Biomedical subjects

Jagdish Singh

Publications and source records attributed to Jagdish Singh.

24 records · Page 2Linked to original sources

PubMed2003

Effect of polyols on the conformational stability and biological activity of a model protein lysozyme.

The purpose of this study was to investigate the stabilizing action of polyols against various protein degradation mechanisms (eg, aggregation, deamidation, oxidation), using a model protein lysozyme. Differential scanning calorimeter (DSC) was used to measure the thermodynamic parameters, mid point transition temperature and calorimetric enthalpy, in order to evaluate conformational stability. Enzyme activity assay was used to corroborate the DSC results. Mannitol, sucrose, lactose, glycerol, and propylene glycol were used as polyols to stabilize lysozyme against aggregation, deamidation, and oxidation. Mannitol was found to stabilize lysozyme against aggregation, sucrose against deamidation both at neutral pH and at acidic pH, and lactose against oxidation. Stabilizers that provided greater conformational stability of lysozyme against various degradation mechanisms also protected specific enzyme activity to a greater extent. It was concluded that DSC and bioassay could be valuable tools for screening stabilizers in protein formulations.

Alcohols↗

PubMed2002

Assessment of protein release kinetics, stability and protein polymer interaction of lysozyme encapsulated poly(D,L-lactide-co-glycolide) microspheres.

Using lysozyme as a model protein, this study investigated protein stability, protein--polymer interaction in different release media and their influence on protein release profile and in vitro--in vivo correlation. Lysozyme was microencapsulated into PLGA 50:50 by a double emulsion--solvent extraction/evaporation method. Protein stability, protein--PLGA adsorption and protein in vitro release were studied in various test media. Differential scanning calorimetry analysis showed lysozyme to be most conformationally stable in pH 4.0 acetate buffer with highest T(m) at 77.2 degree C and DeltaH(cal) 83.1 kcal/mol. Lysozyme exhibited good stability in pH 2.5 glycine buffer with T(m) at 63.8 degree C and DeltaH(cal) 69.9 kcal/mol. In pH 7.4 phosphate-buffered saline (PBS), lysozyme showed a trend toward aggregation when the temperature was elevated. When PLGA polymer was incubated with lysozyme in the various buffers, adsorption was found to occur in PBS only. The adsorption severely limited the amount of lysozyme available for release from microspheres, resulting in slow and incomplete release in PBS. In contrast, the release of the microspheres in acetate and glycine buffers was complete within 40 and 70 days, respectively. Radiolabeled lysozyme blood levels in rats from the microspheres correlated qualitatively well with in vitro release in glycine buffer as a release medium. This study suggests that protein stability and adsorption are critical factors controlling protein release kinetics and in vitro--in vivo correlation of PLGA microspheres.

Adsorption↗

PubMed2002

Effect of tricaprin on the physical characteristics and in vitro release of etoposide from PLGA microspheres.

The purpose of this article is to examine the effects of tricaprin on the physical characteristics and in vitro release of etoposide from poly (lactic-co-glycolic acid) microspheres. The microspheres were synthesized through the use of a single-emulsion solvent-extraction procedure. Samples from each batch of microspheres were then analyzed for size distribution, drug loading efficiency, surface characteristics, in vitro release, and in vitro degradation of microspheres. Microsphere batches were synthesized using three different etoposide concentrations (15%, 10%, and 5% w/w) with tricaprin concentrations of 25% and 50%. The incorporation of 50% tricaprin significantly increased (p<0.05) the size of the microspheres for all three etoposide concentrations in comparison to microspheres prepared without tricaprin (control). The percentage of tricaprin used did not significantly affect the drug loading efficiency of the microspheres. The addition of tricaprin was shown to significantly increase (p<0.05) the in vitro release of etoposide from the microspheres prepared with all three concentrations of etoposide and the two different tricaprin percentages. Examination of the surface characteristics of the tricaprin loaded microspheres showed a dimpled surface with what appeared to be pockets of tricaprin dispersed throughout. In the in vitro degradation study, the tricaprin microspheres grew very porous as the degradation time increased, but they still retained a recognizable structure even after 30 days of degradation.

Biocompatible Materials↗

PubMed2002

Transepidermal transport enhancement of insulin by lipid extraction and iontophoresis.

PURPOSE: To study the effect of Ethyl acetate (EtAc), 1:1 ratio of EtAc and Ethanol (EtOH) and 2:1 ratio of chloroform (C) and methanol (M) on the extent of lipid extraction from the stratum corneum (SC) and in vitro passive and iontophoretic transport of insulin through porcine epidermis. METHODS: The porcine epidermis was pretreated for 40 min with the following solvents: 1) EtAc or EtAc:EtOH (1:1) and 2) C:M (2:1), which is a standard solvent combination for lipid extraction. Franz diffusion cells and Scepter iontophoretic power source were used for the transport studies. Cathodal iontophoresis was performed at 0.2 mA/cm2 current density. Fourier transform infrared spectroscopy (FTIR) studies were performed to assess the extent of lipid extraction. Thin layer chromatography (TLC) and gas chromatography (GC) were used to quantitate the different classes of lipid and identify the composition of the fatty acids, respectively, extracted by solvent(s) treatments. RESULTS: Insulin flux was found to be significantly (P < 0.05) greater through solvent pretreated epidermis compared to untreated controls during both passive and iontophoretic transport. Pretreatment with EtAc:EtOH (1:1) exhibited an insulin flux of 15.29 x 10(-8) nmoles/ cm2/h compared to 52.71 x 10(-8) nmoles/ cm2/h during passive and iontophoretic transport, respectively. The passive and iontophoretic flux of insulin through EtAc:EtOH (1:1) pretreated epidermis was significantly greater (P < 0.05) than EtAc treated epidermis. The SC treated with solvents showed a decrease in peak areas of C-H stretching absorbances in comparison to untreated SC. A greater percent decrease in peak areas was obtained by EtAc:EtOH(1:1), in comparison to EtAc alone. Epidermal resistance measurements revealed its strong correlation with the amount of lipids present in the epidermis. The lipids extracted consisted of six series of ceramides, fatty acids. triglycerides, cholesterol, cholesterol esters, cholesterol sulfate and phospholipids. CONCLUSIONS: The SC lipid extraction using suitable solvents followed by iontophoresis can synergistically enhance the transepidermal transport of insulin.

Acetates↗

PubMed2002

Lysozyme stability in primary emulsion for PLGA microsphere preparation: effect of recovery methods and stabilizing excipients.

PURPOSE: To investigate the conformational stability of a model protein, lysozyme, in the primary emulsion phase of the microsphere preparation process. METHODS: The conformational stability of lysozyme during primary emulsification was assessed by differential scanning calorimetry (DSC) and enzymatic activity assay. PEG 400 was used to separate lysozyme from water-in-oil (w/o) emulsion containing poly(lactideco-glycolide) (PLGA). RESULTS: No significant changes in the recovery of lysozyme were observed due to increasing sonication time from 20 to 60 s at 40 W or increasing intensity from 40 to 60 W for 20 s. By using the method involving PEG 400, lysozyme recovery in the presence of PLGA was increased from 11.8% to 709%. Hydroxypropyl-beta-cyclodextrin (HP-beta-CD) increased lysozyme recovery from 35% to 70% at low lysozyme concentration (20 mg/ml), and from 70% to 77% at high lysozyme concentration (100 mg/ml) in the presence of PLGA. Sugars such as trehalose and mannitol failed to increase lysozyme recovery. DSC results suggested the retention of the conformational structure of the recovered lysozyme, which was supported by an enzymatic activity assay. CONCLUSIONS: HP-beta-CD was found to be a promising stabilizer that protected lysozyme during the primary emulsification. Protein recovery method with the help of PEG 400 allowed the study of protein stability in w/o emulsions in the presence of PLGA. DSC provided supplementary information on the conformational changes of lysozyme during emulsification.

2-Hydroxypropyl-beta-cyclodextrin↗

PubMed2002

In vitro permeability and binding of hydrocarbons in pig ear and human abdominal skin.

Human skin has continual exposure to chemicals due to various occupational activities. Chemicals that get on skin have the potential to be absorbed. Hence, the potential human health hazards of a chemical must include an estimate for percutaneous absorption. An inexpensive, easy, and adequate model for the quantitative measurement of skin penetration of chemicals from JP-8 is absent. Cutaneous penetration studies in vitro through human skin are severely limited due to the lack of availability of the human skin. In this study, we have shown that pig ear skin can be used as a model for risk assessment from the percutaneous absorption of chemicals. We determined flux and permeability coefficient (Kp) of three chemicals--heptane, hexadecane, and xylene--from their permeation profile through porcine and human skin. Binding of these chemicals to porcine stratum corneum (SC) and human SC were also determined. Factors of difference (FOD) in the permeability of pig and human skin were 1.71, 1.28, and 1.16, respectively, for heptane, hexadecane, and xylene. FOD in binding of heptane, hexadecane, and xylene to pig and human SC were found to be 1.04, 0.76, and 1.31, respectively. Since, FOD for permeability and binding parameters were less than 2, hence, we conclude that pig ear skin can be used as model for humans for risk assessment from percutaneous absorption of chemicals.

Alkanes↗