and stigmasterol (ST) loaded LPs (SQ-βS-ST-LPs), were developed based on optimized formulations using an extrusion method and used to encapsulate QU. The applicability of the ML-based liposomal carriers was tested by means of an in-vitro digestion procedure by applying a 2-phase static in- vitro model of the stomach and small intestine allowing for measurement of the bio-accessibility of ingested QU. Stability comparison of the empty liposomal particles, SQ-LPs and SQ-βS-ST-LPs particles was carried out to determine the most appropriate form of QU-loaded ML-based liposomal formulation. Different formulations were observed to possess different amounts of bio-accessible QU, indicating that membrane additives had profound effect on micellization of QU during Gastro-Intestinal Tract (GIT). Bivariate correlation analysis indicated that the %bio-accessibility and %EE of QU were strongly linearly correlated (R2 = 0.989), suggesting that QU bio-accessibility depended strongly on the retaining of QU in the ML bilayer, and entrapment capacity of the system. Our results paint a consistent picture in which the vesicle integrity, membrane fluidity and membrane-stabilizing function varied upon changing the membrane lipid composition in the ML-based-liposomes.

Evaluation of the Effect of Liposome Carriers and Albumin Nanoparticles Containing Activated Melittin on Inhibiting the Growth of Leishmania Major Amastigote in vivo

Evaluation of the Effect of Liposome Carriers and Albumin Nanoparticles Containing Activated Melittin on Inhibiting the Growth of Leishmania Major Amastigote in vivo

Preparation, characterization, pharmacokinetics, and antirenal injury activity studies of Licochalcone A-loaded liposomes.

A liposome of Licochalcone A (LCA-Liposomes) was purposively prepared to ameliorate the low in vivo availability and efficacy of LCA. Physical characterization of LCA-Liposomes was carried out mainly by determining particle size, morphology, zeta potential (Z-potential), and efficiency of LCA encapsulation (EE) via appropriate techniques. Also, the rate of LCA release in vitro and distribution in vivo (plasma and tissues) was evaluated. Evaluation of the antirenal activity of LCA-liposomes was carried out by establishing chronic renal failure (CRF) model in mice through intragastric administration of adenine (200mg/kg) and subsequent determination of biochemical parameters and examination of tissue sections. Respectively, the mean size of liposomal particles, Z-potential and EE of LCA-Liposomes were 71.78±0.99nm, -38.49±0.06mV, and 97.67±1.72%. Pharmacokinetic and tissue distribution studies showed that LCA-Liposomes could improve the availability of LCA in the blood and tissues, whereas during pharmacodynamics studies, the liposome effectively improved the therapeutic effect of LCA on CRF mice by potentially protecting the renal tissues while exhibiting antioxidant activity. In conclusion, LCA-Liposomes could effectively improve the bioavailability of LCA and provide platform for the development of LCA-related functional products. PRACTICAL APPLICATIONS: As a traditional Chinese medicine, licorice is widely used in food and pharmaceutical industries. LCA is a small molecule flavonoid extracted from the root of licorice. In this study, LCA was loaded on liposome carriers, which significantly improved the water solubility and oral bioavailability, and proved that LCA-Liposomes have certain therapeutic effects on chronic renal failure, thereby providing a basis for the development of LCA into drugs or functional food in the future.

Pressure-Biased Nanopores for Excluded Volume Metrology, Lipid Biomechanics, and Cell-Adhesion Rupturing.

Nanopore sensing has been widely used in applications ranging from DNA sequencing to disease diagnosis. To improve these capabilities, pressure-biased nanopores have been explored in the past to-primarily-increase the residence time of the analyte inside the pore. Here, we studied the effect of pressure on the ability to accurately quantify the excluded volume which depends on the current drop magnitude produced by a single entity. Using the calibration standard, the inverse current drop (1/ΔI) decreases linearly with increasing pressure, while the dwell drop reduces exponentially. We therefore had to derive a pressure-corrected excluded volume equation to accurately assess the volume of translocating species under applied pressure. Moreover, a method to probe deformation in nanoliposomes and a single cell is developed as a result. We show that the soft nanoliposomes and even cells deform significantly under applied pressure which can be probed in terms of the shape factor which was introduced in the excluded volume equation. The proposed work has practical applications in mechanobiology, namely, assessing the stiffness and mechanical rigidity of liposomal drug carriers. Pressure-biased pores also enabled multiple observations of cell-cell aggregates as well as their subsequent rupture, potentially allowing for the study of microbial symbioses or pathogen recognition by the human immune system.

Drug development of lyso-thermosensitive liposomal doxorubicin: Combining hyperthermia and thermosensitive drug delivery.

We review the drug development of lyso-thermosensitive liposomal doxorubicin (LTLD) which is the first heat-activated formulation of a liposomal drug carrier to be utilized in human clinical trials. This class of compounds is designed to carry a payload of a cytotoxic agent and adequately circulate in order to accumulate at a tumor that is being heated. At the target the carrier is activated by heat and releases its contents at high concentrations. We summarize the preclinical and clinical experience of LTLD including its successes and challenges in the development process.

Vesicular and Stealth Vesicular Drug Delivery – A Review

Vesicular systems (liposomes), one of several potential novel drug delivery systems, provide an advanced technology for delivering active compounds to the site of action, and numerous formulations are currently in clinical use. Liposome technology has developed from typical vesicles to sterically stabilised vesicles, which produce long-circulating liposomes by varying the lipid content, size, and charge of the vesicle. Several compounds, such as glycolipids, have been used to create liposomes with changed surfaces. The addition of the synthetic polymer poly-(ethylene glycol) (PEG) in liposome composition was a crucial milestone in the creation of long-circulating liposomes. PEG on the liposomal carrier's surface has been found to increase blood circulation time while decreasing mononuclear phagocyte system uptake (Stealth Liposomes). As a consequence of this technique, a vast variety of liposome formulations encapsulating active compounds have been developed, all of which have excellent target efficiency and activity. Stealth liposomes can also be actively targeted with monoclonal antibodies or ligands thanks to a synthetic alteration of the terminal PEG molecule. This article focuses on vesicular drug delivery as well as stealth technology and presents preclinical and clinical data for the most common liposome formulations, as well as discussing developing developments in this promising technology.

Ultrasound-Triggered Delivery of Iproplatin from Microbubble-Conjugated Liposomes.

The PtIV prodrug iproplatin has been actively loaded into liposomes using a calcium acetate gradient, achieving a 3‐fold enhancement in drug concentration compared to passive loading strategies. A strain‐promoted cycloaddition reaction (azide‐ dibenzocyclooctyne) was used to attach iproplatin‐loaded liposomes L(Pt) to gas‐filled microbubbles (M), forming an ultrasound‐responsive drug delivery vehicle [M−L(Pt)]. Ultrasound‐triggered release of iproplatin from the microbubble‐liposome construct was evaluated in cellulo. Breast cancer (MCF‐7) cells treated with both free iproplatin and iproplatin‐loaded liposome−microbubbles [M−L(Pt)] demonstrated an increase in platinum concentration when exposed to ultrasound. No appreciable platinum uptake was observed in MCF‐7 cells following treatment with L(Pt) only or L(Pt)+ultrasound, suggesting that microbubble‐mediated ultrasonic release of platinum‐based drugs from liposomal carriers enables greater control over drug delivery.

Magnetic Resonance Imaging-Guided Multi-Stimulus-Responsive Drug Delivery Strategy for Personalized and Precise Cancer Treatment.

The emergence of nano-targeted controlled release liposomal drug carriers has provided a breakthrough in cancer therapy. However, their clinical efficacy is unsatisfactory, which is related to individualized differences in targeted drugs and poor in vivo release efficiency. In this paper, we prepared a class of personalized targeted and precisely controlled-release therapeutic drug carriers (GF liposomes) by co-assembling targeting and traceable o-nitrobenzyl ester lipids to propose a magnetic resonance imaging (MRI)-guided personalized in vivo targeted drug screening strategy and a multi-stimulus superimposed controlled-release strategy. Furthermore, by following the drug release process of drug-loaded liposomes (GF-D), it was found that these liposomes could rely on energy superposition to achieve more sensitive and efficient controlled drug release. In addition, the indispensable adjustment of liposome formulation for personalized MRI-based targeted therapy was verified by differential cellular uptake and in vivo magnetic resonance imaging. In the end, the 10.22-fold tumor suppression effect in the stimulus superposition group (GF-D-UV) indicates that the multi-stimulus cumulative response strategy and MRI-guided in vivo screening strategy can more effectively treat cancer. This contribution provides a concise and clever design idea for the future development of personalized precise and efficient clinical cancer therapies.

Gold nanoparticle-coated thermosensitive liposomes for the triggered release of doxorubicin, and photothermal therapy using a near-infrared laser

We constructed gold nanoparticle-coated liposome/doxorubicin (GCL/DOX) for the purpose of combination therapy and focused on 1) controlled drug release from the thermosensitive liposomes triggered by near-infrared laser (NIR-laser) irradiation and 2) the photothermal effect of the gold nanoshell-like structures upon absorbing laser light. GCL/DOX was prepared using thiol-group-spiked liposomes containing glutathione conjugated to gold nanoparticles under reducing conditions. The absorption peak of GCL was dependent on the amount of added HAuCl4 and shifted from 636 to 795 nm, appropriate for the absorption of NIR laser light (660 nm) by GCL/DOX. Most of the encapsulated drug was released from GCL/DOX within 1 min (>80%) under NIR laser irradiation. GCL/DOX exhibited a synergistic effect compared to monotherapy (photothermal effect alone or free-DOX treatment alone) on A549 cells. GCL is thus an effective liposome carrier, and in conjunction with NIR laser irradiation its therapeutic modality holds promise for cancer treatment.

Application of Asymmetrical Flow Field-Flow Fractionation for Characterizing the Size and Drug Release Kinetics of Theranostic Lipid Nanovesicles.

Liposome size and in vitro release of the active substance belong to critical quality attributes of liposomal carriers. Here, we apply asymmetric flow field-flow fractionation (AF4) to characterize theranostic liposomes prepared by thin lipid film hydration/extrusion or microfluidics. The vesicles’ size was derived from multi-angle laser light scattering following fractionation (AF4) and compared to sizes derived from dynamic light scattering measurements. Additionally, we adapted a previously developed AF4 method to study zinc phthalocyanine (ZnPc) release/transfer from theranostic liposomes. To this end, theranostic liposomes were incubated with large acceptor liposomes serving as a sink (mimicking biological sinks) and were subsequently separated by AF4. During incubation, ZnPc was transferred from donor to acceptor fraction until reaching equilibrium. The process followed first-order kinetics with half-lives between 119.5–277.3 min, depending on the formulation. The release mechanism was postulated to represent a combination of Fickian diffusion and liposome relaxation. The rate constant of the transfer was proportional to the liposome size and inversely proportional to the ZnPc/POPC molar ratio. Our results confirm the usefulness of AF4 based method to study in vitro release/transfer of lipophilic payload, which may be useful to estimate the unwanted loss of drug from the liposomal carrier in vivo.

Preclinical evaluation of PEGylated liposomal doxorubicin as an effective radiosensitizer in chemoradiotherapy for lung cancer.

Development of asafe and effective systemic chemotherapeutic agent for concurrent administration with definitive thoracic radiotherapy remains amajor goal of lung cancer management. The synergistic effect of PEGylated liposomal doxorubicin and irradiation was evaluated in lung cancer cell lines both in vitro and in vivo. In vitro radiosensitization of A549 and LLC cell lines was evaluated by colony formation assay, γH2AX fluorescent staining and western blot assay, and annexinV staining. Aradiosensitization study with healthy human lung-derived cell line BEAS-2B was performed for comparative purposes. In vivo radiosensitization was evaluated by tumor ectopic growth, cell survival, pharmacokinetics, and biodistribution analyses. Cleaved caspase‑3, the marker for apoptosis, was assessed immunohistochemically in A549 xenograft tumors. Treatment with PEGylated liposomal doxorubicin decreased A549 and LLC cell proliferation in adose-dependent manner. In vitro studies revealed comparable radiosensitizer advantages of PEGylated liposomal doxorubicin and free doxorubicin, showing equivalent DNA double-strand breaks according to γH2AX fluorescent staining and western blot assays, similar numbers of apoptotic cells in the annexin‑V staining assay, and moderately decreased clonogenic survival. In vivo studies demonstrated markedly slow ectopic tumor growth with prolonged survival following treatment with PEGylated liposomal doxorubicin plus irradiation in both A549 and LLC mouse models, suggesting that PEGylated liposomal doxorubicin is more effective as aradiosensitizer than free doxorubicin in vivo. Pharmacokinetics evaluation showed alonger half-life of approximately 40 h for PEGylated liposomal doxorubicin, confirming that the liposomal carrier achieved controlled release. Biodistribution evaluation of PEGylated liposomal doxorubicin confirmed high accumulation of doxorubicin in tumors, indicating the promising drug delivery attributes of PEGylated liposomal doxorubicin. Although free doxorubicin caused histopathologic myocarditis with the cardiac muscle fibers showing varying degrees of damage, PEGylated liposomal doxorubicin caused no such effects. The immunohistochemical expression of cleaved caspase-3-positive cells was greatest expressed in the irradiation and PEGylated liposomal doxorubicin combined treatment group, indicating prolonged tumoricidal effects. Our study provides preclinical in vitro and in vivo evidence of the effectiveness of PEGylated liposomal doxorubicin as aradiosensitizer, supporting its potential clinical development as acomponent of chemoradiotherapy.

Nanocarriers as Prospective Approach in Effective Management of Acne Vulgaris

Acne is a chronic disease associated with the pilosebaceous unit and is affected by Propionibacterium acnes bacteria. Approximately 95% of people worldwide suffer from acne in their life span with a higher prevalence among teenagers (esp. boys). Conventional strategies incorporating antibiotics, steroids, hormones, etc. are recommended orally, systemically, or topically, employed for the treatment of acne but with various side effects (itching, scaling, redness, etc.) and are no more commercially accepted. The literature has been collected using various search engines google scholar, PubMed, science direct, etc. The review highlights the history of acne, its pathophysiology, developmental factors, various treatment evolutions strategies ranging from conventional to novel approach eradicating the secondary effects with enhanced efficacy and safety profile. We discussed various nanotechnological carriers (liposomes, niosomes, solid lipid nanoparticles, nanostructured lipid carriers, microsponges and other nanoscale formulations), which are formulated in the last decade for the effective treatment of acne. Patent literature and marketed formulations are included in the last sections. The encapsulation efficiency of anti-acne drugs in different nanocarriers improves the efficacy as well as minimizes the side effects of the drug. These carriers showed better bioavailability as well as better penetration effects even to the pilosebaceous unit of the skin. Tolerance can be improved by increasing the concentration of anti-acne drugs in the nano-carrier formulation. Conventional treatment strategies for acne had some limitations like scaling, itching, and inflammation which can be overcome by nano-formulations, which exhibit better efficacy. At present, various nano-technological carriers are being used potentially for the treatment of acne. These nano-formulations are also associated with some limitations like drug entrapment, stability issues, but these will be overcome in the upcoming years as long as the research is being continued in this area.

Vascular permeability in chronic rhinosinusitis enhances accumulation and retention of nanoscale pegylated liposomes

Chronic rhinosinusitis (CRS) is a debilitating inflammatory disorder of the sinonasal mucosa that substantially diminishes patient quality of life. Progress surrounding management of this disease has been crippled by a lack of therapeutic innovation. It has been posited that increased vascularity within the diseased sinuses of patients with CRS may allow for improved systemic drug delivery via nanoscale liposomal carriers. Such a system could enhance drug distribution, accumulation, and retention within the sinuses, ultimately leading to improved patient outcomes. PEGylated liposomes loaded with indocyanine green (ICG) were synthesized, characterized and systemically administered in a mouse model of CRS. Accumulation and retention of ICG in sinonasal tissue were evaluated. Compared to healthy controls, CRS mice showed significant sinonasal tissue accumulation and retention of PEGylated liposomal ICG for up to 21 days (P < 0.001). Conversely, free ICG was eliminated from the body after 24 h in both groups.

Development of Vancomycin Delivery Systems Based on Autologous 3D Platelet-Rich Fibrin Matrices for Bone Tissue Engineering.

Autologous platelet-rich fibrin (PRF) is derived from the blood and its use in the bone tissue engineering has emerged as an effective strategy for novel drug and growth factor delivery systems. Studies have approved that combined therapy with PRF ensures higher biological outcomes, but patients still undergo additional treatment with antibiotic drugs before, during, and even after the implantation of biomaterials with PRF. These systematically used drugs spread throughout the blood and lead not only to positive effects but may also induce adverse side effects on healthy tissues. Vancomycin hydrochloride (VANKA) is used to treat severe Staphylococcal infections but its absorption in the target tissue after oral administration is low; therefore, in this study, we have developed and analyzed two kinds of VANKA carriers—liposomes and microparticles in 3D PRF matrices. The adjustment, characterization, and analysis of VANKA carriers in 3D PRF scaffolds is carried out in terms of encapsulation efficiency, drug release kinetics and antibacterial activity; furthermore, we have studied the micro- and macrostructure of the scaffolds with microtomography.

Isolation methods commonly used to study the liposomal protein corona suffer from contamination issues

The liposomal protein corona has been the focus of numerous studies, but there is still no consensus regarding its extent and composition. Rather, the literature is full of conflicting reports on the matter. To elucidate whether there could be a methodological explanation for this, we here scrutinize the efficiency of three commonly used liposome isolation methods at isolating stealth liposomes from human plasma. Firstly, we show that size-exclusion chromatography (SEC) in its standard form is prone to isolating unbound protein material together with the liposomes, but also that the method may be optimized to mitigate this issue. Secondly, we demonstrate that SEC in combination with membrane ultrafiltration is no better at removing the unbound protein material than SEC alone. Thirdly, we show that centrifugation is not able to pellet the liposomes. Overall, our results suggest that previous research on the liposomal protein corona may have suffered from significant methodological problems, in particular related to contaminant proteins interfering with the analysis of the protein corona. We believe that the data presented here may help guide future research around this challenge to reach a converging understanding about the properties of the protein corona on liposomes. Statement of significanceUpon administration into the circulatory system, liposomal drug carriers encounter an environment rich in proteins. These proteins may adsorb to the liposomes to form what is known as the protein corona, potentially governing the interactions of the liposomes with tissues and cells. However, despite decades of intense research efforts, there is currently no clear understanding about the extent and composition of the liposomal protein corona, making it impossible to assess its mechanistic importance. Here we report that the methods commonly used to isolate liposomes from blood plasma or serum to study the protein corona are susceptible to protein contamination. This may be the underlying technical reason for the current confusion about the characteristics of the liposomal protein corona.

Development of a protein replacement therapy for peeling skin syndrome

British Journal of DermatologyVolume 184, Issue 6 p. e197-e197 Plain Language SummaryFree Access Development of a protein replacement therapy for peeling skin syndrome First published: 06 June 2021 https://doi.org/10.1111/bjd.20101AboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onEmailFacebookTwitterLinked InRedditWechat Abstract Linked article: Valentin et al. Br J Dermatol 2021; 184:1123–1131. Peeling skin syndrome type 1 is a severe form of congenital ichthyosis. Patients have severe itch and generalized peeling of the skin. Current therapeutic approaches are very unsatisfactory. The disease is the result of mutations (mistakes) in the CDSN gene, which cause a lack of the protein corneodesmosin (CDSN) in the skin. We wanted to develop the first steps towards a specific protein replacement therapy for CDSN deficiency. The aim is to improve the cell–cell connection in two layers of the epidermis (the granular and corneal layers). Human CDSN was produced in bacterial cells. A liposomal carrier system was developed to transport the CDSN to the outer membrane of the epidermal cells. The liposomal carrier system was checked for stability and toxicity. Also, the effect on epidermal cells and artificial skin models was investigated. The liposomes were found to be successfully present at the epidermal cell membranes. The CDSN-deficient skin models were treated with liposomal CDSN and demonstrated the presence of the replaced protein within the granular layer. Finally, the skin barrier test and microscopy showed improved skin integrity (skin that functioned better), if the skin models were treated with liposomal CDSN. We believe that this study presents helpful laboratory results to develop a protein replacement therapy for patients with peeling skin syndrome and other diseases where there is also a lack of CDSN. Volume184, Issue6June 2021Pages e197-e197 RelatedInformation

Repetitive drug releases from light-activatable micron-sized liposomes.

In this work, a novel light activatable micron-sized liposomal drug carrier that has a unique capability to release drug repetitively in proportion to the cycle number of short irradiation (5 s) of near-infrared (NIR) pulsed laser is reported. We synthesized methotrexate (MTX)-loaded liposomes based on a modified reverse-phase evaporation method. Gold nanorods (AuNR) were attached to the liposomal surfaces, enabling the liposomes to release drug under short NIR irradiation via the photothermal effect. The concentrations of methotrexate (MTX) released from the liposomes were 10.6, 29.8, 43.7 and 65.9 μg/mL after one, two, three or four NIR laser cycles (1.1 W at 1064 nm, 5 s per cycle), respectively. The current finding will provide possible solution to the previously reported inconsistency in drug release from light activatable liposomal drug carriers at each activation cycle. The repeatability of drug release described in this work is believed to be due to reversible nature of the liposomes. The liposomes release drug via lipid bilayer melting when irradiated by laser due to gold nanorods' plasmonic heat on the lipid bilayer surface and quickly regain their original structure once the laser source is removed. We provided evidence of the reversible liposomal structures by monitoring the change of number densities of liposomes using a microelectrode sensor with different laser irradiation durations and powers. We also assessed the micron-sized liposome with respect to long-term stability, drug encapsulation efficiency, and drug-releasing efficiency, demonstrating the possibility of utilizing these liposomes as long-term drug delivery vehicles for various drugs.

Paclitaxel loading in cationic liposome vectors is enhanced by replacement of oleoyl with linoleoyl tails with distinct lipid shapes

Lipid carriers of hydrophobic paclitaxel (PTX) are used in clinical trials for cancer chemotherapy. Improving their loading capacity requires enhanced PTX solubilization. We compared the time-dependence of PTX membrane solubility as a function of PTX content in cationic liposomes (CLs) with lipid tails containing one (oleoyl; DOPC/DOTAP) or two (linoleoyl; DLinPC/newly synthesized DLinTAP) cis double bonds by using microscopy to generate kinetic phase diagrams. The DLin lipids displayed significantly increased PTX membrane solubility over DO lipids. Remarkably, 8 mol% PTX in DLinTAP/DLinPC CLs remained soluble for approximately as long as 3 mol% PTX (the solubility limit, which has been the focus of most previous studies and clinical trials) in DOTAP/DOPC CLs. The increase in solubility is likely caused by enhanced molecular affinity between lipid tails and PTX, rather than by the transition in membrane structure from bilayers to inverse cylindrical micelles observed with small-angle X-ray scattering. Importantly, the efficacy of PTX-loaded CLs against prostate cancer cells (their IC50 of PTX cytotoxicity) was unaffected by changing the lipid tails, and toxicity of the CL carrier was negligible. Moreover, efficacy was approximately doubled against melanoma cells for PTX-loaded DLinTAP/DLinPC over DOTAP/DOPC CLs. Our findings demonstrate the potential of chemical modifications of the lipid tails to increase the PTX membrane loading while maintaining (and in some cases even increasing) the efficacy of CLs. The increased PTX solubility will aid the development of liposomal PTX carriers that require significantly less lipid to deliver a given amount of PTX, reducing side effects and costs.

Personalized Development of Antisense Oligonucleotides for Exon Skipping Restores Type XVII Collagen Expression in Junctional Epidermolysis Bullosa.

Intermediate junctional epidermolysis bullosa caused by mutations in the COL17A1 gene is characterized by the frequent development of blisters and erosions on the skin and mucous membranes. The rarity of the disease and the heterogeneity of the underlying mutations renders therapy developments challenging. However, the high number of short in-frame exons facilitates the use of antisense oligonucleotides (AON) to restore collagen 17 (C17) expression by inducing exon skipping. In a personalized approach, we designed and tested three AONs in combination with a cationic liposomal carrier for their ability to induce skipping of COL17A1 exon 7 in 2D culture and in 3D skin equivalents. We show that AON-induced exon skipping excludes the targeted exon from pre-mRNA processing, which restores the reading frame, leading to the expression of a slightly truncated protein. Furthermore, the expression and correct deposition of C17 at the dermal–epidermal junction indicates its functionality. Thus, we assume AON-mediated exon skipping to be a promising tool for the treatment of junctional epidermolysis bullosa, particularly applicable in a personalized manner for rare genotypes.