Evaluation of antioxidant and antimicrobial potential of a novel Himalayan plant Reinwardtia indica dumort: Scientifically unexplored.

Reinwardtia indica (Lineceae) is a medicinal plant cultivated in the Himalayan region. It is effectively used in folk medicines for the treatment of various health complications. In the present study, the shade dried leaves and stem were extracted in three different solvents such as ethyl acetate, ethanol, and hydro-alcoholic. The antioxidant efficacy of these extracts was confirmed by using different in vitro assays: DPPH free radical scavenging, superoxide radical scavenging, lipid peroxidation, metal ion chelating capability and reducing power determination. Total phenol content was maximum in hydro alcoholic extract of leaf (540.37 mg per g of gallic acid equivalents) and stem (330.51 mg per g of gallic acid equivalents) while flavonoid content was maximum in ethanolic extract of leaf (305 mg per gram of rutin equivalents) and ethyl acetate extract of stem (170.6 mg per gram of rutin equivalents). The antioxidant activity of these extracts was positively correlated with their total phenol and flavonoid content. Among all tested extracts, ethanolic extract of leaf exhibit maximum zone of inhibition against all tested clinical isolates of bacterial (E. coli 11.00 ±â€¯1.73 mm, P. aeurogenosa 11.67 ±â€¯0.58 mm and S. aureus 10.33 ±â€¯1.53 mm) and fungal (C. albicans 11.33 ±â€¯1.10 mm) pathogens, while ethyl acetate extracts of the leaf and stem showed minimum inhibitory concentration against all tested microorganisms. Thus, R. indica extracts can be used as potent natural antioxidant having antifungal and antibacterial action.

Antioxidant, antimicrobial and cytotoxic potential of silver nanoparticles synthesized using flavonoid rich alcoholic leaves extract of Reinwardtia indica.

The present work discusses the establishment of a green route for the rapid synthesis of silver nanoparticles (AgNPs) using an alcoholic extract of Reinwardtia indica (AERI) leaves which act as a reducing as well as a capping agent. The change in color from yellowish green to dark brown confirmed the synthesis of AgNPs. A characteristic surface plasmon resonance (SPR) band at 436 nm advocated the presence of AgNPs. The synthesis process was optimized using one factor at a time approach where 1.0 mM AgNO3 concentration, 5 mL 0.4% (v/v) of AER inoculum dose and 30 min of sunlight exposure were found to be the optimum conditions. The synthesized AgNPs was characterized by several characterizing techniques such as HR- TEM, SAED, HR-SEM, EDX, XRD, FTIR and AFM analysis. For evaluation and comparison of AgNPs with AERI used human pathogen E. coli, P. aeurogenosa, S. aeurus and C. albicans for antimicrobial, for cytotoxicity study SiHa cell line at concentration of (10, 50, 100, 250 and 500 µg mL-1) and for enzymatic assay superoxide dismutase, catalase, malondialdehyde and glutathione peroxidase method were used. The size of nanoparticle in the range of 3-15 nm was confirmed TEM, spherical shape by SEM and crystal lattice nature by XRD. AFM results revealed the 2 D and 3 D pattern of particle scatter nature on the surface. This protocol as simple, rapid, one step, eco-friendly, nontoxic and AgNPs showed strong antimicrobial activity as well as cytotoxic potential in comparison to AERI.

Revalidation of the neuroprotective effects of a United States patented polyherbal formulation on scopolamine induced learning and memory impairment in rats.

OBJECTIVE: Alzheimer's disease (AD) is the most common cause of dementia yet treatment options are extremely limited. The disease is associated with cognitive impairment as well as structural irregularities, accumulation of plaques and neurofibrillary tangles, diminished levels of acetylcholine, oxidative stress, and inflammation in the brain. We have previously reported on the positive effects of a united states patented (US 7,273,626 B2) poly herbal test formulation, consisting of Bacopa monnieri, Hippophae rhamnoides and Dioscorea bulbifera extracts, on cognitive deficits in AD patients. The present study was conducted to investigate the mechanism(s) of action of the formulation using scopolamine treated rats as an AD model. METHOD: The formulation was administered daily along with scopolamine for a period of 14days following which the elevated plus maze, passive avoidance, and Morris water maze tests were performed to assess learning and memory. Rats treated with scopolamine or vehicle only were also included in the experiment. Acetylcholine levels and activities of acetylcholinesterase (AChE) and anti-oxidant enzymes in the brain were also measured at the end of the treatment period. RESULTS: The study demonstrate that scopolamine treatment resulted in learning and memory deficits which were partially and significantly ameliorated by the formulation. The formulation also counteracted scopolamine-induced decreases in acetylcholine levels, increases in AChE activity, and decreases in activities of the antioxidant enzymes. CONCLUSION: The study demonstrates the ability of the test formulation to reverse scopolamine-induced learning and memory deficits in rats which may at least partially be explained by the reversal of scopolamine-induced reductions in brain acetylcholine levels and antioxidant activities by the test formulation.

Nanovesicles of nitrendipine with lipid complex for transdermal delivery: pharmacokinetic and pharmacodynamic studies.

CONTEXT: Vesicular transdermal delivery can enhance the bioavailability of a drug especially affected by first-pass metabolism, e.g. nitrendipine. However effective transdermal delivery employs permeation enhancer, e.g oleic acid (OA) with ceramide 2, stearic acid, behenic acid, and cholesteryl sulfate lipid complex. OBJECTIVE: This study investigated the preparation, characterization of physicochemical properties, ex vivo permeation using human skin, pharmacokinetic parameters and antihypertensive potential in rats, of nitrendipine-loaded nanovesicles of ceramide 2, stearic acid, behenic acid and cholesteryl sulfate containing oleic acid gel (NOVG). MATERIALS AND METHODS: The nanovesicles were made using film hydration method and characterized for physicochemical properties, ex vivo permeation using human skin, pharmacokinetic parameters and antihypertensive potential. RESULTS: Nitrendipine-loaded nanovesicles of ceramide-2 containing oleic acid (NOV-5) have shown fluxes in the range of 4.88-24.72 µg/cm(2)/h nitrendipine oral suspension (NOS) at equal dose. NOVG-5 has shown almost 33% reduction in blood pressure in the first hour and a further decrease of 25% in the second hour to restore the normal pressure. DISCUSSION: The permeation increases with increase in OA content. OA gets integrated in vesicle wall and enhances its permeability, whereas ceramide content makes sure that skin does not become damaged even after permeation. CONCLUSION: NOVG-5 has shown the most favorable physicochemical properties and good permeation through skin providing good management of hypertension during crucial initial hours.

Formulation and characterization of taste masked ondansetron-magnesium aluminum silicate adsorption systems.

CONTEXT: Taste masking greatly influences the acceptability of bitter tasting formulation; moreover, it governs the commercial and therapeutic success of drug products. OBJECTIVE: This work is directed toward masking the bitter taste of ondansetron HCl (ONS) utilizing the excipient, which can delay the reach of drug to the taste buds. MATERIAL AND METHODS: Magnesium aluminum silicate (Veegum F), a clay material having capability to adsorb the drugs onto it, was used. The adsorption systems of ONS with Veegum were obtained by dynamic adsorption technique and examined by scanning electron microscopy, differential scanning calorimetry, Fourier transform infrared (FTIR) spectroscopy, and X-ray diffraction (XRD) for morphology, thermal behavior, and interactions. The taste assessment of prepared systems was done by in vitro method based on drug release. RESULTS: The molecular interaction between ONS and Veegum in the system was revealed by FTIR spectroscopy. A change in thermal behavior of the system was observed owing to interaction or replacement of the cationic groups of Veegum with that of ONS. XRD studies revealed that the prepared system was having lower crystallinity as compared to ONS. The in vitro drug release study showed that ONS release from the system was relatively slow in basic environment than the acidic one. DISCUSSION: Adsorption of ONS on the surface of Veegum was mainly due to electrostatic interactions and hydrogen bonding. CONCLUSION: The experimental results reveal the successful intercalation of ONS into the space available between the layers of Veegum. Furthermore, this resulted in a control on drug release in salivary pH resulting in a concentration lower than bitterness threshold.

Formulation and evaluation of lipid based taste masked granules of ondansetron HCl.

INTRODUCTION AND AIM: Various taste masking approaches comprising the excipients which delay the reach of the drug to taste buds are reported. Lipidic substances can act as release retarding agent and provides a matrix base responsible for suppressing the bitter taste of drug. This work was aimed to study the influence of different proportions of a lipid carrier on the inhibition of bitterness of the drug vis-a-vis in vitro release of drug from the granules. METHODS: The lipid-matrix granules of ondansetron HCl with Geleol pellets (glycerol monostearate) were obtained by manual hot melt fusion technique. The prepared granules were characterized by SEM, DSC and XRD. The taste assessment of prepared granules was done by in vitro method based on drug release. RESULTS: Distribution of drug inside the lipid-matrix granules was not properly analyzed by DSC and XRD, moreover these studies revealed no interaction between the drug and lipid. The dissolution tests displayed the significant retardation of drug release from the granules compared to pure drug and additionally indicated the attainment of matrix system via appearance of unbroken granules during in vitro testing. Higuchi relationship for drug release was obtained by drug release kinetics, which also revealed the functioning drug release mechanism, as diffusion controlled but the addition of hydrophilic substance (Cab-o-sil) has changed the mechanism of drug release. CONCLUSION: The proportions of Geleol and Cab-o-sil taken in granules had affected the dissolution profile. Higher amount of GE resulted in high taste masking ability.

Development and characterization of stable nanovesicular carrier for drug delivery.

Lipid vesicles are an important drug carrier which can serve for controlled delivery of drugs; however, these vesicles are quite unstable at ambient temperature and require stringent storage condition. Present work was done to develop a stable vesicular system for drug delivery. Vesicles of ceramide-2, cholesterol, cholesterol sulfate, and palmitic acid were prepared and compared with phosphatidylcholine vesicles for physicochemical parameters and accelerated stability. Diclofenac sodium was used as a model drug. Based on physicochemical parameter and in vitro release PCV-3 and CV-3 were selected for further studies in three different accelerated stability conditions. PCV-3 showed moderate changes at 4°C but was severely affected at 25°C and 40°C. CV-3 showed stable characteristics at 4°C and 25°C whereas at 40°C, CV-3 showed signs of slight modification owing to moisture absorption. Based on the study, CV-3 containing highest content of palmitic acid was found to be most stable.

Preparation, characterization and permeation studies of a nanovesicular system containing diclofenac for transdermal delivery.

CONTEXT: Transdermal formulations contain permeation enhancer which causes skin damage. Ceramide 2 is natural lipid found in stratum corneum (SC). OBJECTIVE: Drug-loaded nanovesicles of ceramide-2, cholesterol, palmitic acid, cholesteryl sulfate were formulated and analyzed for physical and biological properties. Diclofenac was used as a model drug. MATERIALS AND METHOD: The vesicles were prepared using the film hydration method and characterized for physical parameters, in vitro drug release, accelerated stability studies and formulated into gel. Respective gels were compared with a commercial formulation (CEG) and plain carbopol gel (CG) containing drug for ex vivo, in vivo drug permeation and anti-inflammatory activity. RESULTS: The vesicles were stable with optimum physical parameters. DCG-1 showed 92.89% in vitro drug release. Ceramide vesicles showed drug release between 18 and 25 µg/cm(2) whereas CG and CEG released 0.33 and 1.35 µg/cm(2) drug, respectively. DCG-1 and CEG showed corresponding Cmax at 6 and 4 h, respectively. DCG-1 showed six times AUC than CEG. DCG-1 inhibited edema by 86.37% by 4th hour of application. DISCUSSION: The presence of ceramide 2 specifically promotes the drug permeation through SC and dermis and also contribute towards stability and non-irritancy. CONCLUSION: The composition of the nanovesicle played an important role in physical properties and drug permeation.

Ceramide-2 nanovesicles for effective transdermal delivery: development, characterization and pharmacokinetic evaluation.

UNLABELLED: Abstract Context: The vesicles based on skin lipid have a drug localization effect and its main lipid, ceramide provides protective and regenerative effects while oleic acid (OA) is a penetration enhancer, however, it causes slight irritation, so we have formulated formulation incorporating both of these to develop a transdermal formulation for better permeation. OBJECTIVE: Present study investigated the preparation and characterization of physicochemical properties and permeation of nanovesicles of ceramide-2 containing OA and palmitic acid (PA) respectively and a commercial gel. MATERIALS AND METHODS: The vesicles were made using ceramide 2, cholesterol (Chol), cholesteryl sulfate (CS) and OA or PA, respectively, using film hydration method. The vesicles were characterized for physicochemical properties, ex vivo permeation using human skin and pharmacokinetic parameters and anti-inflammatory activity in rats. RESULTS: The vesicles showed size at 102-125 nm while PDI was 0.11-0.13 and negative zeta potential. OV-3 showed highest entrapment efficiency. The drug fluxes were 92.02 and 8.920 µg/cm(2)/h, respectively, for OV-3 and PV-1. The Cmax were 7.91 and 4.01 µg/ml at 4 and 6 h for OV-3 (2.5 mg) and PV-1 (10 mg), respectively. OV-3 and PV-1 showed 98.8% and 77.36% edema inhibition, respectively, at 3 h. DISCUSSION: Both formulations showed similar physical parameters and different permeation since OA get incorporated in vesicles and increases its permeability and ceramide makes sure that vesicles can rapidly traverse the stratum corneum. CONCLUSION: OV-3 containing 3% OA showed optimum physical parameters and good permeation with maximum anti-inflammatory activity.

Development of a new nanovesicle formulation as transdermal carrier: formulation, physicochemical characterization, permeation studies and anti-inflammatory activity.

CONTEXT: Ibuprofen is an important NSAID, however, it can cause GI disturbances when given orally, and employment of transdermal route will require permeation enhancer causing skin injury. OBJECTIVE: Drug-loaded nanovesicles of ceramide-2, cholesterol, palmitic acid, and cholesteryl sulfate (ICVG) were formulated and analyzed for physicochemical and permeation properties. MATERIALS AND METHOD: Vesicles were formulated using film hydration method and physicochemical parameters, in vitro drug release, and stability were assessed. Further, nanovesicle gels were evaluated against plain gel containing drug (CG) for ex vivo/in vivo drug permeation and anti-inflammatory activity. RESULTS: The developed formulations showed optimal physicochemical profile and ICV-1 gave 97.24% drug release. Drug permeation was between 17.32 and 33.12 µg/cm(2) for ICVG formulations and 0.27 µg/cm(2) for CG. ICVG-1 and CG showed Cmax of 9.6 and 0.7 µg/ml at 8 and 4 h. ICVG-1 showed 19.9 times higher AUC than CG. Edema inhibition was 57.98% during initial hours by ICVG-1. DISCUSSION: Ratio of ceramide 2 and palmitic acid plays a critical role in drug permeation through stratum corneum. The stability and protective effect of the formulations were due to ceramide content. CONCLUSION: The composition has an important role in physicochemical properties and drug permeation thereby generating an optimum formulation.

Formulation, evaluation and 3(2) full factorial design-based optimization of ondansetron hydrochloride incorporated taste masked microspheres.

CONTEXT: Masking the bitter taste of Ondansetron hydrochloride (ONS) may improve palatability, acceptance and compliance of ONS products. OBJECTIVE: ONS-loaded, taste-masked microspheres were prepared with a polycationic pH-sensitive polymer and 3(2) full factorial design (FFD) was applied to optimize microsphere batches. MATERIALS AND METHODS: Solvent evaporation, in acetone--methanol/liquid paraffin system, was used to prepare taste-masked ONS microspheres. The effect of varying drug/polymer (D/P) ratios on microspheres characteristics were studied by 3(2) FFD. Desirability function was used to search the optimum formulation. Microspheres were evaluated by FTIR, XRD and DSC to examine interaction and effect of microencapsulation process. In vitro taste assessment approach based on bitterness threshold and drug release was used to assess bitterness scores. RESULTS: Prepared ONS microspheres were spherical and surface was wrinkled. ONS was molecularly dispersed in microspheres without any incompatibility with EE100. In hydrochloric acid buffer pH 1.2, ONS released completely from microsphere in just 10 min. Contrary to this, ONS release at initial 5 min from taste-masked microspheres was less than the bitterness threshold. CONCLUSION: Full factorial design and in vitro taste assessment approach, coupled together, was successfully applied to develop and optimize batches of ONS incorporated taste-masked microspheres.

Solid lipid nanoparticles of guggul lipid as drug carrier for transdermal drug delivery.

Diclofenac sodium loaded solid lipid nanoparticles (SLNs) were formulated using guggul lipid as major lipid component and analyzed for physical parameters, permeation profile, and anti-inflammatory activity. The SLNs were prepared using melt-emulsion sonication/low temperature-solidification method and characterized for physical parameters, in vitro drug release, and accelerated stability studies, and formulated into gel. Respective gels were compared with a commercial emulgel (CEG) and plain carbopol gel containing drug (CG) for ex vivo and in vivo drug permeation and anti-inflammatory activity. The SLNs were stable with optimum physical parameters. GMS nanoparticle 1 (GMN-1) and stearic acid nanoparticle 1 (SAN-1) gave the highest in vitro drug release. Guggul lipid nanoparticle gel 3 (GLNG-3) showed 104.68 times higher drug content than CEG in receptor fluid. The enhancement ratio of GLNG-3 was 39.43 with respect to CG. GLNG-3 showed almost 8.12 times higher C(max) than CEG at 4 hours. The AUC value of GLNG-3 was 15.28 times higher than the AUC of CEG. GLNG-3 showed edema inhibition up to 69.47% in the first hour. Physicochemical properties of major lipid component govern the properties of SLN. SLN made up of guggul lipid showed good physical properties with acceptable stability. Furthermore, it showed a controlled drug release profile along with a promising permeation profile.

Development of aceclofenac nanovesicular system using biomaterial for transdermal delivery: physical characterization, ex vivo, in vivo, and anti-inflammatory studies.

CONTEXT: Aceclofenac is an important NSAID; however, it causes GI disturbances whereas employing transdermal route would require permeation enhancer for systemic application, thereby causing skin damage. Ceramide 2 is a natural lipid having an important role in the maintenance of skin. OBJECTIVE: Aceclofenac-loaded nanovesicles of ceramide-2, cholesterol, palmitic acid, and cholesteryl sulfate were formulated and analyzed for physical and biological properties. MATERIALS AND METHOD: Film hydration method was used to prepare the vesicles and physical parameters, in vitro drug release and stability were evaluated. Then, they were formulated into gel and evaluated against a commercial formulation (CF) and gel-containing plain drug (CPG) for ex vivo, in vivo drug permeation, and anti-inflammatory activity. RESULTS: The developed formulations showed best physical profile and ACV-1 gave 92.89% drug release in in vitro studies. Ex vivo studies showed drug permeation between 15.32-31.12 µg/cm(2), whereas CPG and CF released 0.47 and 2.81 µg/cm(2), respectively. ACVG-1 and CF showed Cmax of 8.1 and 1.2 µg/ml at 8 and 4 h, respectively. ACVG-1 showed 11.6 times AUC than CF. ACVG-1 inhibited edema by 44% in first hour itself. DISCUSSION: Ceramide 2 and palmitic acid played an important role in the formulation and promotes the drug permeation through stratum corneum and dermis. Ceramide content of the formulation also contributes towards stability and skin protection. CONCLUSION: The composition of the vesicle formulation performs an important role in physical properties and drug permeation, thereby producing an optimum formulation.

Targeted drug delivery of Rifampicin to the lungs: formulation, characterization, and stability studies of preformed aerosolized liposome and in situ formed aerosolized liposome.

PURPOSE: This study aimed at the preparation and characterization of preformed and in situ formed liposomes for sustained delivery to the lungs. METHODS: Two different liposome formulations were prepared and subjected to characterization of physical parameters and drug release profile (% cumulative drug release and % drug retained). Formulations were then subjected to accelerated stability studies as per ICH guidelines. RESULTS: In situ formed liposome showed better sustained release profile than the preformed liposome as it released sufficient amount of drug while retaining considerable amount of drug. Upon subjection to accelerated conditions for 60 days, preformed liposome lost the objective of being controlled release formulation.

In-situ formation of liposome of rifampicin: better availability for better treatment.

This study aimed at the preparation and characterization of preformed and in-situ formed liposomes for controlled delivery to the lungs. Two different liposome formulations were prepared and subjected to characterization of physical parameters (vesicle size, appearance, discharge rate, spray pattern and internal pressure) and drug release profile (% cumulative drug release and % drug retained). Formulations were then subjected to accelerated stability studies as per ICH guidelines. It was observed that vesicle size ranged between 2.35 +/- 0.23 - 2.41+/- 0.3 microm and 1.22 +/- 0.22- 1.34 +/- 0.15 microm respectively for preformed and in-situ formed liposomes. The discharge rate and spray area were in ranges of 110 +/- 2.4 -123 +/- 2.6 & 115+/- 2.4 -127+/-2.3 mg/ actuation & 12.5 +/-1.7 -14.1+/- 2.2 and 13.9 +/- 2.1- 14.7+/- 2.0 cm(2) respectively. In-situ liposomes showed better controlled release profile then preformed liposomes as it released 76.5+/- 2.4 - 58.5 +/- 1.8 % drug in 12 hour while retaining 23.5 +/- 1.9- 41.5 +/- 1.6 % drug whereas preformed liposomes showed cumulative release of 66.5 +/- 2.2- 84.5 +/- 2.3 % and 15.5 +/- 2.4- 33.5+/- 2.1 % fraction retained. Upon subjection to accelerated conditions for a period of 60 days, preformed liposome completely lost the objective of being controlled release formulation as they showed 92+/- 2.4 % cumulative release and only 08 +/- 1.6 % fraction retained whereas in-situ formulation showed 62 +/- 2.3 % cumulative release and 38 +/- 2.1 % fraction retained.