Cryoprotective Hydrogels with Stabilizing and Tunable Properties

NU2025-166

INVENTORS

• Songi Han*
• Shiny Maity
• Pravin Taware
• Syeda Hasany
• Karen Tsay
• Maria-Jose Ferrer
• Candice Crilly
• Warren Kincaid
• Lokeswara Potnuru

SHORT DESCRIPTION

This invention describes a novel hydrogel made from poly(vinyl alcohol) PVA and trehalose that protects biological materials against ice damage. It works by controlling the size of the growing ice and preventing ice damage. This hydrogel can cryopreserve cells and tissues, stabilize sensitive biomolecules, and extend the shelf life of biopharmaceutical products, offering a non-toxic solution for bio-storage.

BACKGROUND

The preservation of biological materials is severely hampered by challenges inherent in current cryopreservation and storage methods. A primary concern is the extensive damage caused by ice formation and recrystallization during freezing and thawing, leading to mechanical injury, osmotic stress, and reduced post-thaw viability and function. Cryoprotectants such as dimethyl sulfoxide (DMSO), glycerol, ethylene glycol, propylene glycol, and trehalose are widely used in cryopreservation mixtures to mitigate intracellular ice formation and protect cells and tissues from damage. However, at the concentrations necessary for efficacy, often exceeding 10% by volume (v/v), these agents can induce cytotoxicity and oxidative damage. While some non-toxic solutions have been widely used, their efficiency to inhibit ice recrystallization is very limited at practical concentrations. Thus, existing approaches fail to simultaneously inhibit ice recrystallization, encapsulate and protect sensitive bioactive molecules, and extend shelf life in a single, integrated, and non-toxic platform.

ABSTRACT

This invention provides a multifunctional PVA-trehalose hydrogel platform. The synergistic interaction between PVA and trehalose stabilizes a non-freezing hydration layer of water at the PVA-ice interface. This unique mechanism inhibits ice recrystallization by structuring interfacial water instead of directly binding to ice. The resulting gel can reduce the mechanical and osmotic damage during freeze-thaw cycles. The resulting hydrogel matrix can physically encapsulate sensitive bioactive molecules, offering a hydrated protective environment that prevents denaturation, aggregation, and degradation. Furthermore, by reducing water activity and mitigating ice- or dehydration-induced damage, the hydrogel extends the functional stability and shelf life of biological and pharmaceutical materials, offering a tunable and non-toxic solution.

APPLICATIONS

• Non-toxic cryopreservation of biological samples, including cells and tissues
• Encapsulation and controlled release of proteins, nucleic acids, vaccines, and other biotherapeutics
• Long-term storage and transportation of biological and pharmaceutical products

ADVANTAGES

• Triple-purpose functionality in a single product:
  1. Effective biocompatible cryopreservation without cytotoxicity – does not compromise cell viability or function, unlike traditional cryoprotectants such as DMSO and glycerol
  2. Improved stabilization of bioactive molecules – encapsulates and protects sensitive molecules from denaturation and aggregation.
  3. Extended shelf life – minimizes ice-induced damage to surpass the shelf life of existing cryoprotectants.
• Water-structuring mechanism – hydration shell engineering differs from traditional methods that rely on direct ice binding.
• Scalable and versatile properties – the hydrogel’s physical properties can be adjusted via annealing conditions, PVA concentration and different co-solutes.

IP STATUS

A provisional patent has been filed.