Demonstration of the cryoprotective properties of the fucose-containing polysaccharide FucoPol

Bruno M Guerreiro; Filomena Freitas; João C Lima; Jorge C Silva; Madalena Dionísio; Maria A M Reis

doi:10.1016/j.carbpol.2020.116500

Demonstration of the cryoprotective properties of the fucose-containing polysaccharide FucoPol

Carbohydr Polym. 2020 Oct 1:245:116500. doi: 10.1016/j.carbpol.2020.116500. Epub 2020 Jun 3.

Authors

Bruno M Guerreiro¹, Filomena Freitas², João C Lima³, Jorge C Silva⁴, Madalena Dionísio⁵, Maria A M Reis⁶

Affiliations

¹ UCIBIO-REQUIMTE, Chemistry Department, Faculty of Sciences and Technology, Nova University of Lisbon, Portugal; LAQV-REQUIMTE, Chemistry Department, Faculty of Sciences and Technology, Nova University of Lisbon, Portugal; CENIMAT/I3N, Physics Department, Faculty of Sciences and Technology, Nova University of Lisbon, Portugal. Electronic address: bm.guerreiro@campus.fct.unl.pt.
² UCIBIO-REQUIMTE, Chemistry Department, Faculty of Sciences and Technology, Nova University of Lisbon, Portugal. Electronic address: a4406@fct.unl.pt.
³ LAQV-REQUIMTE, Chemistry Department, Faculty of Sciences and Technology, Nova University of Lisbon, Portugal. Electronic address: lima@fct.unl.pt.
⁴ CENIMAT/I3N, Physics Department, Faculty of Sciences and Technology, Nova University of Lisbon, Portugal. Electronic address: jcs@fct.unl.pt.
⁵ LAQV-REQUIMTE, Chemistry Department, Faculty of Sciences and Technology, Nova University of Lisbon, Portugal. Electronic address: madalena.dionisio@fct.unl.pt.
⁶ UCIBIO-REQUIMTE, Chemistry Department, Faculty of Sciences and Technology, Nova University of Lisbon, Portugal. Electronic address: amr@fct.unl.pt.

PMID: 32718611
DOI: 10.1016/j.carbpol.2020.116500

Abstract

We report the cryoprotective potential of FucoPol, a fucose-containing bacterial exopolysaccharide produced by Enterobacter A47. In vitro cryopreservation assays of Vero, Saos-2, HFFF2 and C2C12 cell lines exposed to a validated non-cytotoxic 2.5 mg/mL FucoPol concentration demonstrated a consistent post-thaw metabolic viability increase. Calorimetric analysis showed a non-colligative, FucoPol concentration-dependent increase of the freezing point (T_f), with minimal change in melting point (T_m). Freezing point variation was corroborated by Polarized Optical Microscopy studies, also showing a reduction of ice crystal dimensions. Its proven shear-thinning behaviour and polyanionicity favour interactivity between the polysaccharide and the water-ice interface, resulting in ice growth inhibition. These findings demonstrate FucoPol's high promise as a bio-based, biodegradable approach to be implemented into cryopreservation formulations.

Keywords: Calorimetry; Cryopreservation; Cytotoxicity; FucoPol; Non-colligative; Polysaccharide.

MeSH terms

Animals
Antifreeze Proteins / chemistry
Calorimetry, Differential Scanning
Cell Line, Tumor
Cell Survival / drug effects
Chlorocebus aethiops
Cryopreservation / methods*
Cryoprotective Agents / chemistry*
Cryoprotective Agents / pharmacology*
Crystallization
Enterobacter / chemistry
Freezing
Fucose / chemistry*
Fucose / pharmacology*
Humans
Ice
Mice
Polyelectrolytes / chemistry
Polysaccharides, Bacterial / chemistry*
Polysaccharides, Bacterial / pharmacology*
Vero Cells
Water / chemistry

Substances

Antifreeze Proteins
Cryoprotective Agents
Ice
Polyelectrolytes
Polysaccharides, Bacterial
polyanions
Water
Fucose