Recent advances in drug delivery and targeting for the treatment of pancreatic cancer.

Despite significant treatment efforts, pancreatic ductal adenocarcinoma (PDAC), the deadliest solid tumor, is still incurable in the preclinical stages due to multifacet stroma, dense desmoplasia, and immune regression. Additionally, tumor heterogeneity and metabolic changes are linked to low grade clinical translational outcomes, which has prompted the investigation of the mechanisms underlying chemoresistance and the creation of effective treatment approaches by selectively targeting genetic pathways. Since targeting upstream molecules in first-line oncogenic signaling pathways typically has little clinical impact, downstream signaling pathways have instead been targeted in both preclinical and clinical studies. In this review, we discuss how the complexity of various tumor microenvironment (TME) components and the oncogenic signaling pathways that they are connected to actively contribute to the development and spread of PDAC, as well as the ways that recent therapeutic approaches have been targeted to restore it. We also illustrate how many endogenous stimuli-responsive linker-based nanocarriers have recently been developed for the specific targeting of distinct oncogenes and their downstream signaling cascades as well as their ongoing clinical trials. We also discuss the present challenges, prospects, and difficulties in the development of first-line oncogene-targeting medicines for the treatment of pancreatic cancer patients.

Gemcitabine-Lipid Conjugate and ONC201 Combination Therapy Effectively Treats Orthotopic Pancreatic Tumor-Bearing Mice.

Gemcitabine (GEM) is a nucleoside analogue approved as a first line of therapy for pancreatic ductal adenocarcinoma (PDAC). However, rapid metabolism by plasma cytidine deaminase leading to the short half-life, intricate intracellular metabolism, ineffective cell uptake, and swift development of chemoresistance downgrades the clinical efficacy of GEM. ONC201 is a small molecule that inhibits the Akt and ERK pathways and upregulates the TNF-related apoptosis-inducing ligand (TRAIL), which leads to the reversal of both intrinsic and acquired GEM resistance in PDAC treatment. Moreover, the pancreatic cancer cells that were able to bypass apoptosis after treatment of ONC201 get arrested in the G1-phase, which makes them highly sensitive to GEM. To enhance the in vivo stability of GEM, we first synthesized a disulfide bond containing stearate conjugated GEM (lipid-GEM), which makes it sensitive to the redox tumor microenvironment (TME) comprising high glutathione levels. In addition, with the help of colipids 1,2-dioleoyl-glycero-3-phosphocholine (DOPC), cholesterol, and 1,2-distearoyl-glycero-3-phosphoethanolamine-poly(ethylene glycol)-2000 (DSPE-PEG 2000), we were able to synthesize the lipid-GEM conjugate and ONC201 releasing liposomes. A cumulative drug release study confirmed that both ONC201 and GEM showed sustained release from the formulation. Since MUC1 is highly expressed in 70-90% PDAC, we conjugated a MUC1 binding peptide in the liposomes which showed higher cytotoxicity, apoptosis, and cellular internalization by MIA PaCa-2 cells. A biodistribution study further confirmed that the systemic delivery of the liposomes through the tail vein resulted in a higher accumulation of drugs in orthotopic PDAC tumors in NSG mice. The IHC of the excised tumor grafts further confirmed the higher apoptosis and lower metastasis and cell proliferation. Thus, our MUC1 targeting binary drug-releasing liposomal formulation showed higher drug payload, enhanced plasma stability, and accumulation of drugs in the pancreatic orthotopic tumor and thus is a promising therapeutic alternative for the treatment of PDAC.

Surface-Coated Cerium Nanoparticles to Improve Chemotherapeutic Delivery to Tumor Cells.

The antioxidant property of cerium oxide nanoparticles has increased their demand as a nanocarrier to improve the delivery and therapeutic efficacy of anticancer drugs. Here, we report the synthesis of alginate-coated ceria nanoformulations (ceria NPs) and characterization using FTIR spectroscopy, Raman microscopy, and X-ray diffraction. The synthesized ceria NPs show negligible inherent in vitro toxicity when tested on a MDA-MB-231 breast cancer cell line at higher particle concentrations. Upon loading these particles with doxorubicin (Dox) and paclitaxel (PTX) drugs, we observe a potential synergistic cytotoxic effect mediated by the drug and the ceria NPs, resulting in the better killing capacity as well as suppression of cell migration against the MDA-MB-231 cell line. Further, to verify the immune-escaping capacity before targeting cancer cells, we coated the drug-loaded ceria NPs with the membrane of MDA-MB-231 cells using an extrusion method. The resultant delivery system exhibited in vitro preferential uptake by the MDA-MB-231 cell line and showed reduced uptake by the murine macrophage cell line (RAW 264.7), assigning its potential application as non-immunogenic personalized therapy in targeting and killing of cancer cells.

A Composite of Hyaluronic Acid-Modified Graphene Oxide and Iron Oxide Nanoparticles for Targeted Drug Delivery and Magnetothermal Therapy.

Graphene oxide (GO) nanoparticles have been developed for a variety of biomedical applications as a number of different therapeutic modalities may be added onto them. Here, we report the development and testing of such a multifunctional GO nanoparticle platform that contains a grafted cell-targeting functionality, active pharmaceutical ingredients, and particulates that enable the use of magnetothermal therapy. Specifically, we demonstrate the ability to covalently attach hyaluronic acid (HA) onto GO, and the resultant nanoparticulates (GO-HA) exhibited low inherent toxicity toward two different breast cancer cell lines, BT-474 and MDA-MB-231. Doxorubicin (Dox) and paclitaxel (Ptx) were successfully loaded onto GO-HA with high and moderate efficiencies, respectively. A GO-HA-Dox/Ptx system was significantly better than the GO-Dox/Ptx system at specifically killing CD44-expressing MDA-MB-231 cells but not BT-474 cells that do not express CD44. Further, modified iron oxide nanoparticles were loaded onto the GO-HA-Dox system, enabling the use of magnetic hyperthermia. Hyperthermia in combination with Dox treatment through the GO-HA system showed significantly better performance in reducing viable tumor cell numbers when compared to the individual systems. In summary, we showcase a multifunctional GO nanoparticle system that demonstrates improved efficacy in killing tumor cells.

Polyhydroxybutyrate-co-hydroxyvalerate copolymer modified graphite oxide based 3D scaffold for tissue engineering application.

In this study, we have fabricated the PHBV functionalized graphite oxide using freeze drying technique, followed by 'in situ' pay loading of Fe3O4 nanoparticles onto the hydrophobic plate of the composite basal plane; thereby, mechanically and thermally stable, bio-imaging Fe3O4/GO-g-PHBV composites have been developed. The synthesis of Fe3O4/GO-g-PHBV composite was confirmed by field emission SEM and TEM analyses, X-ray diffraction and Fourier transform infrared spectroscopy. The wrapping of PHBV copolymer into the graphene layers was investigated by atomic force microscopy and Raman spectral analyses which provided the shifting of the 2D band with low signal intensity in the range of 2600-3000 cm-1. The bactericidal activities of the Fe3O4/GO-g-PHBV composite films were found to exhibit more efficiency against Gram-negative bacteria strains compared to Gram-positive strains. In vibrating sample magnetometer (VSM) analysis, the zero value of coercivity revealed the super-paramagnetic nature of the Fe3O4/GO-g-PHBV composites. The Phantom agar magnetic resonance imaging analysis revealed the efficiency of Fe3O4 nanoparticles as a negative contrast (T2 contrast) along with higher relaxivity value. The significant fibroblast cell (NIH 3T3) adhesion and proliferation (85%) on the Fe3O4/GO-g-PHBV composite surface indicated the physiological and biocompatible stability of that composite along with the presence of large π conjugated aromatic domain.

Optimized peptide based inhibitors targeting the dihydrofolate reductase pathway in cancer.

We report the first peptide based hDHFR inhibitors designed on the basis of structural analysis of dihydrofolate reductase (DHFR). A set of peptides were rationally designed and synthesized using solid phase peptide synthesis and characterized using nuclear magnetic resonance and enzyme immunoassays. The best candidate among them, a tetrapeptide, was chosen based on molecular mechanics calculations and evaluated in human lung adenocarcinoma cell line A549. It showed a significant reduction of cell proliferation and an IC50 of 82 µM was obtained. The interaction of the peptide with DHFR was supported by isothermal calorimetric experiments revealing a dissociation constant Kd of 0.7 µM and ΔG of -34 ± 1 kJ mol-1. Conjugation with carboxylated polystyrene nanoparticles improved further its growth inhibitory effects. Taken together, this opens up new avenues to design, develop and deliver biocompatible peptide based anti-cancer agents.

Organically modified clay supported chitosan/hydroxyapatite-zinc oxide nanocomposites with enhanced mechanical and biological properties for the application in bone tissue engineering.

The objective of this study is to design biomimetic organically modified montmorillonite clay (OMMT) supported chitosan/hydroxyapatite-zinc oxide (CTS/HAP-ZnO) nanocomposites (ZnCMH I-III) with improved mechanical and biological properties compared to previously reported CTS/OMMT/HAP composite. Fourier transform infrared spectroscopy, powder X-ray diffraction, scanning electron microscopy and transmission electron microscopy were used to analyze the composition and surface morphology of the prepared nanocomposites. Strong antibacterial properties against both Gram-positive and Gram-negative bacterial strains were established for ZnCMH I-III. pH and blood compatibility study revealed that ZnCMH I-III should be nontoxic to the human body. Cytocompatibility of these nanocomposites with human osteoblastic MG-63 cells was also established. Experimental findings suggest that addition of 5wt% of OMMT into CTS/HAP-ZnO (ZnCMH I) gives the best mechanical strength and water absorption capacity. Addition of 0.1wt% of ZnO nanoparticles into CTS-OMMT-HAP significantly enhanced the tensile strengths of ZnCMH I-III compared to previously reported CTS-OMMT-HAP composite. In absence of OMMT, control sample (ZnCH) also showed reduced tensile strength, antibacterial effect and cytocompatibility with osteoblastic cell compared to ZnCMH I. Considering all of the above-mentioned studies, it can be proposed that ZnCMH I nanocomposite has a great potential to be applied in bone tissue engineering.

Development of bone-like zirconium oxide nanoceramic modified chitosan based porous nanocomposites for biomedical application.

Here, zirconium oxide nanoparticles (ZrO2 NPs) were incorporated for the first time in organic-inorganic hybrid composites containing chitosan, poly(ethylene glycol) and nano-hydroxypatite (CS-PEG-HA) to develop bone-like nanocomposites for bone tissue engineering application. These nanocomposites were characterized by FT-IR, XRD, TEM combined with SAED. SEM images and porosity measurements revealed highly porous structure having pore size of less than 1µm to 10µm. Enhanced water absorption capacity and mechanical strengths were obtained compared to previously reported CS-PEG-HA composite after addition of 0.1-0.3wt% of ZrO2 NPs into these nanocomposites. The mechanical strengths and porosities were similar to that of human spongy bone. Strong antimicrobial effects against gram-negative and gram-positive bacterial strains were also observed. Along with getting low alkalinity pH (7.4) values, similar to the pH of human plasma, hemocompatibility and cytocompatibility with osteoblastic MG-63 cells were also established for these nanocomposites. Addition of 15wt% HA-ZrO2 (having 0.3wt% ZrO2 NPs) into CS-PEG (55:30wt%) composite resulted in greatest mechanical strength, porosity, antimicrobial property and cytocompatibility along with suitable water absorption capacity and compatibility with human pH and blood. Thus, this nanocomposite could serve as a potential candidate to be used for bone tissue engineering.

Aromatic π-Conjugated Curcumin on Surface Modified Polyaniline/Polyhydroxyalkanoate Based 3D Porous Scaffolds for Tissue Engineering Applications.

Curcumin-entrapped polyaniline (PAni)-conjugated poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) electroconductive porous scaffolds were fabricated for application in tissue engineering. The physical and chemical characterizations of the as-prepared biomaterials were performed by UV-vis and ATR/FT-IR spectrophotometric, thermogravimetric, fluorescence microscopic, and X-ray diffractometric analyses. It was observed that compared to the pure PHBV copolymer, which is an insulator, the electroconductivity of the PAni-modified PHBV copolymer increased up to the value of 5.78 × 10-5 S cm-1. An antimicrobial study revealed that the curcumin-loaded biomaterials exhibited better bactericidal effect against Gram-positive bacterial strains compared to Gram-negative strains. The composite also demonstrated significant compatibility toward blood and fibroblast cells and exhibited the maximum cell viability (90% to 80%). Cell migration and proliferation on the injured tissues were found to occur at a faster rate, resulting in faster repair, in the presence of anti-inflammatory and anticancer curcumin drug loaded composites compared to that of the pure PHBV copolymer.

Multifunctional zirconium oxide doped chitosan based hybrid nanocomposites as bone tissue engineering materials.

This paper reports the development of multifunctional zirconium oxide (ZrO2) doped nancomposites having chitosan (CTS), organically modified montmorillonite (OMMT) and nano-hydroxyapatite (HAP). Formation of these nanocomposites was confirmed by various characterization techniques such as Fourier transform infrared spectroscopy and powder X-ray diffraction. Scanning electron microscopy images revealed uniform distribution of OMMT and nano-HAP-ZrO2 into CTS matrix. Powder XRD study and TEM study revealed that OMMT has partially exfoliated into the polymer matrix. Enhanced mechanical properties in comparison to the reported literature were obtained after the addition of ZrO2 nanoparticle into the nanocomposites. In rheological measurements, CMZH I-III exhibited greater storage modulus (G') than loss modulus (G″). TGA results showed that these nanocomposites are thermally more stable compare to pure CTS film. Strong antibacterial zone of inhibition and the lowest minimum inhibition concentration (MIC) value of these nanocomposites against bacterial strains proved that these materials have the ability to prevent bacterial infection in orthopedic implants. Compatibility of these nanocomposites with pH and blood of human body was established. It was observed from the swelling study that the swelling percentage was increased with decreasing the hydrophobic OMMT content. Human osteoblastic MG-63 cell proliferations were observed on the nanocomposites and cytocompatibility of these nanocomposites was also established. Moreover, addition of 5wt% OMMT and 5wt% nano-HAP-ZrO2 into 90wt% CTS matrix provides maximum tensile strength, storage modulus, aqueous swelling and cytocompatibility along with strong antibacterial effect, pH and erythrocyte compatibility.

Potential use of curcumin loaded carboxymethylated guar gum grafted gelatin film for biomedical applications.

Present study describes the synthesis of carboxylmethyl guar gum (CMGG) from the native guar gum (GG). Further, the prepared CMGG is grafted with gelatin to form CMGG-g-gelatin and then mixed with curcumin to prepare a biomaterial. The resultant biomaterial is subjected to the analysis of (1)H NMR, ATR-FTIR, TGA, SEM and XRD ensure the carboxymethylation and grafting. The results reveal that 45% of the amine groups of gelatin have been reacted with the--COOH group of CMGG and 90-95% of curcumin is released from CMGG-g-gelatin after 96h of incubation in the phosphate buffer at physiological pH. In vitro cell line studies reveal the biocompatibility of the biomaterial and the antimicrobial studies display the growth inhibition against gram +ve and gram -ve organisms at a considerable level. Overall, the study indicates that the incorporation of curcumin into CMGG-g-gelatin can improve the functional property of guar gum as well as gelatin.

Microbial degradation of linseed oil-based elastomer and subsequent accumulation of poly(3-hydroxybutyrate-co-3-hydroxyvalerate) copolymer.

The microbial synthesis of environment-friendly poly(3-hydroxybutyrate--co-3-hydroxyvalerate), PHBV, has been performed by using an alkaliphilic microorganism, Alkaliphilus oremlandii OhILAs strain (GenBank Accession number NR_043674.1), at pH 8 and at a temperature of 30-32 °C through the biodegradation of linseed oil-based elastomer. The yield of the copolymer on dry cell weight basis is 90 %. The elastomers used for the biodegradation have been synthesized by cationic polymerization technique. The yield of the PHBV copolymer also varies with the variation of linseed oil content (30-60 %) in the elastomer. Spectroscopic characterization ((1)H NMR and FTIR) of the accumulated product through biodegradation of linseed oil-based elastomers indicates that the accumulated product is a PHBV copolymer consisting of 13.85 mol% of 3-hydroxyvalerate unit. The differential scanning calorimetry (DSC) results indicate a decrease in the melting (T m) and glass transition temperature (T g) of PHBV copolymer with an increase in the content of linseed oil in the elastomer, which is used for the biodegradation. The gel permeation chromatography (GPC) results indicate that the weight average molecular weight (M w) of PHBV copolymer decreases with an increasing concentration of linseed oil in the elastomer. The surface morphology of the elastomer before and after biodegradation is observed under scanning electron microscope (SEM) and atomic force microscope (AFM); these results indicate about porous morphology of the biodegraded elastomer.