# Ice-free vitrification and nano warming technology for banking of cardiovascular structures.

> **NIH NIH R44** · TISSUE TESTING TECHNOLOGIES, LLC · 2022 · $124,317

## Abstract

ABSTRACT
This proposal focuses on translation of ice-free cryopreservation by vitrification employing a novel approach of
volumetric heating by nanowarming using Fe nanoparticles in an alternating electromagnetic ?eld. Vitrification,
sub-zero storage below the glass transition temperature in a “glassy” rather than a crystalline frozen phase, is
a form of cryopreservation that avoids ice formation. Vitri?cation can be achieved by quickly cooling the
material to cryogenic storage temperatures, where ice cannot form. Vitri?cation can be maintained at the end
of the cryogenic protocol by quickly rewarming the tissue to temperatures above the temperatures where ice
nucleation may occur. The magnitude of the rewarming rates necessary to maintain vitri?cation is much higher
than the magnitude of the cooling rates that are required to achieve it in the ?rst place. The most common
approach to achieve the required cooling and rewarming rates is by convection based boundary warming in
which the the specimen's surface is exposed to a temperature controlled environment, such as a fluid bath.
Due to the underlying principles of heat transfer, there is a size limit in the case of surface boundary heating
beyond which crystallization cannot be prevented at the center of the specimen. Furthermore, due to the
underlying principles of solid mechanics, there is also a size limit beyond which thermal expansion in the
specimen can lead to structural damage and fractures. Volumetric heating by nanowarming during the
rewarming phase of the cryogenic protocol can alleviate these size limitations. Vitrification is already an
important enabling approach for reproductive medicine with the potential to permit storage and transport of
cells, tissues and organs for a great variety of biomedical uses. Unfortunately, practical application of
vitrification has been limited to smaller systems such as cells and thin tissues due to diffusive and phase
change limitations that preclude use for blood vessels, larger tissues and organs. To circumvent this problem
we demonstrated that nanowarming effectively rewarms blood vessels in our preliminary research. Our
experiments demonstrated that this innovative rewarming technique rewarmed vitrified femoral and carotid
arteries in volumes ranging from 1 to 50mL with retention of cell viability and physiologic function.
However, warming of thick arteries was suboptimal. We propose using large animal blood vessel, models
for further optimization and evaluation of nanowarmed vessels using a combination of in vitro and in vivo
studies. In Phase 1 in a single specific aim we will optimize ice-free vitrification of thick walled arteries,
aorta and pulmonary, with a go/no go objective of achieving > 90% viability for progression to Phase 2. In
Phase 2 specific aims, we propose using porcine vascular models in a combination of ex vivo and in vivo
studies. The magnetic nanoparticles will be distributed around and within the internal spaces of vessels...

## Key facts

- **NIH application ID:** 10587348
- **Project number:** 3R44HL142455-03S1
- **Recipient organization:** TISSUE TESTING TECHNOLOGIES, LLC
- **Principal Investigator:** Kelvin G.M. Brockbank
- **Activity code:** R44 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2022
- **Award amount:** $124,317
- **Award type:** 3
- **Project period:** 2020-11-01 → 2024-03-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10587348

## Citation

> US National Institutes of Health, RePORTER application 10587348, Ice-free vitrification and nano warming technology for banking of cardiovascular structures. (3R44HL142455-03S1). Retrieved via AI Analytics 2026-05-22 from https://api.ai-analytics.org/grant/nih/10587348. Licensed CC0.

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