Crab (Charybdis natator) exoskeleton derived chitosan nanoparticles for the in vivo delivery of poorly water-soluble drug: Ibuprofen.

The study aims to extract and purify chitosan (CS) from the exoskeleton of crab (C. natator) and develop ibuprofen (IBU) encapsulated CS nanoparticles (IBU-CSNPs). Analysis of purified CS revealed characteristic functional and crystallinity peaks. Moreover, morphological analysis of prepared IBU-CSNPs showed uniform spherical shape with a size range of 40-100 nm whereas encapsulation efficiency (EE%) and loading capacity (LC%) were estimated to be 68.94 ± 1.61% and 28 ± 1.18% respectively. Further, in vitro release profile of IBU from IBU-CSNPs was observed to be in biphasic form with initial release up to 15 h followed by the sustained release in different test conditions. Further, the effects of purified CS on the viability of RAW264.7 cells exhibited no toxic effects in higher concentrations. Furthermore, fluorescein isothiocyanate (FITC) conjugated nanoparticles (FITC-IBU-CSNPs) were investigated on in vivo model of adult zebrafish for time-dependent circulation and accumulation of the drug through the nano-carrier system. It was observed that the drug diffusion from the nanoparticles was in a sustained manner throughout the gastrointestinal region which resulted in suppression of inflammation. Overall, this study provides an effective and facile process for preparing a crab CS-based nano-carrier system used for the delivery of IBU in vivo which may help in the curing of prolonged chronic inflammatory diseases. Moreover, it may also help to reduce adverse effects of these drugs in the gastrointestinal tract such as ulcers and bleeding.

Angiotensin I-Converting Enzyme (ACE-I) Inhibition and Antioxidant Peptide from a Squilla Species.

BACKGROUND: Oratosquilla woodmasoni is one of the marine squilla species, which is found in the entire Asia-Pacific region. This current study assesses the species as the main basis of both ACEi and antioxidant peptide. OBJECTIVE: To isolate the ACEi peptide derived from O. woodmasoni and examine its ACE inhibition along with antioxidant potential. MATERIALS AND METHODS: The squilla muscle protein was hydrolysed using alcalase and trypsin enzymes for 12 hours and tested for DH. The hydrolysates were examined for their ACEi activity and then the best hydrolysate was sequentially purified in various chromatographical methods. The purified peptide was studied for anti-oxidant and functional properties, followed by amino acid sequencing. The purified peptide was also evaluated for its toxicity by in vitro cell viability assay. RESULTS: The DH% was found to be 47.13 ± 0.72% and 89.43 ± 2.06% for alcalase and trypsin, respectively. The alcalase 5th-hour hydrolysate was detected with potent activity (65.97 ± 0.56%) using ACEi assay and was primarily fractionated using ultrafiltration; the maximum inhibitory activity was found with 77.04 ± 0.52% in 3-10 kDa fraction. Subsequently, the fraction was purified using IEC and GFC, in which the AC1-A2 fraction had higher antihypertensive activity (70.85 ± 0.78%). The non-toxic fraction showed hexapeptide HVGGCG with molecular weight 529 Da with great potential of antioxidant activity along with functional property. CONCLUSION: This peptide could be developed as a potential ACE-inhibitory and antioxidant agent.