Building and behavior of a pH-stimuli responsive chitosan nanoparticles loaded with folic acid conjugated gemcitabine silver colloids in MDA-MB-453 metastatic breast cancer cell line and pharmacokinetics in rats.

The pH-stimuli release behavior of nanoformulations may enhance the success rate of chemotherapeutic drugs in cancers by site-specific delivery of drugs to cancer tissues. The aim of the present study was to prepare chitosan (CS) nanoparticles (NPs) with previously synthesized folic acid (FA) capped silver nanoparticles (AgNPs) loaded with the anti-cancer drug gemcitabine (GEM) (FA-GEM-AgNPs). The CS-FA-GEM-AgNPs (CS-NPs) were characterized with dynamic light scattering (DLS), transmission electron microscopy (TEM), energy dispersive x-ray analysis (EDAX), selected area electron diffraction (SAED), and differential scanning calorimetric (DSC) analyses. The in-vitro drug release of GEM was evaluated in media of different pH. The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay was carried out to determine the cytotoxic effects of the prepared nanoformulations in media with various pH. The time- and pH-dependent apoptotic cell death induced by CS-NPs with MDA-MB-453 human breast cancer cell line was observed using acridine orange (AO)/ethidium bromide (EtBr) staining. The pharmacokinetic parameters were studied with high-performance liquid chromatography (HPLC) and atomic absorption spectroscopy (AAS). Two batches of CS-NPs formulations were prepared, one with AgNPs of particle size 143 nm and the other with 244 nm. The particle size for CS-NPs-I (FA-GEM-AgNPs-143 nm) and CS-NPs-II (FA-GEM-AgNPs-244 nm) was found to be 425 and 545 nm, respectively. The zeta potential was found to be 36.1 and 37.5 mV for CS-NPs-I and CS-NPs-II, respectively. CS-NPs-I and CS-NPs-II showed a polydispersity index (PDI) of 0.240 and 0.261, respectively. A TEM study confirmed the spherical nature of the NPs. The nanoformulations exerted pH-dependant effect against MDA-MB-453 cells with relatively higher cytotoxicity at the lower pH than at higher pH levels. The pharmacokinetic profile and tissue distribution of CS-NPs in rats exerted drug release in a pH-dependent manner with enhanced excretion of Ag+. An optimized nanoformulation for pH-stimuli responsive release of GEM was successfully developed for future therapeutic exploration.

Ameliorative potential of flavonoids of Aegle marmelos in vincristine-induced neuropathic pain and associated excitotoxicity.

Introduction: The objective of the study is to elucidate the effect of Aegle marmelos bark hydroalcoholic extract (AMHE) and the role of its constituents marmelosin, umbelliferone, and Para-coumaric acid in attenuating neuropathic pain.Methodology: Peripheral neuropathy was induced by vincristine 100 µg/ml. AMHE was administered in three dose levels (100, 200 and 300 mg/kg) for 21 days. Mechanical hyperalgesia and allodynia were assessed by Randall Sellitto and electronic Von-Frey test, respectively. Functional loss and recovery of the nerve were assessed by sciatic functional index test. The nerve conduction velocity and formalin test were done to assess the peripheral and central response of the extract. Inflammatory mediators in both sciatic nerve and brain and neurotransmitters glutamate and aspartate were measured to support the data.Results and discussion: The inflammatory mediators in both sciatic nerve and brain (TNF-α, IL-1ß, and IL-6) were found to be attenuated with AMHE-treated group in comparison to the group treated only with vincristine, which indicates the extract has anti-inflammatory property. AMHE treated rats were found to be active in all the behavioural tests, suggesting its activity could be mediated through a central and peripheral mechanism to attenuate the pain response. The levels of excitatory neurotransmitters were found to be reduced with AMHE treatment.Conclusion: It could be concluded that AMHE is active in attenuating the neuropathic pain caused by vincristine. The peripheral action would have mediated through lowering the inflammatory mediators as well as the excitotoxicity caused due to peripheral neuropathy and neuroinflammation.

Saponins of Tribulus terrestris attenuated neuropathic pain induced with vincristine through central and peripheral mechanism.

The study comprises exploring the effects of saponins from Tribulus terrestris (TT) in attenuating the neuropathic pain caused by vincristine (100 µg/ml i.p.) for 10 days (in two 5 day cycles with 2 days pause). Mechanical hyperalgesia and allodynia were assessed by Randall-Sellitto and electronic von Frey tests, respectively. Chemical- induced nociception was assessed by formalin test. Neurophysiological effect of the extract was evaluated by recording sciatic functional index (SFI) on the test days (7, 10, 14, and 21) and sciatic nerve conduction velocity test (SNCV) on the last day. Inflammatory mediators (TNF-α, IL-1ß, and IL-6) in both sciatic nerve and brain and brain neurotransmitters, glutamate and aspartate, were measured to support the behavioral response. The saponins of TT-treated group were found to be effective in the behavioral experiments, implying its activity both centrally and peripherally in attenuating pain. The inflammatory mediators in both sciatic nerve and brain (TNF-α, IL-1ß, and IL-6) were found to be attenuated with TT saponin treatment in comparison to vincristine-treated group, indicating its anti-inflammatory property. The excitatory neurotransmitters, L-glutamic acid and L-aspartic acid, were also found to be attenuated with TT saponins, implying restoration of neuronal damage and synaptic activity caused by high amount of glutamate due to excess TNF-α in brain and reversing the nociceptive threshold lowered due to aspartate. Thus, TT(S) is peripherally and centrally active in lowering the inflammatory mediators, reversing the neuronal damage and increasing the nociceptive threshold caused due to peripheral neuropathy.