Cisplatin-loaded albumin nanoparticle and study their internalization effect by using ß-cyclodextrin.

The present study with aim at enhancing the therapeutic and anti-cancer properties of cisplatin (CPT)-loaded bovine serum albumin (BSA) nanoparticles. The BSA nanoparticles containing CPT (CPT-BSANPs) were successfully prepared by the desolvation technique. The physicochemical characterization of the CPT-BSANPs were used by Fourier transformed infrared spectroscopy (FTIR), scanning electron microscopy (SEM) and atomic force microscopy (AFM). The particle size of CPT-BSANPs was found less than 200 nm with 75.02 ± 0.15% entrapment efficiency (EE), while zeta potential and PDI were -17.6 mV and 0.2, respectively. In vitro release behavior of the CPT from the carrier suggests that about 64% of the drug gets released after 48 hrs. The anti-cancer activities of the CPT-BSANPs were tested on MCF-7 cell lines. Our studies show that CPT-BSANPs nanoparticles showed specific targeting and enhanced cytotoxicity to MCF-7 cells when compared to the bare CPT. Thus results suggest that CPT-BSANPs fallowed caveolae-mediated endocytosis, it may become better option for intracellular delivery of anticancer drug.

Molecular determinants as therapeutic targets in cancer chemotherapy: An update.

It is well known that cancer cells are heterogeneous in nature and very distinct from their normal counterparts. Commonly these cancer cells possess different and complementary metabolic profile, microenvironment and adopting behaviors to generate more ATPs to fulfill the requirement of high energy that is further utilized in the production of proteins and other essentials required for cell survival, growth, and proliferation. These differences create many challenges in cancer treatments. On the contrary, such situations of metabolic differences between cancer and normal cells may be expected a promising strategy for treatment purpose. In this article, we focus on the molecular determinants of oncogene-specific sub-organelles such as potential metabolites of mitochondria (reactive oxygen species, apoptotic proteins, cytochrome c, caspase 9, caspase 3, etc.), endoplasmic reticulum (unfolded protein response, PKR-like ER kinase, C/EBP homologous protein, etc.), nucleus (nucleolar phosphoprotein, nuclear pore complex, nuclear localization signal), lysosome (microenvironment, etc.) and plasma membrane phospholipids, etc. that might be exploited for the targeted delivery of anti-cancer drugs for therapeutic benefits. This review will help to understand the various targets of subcellular organelles at molecular levels. In the future, this molecular level understanding may be combined with the genomic profile of cancer for the development of the molecularly guided or personalized therapeutics for complete eradication of cancer.

Hyaluronic acid modified pH-sensitive liposomes for targeted intracellular delivery of doxorubicin.

CONTEXT: Surface-modified pH-sensitive liposomal system may be useful for intracellular delivery of chemotherapeutics. OBJECTIVE: Achieving site-specific targeting with over-expressed hyaluronic acid (HA) receptors along with using pH sensitive liposome carrier for intracellular drug delivery was the aim of this study. MATERIALS AND METHODS: Stealth HA-targeted pH-sensitive liposomes (SL-pH-HA) were developed and evaluated to achieve effective intracellular delivery of doxorubicin (DOX) vis-a-vis enhanced antitumor activity. RESULTS: The in vitro release studies demonstrated that the release of DOX from SL-pH-HA was pH-dependent, i.e. faster at mildly acidic pH ∼5, compared to physiological pH ∼7.4. SLpH-HA was evaluated for their cytotoxicity potential on CD44 receptor expressing MCF-7 cells. The half maximal inhibitory concentration (IC50) of SL-pH-HA and SL-HA were about 1.9 and 2.5 µM, respectively, after 48 h of incubation. The quantitative uptake study revealed higher localization of targeted liposomes in the receptor positive cells, which was further confirmed by fluorescent microscopy. The antitumor efficacy of the DOX-loaded HA-targeted pH-sensitive liposomes was also verified in a tumor xenograft mouse model. DISCUSSION: DOX was efficiently delivered to the tumor site by active targeting via HA and CD44 receptor interaction. The major side-effect of conventional DOX formulation, i.e. cardiotoxicity was also estimated by measuring serum enzyme levels of LDH and CPK and found to be minimized with developed formulation. Overall, HA targeted pH-sensitive liposomes were significantly more potent than the non-targeted liposomes in cells expressing high levels of CD44. CONCLUSION: Results strongly implies the promise of such liposomal system as an intracellular drug delivery carrier developed for potential anticancer treatment.

Capsaicin-loaded vesicular systems designed for enhancing localized delivery for psoriasis therapy.

The aim of the current investigation is to evaluate the potential of capsaicin (CAP)-containing liposomes, niosomes and emulsomes in providing localized and controlled delivery, to improve the topical delivery of drug. CAP-bearing systems were prepared by the film hydration method and compared through various in vitro and in vivo parameters. The TEM photographs suggested that the carrier systems were spherical in shape and nanometric in size range. Skin retention studies of CAP from in vitro and in vivo experiments revealed significantly higher accumulation of drug in the case of the emul-gel formulation. Based on the results, we concluded that the emul-gel may be a potential approach for the topical delivery of CAP, for an effective therapy for psoriasis.

Mannosylated chitosan nanoparticles for delivery of antisense oligonucleotides for macrophage targeting.

The therapeutic potential of antisense oligonucleotides (ASODN) is primarily dependent upon its safe and efficient delivery to specific cells overcoming degradation and maximizing cellular uptake in vivo. The present study focuses on designing mannosylated low molecular weight (LMW) chitosan nanoconstructs for safe ODNs delivery by macrophage targeting. Mannose groups were coupled with LMW chitosan and characterized spectroscopically. Mannosylated chitosan ODN nanoparticles (MCHODN NPs) were formulated by self-assembled method using various N/P ratio (moles of amine groups of MCH to phosphate moieties of ODNs) and characterized for gel retardation assay, physicochemical characteristics, cytotoxicity and transfection efficiency, and antisense assay. Complete complexation of MCH/ODN was achieved at charge ratio of 1:1 and above. On increasing the N/P ratio of MCH/ODN, particle size of the NPs decreased whereas zeta potential (ZV) increased. MCHODN NPs displayed much higher transfection efficiency into Raw 264.7 cells (bears mannose receptors) than Hela cells and no significant toxicity was observed at all MCH concentrations. Antisense assay revealed that reduction in lipopolysaccharide (LPS) induced serum TNF-α is due to antisense activity of TJU-2755 ODN (sequence complementary to 3'-UTR of TNF-α). These results suggest that MCHODN NPs are acceptable choice to improve transfection efficiency in vitro and in vivo.

Dual targeted polymeric nanoparticles based on tumor endothelium and tumor cells for enhanced antitumor drug delivery.

Some specific types of tumor cells and tumor endothelial cells represented CD13 proteins and act as receptors for Asn-Gly-Arg (NGR) motifs containing peptide. These CD13 receptors can be specifically recognized and bind through the specific sequence of cyclic NGR (cNGR) peptide and presented more affinity and specificity toward them. The cNGR peptide was conjugated to the poly(ethylene glycol) (PEG) terminal end in the poly(lactic-co-glycolic) acid PLGA-PEG block copolymer. Then, the ligand conjugated nanoparticles (cNGR-DNB-NPs) encapsulating docetaxel (DTX) were synthesized from preformed block copolymer by the emulsion/solvent evaporation method and characterized for different parameters. The various studies such as in vitro cytotoxicity, cell apoptosis, and cell cycle analysis presented the enhanced therapeutic potential of cNGR-DNB-NPs. The higher cellular uptake was also found in cNGR peptide anchored NPs into HUVEC and HT-1080 cells. However, free cNGR could inhibit receptor mediated intracellular uptake of NPs into both types of cells at 37 and 4 °C temperatures, revealing the involvement of receptor-mediated endocytosis. The in vivo biodistribution and antitumor efficacy studies indicated that targeted NPs have a higher therapeutic efficacy through targeting the tumor-specific site. Therefore, the study exhibited that cNGR-functionalized PEG-PLGA-NPs could be a promising approach for therapeutic applications to efficient antitumor drug delivery.

In vivo immunomodulatory activities of the aqueous extract of bonduc nut Caesalpinia bonducella seeds.

This study evaluated the in vivo immunomodulatory activities of the aqueous extract of Caesalpinia bonducella Fleming (Caesalpiniaceae) seeds. C. bonducella is a plant widely used in the traditional medicinal systems of India. In the present investigation, the aqueous extract of C. bonducella seeds was tested for its effect on cell mediated and humoral components of the immune system in rats. Administration of C. bonducella seed extract produced an increase of 93.03 +/- 4 mean hemagglutinating antibody (HA) titer and a change of 0.56 +/- 0.058 mm in delayed type hypersensitivity (DTH) as compared to control at a dose of 400 mg/kg body weight. Thus, the results of this study indicate that C. bonducella extract could be a promising immunostimulatory agent.

Mannosylated liposomes for targeted vaccines delivery.

Mannosylated liposomes appear to be a promising and potential carrier system for delivery of proteins, peptides, or nucleic acids. The present chapter describes novel mannosylated liposomes, which increase the intracellular targeting of immunogen to dendritic cells and macrophages possessing the specific receptors. The liposomes used in the present investigation were prepared by hand-shaken method and characterized for size, shape, surface charge, encapsulation efficiency, ligand binding, and specificity and uptake studies. The immune-stimulating activity of the liposomes was studied by measuring antigen-specific antibody titer following subcutaneous administration of different liposomal formulations in BALB/c mice. It was found that O-palmitoyl mannan (OPM)-coated liposomes showed better uptake efficiency. In vivo studies revealed that the OPM-coated liposomes exhibited significant higher serum antibody response and stronger TH1/TH2-based cellular responses. In conclusion, novel vesicular constructs are useful nanosized carriers having superior surface characteristics--for active interaction with the antigen-presenting cells and subsequent processing and presentation of antigen.

Studies on anti-inflammatory, antipyretic and analgesic properties of Caesalpinia bonducella F. seed oil in experimental animal models.

Caesalpinia bonducella FLEMING (Caesalpiniaceae) plant is well known for its medicinal and therapeutic values in Indian Ayurveda. However, to be clinically useful, more scientific data are needed. Therefore, in the present study, we investigated the effects of C. bonducella seed oil on acute and chronic inflammation. To assess the anti-inflammatory, antipyretic and analgesic activities, varied concentrations of the seed oil of C. bonducella (100, 200 and 400mg/kg orally) were tested in carrageenan-induced rat paw oedema, brewer's yeast-induced pyrexia, acetic acid-induced writhing and hot plate reaction time in experimental rats. The paw volumes, pyrexia and writhes in experimental rats were reduced significantly (p<0.05) as compared to that of control, and hot plate test showed significant licking effect in rats. These results clearly indicate that the oil of C. bonducella seeds could be a potential source for using as anti-inflammatory, antipyretic and analgesic agent.

Immunomodulatory activities of the ethanolic extract of Caesalpinia bonducella seeds.

UNLABELLED: Caesalpinia bonducella FLEMING (Caesalpiniaceae) is a plant well known for its medicinal value in Indian Ayurveda. However, to prove its efficiency for the clinical utilization, more experimental data will be beneficial. AIMS OF THE STUDY: The present study involved the investigation of immunomodulatory activities of ethanolic extract of Caesalpinia bonducella seeds. MATERIALS AND METHODS: Neutrophil adhesion test, haemagglutinating antibody (HA) titre, delayed-type hypersensitivity (DTH) response, phagocytic activity and cyclophosphamide-induced myelosuppression were determined by in vivo experiments. RESULTS: The evaluation of immunomodulatory potential by oral administration of ethanolic seed extract of Caesalpinia bonducella (200-500 mg/kg) evoked a significant increase in percent neutrophil adhesion to nylon fibers as well as a dose-dependent increase in antibody titre values, and potentiated the delayed-type hypersensitivity reaction induced by sheep red blood cells. Also it prevented myelosuppression in cyclophosphamide drug treated rats and good response towards phagocytosis in carbon clearance assay. CONCLUSIONS: The results obtained in this study indicate that Caesalpinia bonducella possesses potential immunomodulatory activity and has therapeutic potential for the prevention of autoimmune diseases.

Antioxidant activity and total phenolic content of ethanolic extract of Caesalpinia bonducella seeds.

The aim of this study was to assess the in vitro potential of ethanolic extract of Caesalpinia bonducella seeds as a natural antioxidant. The DPPH activity of the extract (20, 40, 50, 100 and 200 microg/ml) was increased in a dose dependent manner, which was found in the range of 38.93-74.77% as compared to ascorbic acid (64.26-82.58%). The IC(50) values of ethanolic extract and ascorbic acid in DPPH radical scavenging assay were obtained to be 74.73 and 26.68 microg/ml, respectively. The ethanolic extract was also found to scavenge the superoxide generated by EDTA/NBT system. Measurement of total phenolic content of the ethanolic extract of C. bonducella was achieved using Folin-Ciocalteau reagent containing 62.50mg/g of phenolic content, which was found significantly higher when compared to reference standard gallic acid. The ethanolic extract also inhibited the hydroxyl radical, nitric oxide, superoxide anions with IC(50) values of 109.85, 102.65 and 89.84 microg/ml, respectively. However, the IC(50) values for the standard ascorbic acid were noted to be 70.79, 65.98 and 36.68 microg/ml respectively. The results obtained in this study clearly indicate that C. bonducella has a significant potential to use as a natural antioxidant agent.

Enhancement of T-helper type I immune responses against hepatitis B surface antigen by LPS derivatives adjuvanted liposomes delivery system.

Currently, there is a clinical need for more effective vaccine for hepatitis B that induces robust cell-mediated immune response capable of viral clearance in chronic hepatitis B infection. In the present study, hepatitis B vaccines formulations were designed by loading the hepatitis B surface antigen into liposomes adjuvanted with rough lipopolysaccharide (Re-LPS) and lpxL1 LPS using conventional rotatory evaporation method and were characterized for various parameters, such as vesicle shape and surface morphology, size and size distribution, entrapment efficiency, turbidity, and in vitro release pattern. The immunoreactivity in mice was evaluated by measuring anti-HBs IgG titer and compared with alum-adsorbed HBsAg solution, plain HBsAg, and liposomal HBsAg formulations. The formulations were also evaluated for cell-mediated immune response by HBsAg specific proliferation of spleenocytes after secondary immunization and re-stimulation in vitro with the same antigen. Simultaneous estimation of cytokines (IL-4, IFN-gamma) was also carried out. Ex vivo cellular uptake study was performed by fluorescence microscopy. Results indicate that the serum IgG titer obtained after i.m administration of Re-LPS- and lpxL1 LPS-adjuvanted liposomal HBsAg formulation was equivalent to alum-adsorbed HBsAg formulation but was more responsive, sustained, and significantly higher than the corresponding liposomal HBsAg and plain HBsAg formulations. Incorporation of lpxL1 LPS into the liposomal HBsAg increased the stimulation index (SI) 6-10 times as compared with plain HBsAg. Re-LPS- and lpxL1 LPS-adjuvanted liposomal HBsAg formulations induced stronger cellular immune response with a predominant Interferon-gamma (IFN-gamma) level than those induced by free HBsAg alone, alum-adsorbed HBsAg, and non-adjuvanted liposomal HBsAg. Probably, the possible mechanism for the enhancement of cellular immunity in addition to humoral immunity by LPS-adjuvanted liposomal HBsAg formulation is due to marked enhancement of immunological presentation and recruitment of antigen via macrophage and antigen-presenting cells (APCs).

Aquasomes--a nanoparticulate approach for the delivery of antigen.

The development of compound that enhances immune responses to recombinant or synthetic epitopes is of considerable importance in vaccine research. Of the many different types of immunopotentiating compounds that have been researched, aquasomes are of considerable promise, because of their potency and adjuvanticity. Aquasomes were prepared by self-assembling of hydroxyapatite by co-precipitation method and thereafter preliminary coated with polyhydroxyl oligomers (cellobiose and trehalose) and subsequently adsorbed with bovine serum albumin (BSA) as a model antigen. The prepared systems were characterized for size, shape, antigen-loading efficiency, in vitro antigen stability, and in vivo performance. BSA-immobilized aquasomes were around 200 nm in diameter and spherical in shape and had approximately 20-30% BSA-loading efficiency. The immunological activity of the formulated aquasomes was compared with plain BSA and better results were observed. Studies also indicated that aquasome formulations could elicit combined T-helper 1 (Th1) and Th2 immune response.