Liposome-based codelivery of celecoxib and doxorubicin hydrochloride as a synergistic dual-drug delivery system for enhancing the anticancer effect.

Combination therapy with conventional chemotherapeutic drugs strongly demonstrates a good approach to reduce cytotoxicity, resistance, and the dose of the potent anticancer drugs. The purpose of this research was to design and characterize liposome incorporating celecoxib (CEL) and doxorubicin hydrochloride (DOX) and investigate the anti-tumor efficacy of this combination on different tumor cells. A simple comparison study had been performed for liposomes formulation using thin-film hydration method and pH-gradient method. HSPC-incorporated liposomes were chosen for encapsulation of both CEL and DOX. The formulations showed small particle size and polydispersity index with high encapsulation efficiency. DOX/CEL liposomes displayed the strongest cytotoxicity against B16 and MGC80-3 cells in comparison to the corresponding drug solutions. By incorporation of both agents, a significant reduction in IC50 from 0.927 to 0.198 µg/ml and from 0.81 to 0.535 µg/ml against B16 cells and MGC80-3 cells, respectively, was observed. CEL also significantly improved the intracellular retention and accumulation of DOX in vitro. Our data suggest that the developed liposomal formulation proved to be the most effective formulative strategy as a dual drug delivery system for incorporation of both doxorubicin HCL and CEL and could be considered a useful tool for enhancing the therapeutic efficacy of the anticancer drug.

Implication of CDKAL1 single-nucleotide polymorphism rs 9465871 in obese and non-obese Egyptian children.

INTRODUCTION: CDKAL1 single-nucleotide polymorphism rs 9465871variant is a risk locus for Type 2 Diabetes (T2DM).The study evaluated the associations of CDKAL1- rs9465871 with glycosylated hemoglobin A1C Level (HbA1c), fasting insulin level, insulin resistance and metabolic syndrome among obese and non- obese Egyptian children. MATERIALS AND METHODS: The study included 43 obese children and 40 normal weight children. Anthropometric body measurements, bio-specimen and biochemistry assays were done. Genotyping of rs9465871 (CDKAL1) was conducted. RESULTS: The percentages of the CC, CT, and TT genotypes of rs9465871in the lean children were 15%, 42.5%, and 42.5%, respectively. Regarding obese children, the frequencies were 18.6%, 58.1% and 23.3% respectively with no significant statistical difference. Comparison between the CDKAL1 rs 9465871 polymorphism showed that the highest value of fasting insulin was recorded in CC genotype (22.80± 15.18 [uIU/mL] P<.014). Levels of HOMA-IR, FBS and HBA1C were highest in CC group with no statistical significant differences. However, fasting insulin level was higher in the CC group than in the TT+ CT group (P<.01). A higher level of HbA1c was found among CC group at CDKAL1-rs9465871 (5.9%) than TT+CT genotype group (5.6%), with no statistical significant difference. There was increase in the risk of type 2 diabetes the percentages were 78.6% and 46.4% respectively when comparing CC with TT+CT genotype groups ( P<.039). CC group was not associated with significant increase in metabolic syndrome. CONCLUSION: There is a significant risk association between CDKAL1-rs9465871polymorphism and development of T2DM in a subset of the Egyptian children.

Stimuli-responsive, dual-function prodrug encapsulated in hyaluronic acid micelles to overcome doxorubicin resistance.

Multidrug resistance (MDR) is the main challenge faced by cancer chemotherapy. Drug-conjugate offers a promising strategy for breast cancer therapy. In this regard, we developed a DNVM multifunctional drug delivery system by crosslinking doxorubicin (DOX) and vitamin E succinate (VES) with a pH-sensitive hydrazone bond and then encapsulated the DOX-NN-VES prodrug into pH-sensitive hyaluronic acid-2-(octadecyloxy)-1,3-dioxan-5-amine (HOD) micelles. DOX resistant MCF-7/ADR cell were adopted as a model to study the capability and mechanism of MDR reversal. DNVM exhibited much higher cytotoxicity and cell uptake efficiency compared with that of acid-insensitive DOX-VES loaded HOD micelles (DVSM) and DOX loaded HOD micelles (DOXM), indicating the better capacity of DNVM for the reversal of MDR. Moreover, DNVM prevented drug efflux more effectively, inhibited the expression of P-gp, induced excessive production of reactive oxygen species and affected the expression of apoptosis-related proteins. In vivo experiments showed that DNVM significantly inhibited the tumor growth with no obvious changes in the body weight of MCF-7/ADR cells-bearing nude mice. The results suggested that the "double gain" DNVM can synergistically enhance the efficacy of chemotherapeutics for DOX resistant tumor cells and has the potential to overcome tumor MDR. STATEMENT OF SIGNIFICANCE: A dual-functional pH-sensitive doxorubicin - vitamin E succinate prodrug was developed and loaded into tumor microenvironment-sensitive hyaluronic acid-2-(octadecyloxy)-1,3-dioxan-5-amine micelle system (DNVM) for sequencing stimuli-release and overcoming doxorubicin resistance. The "double gain" DNVM can synergistically enhance the efficacy of chemotherapeutics for doxorubicin resistant tumor cells and has the potential to overcome tumor multiple drug resistance.

Dual-Functional Peptide Driven Liposome Codelivery System for Efficient Treatment of Doxorubicin-Resistant Breast Cancer.

BACKGROUND: The active-targeted drug delivery systems had attracted more and more attention to efficiently overcome multidrug resistance (MDR) in cancer treatments. The aim of the work was to develop a multifunctional nano-structured liposomal system for co-delivery of doxorubicin hydrochloride (DOX) and celecoxib (CEL) to overcome doxorubicin resistance in breast cancer. METHODS: A functional hybrid peptide (MTS-R8H3) with unique cellular penetrability, endo-lysosomal escape and mitochondrial targeting ability was successfully synthesized using solid phase synthesis technology. The peptide modified targeted liposomes (DOX/CEL-MTS-R8H3 lipo) for co-delivery of DOX and CEL were formulated to overcome the chemoresistance in MCF/ADR cells. RESULTS: DOX/CEL-MTS-R8H3 lipo showed nanosized shape and displayed high stability for one month. The cytotoxicity effect of the co-delivery of DOX and CEL through peptide modified liposomes had remarkable treatment efficacy on killing MCF/ADR cells. Targeted liposome exhibited greater cellular entry ability about 5.72-fold stronger than DOX solution. Moreover, as compared with unmodified liposomes, the presence of MTS-R8H3 peptide entity on liposome surface enhanced the mitochondrial-targeting ability and achieved effective reactive oxygen species (ROS) production with significant inhibition of P-gp efflux activity. CONCLUSION: The study suggested that the DOX/CEL-MTS-R8H3 lipo is a promising strategy for overcoming drug resistance in breast cancer treatments with high targeting inhibition efficiency.

Redox-responsive biocompatible nanocarriers based on novel heparosan polysaccharides for intracellular anticancer drug delivery.

Heparosan is a natural precursor of heparin biosynthesis in mammals. It is stable in blood circulation but can be degraded in lysosomes, showing good biocompatibility and long circulation features. So heparosan can be designed as anticancer drug carriers to increase tumor selectivity and improve the therapeutic effect. A novel redox-sensitive heparosan-cystamine-vitamin E succinate (KSV) micelle system was constructed for intracellular delivery of doxorubicin (DOX). Simultaneously, the redox-insensitive heparosan-adipic acid dihydrazide-vitamin E succinate copolymer (KV) was synthesized as control. DOX-loaded micelles (DOX/KSV) with an average particle size of 90-120 nm had good serum stability and redox-triggered depolymerization. In vitro drug release test showed that DOX/KSV micelles presented obvious redox-triggered release behavior compared with DOX/KV. Cytotoxicity and cell uptake were investigated using MGC80-3 tumor cells and COS7 fibroblast-like cells. The cell survival rate of blank micelles was more than 90%, and the cytotoxicity of DOX/KSV in MGC80-3 cells was higher than in COS7 cells, indicating that the carrier has better biocompatibility and less toxicity side effect. The cytotoxicity of DOX/KSV against MGC80-3 cells was significantly greater than that of free DOX and DOX/KV. Furthermore, compared with DOX/KV in MGC80-3 cells, DOX/KSV micelles uptook more anticancer drugs and then released DOX faster into the cell nucleus. The micelles were endocytosed by multiple pathways, but clathrin-mediated endocytosis was the main pathway. Therefore, heparosan polysaccharide could be a potential option as anticancer carrier for enhancing efficacy and mitigating toxicity.

Derma roller® microneedles-mediated transdermal delivery of doxorubicin and celecoxib co-loaded liposomes for enhancing the anticancer effect.

The topical delivery of chemotherapeutics is a promising approach for the management of skin disorders. However, diverse pharmaceutical strategies are essential to allow penetration of large quantities of drugs to tumor tissue. Herein, an attempt was made to investigate the use of Derma roller® microneedles in combination with doxorubicin HCl (DOX) and celecoxib (CEL) co-loaded liposomes as a potential therapeutic approach for the management of melanoma. DOX/CEL co-loaded liposomes/Gels were prepared and characterized. The results showed that microneedle pretreatment with liposomes gel increased DOX penetration into the skin approximately 2-fold compared with the passive delivery. Both CEL liposomes and DOX liposomes caused significant growth inhibition on B16 cells. Besides, DOX/CEL co-loaded liposome was found more cytotoxic than DOX/CEL solution and single drug loaded liposome. The transdermal delivery of DOX/CEL co-loaded liposome successfully inhibited subcutaneous melanoma in female BALB/nude mice, and the co-administration of DOX/CEL with liposomes was better and significantly enhanced the antitumor effect more than single-drug-loaded liposomes. Furthermore, Dermarollers treatment prior to gel application strongly improved the tumor inhibition rate. DOX/CEL co-loaded liposome delivery via microneedles is a promising strategy for skin tumor treatment with targeting inhibition efficiency and negligible side effects.

Fabrication and characterization of collagen-heparin-polypyrrole composite conductive film for neural scaffold.

In this work, a conductive film consisted of polypyrrole-heparin-collagen (PHC film) was fabricated as a potential neural scaffold. Heparin was initially modified with pyrrole, which was further polymerized with pyrrole monomer under the catalysis of ferric trichloride. Then collagen was added and crosslinked through amide bond, as well as physical interaction with pyrrole through hydrogen bond. In this system, heparin and collagen contributed to improving the biocompatibility, because they were the major component of the extracellular matrix. Additionally, heparin was verified to promote nerve cells growth. The physicochemical properties of PHC film were verified, including structure, morphological analysis, degradation, swelling, electrical properties and so on. Combined with the promotion results of pheochromocytoma cells growing, this PHC film is expected to be a promising alternative for nerve regeneration.

Liposome: composition, characterisation, preparation, and recent innovation in clinical applications.

In the last decades, pharmaceutical interested researches aimed to develop novel and innovative drug delivery techniques in the medical and pharmaceutical fields. Recently, phospholipid vesicles (Liposomes) are the most known versatile assemblies in the drug delivery systems. The discovery of liposomes arises from self-forming enclosed phospholipid bilayer upon coming in contact with the aqueous solution. Liposomes are uni or multilamellar vesicles consisting of phospholipids produced naturally or synthetically, which are readily non-toxic, biodegradable, and are readily produced on a large scale. Various phospholipids, for instance, soybean, egg yolk, synthetic, and hydrogenated phosphatidylcholine consider the most popular types used in different kinds of formulations. This review summarises liposomes composition, characterisation, methods of preparation, and their applications in different medical fields including cancer therapy, vaccine, ocular delivery, wound healing, and some dermatological applications.

Elucidation of cellular uptake and intracellular trafficking of heparosan polysaccharide-based micelles in various cancer cells.

Heparosan polysaccharide, known as a heparin precursor, can be used in drug delivery systems due to its good biocompatibility and anti-cancer effect. But few studies on the cellular uptake mechanism of heparosan polysaccharide-based nanocarrier have been investigated. Therefore, the intracellular trafficking and the uptake mechanism of heparosan-based micelles by different tumor cells were investigated in this study. Heparosan-cholesterol amphipathic conjugates (KC) were constructed and doxorubicin (DOX) was loaded to prepare DOX/KC micelles. Different cancer cells were selected to find out the influence on DOX/KC uptake. There was an obvious difference in cytotoxicity and cellular uptake of DOX/KC in various cancer cells. Interestingly, DOX/KC micelles exhibited the strongest cytotoxicity against MGC80-3 cells and displayed highly cellular uptake by B16 cells. The results of the uptake mechanism showed that the internalization of DOX/KC micelles into MGC80-3 cells and A549 cells was mainly through clathrin-mediated endocytosis and macropinocytosis, while micropinocytosis, clathrin-mediated endocytosis and clathrin/caveolae -independent multi-pathways all contributed to the uptake of micelles in B16 cells. Furthermore, after being internalized by MGC80-3 cells, DOX/KC could escape from the lysosome and simultaneously be transported into the nucleus and mitochondria resulting in the greatest cytotoxicity. These results indicate that heparosan-based drug delivery systems may have different uptake and subcellular distribution behavior in tumor cells, and they will achieve the maximum efficacy only in specific kind of cancers.