Journal
JOURNAL OF PHARMACY AND PHARMACOLOGY
Volume 71, Issue 6, Pages 920-928Publisher
WILEY
DOI: 10.1111/jphp.13082
Keywords
chitosan; endotoxin contamination; purification
Categories
Funding
- COMPETE 2020 - Operational Programme for Competitiveness and Internationalisation
- Portuguese national funds via FCT - Fundacao para a Ciencia e a Tecnologia, I.P. [PROSAFE/0001/2016, POCI-01-0145-FEDER-030331, POCI-01-0145-FEDER-007440 (UID/NEU/04539/2013)]
- FCT [SFRH/BD/64046/2009]
- GlaxoSmithKline Portuguese Foundation
- Fundação para a Ciência e a Tecnologia [ProSafe/0001/2016, SFRH/BD/64046/2009] Funding Source: FCT
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Objectives The cationic biopolymer chitosan (CH) has emerged as a promising candidate adjuvant due to its safety profile and immunostimulatory properties. The presence of endotoxin contamination in biomaterials is generally underappreciated and can generate misleading results. It is important to establish a convenient methodology to obtain large amounts of high quality chitosan nanoparticles for biomedical applications. Methods We developed an easy method to generate endotoxin-free chitosan and assessed its purity using the Limulus amebocyte lysate assay and by measuring dendritic cell activation. Key findings Purified chitosan-based formulations alone failed to induce production of the proinflammatory cytokines tumour necrosis factor alpha (TNF-alpha) and interleukin (IL)-6 in bone marrow-derived dendritic cells (BMDCs) generated from C57BL/6 mice, while maintaining its ability to promote IL-1 beta secretion in combination with the Toll-like receptor (TLR)-9 agonist, CpG. Moreover, BMDCs from C3H/HeN and TLR4-deficient mice, C3H/HeJ were stimulated with endotoxin-free chitosan-based formulations and no differences were observed in IL-6 and IL-1 beta secretion, excluding the involvement of TLR-4 in the immunomodulatory effects of chitosan. Conclusions The developed method provides simple guidelines for the production of endotoxin-free chitosan, ideal for biomedical applications.
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