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Effective Endotoxin Removal from Chitosan That Preserves Chemical Structure and Improves Compatibility with Immune
Sophie L Reay1, Emma L Jackson2, Daniel Salthouse1
1School of Engineering, Newcastle University, Newcastle upon Tyne NE1 7RU, UK.
Polymers
|April 13, 2023
Summary
NaOH treatment effectively removes endotoxins from chitosan, preserving its structure for biomedical applications. This method prevents inflammatory responses, unlike heat treatment, ensuring reliable in vivo performance.
Area of Science:
- Biomaterials Science
- Polymer Chemistry
- Immunology
Background:
- Chitosan is a widely studied biopolymer for healthcare but is prone to endotoxin contamination.
- Endotoxins trigger inflammatory responses, compromising chitosan's efficacy and research validity for in vivo applications.
- Effective endotoxin removal is crucial for safe and accurate use of chitosan in medical devices and research.
Purpose of the Study:
- To investigate heat and mild sodium hydroxide (NaOH) treatments for facile endotoxin removal from chitosan.
- To evaluate the impact of these treatments on chitosan's structural integrity and chemical properties.
- To assess the biocompatibility and inflammatory potential of treated chitosan for in vivo applications.
Main Methods:
- Chitosan samples were treated with heat and mild NaOH to remove endotoxins.
- Endotoxin levels were quantified to ensure they were below the FDA limit (<0.5 EU/mL).
- Fourier-transform infrared-attenuated total reflectance (FTIR-ATR) spectroscopy analyzed chemical changes.
- Lysozyme degradation studies assessed hydrogel stability.
- In vitro co-culture studies with peripheral blood mononuclear cells (PBMCs) and monocyte-derived dendritic cells (moDCs) evaluated inflammatory responses and cell viability.
Main Results:
- Both heat and NaOH treatments reduced endotoxins below the FDA limit.
- NaOH-treated chitosan maintained chemical and physical properties similar to native chitosan.
- Heat-treated chitosan showed changes due to the Maillard reaction, indicating structural alteration.
- NaOH-treated chitosan hydrogels exhibited similar lysozyme degradation rates as native chitosan hydrogels.
- NaOH-treated chitosan did not induce pro-inflammatory cytokine release (TNF-α) or negatively impact moDC viability and maturation in vitro.
Conclusions:
- Mild NaOH treatment is a superior method for endotoxin removal from chitosan compared to heat treatment.
- NaOH treatment effectively purifies chitosan without compromising its structural integrity or biocompatibility.
- This optimized endotoxin removal strategy is vital for the reliable application of chitosan-based biomaterials in healthcare and biomedical research.
