Olive Oil-Based Reverse Microemulsion for Stability and Topical Delivery of Methotrexate: In Vitro.

Hydrolysis of pharmaceutically active molecules can be in control under a confined environment of water-in-oil microemulsion. Stability of model drug methotrexate (MTX) in a sodium bis(2-ethylhexyl) sulfosuccinate (AOT) and olive oil microemulsion system has been evaluated. The physicochemical properties of AOT-MTX-water-olive oil reverse microemulsion (MTX-RM) were examined by UV-vis, Fourier transform infrared, and X-ray diffraction techniques, and the hydrodynamic size was determined by dynamic light scattering techniques and morphologies were characterized by a transmission electron microscope and atomic force microscope. In vitro permeation of MTX-RM through treated skin and its mechanism are evaluated by a UV-visible spectrophotometer, confocal laser scanning microscope, differential scanning calorimeter, and attenuated total reflecting infrared spectroscopy (ATR). The interaction of MTX with the AOT headgroup in confined environment RM enhanced the stability of MTX without affecting the molecular integrity at room temperature. Chemical stability of MTX in MTX-RM (W0 = 5) is significantly higher (∼97%) at room temperature for the study period of 1 year than in MTX-RM (W0 = 15) (∼72%). Interaction of MTX with the AOT headgroup is also visualized by a high-resolution transmission electron microscope and is in correlation with FT-IR data of MTX-RM. The skin fluxes of MTX are 15.1, 19.75, and 22.75 times higher at water content (W0) of 5, 10, and 15, respectively, in MTX-RM in comparison to aqueous solution of MTX. The enhanced amounts of the MTX were detected using CLSM in hair follicles, sweat glands, and epidermis layer of the skin. Merging of T2, T3, and T4 thermal peaks in one broad peak in treated skin endothermograph shows that carrier MTX-RM affects the lipid as well protein structure of the treated skin. ATR data of treated skin showed an increase in the intensity of the carbonyl peak at 1750 cm-1 (lipid), shifting of the amide II peaks, and separation of peaks in the range of 1060 to 1000 cm-1 (vibration mode of -CH2OH, C-O stretching, and C-OH bending peak of the carbohydrate) in comparison to control skin, which indicates that MTX-RM interacts with glycolipid and glycoprotein through carbohydrate hydroxy groups.

Ocimum sanctum L water extract: In-situ green synthesis of zinc oxide nanoparticles for treatment of rheumatoid arthritis: Preclinical study.

BACKGROUND AND OBJECTIVE: Rheumatoid arthritis is a chronic progressive autoimmune disorder, characterised by destruction of cartilage and under line bones. Though exact etiology of rheumatoid arthritis (RA) remains unknown. It is believed that alteration in control of cellular or molecular responses is involved in the chronic inflammation. Earlier in RA patients it was observed the circulating RA specific biomarkers and immunoglobulin deposits in the synovial joints. Zinc Oxide Nanoparticles (ZnO NPs) is used as an anti-inflammatory and anticancer agent, however there is nil/very less scientific data shows the anti-arthritic activity of green synthesis ZnO nanoparticles (Ocimum sanctum water extract in-situ synthesis of ZnO NPs having active compound Caffeic acid and Rosmerinic acid). Hence, the present activity was planned to assess the anti-arthritic activity of ZnO NPs in CIA rats. METHODS: Arthritis in rats were induced by subcutaneous injection of collagen type II (CII) (200 µl) at the base of tail on day 0 followed by booster dose on day 14. ZnO NPs were given (2 mg/kg b.wt./day) orally for 20 days. At the end of the study serum, joint homogenate was used to assess the level of biomarkers (RF, a-CCP, a-CII and CRP) and inflammatory mediators. In addition, m-RNA expression of various genes such as Nuclear factor-kB (NF-kB), inflammatory mediators like tumour necrosis factor-α (TNF-α), cyclooxygenase-2 (COX-2) etc. were assayed in joint tissue. Finally all these biochemical and molecular results were confirmed by microscopic study of joint tissue. RESULTS: ZnO NPs, treated rats showed decrease in inflammation and clinical severity. This was related with decrease in the level of biomarkers (like RF, a-CCP and CRP), inflammatory mediators (TNF-α, COX-2) and activity of transcription factor NF-kB. All these findings were positively correlated with microscopic analysis of joint tissue that showed reduced inflammation and bone erosion in treated group. CONCLUSION: This study validates the anti-arthritic activity of ZnO NPs as it mitigates the arthritis related symptoms in CIA rats.

Synergic effect of aqueous extracts of Ocimum sanctum and Trigonella foenum-graecum L on the in situ green synthesis of silver nanoparticles and as a preventative agent against antibiotic-resistant food spoiling organisms.

The combination of Ocimum sanctum and Trigonella foenum-graecum L leaf water extract synergistically acts as a reducing and capping agent for the synthesis of narrow polydisperse silver nanoparticles (Ag NPs) with controlled sizes depending on the precursor (AgNO3) concentration in the plant extract. The toxicity of 40 nm-sized green synthesized Ag NPs is less than that of 10 nm-sized NPs. The Ag NP solution in Ocimum sanctum and Trigonella foenum-graecum L leaf water extract shows synergic antibacterial effect on Gram-negative bacteria by effecting the ester group of the lipids (hydrolysis) and also breaking the amide bonds of the bacterial chemical constituents, which leads to their rapid death.

Polyaspartic acid, 2-acrylamido-2-Methyl propane sulfonic acid and sodium alginate based biocompatible stimuli responsive polymer gel for controlled release of GHK-Cu peptide for wound healing.

Stimuli responsive polymer based on Polyaspartic acid, 2-Acrylamido-2-methylpropane sulfonic acid and sodium alginate (NaAlg) were synthesized using two cross-linkers Ethylene glycol dimethacrylate (EGDMA) and TMPTA (Trimethylolpropane triacrylate). The polymers were standardized and optimized to obtain a polymer with maximum swelling in distilled water, saline, glucose and solutions of varying pH. The synthesized polymer swelled well in distilled water, glucose solution and acidic- alkaline medium. The biocompatibility of the polymer was evaluated for blood compatibility and protein adsorption. The polymer with maximum swelling property was used for peptide release studies. The polymer was further used to study the peptide encapsulation and release efficiency of the polymeric material which was confirmed by FTIR, Scanning Emission Microscope and EDX. The encapsulation efficiency of the polymer for encapsulating (glycyl-l-histidyl-l-lysine-copper) GHK-Cu was observed to be 55.26% and peptide release of 51.84% was observed for Ethylene glycol dimethacrylate based polymer after 24 h whereas for Trimethylolpropane triacrylate based polymer the encapsulation efficiency was observed to be 49.6% and release was 39.01%. The EGDMA based polymer was further examined under in vivo studies in order to evaluate the efficiency of the synthesized polymer. The in vivo studies include wound closure, histopathological analysis, biochemical and toxicity assay. The material has shown promising results for both in vivo and in vitro studies.

Fabrication and evaluation of antimicrobial biomimetic nanofiber coating for improved dental implant bioseal: An in vitro study.

BACKGROUND: A weak implant-soft tissue interface may lead to bacterial ingression, breakdown of underlying tissues, and eventually implant failure. This study proposes a surface modification technique of titanium alloy (Ti), using a nano-biopolymer scaffold to enhance soft tissue attachment in dental implants. METHODS: Gelatin (20% w/v) embedded with 10 ± 2 nm silver nanoparticles (AgNPs) was electrospun to form a gelatin electrospun mat (GEM) scaffold, bonded to Ti alloy surface using chemical surface functionalization. Antimicrobial activity of AgNPs was tested against representative Gram-positive (Staphylococcus aureus) and Gram-negative bacteria (Escherichia coli) at 4, 24, and 48 hours and after embedding in scaffold at 48 hours. Cytotoxicity analysis (MTT assay) was performed using the 3T3 mouse fibroblast cell line at 24 and 72 hours for two groups: control (unmodified Ti disc) and experimental (GEM embedded with AgNPs); and further validated by scanning electron microscopy. RESULTS: The AgNPs-embedded GEM showed good antimicrobial activity at 48 hours, with the AgNPs showing complete (99.99%) inhibition of bacterial colony counts at 24 and 48 hours. Cell viability and proliferation over the GEM modified Ti discs were seen to be significantly increased (P < 0.05) at 72 hours as compared with control. SEM images revealed intimate spreading of fibroblasts, with differentiated cell morphology and pseudopodial processes, indicative of enhanced fibroblastic adhesion, growth, and differentiation over the scaffold. CONCLUSION: Results show good antifouling properties and biocompatibility of the fabricated coating, making it a promising strategy to reduce postoperative infections and peri-implant diseases in Ti dental implants.

A combinatorial approach to modulate microenvironment toward regeneration and repair after spinal cord injury in rats.

Traumatic spinal cord injury (SCI) is a devastating condition of CNS which leads to loss of sensory as well as motor functions. Secondary damage after SCI initiates cascade of events that creates an inhibitory milieu for axonal growth and repair. Combinatorial therapies are the hope to attenuate secondary injury progression and make the microenvironment growth and repair friendly for the neurons. We fabricated gelatin- genipin hydrogel system which was impregnated with IONPs and injected at the lesion site in a clinically relevant contusion rat model of SCI. 24 h later, the rats were exposed to magnetic fields (17.96 µT, 50 Hz uniform EMF) for 2 h/day for 5 weeks. A significant (P < 0.001) improvement in Basso, Beattie and Bresnahan (BBB) locomotor score, amplitude and threshold of spinally mediated reflexes and motor and somatosensory evoked potentials (MEP & SSEP) was observed following IONPs implantation and EMF exposure. Moreover, retrograde tracing showed a higher level of neuronal connectivity and survival after the intervention. There was also a reduction in activated microglia and lesion volume which attenuate secondary damage as evident by reduction in the scaring following intervention for 5 weeks. Moreover, we observed increase in the neuronal growth cone marker, GAP-43, growth promoting neurotrophins (GDNF, BDNF & NT-3) and reduction in the inhibitory molecule (Nogo-A) after this combinatorial therapy. We obsrvered that a significant improvement in behavioral, electrophysiological and morphological parameters was due to an alteration in neurotrophin levels, reduction in activated microglia and increase in GAP-43 expression after the combinatorial therapy. We propose that implantation of IONPs embedded gelatin-genipin hydrogel system along with MF exposure modulated the microenvironment, making it conducive for neural repair and regeneration.

Solution-State Long-Range Molecular Ordering in Poly(3-hexylthiophene).

A blend of poly(3-hexylthiophene) (P3HT) and poly(n-hexyl isocyanate-block-2-vinylpyridine) (PHIC-b-P2VP) in a common solvent shows the formation of long-range (micrometer-scale) nanowires of P3HT through hydrophobic interactions between the hexyl arms of P3HT and PHIC in a parallel way, which increase the planarity that leads to the generation of vibration bands with a lower free exciton bandwidth (W = 67 meV) in the solution state, which is further decreased to 9 meV after 48 h annealing of the blend film. The resulting nanowires of the P3HT show a 100-fold increase in current in comparison to pristine P3HT.

In Vitro and in Vivo Studies of pH-Sensitive GHK-Cu-Incorporated Polyaspartic and Polyacrylic Acid Superabsorbent Polymer.

The main aim of this study was to evaluate the in vitro and in vivo efficiency of the polyaspartic acid- and acrylic acid-based superabsorbent polymer. The synthesized polymer was first investigated to check the blood compatibility by protein adsorption and blood clotting tests. Further, the GHK-Cu peptide was incorporated within the polymer and release studies were performed to evaluate the drug-delivery efficiency of the superabsorbent polymer. The polymer with best peptide release results were further used for in vivo analysis for wound healing. The healing efficiency of polymer with and without peptide was analyzed using wound closure, biochemical assay, histopathological, and toxicity studies.

Synthesis and Characterization of Zero-Valent Iron Nanoparticles, and the Study of Their Effect against the Degradation of DDT in Soil and Assessment of Their Toxicity against Collembola and Ostracods.

Of late, novel magnetic nanomaterials have drawn worldwide attention because of the uniqueness in their properties and uses. In our studies, we have prepared nearly monodisperse zero-valent iron nanoparticles (nZVIs) of diameter of less than 60 nm in aqueous medium by a reductive precipitation process and pectin as stabilizing agent. The characterization of these nanoparticles was done by dynamic light scattering and transmission electron microscopy (TEM) techniques. The TEM images confirmed that the average size of the nZVIs was about 25 nm. The resultant nZVIs were then employed to degrade DDT (dichlorodiphenyltrichloroethane) in spiked soil, and their toxicity toward Collembola (Folsomia candida) and Ostracods (Heterocypris incongruens) was measured. The fabricated nZVIs degraded DDT in soil quite effectively. Further, the effects of nZVIs on Collembola and Ostracods were found to be negative. This was due to the oxidation of nZVIs and creation of anoxic conditions thereupon, and the generation of excess Fe(II) in soil. In addition, the negative effects of DDT on ostracod development and Collembola reproduction were found to be quite weak.

Development of polymeric nanopaclitaxel and comparison with free paclitaxel for effects on cell proliferation of MCF-7 and B16F0 carcinoma cells.

Paclitaxel is hydrophobic in nature and is recognized as a highly toxic anticancer drug, showing adverse effects in normal body sites. In this study, we developed a polymeric nano drug carrier for safe delivery of the paclitaxel to the cancer that releases the drug in a sustained manner and reduces side effects. N-isopropylacrylamide/ vinyl pyrrolidone (NIPAAm/VP) nanoparticles were synthesized by radical polymerization. Physico- chemical characterization of the polymeric nanoparticles was conducted using dynamic light scattering, transmission electron microscopy, scanning electron microscopy and nuclear magnetic resonance, which confirmed polymerization of formulated nanoparticles. Drug release was assessed using a spectrophotometer and cell viability assays were carried out on the MCF-7 breast cancer and B16F0 skin cancer cell lines. NIPAAm/ VP nanoparticles demonstrated a size distribution in the 65-108 nm range and surface charge measured -15.4 mV. SEM showed the nanoparticles to be spherical in shape with a slow drug release of ~70% in PBS at 38° over 96 h. Drug loaded nanoparticles were associated with increased viability of MCF-7 and B16F0 cells in comparison to free paclitaxel. Nano loaded paclitaxel shows high therapeutic efficiency by sustained release action for the longer period of time, i increasing its efficacy and biocompatibility for human cancer therapy. Therefore, paclitaxel loaded (NIPAAm/VP) nanoparticles may provide opportunities to expand delivery of the drug for clinical selection.

In vitro toxicity and antidiabetic activity of a newly developed polyherbal formulation (MAC-ST/001) in streptozotocin-induced diabetic Wistar rats.

The present study was designed to investigate the hypoglycemic effect of an aqueous extract of MAC-ST/001 (a new polyherbal formulation) which was given once daily to rats at different doses. The animals were divided into diabetic and nondiabetic control groups. The duration of each experiment lasted from 1 week to 1 month, and the results were compared with that of the standard hypoglycemic drug glibenclamide (10 mg/kg), which was given once daily. In this study, biochemical and histopathological parameters were studied in streptozotacin (STZ) (single intraperitoneal injection of 55 mg/kg)-induced diabetic rats. The diabetic rats showed a significant (p < 0.05 and p < 0.01) decrease in their body weight and serum amylase with marked elevation in blood glucose, serum cholesterol, blood urea nitrogen, creatinine, alkaline phosphatase, and serum transaminases (AST and ALT) after 1 week till the 28th day of diabetes. Cytotoxicity of MAC-ST/001 formulation was also studied on C2C12, 3T3-L1, and HepG2 cells through MTT assay. Histological examination of the liver and pancreas of normal control, diabetic control, and drug-treated rats revealed significant results. Finally, it was concluded that administration of this MAC-ST/001 extract reversed most blood and tissue changes caused by STZ-induced diabetes in rats.