Hybrid nanosystems based on natural polymers as protein carriers for respiratory delivery: Stability and toxicological evaluation

Susana Rodrigues,Clara Cordeiro,Begoña Seijo,Carmen Remuñán-López,Ana Grenha

doi:10.1016/j.carbpol.2015.01.048

Abstract

Chitosan/carrageenan/tripolyphosphate nanoparticles were previously presented as holding potential for an application in transmucosal delivery of macromolecules, with tripolyphosphate demonstrating to contribute for both size reduction and stabilisation of the nanoparticles. This work was aimed at evaluating the capacity of the nanoparticles as protein carriers for pulmonary and nasal transmucosal delivery, further assessing their biocompatibility pattern regarding that application. Nanoparticles demonstrated stability in presence of lysozyme, while freeze-drying was shown to preserve their characteristics when glucose or sucrose were used as cryoprotectants. Bovine serum albumin was associated to the nanoparticles, which were successfully microencapsulated by spray-drying to meet the aerodynamic requirements inherent to pulmonary delivery. Finally, a satisfactory biocompatibility profile was demonstrated upon exposure of two respiratory cell lines (Calu-3 and A549 cells) to the carriers. A negligible effect on cell viability along with no alterations on transepithelial electrical resistance and no induction of inflammatory response were observed.

Highlights

Several physicochemical and biopharmaceutical issues make systemic administration of biomolecules a challenging task (Gupta & Sharma, 2009), compelling the need to develop adequate drug delivery systems that overcome inherent limitations
Materials Chitosan, pentasodium tripolyphosphate, bovine serum albumin (BSA), phosphate buffer saline (PBS) tablets pH 7.4, Dulbecco’s modified Eagle’s medium (DMEM), penicillin/streptomycin (10000 units/mL, 10000 μg/mL), non-essential amino acids, L-glutamine 200 mM, trypsin–EDTA solution (2.5 g/L trypsin, 0.5 g/L EDTA), trypan blue solution (0.4%), thiazolyl blue tetrazolium bromide (MTT), sodium dodecyl sulphate (SDS), N,N-dimethylformamyde (DMF), glycerol, lipopolysacharide (LPS), dimethyl sulfoxide (DMSO), lysozyme and glacial acetic acid were supplied by Sigma Chemicals (Germany). k-carrageenan was obtained from FMC Biopolymer (Norway) and fetal bovine serum (FBS) was from Gibco (USA)
Nanoparticles have been reported to avoid or delay mucociliary clearance, this ability is highly dependent on their size and is more relevant for particles below 100 nm (Oberdörster, Oberdörster & Oberdörster, 2005; Sung, 380 Pulliam & Edwards, 2007)

Summary

Introduction

Several physicochemical and biopharmaceutical issues make systemic administration of biomolecules a challenging task (Gupta & Sharma, 2009), compelling the need to develop adequate drug delivery systems that overcome inherent limitations. These carriers are mainly developed to avoid the need of parenteral administration, instead permitting effective transmucosal delivery. A previous study demonstrated the ability of two natural polymers, chitosan (CS) and κ-carrageenan (CRG), to assemble into nanoparticles of 400-600 nm by simple polyelectrolyte complexation (Grenha et al, 2010). Apart from the advantage of using natural materials, which facilitates biocompatibility (Malafaya, Silva & Reis, 2007), the polyelectrolyte complexation between CS and CRG uses very mild conditions, occurring in hydrophilic medium (Grenha, 2012). More recently we associated tripolyphosphate (TPP) to the CS/CRG nanoparticle formulation as a cross-linking agent, providing a size decrease and obtaining more stable nanoparticles (Rodrigues, Rosa da Costa & Grenha, 2012)

Objectives

Methods

Results

Conclusion