Statistical Optimisation of Fermentation Media for Novel Clostridium novyi -NT Using Response Surface Methodology

ABSTRACT Clostridial collagenase is recognized as one of the important proteolytic enzymes used in treatment of varieties of fibro-proliferative disorders. The present work aimed to optimise the biomass concentration and collagenase enzyme activity of novel bacilli Clostridium novyi-NT. The response surface methodology tool was used to identify the optimal fermentation parameters. Central composite design (CCD) was applied with respect to three influencing factors - pH, proteose peptone and trypticase soya broth. These factors showed significant effect on the overall biomass concentration and collagenase enzyme activity (p<0.05). The maximum biomass concentration in terms of absorbance and collagenase activity (of crude enzyme) was achieved and recorded as 0.8309±0.0012 and 298.88±1.36 units/mg respectively after 22 hours of fermentation period while optimisation of media factors with help of response surface quadratic model. This is the first study to report maximum biomass concentration and collagenase activity by Clostridium novyi-NT till date by combining statistical designs.

Formulation and Evaluation of Nanoscale Solid Lipid Particles Containing a Hydrophilic Drug-Rasagiline Mesylate.

The present study was to prepare and characterize Rasagiline mesylate loaded nanoscale solid lipid particles and evaluate its In-vitro release. Rasagiline mesylate loaded solid lipid nanoparticles was fabricated by microemulsion technique and then characterized with respect to particle size, polydispersity index and zeta potential were measured by photon correlation spectroscopy. Morphological examination was visualized by transmission electron microscopy. The release of Rasagiline mesylate from solid lipid nanoparticles was performed by using the incubator shaking technique. The fabricated formulations particle size ranged from 160.20±3.2 to 210.12±5.3 nm and the zeta potential measurements indicates a narrow size distribution. The Polydispersity index values shown narrow that means the nanoparticles are homogenous in nature. Drug loading and entrapment efficiency of RMSLN-I & RMSLN-II were 33.34±1.45 % & 76.24±1.2 % and 16.67±0.86 % & 73.75±1.02 %, respectively. TEM revealed the particles were monodisperse, uniform size and quasispherical shape. Release of Rasagiline mesylate loaded nanoscale solid lipid particles were shown efficient prolonged drug release and followed by Fickian diffusion mechanism. Furthermore, RM loaded SLNs were found to be stable, after 6 months of storage at different conditions. The developed solid lipid nanoparticles were able to control the drug release for a prolonged period of time.

Fabrication, Physicochemical Characterization and Evaluation of In vitro Anticancer Efficacy of a Novel pH Sensitive Polymeric Nanoparticles for Efficient Delivery of Hydrophobic Drug against Colon Cancer.

Objective: The present investigation was aimed to overcome the limitations and to enhance the incorporation of the hydrophobic drug into polymeric nanoparticles and characterize the prepared nanoparticles and also to evaluate the in vitro anticancer efficacy of prepared nanoparticles. Method: Nanoprecipitation method was used to prepare plain and hydrophobic drug (Camptothecin) loaded polymeric nanoparticles. Prepared nanoformulations were evaluated for average particle size, particle size uniformity, surface area, zeta potential, surface morphology, drug content, encapsulation efficiency, drug loading, in vitro release, anticancer activity and stability studies at long term and accelerated storage conditions. Results: Plain and Camptothecin loaded polymeric nanoparticles were successfully prepared by nanoprecipitation method using stirring technique. Prepared Camptothecin encapsulated polymeric nanoparticles were (a) spherical in shape with size < 100 nm, displayed excellent uniformity with <0.3 and zeta potential > 20 mV; (b) showed > 95% release in colonic environment; (c) demonstrated enhanced anticancer activity than pure Camptothecin; and (d) extremely stable at both long term and accelerated storage conditions. Conclusion: In summary, the investigation concluded that the prepared Camptothecin encapsulated polymeric nanoformulations may be considered as an attractive and promising formulation which significantly overcome the limitations of Camptothecin and synergistically enhance its anticancer activity.