Self-assembled lyotropic liquid crystal gel for osteoarthritis treatment via anti-inflammation and cartilage protection.

Osteoarthritis (OA) is a chronic joint disease with occurrence of articular inflammation and cartilage degeneration. An ideal drug delivery system for effective treatment of OA should integrate inflammation alleviation with cartilage protection. Herein, a lyotropic liquid crystal (LLC) precursor co-loading hyaluronic acid (HA) and celecoxib, formulated as the HLC precursor, was developed for the combined therapeutic efficacy. The in situ gelling property of the HLC precursor effectively prolongs drug retention in the articular cavity to achieve a long-term anti-inflammation effect. Based on the rheological tests, HLC gel with a cubic lattice structure endows it with a spring-like effect to buffer joint shock and shows great potential in providing cartilage protection by resisting mechanical destruction, lubricating joint, and decomposing intensive stress (about 50%). Meanwhile, the pharmacodynamics study on the OA-induced SD rats demonstrated that HLC gel was the most effective to reduce inflammation levels and to protect the cartilage against abrasion and degeneration. Furthermore, the in vivo degradation behavior and the intra-articular irritation results of LLC/HLC gel demonstrated that it was biodegradable and biocompatible. These results collectively demonstrated that HLC gel with anti-inflammation and cartilage protection performance provides a useful approach to treat OA.

EVI1 induces autophagy to promote drug resistance via regulation of ATG7 expression in leukemia cells.

Ecotropic viral integration site 1 (EVI1) is an oncogenic transcription factor, which is abnormally expressed in myeloid leukemia and other several solid cancers. It is associated with short survival as well as anticancer drug resistance. Autophagy is a protective mechanism that promotes cancer cell growth and survival under stressed conditions including clinical drug treatment. Here evidences are provided that EVI1 induces autophagy and mediated drug resistance in myeloid leukemia cells. Both knockdown using RNAi and pharmacological inhibition of autophagy significantly increase sensitivity to cytotoxic drug treatment in EVI1high cells. Mechanistic studies revealed that EVI1 regulated autophagy by directly binding to autophagy-related gene autophagy related 7 (ATG7) promoter and transcriptionally upregulating its expression. Notably, ATG7 expression was positively correlated with EVI1 in bone marrow mononuclear cells from myeloid leukemia patients. Acute myeloid leukemia patients with high level of EVI1 are associated with unfavorable overall survival, which was aggravated by simultaneous high expression of ATG7 in these patients. Furthermore, ChIP and firefly luciferase reporter assay identified an EVI1-binding site at 227 upstream promoter region of ATG7 which regulated its transcription. In addition, enforced expression of EVI1 also increased intracellular reactive oxygen species and ATG7 mRNA levels as well as autophagy activity, whereas the increase was attenuated after treatment with reactive oxygen species scavenger, suggesting the involvement of reactive oxygen species in EVI1-induced autophagy. These findings demonstrate that EVI protects myeloid leukemia cell from anticancer drug treatment by inducing autophagy through dual control of ATG7. These results might present a new therapeutic approach for improving treatment outcome in myelogenous leukemia with EVI1high.

Injectable in situ forming gel based on lyotropic liquid crystal for persistent postoperative analgesia.

Local anesthetics have been widely used for postoperative analgesia. However, multiple injections or local infiltration is required due to the short half-lives of local anesthetics after single injection, which results in poor compliance and increasing medical expense. In this study, an in situ forming gel (ISFG) based on lyotropic liquid crystal was developed to deliver bupivacaine hydrochloride (BUP) for long-acting postoperative analgesia. BUP-ISFG was designed to be administrated as a precursor solution which would spontaneously transform into gel with well-defined internal nanostructures for sustained drug release at the site of administration when exposed to physiological fluid. A lamellar-hexagonal-cubic phase transition occurred during the in situ gelation. The lamellar phase of the precursor solution endows it with low viscosity for good syringeability while the unique nanostructures of hexagonal and cubic phases of the in situ gel provide sustained drug release. Persistent analgesia effect in vivo was achieved with BUP-ISFG, and the plasma BUP concentration was found to be steadier compared to commercially available BUP for injection. In addition, the ISFG displayed acceptable biocompatibility and good biodegradability. The findings are positive about ISFG as a sustained release system for persistent postoperative analgesia. STATEMENT OF SIGNIFICANCE: To address the issue of insufficient postoperative analgesia associated with short half-lives of local anesthetics after single injection, an in situ forming gel (ISFG) based on lyotropic liquid crystal was developed to deliver bupivacaine hydrochloride (BUP) for postoperative analgesia over three days. The results demonstrated that persistent analgesia effect in vivo was achieved with single injection of BUP-ISFG, and the plasma BUP concentration was found to be steadier compared to commercially available BUP injection. The BUP-ISFG possessed a lamellar-hexagonal-cubic phase transition with corresponding crystal change in 3D nanostructure during the in situ gelation. The relationship between crystal nanostructure and carrier function, might provide some insights to the design and clinical applications of the drug delivery systems based on lyotropic liquid crystal.

Expansible thermal gelling foam aerosol for vaginal drug delivery.

Vaginal delivery of antimicrobial drugs is the most effective method for the local treatment of the vaginal infections. However, current vaginal drug delivery systems (VDDS), including gel, lotion, aerosol and cream, are suffering from low penetration in the deep vaginal rugae and easy elimination by self-cleaning of vaginal canal. To address these issues, a foam aerosol based on the thermal transformation was designed to improve penetration efficiency and achieve the extended retention. The expansible thermal gelling foam aerosol (ETGFA) consisting of thermal sensitive matrix, silver nanoparticle, adhesive agent and propellant, was optimized by evaluations of precursor viscosity, foam expansion, thermal gelation, gel adhesiveness, antimicrobial effects and tissue irritation. The ETGFA would penetrate to the deep vaginal rugae to cover the infectious sites by foam expansion. Drug leakage was intended to be avoided by the thermal gelation at physiological temperature before foam collapse. The gel could be retained in the vaginal canal for extended time due to its superior adhesiveness when compared to the commercial gel Asimi®. The ETGFA provided extended drug release for over 4 h and maintained effective drug concentrations at the infectious sites. The ETGFA containing silver nanoparticles showed dose-dependent antimicrobial effects on the vaginal floras and irritation reduction to the vaginal tissues. The results demonstrated that the ETGFA could overcome the limitations of conventional dosage forms, including poor drug penetration, carrier retention and patient compliance and satisfied the requirements for vaginal drug delivery.

An injectable in situ gel with cubic and hexagonal nanostructures for local treatment of chronic periodontitis.

Periodontitis is a chronic bacterial infection, and its effective treatment is dependent on the retention of antibiotics of effective concentrations at the periodontal pockets. In this study, a solution-gel based inverse lyotropic liquid crystalline (LLC) system was explored to deliver metronidazole to the periodontal pockets for local treatment of periodontitis. It was found that the metronidazole-loaded LLC precursor spontaneously transformed into gel in the presence of water in the oral cavity. The low viscosity of the precursor would allow its penetration to the rather difficult to reach infection sites, while the adhesiveness and crystalline nanostructures (inverse bicontinuous cubic Pn3m phase and inverse hexagonal phase) of the formed gel would permit its firm adhesion to the periodontal pockets. The LLC system provided sustained drug release over one week in vitro. Results from in vivo study using a rabbit periodontitis model showed that the LLC system was able to maintain the metronidazole concentrations in the periodontal pockets above the minimum inhibition concentration for over 10 days without detectable drug concentration in the blood. Owing to the spontaneous solution-gel transition in the periodontal pockets and unique liquid crystalline nanostructures, the LLC in situ gel provided effective treatment of periodontitis for a prolonged period of time with reduced systematic side effects, compared to metronidazole suspension which was effective for 24 h with detectable metronidazole concentrations in the blood after 6 h.

A novel design for stable self-assembly cubosome precursor-microparticles enhancing dissolution of insoluble drugs.

Cubosomes have been presented to enhance dissolution of insoluble drugs, but their applications are limited by the practical hurdles associated with both preparation and storage instability, resulting in drug delivery failure. In the present study, an innovative cubosome precursor-microparticles (CPMs) spray dried from an aqua-free precursor solution was developed to improve cubosome stability during both preparation and storage as well as to enhance the dissolution of insoluble drugs. These CPMs spontaneously self-assembled in situ forming homogeneous cubosome dispersion by hydration and disintegration after exposure to the aqueous medium. The stable cubosome dispersion was obtained from self-assembly (SA) of CPMs after administration instead of fragmentation of bulk cubic phase gel into cubosomes, which settled the preparation instability due to avoidance of high energy fragmentation (e.g. ultrasonic effect, high speed shear and high pressure homogenization). Also, the subsequent storage instability issue can be excluded as the CPMs were stored in a solid stable form. The CPMs disintegration and cubosome SA were demonstrated by the notable morphology variation and the distinct microparticle size decrease from CPMs (10-20 µm) to SA-cubosomes (150-200 nm). The cumulative release of docetaxel (DTX, model insoluble drug) incorporated in CPMs increased to 96.4% within 120 minutes compared with only 75.2% for blank CPMs and DTX physical mixture, demonstrating that CPMs significantly enhanced the dissolution extent of insoluble drug. The SA-cubosomes possessed quite high drug entrapment efficiency (>95%) and an integrated drug dissolution content, which significantly increased the drug utilization rate.