Exosome-sheathed porous silica nanoparticle-mediated co-delivery of 3,3'-diindolylmethane and doxorubicin attenuates cancer stem cell-driven EMT in triple negative breast cancer.

BACKGROUND: Therapeutic management of locally advanced and metastatic triple negative breast cancer (TNBC) is often limited due to resistance to conventional chemotherapy. Metastasis is responsible for more than 90% of breast cancer-associated mortality; therefore, the clinical need to prevent or target metastasis is immense. The epithelial to mesenchymal transition (EMT) of cancer stem cells (CSCs) is a crucial determinant in metastasis. Doxorubicin (DOX) is the frequently used chemotherapeutic drug against TNBC that may increase the risk of metastasis in patients. After cancer treatment, CSCs with the EMT characteristic persist, which contributes to advanced malignancy and cancer recurrence. The latest developments in nanotechnology for medicinal applications have raised the possibility of using nanomedicines to target these CSCs. Hence, we present a novel approach of combinatorial treatment of DOX with dietary indole 3,3'-diindolylmethane (DIM) which is an intriguing field of research that may target CSC mediated EMT induction in TNBC. For efficient delivery of both the compounds to the tumor niche, advance method of drug delivery based on exosomes sheathed with mesoporous silica nanoparticles may provide an attractive strategy. RESULTS: DOX, according to our findings, was able to induce EMT in CSCs, making the breast cancer cells more aggressive and metastatic. In CSCs produced from spheres of MDAMB-231 and 4T1, overexpression of N-cadherin, Snail, Slug, and Vimentin as well as downregulation of E-cadherin by DOX treatment not only demonstrated EMT induction but also underscored the pressing need for a novel chemotherapeutic combination to counteract this detrimental effect of DOX. To reach this goal, DIM was combined with DOX and delivered to the CSCs concomitantly by loading them in mesoporous silica nanoparticles encapsulated in exosomes (e-DDMSNP). These exosomes improved the specificity, stability and better homing ability of DIM and DOX in the in vitro and in vivo CSC niche. Furthermore, after treating the CSC-enriched TNBC cell population with e-DDMSNP, a notable decrease in DOX mediated EMT induction was observed. CONCLUSION: Our research seeks to propose a new notion for treating TNBC by introducing this unique exosomal nano-preparation against CSC induced EMT.

Essential oil impregnated luminescent hydroxyapatite: Antibacterial and cytotoxicity studies.

In this study, porous fluorescent nanocrystalline erbium doped hydroxyapatite (eHAp) was synthesized via hydrothermal assisted co-precipitation method. Eucalyptus oil (EU), frankincense oil (FO), Tea tree oil (TTO), wintergreen oil (WO) were successfully absorbed into eHAp pellet by vacuum filtration technique using Buckner funnel. Phase crystallization, fluorescence property and microstructure of eHAp were confirmed by X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FTIR), Photoluminiscence spectroscopy (PL) and Field emission scanning electron microscopy (FESEM). Strong antimicrobial activity was observed for EU, TTO and WO on both E. coli and S. aureus mediated by cell membrane damage and leakage of cytoplasmic components. The oil absorbed eHAp nanocomposites were found to be moderately biocompatible with normal WI-38 cells up to MIC concentration various time scale. The nanocomposites showed significant cytotoxic activity on breast cancer cell line MDA-MB 468 and the fluorescent property of the eHAp was utilized to visualize internalization of particles in the cells. The release profile of the oils from the eHAp matrix showed pH dependent release indicated that the porous matrix can be used as a suitable carrier for modulated and sustained release of bioactive components. Thus, given the multifunctional attributes these natural essential oil-based nanocomposites show great promise as an alternative to conventional therapeutic treatments.

Antimicrobial and biocompatible fluorescent hydroxyapatite-chitosan nanocomposite films for biomedical applications.

Development of fluorescent erbium doped hydroxyapatite (eHAp)-chitosan nanocomposite film is reported. Nanocrystalline eHAp has been synthesized by hydrothermal assisted precipitation method using erbium (III) ions as dopant. Physico-chemical characterization by UV/Visible spectroscopy, X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FTIR), photoluminiscence spectroscopy (PL) and Field emission scanning electron microscopy(FESEM) confirmed incorporation and uniform distribution of eHAp in the chitosan films. Strong antimicrobial activity was observed using eHAp incorporated chitosan films against E. coli and S. aureus by contact inhibition on agar plates. On the other hand, excellent biocompatibility was observed with human lung fibroblast cells (WI-38) which showed strong attachment and proliferation on the chitosan films with minimal cytotoxicity. Moreover, the doped films showed good biodegradation and mineralization behavior after 2 weeks in simulated body fluid. Thus the doped fluorescent chitosan films with multifunctional attributes can be a strong candidate for diverse applications like in antimicrobial treatments, wound healing, tissue scaffolds and bioimaging.

Anti-SSTR2 peptide based targeted delivery of potent PLGA encapsulated 3,3'-diindolylmethane nanoparticles through blood brain barrier prevents glioma progression.

Current therapy for Glioblastoma is insufficient because of the presence of blood brain barrier. It limits the transport of essential drugs to the tumor sites. To overcome this limitation we strategized the delivery of an anticancer compound 3,3'-diindolylmethane by encapsulation in poly (lactic-co-glycolic acid) nanoparticles. These nanoparticles were tagged with a novel peptide against somatostatin receptor 2 (SSTR2), a potential target in glioma. The nanoformulation (27-87nm) had loading and encapsulation efficiency of 7.2% and 70% respectively. It was successfully internalized inside the glioma cells resulting in apoptosis. Furthermore, an in vivo bio-distribution study revealed the selective accumulation of the nanoformulation into rat brain tumor sites by crossing the blood brain barrier. This resulted in abrogation of epidermal growth factor receptor pathway activation in glioma cells. Our novel nanopreparation therefore shows great promise to serve as a template for targeted delivery of other therapeutics in treating GBM.

Synthesis of eucalyptus/tea tree oil absorbed biphasic calcium phosphate-PVDF polymer nanocomposite films: a surface active antimicrobial system for biomedical application.

A biocompatible poly(vinylidene) difluoride (PVDF) based film has been prepared by in situ precipitation of calcium phosphate precursors. Such films were surface absorbed with two essential oils namely eucalyptus and tea tree oil. Physico-chemical characterization of the composite film revealed excellent stability of the film with 10% loading of oils in the PVDF matrix. XRD, FTIR and FESEM measurements confirmed the presence of hydroxyapatite and octacalcium phosphate in the PVDF matrix which showed predominantly ß phase. Strong bactericidal activity was observed with very low minimum bactericidal concentration (MBC) values on both E. coli and S. aureus. The composite films also resisted biofilm formation as observed by FESEM. The release of essential oils from the film showed an initial burst followed by a very slow release over a period of 24 hours. Antibacterial action of the film was found to be primarily due to the action of essential oils which resulted in leakage of vital fluids from the microorganisms. Both necrotic and apoptotic morphologies were observed in bacterial cells. Biocompatibility studies with the composite films showed negligible cytotoxicity to mouse mesenchymal and myoblast cells at MBC concentration.

Formulation and antitumorigenic activities of nanoencapsulated nifetepimine: A promising approach in treating triple negative breast carcinoma.

Triple negative breast cancer (TNBC) is one of the most common invasive malignancies among women, associated with poor prognosis. Standard chemotherapy targets all dividing cells, resulting in dose-limiting toxicities. In this study, we demonstrated a strategy of encapsulating a hydrophobic synthetic compound, nifetepimine, having anticancer properties, in poly (lactic-co-glycolic acid) nanoparticles to increase selectivity of drug to cancerous cells with minimum toxicity towards normal cells. Nanoencapsulated nifetepimine (30-100nm) having loading and encapsulation efficiency of 7.45% and 75% respectively, was successfully internalized inside TNBC cells upon sustained release resulting in apoptosis. An in vivo bio-distribution study indicated that nanonifetepimine selectively accumulated into breast tumor sites of mice, primarily due to prolonged blood circulation time and binding of nifetepimine to epidermal growth factor receptor that remains overexpressed in most of the TNBC tumors. Moreover, we observed significant reduction in breast tumor volume with improved survival implying high tumor targetability of nanonifetepimine.

Pyruvate dehydrogenase complex deficiency and its relationship with epilepsy frequency--An overview.

The pyruvate dehydrogenase complex (PDHc) is a member of a family of multienzyme complexes that provides the link between glycolysis and the tricarboxylic acid (TCA) cycle by catalyzing the physiologically irreversible decarboxylation of various 2-oxoacid substrates to their corresponding acyl-CoA derivatives, NADH and CO2. PDHc deficiency is a metabolic disorder commonly associated with lactic acidosis, progressive neurological and neuromuscular degeneration that vary with age and gender. In this review, we aim to discuss the relationship between occurrence of epilepsy and PDHc deficiency associated with the pyruvate dehydrogenase complex (E1α subunit (PDHA1) and E1ß subunit (PDHB)) and PDH phosphatase (PDP) deficiency. PDHc plays a crucial role in the aerobic carbohydrate metabolism and regulates the use of carbohydrate as the source of oxidative energy. In severe PDHc deficiency, the energy deficit impairs brain development in utero resulting in physiological and structural changes in the brain that contributes to the subsequent onset of epileptogenesis. Epileptogenesis in PDHc deficiency is linked to energy failure and abnormal neurotransmitter metabolism that progressively alters neuronal excitability. This metabolic blockage might be restricted via inclusion of ketogenic diet that is broken up by ß-oxidation and directly converting it to acetyl-CoA, and thereby improving the patient's health condition. Genetic counseling is essential as PDHA1 deficiency is X-linked. The demonstration of the X-chromosome localization of PDHA1 resolved a number of questions concerning the variable phenotype displayed by patients with E1 deficiency. Most patients show a broad range of neurological abnormalities, with the severity showing some dependence on the nature of the mutation in the Elα gene, while PDHB and PDH phosphatase (PDP) deficiencies are of autosomal recessive inheritance. However, in females, the disorder is further complicated by the pattern of X-chromosome inactivation, i.e., unfavorable lyonization. Furthermore research should focus on epileptogenic animal models; this might pave a new way toward identification of the pathophysiology of this challenging disorder.

Formulation of nanotized curcumin and demonstration of its antimalarial efficacy.

AIM: The present study was conducted to overcome the disadvantages associated with the poor water solubility and low bioavailability of curcumin by synthesizing nanotized curcumin and demonstrating its efficacy in treating malaria. MATERIALS AND METHODS: Nanotized curcumin was prepared by a modified emulsion-diffusion-evaporation method and was characterized by means of transmission electron microscopy, atomic force microscopy, dynamic light scattering, Zetasizer, Fourier transform infrared spectroscopy, and differential thermal analysis. The novelty of the prepared nanoformulation lies in the fact that it was devoid of any polymeric matrices used in conventional carriers. The antimalarial efficacy of the prepared nanotized curcumin was then checked both in vitro and in vivo. RESULTS: The nanopreparation was found to be non-toxic and had a particle size distribution of 20-50 nm along with improved aqueous dispersibility and an entrapment efficiency of 45%. Nanotized curcumin (half maximal inhibitory concentration [IC50]: 0.5 µM) was also found to be ten-fold more effective for growth inhibition of Plasmodium falciparum in vitro as compared to its native counterpart (IC50: 5 µM). Oral bioavailability of nanotized curcumin was found to be superior to that of its native counterpart. Moreover, when Plasmodium berghei-infected mice were orally treated with nanotized curcumin, it prolonged their survival by more than 2 months with complete clearance of parasites in comparison to the untreated animals, which survived for 8 days only. CONCLUSION: Nanotized curcumin holds a considerable promise in therapeutics as demonstrated here for treating malaria as a test system.

In situ synthesis and antibacterial activity of copper nanoparticle loaded natural montmorillonite clay based on contact inhibition and ion release.

Copper nanoparticle based clay composite has been synthesized by in situ reduction of a copper ammonium complex ion and characterized by different analytical instruments. The copper nanoparticles were both intercalated and adsorbed on the surface with diameters of <5nm (for intercalated) and 25-30nm (for adsorbed). The composite showed good stability for over 3 months in air. Excellent antimicrobial activity of the composite was observed on Escherichia coli, Staphylococcus aureus, Pseudomonas aeruginosa and Enterococcus faecalis with mortality rates >90% after 12h. Cellular membrane damage permeated by direct attachment of the composite and indirect damage caused by released copper ion are the primary sources of antibacterial action. Cytotoxicity measurements showed minimal adverse effect on the two human cell lines beyond the M.B.C. value for the microorganisms studied. In the present form the clay composite shows good promise for use in therapeutic applications.

Degradation of bacterial DNA by a natural antimicrobial agent with the help of biomimetic membrane system.

The antimicrobial efficacy of methylglyoxal (MG) against several gram-negative bacteria including Escherichia coli has been reported. To determine the mechanism of action of MG, molecular interactions between lipid and MG within the liposomal membrane were also investigated. Multilamellar and unilamellar vesicles were prepared from 1, 2-dipalmitoyl-snglycero-3-phosphocholine (DPPC). The effect of MG on DPPC liposomal membrane was studied by fluorescence spectroscopy and differential scanning calorimetry. The results indicate that MG interacts mainly with the DPPC head group that produces a significant increase in the fluidity of liposomal vesicles, which could be the cause of a fusion/aggregation effect in microbial cells. The agarose gel electrophoresis study with the genomic DNA extracted from E. coli ATCC 25922 revealed that addition of MG could completely degrade this DNA within 1 h, pointing out to their distinctly high degree of sensitivity towards MG. Further, the drug was able to cross the cell membranes, penetrating into the interior of the cell and interacting with DNA for demonstrating antibacterial activity of MG.

Antimicrobial efficacy and biocompatibility study of copper nanoparticle adsorbed mullite aggregates.

A mullite based antimicrobial ceramic composite has been developed by simple adsorption of copper nano particle suspension. The physico-chemical properties of samples were characterized by different instruments which showed that the composite is well crystalline with homogeneous distribution of copper nanoparticles on the surface. Antimicrobial study was performed by plate count technique which showed >99% mortality for all the bacterial species studied after 24h of incubation. Minimum inhibitory concentration (MIC) values determined by batch culture process showed considerably low values (in terms of copper content) indicating that mullite matrix plays a role in enhancing the antimicrobial efficacy of the composite. Biocompatibility studies on human cancer cell lines indicated that the composite had negligible toxicity below 100µg/mL of Cu content. Thus the composite can be suitable for developing antimicrobial ceramic wares and therapeutic purposes like treatment of variety of microbial infections.