TY - JOUR
T1 - Diffusion Limited Cryopreservation of Tissue with Radiofrequency Heated Metal Forms
AU - Han, Zonghu
AU - Sharma, Anirudh
AU - Gao, Zhe
AU - Carlson, Timothy W.
AU - O'Sullivan, Gerard
AU - Finger, Erik B.
AU - Bischof, John C.
N1 - Publisher Copyright:
© 2020 The Authors. Published by Wiley-VCH GmbH
PY - 2020/10/1
Y1 - 2020/10/1
N2 - Cryopreserved tissues are increasingly needed in biomedical applications. However, successful cryopreservation is generally only reported for thin tissues (≤1 mm). This work presents several innovations to reduce cryoprotectant (CPA) toxicity and improve tissue cryopreservation, including 1) improved tissue warming rates through radiofrequency metal form and field optimization and 2) an experimentally verified predictive model to optimize CPA loading and rewarming to reduce toxicity. CPA loading is studied by microcomputed tomography (µCT) imaging, rewarming by thermal measurements, and modeling, and viability is measured after loading and/or cryopreservation by alamarBlue and histology. Loading conditions for three common CPA cocktails (6, 8.4, and 9.3 m) are designed, and then fast cooling and metal forms rewarming (up to 2000 °C min−1) achieve ≥90% viability in cryopreserved 1–2 mm arteries with various CPAs. Despite high viability by alamarBlue, histology shows subtle changes after cryopreservation suggesting some degree of cell damage especially in the central portions of thicker arteries up to 2 mm. While further studies are needed, these results show careful CPA loading and higher metal forms warming rates can help reduce CPA loading toxicity and improve outcomes from cryopreservation in tissues while also offering new protocols to preserve larger tissues ≥1 mm in thickness.
AB - Cryopreserved tissues are increasingly needed in biomedical applications. However, successful cryopreservation is generally only reported for thin tissues (≤1 mm). This work presents several innovations to reduce cryoprotectant (CPA) toxicity and improve tissue cryopreservation, including 1) improved tissue warming rates through radiofrequency metal form and field optimization and 2) an experimentally verified predictive model to optimize CPA loading and rewarming to reduce toxicity. CPA loading is studied by microcomputed tomography (µCT) imaging, rewarming by thermal measurements, and modeling, and viability is measured after loading and/or cryopreservation by alamarBlue and histology. Loading conditions for three common CPA cocktails (6, 8.4, and 9.3 m) are designed, and then fast cooling and metal forms rewarming (up to 2000 °C min−1) achieve ≥90% viability in cryopreserved 1–2 mm arteries with various CPAs. Despite high viability by alamarBlue, histology shows subtle changes after cryopreservation suggesting some degree of cell damage especially in the central portions of thicker arteries up to 2 mm. While further studies are needed, these results show careful CPA loading and higher metal forms warming rates can help reduce CPA loading toxicity and improve outcomes from cryopreservation in tissues while also offering new protocols to preserve larger tissues ≥1 mm in thickness.
KW - 2 mm thick tissue systems
KW - porcine aortas
KW - tissue cryopreservation
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U2 - 10.1002/adhm.202000796
DO - 10.1002/adhm.202000796
M3 - Article
C2 - 32875732
AN - SCOPUS:85090062038
SN - 2192-2640
VL - 9
JO - Advanced Healthcare Materials
JF - Advanced Healthcare Materials
IS - 19
M1 - 2000796
ER -