SCI和EI收录∣中国化工学会会刊

Chinese Journal of Chemical Engineering ›› 2024, Vol. 72 ›› Issue (8): 1-9.DOI: 10.1016/j.cjche.2024.03.006

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Vitrification cryopreservation of ligaments based on zwitterionic betaine

Liming Zhang1, Xinmeng Liu1, Haoyue Li1, Lei Zhang1,2   

  1. 1 Department of Biochemical Engineering, Frontier Science Center for Synthetic Biology and Key Laboratory of Systems Bioengineering (MOE), School of Chemical Engineering and Technology, Tianjin University, Tianjin 300350, China;
    2 Haihe Laboratory of Sustainable Chemical Transformations, Tianjin 300192, China
  • Received:2023-12-13 Revised:2024-03-09 Online:2024-10-17 Published:2024-08-28
  • Contact: Lei Zhang,E-mail:lei_zhang@tju.edu.cn
  • Supported by:
    This article is supported by the National Natural Science Foundation of China (22078238, U23B20121).

Vitrification cryopreservation of ligaments based on zwitterionic betaine

Liming Zhang1, Xinmeng Liu1, Haoyue Li1, Lei Zhang1,2   

  1. 1 Department of Biochemical Engineering, Frontier Science Center for Synthetic Biology and Key Laboratory of Systems Bioengineering (MOE), School of Chemical Engineering and Technology, Tianjin University, Tianjin 300350, China;
    2 Haihe Laboratory of Sustainable Chemical Transformations, Tianjin 300192, China
  • 通讯作者: Lei Zhang,E-mail:lei_zhang@tju.edu.cn
  • 基金资助:
    This article is supported by the National Natural Science Foundation of China (22078238, U23B20121).

Abstract: Ligament cryopreservation enables a prolonged shelf life of allogeneic ligament grafts, which is fundamentally important to ligament reconstruction. However, conventional cryopreservation techniques fail to eliminate the damage caused by ice crystal growth and the toxicity of cryopreservation agents (CPAs). Here, we report a novel CPA vitrification formulation primarily composed of betaine for ligament cryopreservation. Comprehensive optimization was conducted on the methods for vitrification and rewarming, as well as the loading and unloading conditions, based on the critical cooling rate (CCR), critical warming rate (CWR), and permeation properties of the CPA. Using biomechanical and histological level tests, we demonstrate the superior performance of our method in ligament cryopreservation. After 30 days of vitrification cryopreservation, parameters such as the Young's modulus, tensile stress, denaturation temperature, and glycosaminoglycans content of the ligament remained essentially unchanged. This work pioneers the application of ice-free cryopreservation for ligament and holds great potential for improving the long-term storage of ligament, providing valuable insights for future cryopreservation technique development.

Key words: Ligament, Betaine, Vitrification, Cryopreservation

摘要: Ligament cryopreservation enables a prolonged shelf life of allogeneic ligament grafts, which is fundamentally important to ligament reconstruction. However, conventional cryopreservation techniques fail to eliminate the damage caused by ice crystal growth and the toxicity of cryopreservation agents (CPAs). Here, we report a novel CPA vitrification formulation primarily composed of betaine for ligament cryopreservation. Comprehensive optimization was conducted on the methods for vitrification and rewarming, as well as the loading and unloading conditions, based on the critical cooling rate (CCR), critical warming rate (CWR), and permeation properties of the CPA. Using biomechanical and histological level tests, we demonstrate the superior performance of our method in ligament cryopreservation. After 30 days of vitrification cryopreservation, parameters such as the Young's modulus, tensile stress, denaturation temperature, and glycosaminoglycans content of the ligament remained essentially unchanged. This work pioneers the application of ice-free cryopreservation for ligament and holds great potential for improving the long-term storage of ligament, providing valuable insights for future cryopreservation technique development.

关键词: Ligament, Betaine, Vitrification, Cryopreservation