Apremilast loaded lyotropic liquid crystalline nanoparticles embedded hydrogel for improved permeation and skin retention: An effective approach for psoriasis treatment.

The present work aimed to prepare and evaluate Apremilast loaded lyotropic liquid crystalline nanoparticles (LCNPs) formulation for skin delivery to enhance the efficacy with reduced adverse effects of the oral therapy in psoriasis treatment. The LCNPs were prepared using the emulsification using a high shear homogenizer for size reduction and optimized with Box Behnken design to achieve desired particle size and entrapment efficiency. The selected LCNPs formulation was evaluated for in-vitro release, in-vitro psoriasis efficacy, skin retention, dermatokinetic, in-vivo skin retention, and skin irritation study. The selected formulation exhibited 173.25 ± 2.192 nm (polydispersity 0.273 ± 0.008) particle size and 75.028 ± 0.235% entrapment efficiency. The in-vitro drug release showed the prolonged-release for 18 h. The ex-vivo studies revealed that LCNPs formulation exhibited drug retention up to 3.2 and 11.9-fold higher, in stratum corneum and viable epidermis compared to conventional gel preparation. In-vitro cell line studies performed on immortal keratinocyte cells (HaCaT cells) demonstrated non-toxicity of selected excipients used in designed LCNPs. The dermatokinetic study revealed the AUC0-24 of the LCNPs loaded gel was 8.4 fold higher in epidermis and 2.06 fold in dermis, respectively compared to plain gel. Further, in-vivo animal studies showed enhanced skin permeation and retention of Apremilast compared to conventional gel.

Lipid Liquid Crystal Nanoparticles: Promising Photosensitizer Carriers for the Treatment of Infected Cutaneous Wounds.

Cutaneous chronic wounds impose a silent pandemic that affects the lives of millions worldwide. The delayed healing process is usually complicated by opportunistic bacteria that infect wounds. Staphylococcus aureus is one of the most prevalent bacteria in infected cutaneous wounds, with the ability to form antibiotic-resistant biofilms. Recently, we have demonstrated the potential of gallium protoporphyrin lipid liquid crystalline nanoparticles (GaPP-LCNP) as a photosensitizer against S. aureus biofilms in vitro. Herein, we investigate the potential of GaPP-LCNP using a pre-clinical model of infected cutaneous wounds. GaPP-LCNP showed superior antibacterial activity compared to unformulated GaPP, reducing biofilm bacterial viability by 5.5 log10 compared to 2.5 log10 in an ex vivo model, and reducing bacterial viability by 1 log10 in vivo, while unformulated GaPP failed to reduce bacterial burden. Furthermore, GaPP-LCNP significantly promoted wound healing through reduction in the bacterial burden and improved early collagen deposition. These findings pave the way for future pre-clinical investigation and treatment optimizations to translate GaPP-LCNP towards clinical application.

Gallium Protoporphyrin Liquid Crystalline Lipid Nanoparticles: A Third-Generation Photosensitizer against Pseudomonas aeruginosa Biofilms.

The looming antimicrobial resistance pandemic has encouraged the investigation of antimicrobial photodynamic therapy (aPDT) as a promising technology to combat recalcitrant bacterial infections caused by antibiotic resistant strains. Here, we report on the optimization and effective application of gallium protoporphyrin liquid crystalline lipid nanoparticles (GaPP-LCNP) as a photosensitizer for aPDT against the Gram-negative bacteria P. aeruginosa in both planktonic and biofilm modes of growth. LCNP significantly enhanced the performance of GaPP as photosensitizer by two-fold, which was correlated with higher antibacterial activity, reducing the viability of planktonic P. aeruginosa by 7 log10 using 0.8 µM GaPP-LCNP and a light dose of 17 J.cm-2. Importantly, GaPP-LCNP also reduced the viability of biofilms by 6 log10 at relatively low light dose of 34.2 J.cm-2 using only 3 µM GaPP-LCNP. The high antibiofilm activity of GaPP-LCNP at low GaPP-LCNP dose indicated the high efficiency and safety profile of GaPP-LCNP as a promising platform for photodynamic inactivation of recalcitrant infections.

Liquid Crystal Nanoparticle Conjugates for Scavenging Reactive Oxygen Species in Live Cells.

The elevated intracellular production of or extracellular exposure to reactive oxygen species (ROS) causes oxidative stress to cells, resulting in deleterious irreversible biomolecular reactions (e.g., lipid peroxidation) and disease progression. The use of low-molecular weight antioxidants, such as 4-amino-2,2,6,6-tetramethylpiperidine-1-oxyl (TEMPO), as ROS scavengers fails to achieve the desired efficacy because of their poor or uncontrolled cellular uptake and off-target effects, such as dysfunction of essential redox homeostasis. In this study, we fabricated a liquid crystal nanoparticle (LCNP) conjugate system with the fluorescent dye perylene (PY) loaded in the interior and poly (ethylene glycol) (PEG) decorated on the surface along with multiple molecules of TEMPO (PY-LCNP-PEG/TEMPO). PY-LCNP-PEG/TEMPO exhibit enhanced cellular uptake, and efficient ROS-scavenging activity in live cells. On average, the 120 nm diameter PY-LCNPs were conjugated with >1800 molecules of TEMPO moieties on their surface. PY-LCNP-PEG/TEMPO showed significantly greater reduction in ROS activity and lipid peroxidation compared to free TEMPO when the cells were challenged with ROS generating agents, such as hydrogen peroxide (H2O2). We suggest that this is due to the increased local concentration of TEMPO molecules on the surface of the PY-LCNP-PEG/TEMPO NPs, which efficiently bind to the plasma membrane and enter cells. Overall, these results demonstrate the enhanced capability of TEMPO-conjugated LCNPs to protect live cells from oxidative stress by effectively scavenging ROS and reducing lipid peroxidation.

Thiol-responsive lyotropic liquid crystals exhibit triggered phase re-arrangement and hydrogen sulfide (H2S) release.

Hydrogen sulfide (H2S) is an important signalling molecule with potential pharmaceutical applications. In pursuit of a suitable delivery system for H2S, herein we apply an amphiphilic trisulfide to concomitantly alter the mesophase behaviour of dispersed lipid particles and enable triggered H2S release. Amperometric release studies indicate the trisulfide acts as a sustained H2S donor, with inclusion into the mesophase attenuating release vs neat dispersed trisulfide. Taken together the results highlight the potential for including trisulfide-based additives in stimuli-responsive drug delivery vehicles.

Spectrophotometric method to quantify tofacitinib in lyotropic liquid crystalline nanoparticles and skin layers: Application in ex vivo dermal distribution studies.

Tofacitinib is an oral Janus kinase inhibitor used in the treatment of Rheumatoid arthritis. The topical delivery of novel Tofacitinib loaded liquid crystal nanoparticles (LCNPs) can provide a controlled release, and also targeted drug delivery to inflamed synovium. There is need of UV spectroscopic method which can determine Tofacitinib in designed nanocarriers like LCNPs, that can be applied to evaluate entrapment efficiency, in vitro drug release, and ex vivo skin studies. In the present study, we have developed and validated a simple and sensitive spectrophotometric method for the quantitative determination of Tofacitinib in methanol and phosphate buffer saline. The linearity range in both the media was 5-30 µg/mL (methanol) and 5-40 µg/ mL (phosphate buffer saline) with high correlation coefficient value (>0.9998). This indicates the clear correlation between Tofacitinib concentrations and their absorbance within the test ranges. The repeatability and intermediate precision articulated by the relative standard deviation were less than 2% in the developed method. The method specificity and applicability were also ascertained by performing recovery studies by spiking method, which was 95.85 ± 1.98% with % RSD 1.24 ± 0.045. The method developed in methanol was successfully applied to determine the entrapment efficiency of Tofacitinib in developed LCNPs formulation and skin retention (dermatokinetics). The method developed in pH 7.4 phosphate buffer saline was applied to quantify Tofacitinib from LCNPs in in vitro and ex vivo drug release samples. In conclusion, a simple, sensitive, accurate, and precise UV spectrophotometric method was established to determine Tofacitinib in in vitro and ex vivo skin studies.

Reprogramming Cancer Stem-like Cells with Nanoforskolin Enhances the Efficacy of Paclitaxel in Targeting Breast Cancer.

Cancer stem-like cells (CSCs) have emerged as an important target for breast cancer therapy owing to their self-renewability, proliferation, and elevated chemoresistance properties. Here, we present a strategy of eliminating CSCs by differentiation therapy where "forced differentiation" reprograms CSCs so that they lose their intrinsic properties and become susceptible for conventional chemotherapeutic drugs. In this study, we report that a conventional chemotherapeutic paclitaxel enhances the stemness of CSCs, while a phytochemical forskolin being essentially nontoxic to CSCs possesses the intrinsic ability to reprogram them. To achieve simultaneous targeting of CSCs and bulk tumor cells, we used a co-delivery system where liquid crystal nanoparticles (LCN) were co-encapsulated with both paclitaxel and forskolin. LCN showed higher uptake, retention, and penetration potential in CSCs overcoming their high drug efflux property. Moreover, LCN improved the pharmacokinetic parameters of forskolin, which otherwise had very low retention and bioavailability. Forskolin-loaded LCN forced CSCs to exit from their mesenchymal state, which reduced their stemness and chemosensitized them while inhibiting E-cadherin-mediated survival and tumor-initiating potential as well as reversing paclitaxel-induced stemness. We further showed that upon administration of paclitaxel and forskolin co-loaded LCN to an orthotropic xenograft mouse model, the nanomedicine showed enhanced passive tumor targeting capability with very potent antitumor activity that eradicated small solid tumor in a single dose and showed no sign of tumor relapse or systemic toxicity over a long period. Overall, these findings give a proof of concept that co-delivery of forskolin and paclitaxel in a single nanoformulation can achieve overall tumor targeting where forskolin can efficiently reprogram/differentiate CSCs and paclitaxel can induce cytotoxicity in both differentiated CSCs and bulk tumor cells simultaneously. Hence, this study can provide a nanoformulation that can offer an efficient strategy for cancer therapy.

Dynamics and Pretransitional Effects in C60 Fullerene Nanoparticles and Liquid Crystalline Dodecylcyanobiphenyl (12CB) Hybrid System.

The report shows the strong impact of fullerene C60 nanoparticles on phase transitions and complex dynamics of rod-like liquid crystal dodecylcyanobiphenyl (12CB), within the limit of small concentrations. Studies were carried out using broadband dielectric spectroscopy (BDS) via the analysis of temperature dependences of the dielectric constant, the maximum of the primary loss curve, and relaxation times. They revealed a strong impact of nanoparticles, leading to a ~20% change of dielectric constant even at x = 0.05% of C60 fullerene. The application of the derivative-based and distortion-sensitive analysis showed that pretransitional effects dominate in the isotropic liquid phase up to 65 K above the clearing temperature and in the whole Smectic A mesophase. The impact of nanoparticles on the pretransitional anomaly appearance is notable for the smectic-solid phase transition. The fragility-based analysis of relaxation times revealed the universal pattern of its temperature changes, associated with scaling via the "mixed" ("activated" and "critical") relation. Phase behavior and dynamics of tested systems are discussed within the extended Landau-de Gennes-Ginzburg mesoscopic approach.

Research Status,Problems and Countermeasures of Lyotropic Liquid Crystal in New Drug Delivery Systems of Traditional Chinese Medicine / 中国实验方剂学杂志

Lyotropic liquid crystal is an ordered system with various geometric shapes or three-dimensional structures formed by the interaction of amphiphilic molecules dissolved in polar solvents, and has the characteristics of excellent drug applicability, high drug loading, good bioadhesive property and high transdermal permeability. Through a comprehensive analysis of the research results in this research group and the related research reports of LLC drug delivery system, the authors systematically discussed the research value, development potential and research status of LLC in the field of new drug delivery system of traditional Chinese medicine(TCM), especially in the percutaneous and mucosal drug delivery system and the oral microparticle drug delivery system of TCM. At the same time, due to the current research field of TCM-LLC drug delivery system starts late, many of the basic research problems to be further perfect, this paper had carried on the induction summary to these problems, and put forward the research countermeasures:①the basic research of TCM-LLC drug delivery system should be strengthened by referencing the research experience and methods of LLC drug delivery system of single chemical component combined with TCM characteristics, ②the research on the release mechanism of the chemical components in TCM should be strengthened, and the basic research on the LLC drug delivery system of synchronized sustained release TCM should be carried out, ③development of new LLC materials applicable to TCM, ④the quality evaluation system of TCM-LLC should be improved, ⑤to explore the LLC preparation process suitable for industrialization.

Towards topical microRNA-directed therapy for epidermal disorders.

There remains an unmet dermatological need for innovative topical agents that achieve better longterm outcomes with fewer side effects. Modulation of the expression and activity of microRNA (miRNAs) represents an emerging translational framework for the development of such innovative therapies because changes in the expression of one miRNA can have wide-ranging effects on diverse cellular processes associated with disease. In this short review, the roles of miRNA in epidermal development, psoriasis, cutaneous squamous cell carcinoma and re-epithelisation are highlighted. Consideration is given to the delivery of oligonucleotides that mimic or inhibit miRNA function using vehicles such as cell penetrating peptides, spherical nucleic acids, deformable liposomes and liquid crystalline nanodispersions. Formulation of miRNA-directed oligonucleotides with such skin-penetrating epidermal agents will drive the development of RNA-based cutaneous therapeutics for deployment as primary or adjuvant therapies for epidermal disorders.