Amino acid-functionalised trehalose for multifunctional cryopreservation

Area of Science:

• Biochemistry
• Materials Science
• Cell Biology

Background:

• Cryopreservation is vital in medicine, but conventional cryoprotective agents (CPAs) like dimethyl sulfoxide (DMSO) exhibit toxicity.
• Natural cryoprotectants, such as trehalose and amino acids, inspire strategies to mitigate cryoinjury.

Purpose of the Study:

• To synthesize and evaluate novel 6,6'-trehalose diesters of amino acids (glycine, L-alanine, L-proline) as potential CPAs.
• To assess their cryoprotective efficacy, ice recrystallization inhibition, and cytocompatibility.
• To explore their potential as DMSO-free alternatives in cryopreservation.

Main Methods:

• Differential scanning calorimetry (DSC) for thermal properties and vitrification capacity.
• Ice recrystallization inhibition (IRI) assays and freeze-float assays for ice formation control.
• Radical-scavenging activity assays and molecular dynamics (MD) simulations for mechanistic insights.
• Cytotoxicity and cryopreservation trials using mammalian HaCaT cells.

Main Results:

• Synthesized trehalose diesters (1a-c) exhibited favorable glass transition temperatures (up to -25 °C) and comparable IRI to trehalose.
• Controlled ice nucleation was observed, and diesters showed radical-scavenging activity, unlike native trehalose.
• MD simulations revealed localized hydration shells disrupting water networks.
• In cell-based assays, diesters demonstrated lower cytotoxicity than DMSO and trehalose, conferring measurable cryoprotection.

Conclusions:

• Bio-inspired functionalization of trehalose offers a promising route to developing safer, effective CPAs.
• The novel trehalose diesters present a viable strategy for DMSO-free cryopreservation with reduced toxicity.
• Further research into these compounds could advance cryopreservation techniques in various medical applications.