JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES) ›› 2016, Vol. 54 ›› Issue (8): 44-49.doi: 10.6040/j.issn.1671-7554.0.2015.1185

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Effects of different cryoprotectants and cooling methods on the histology and immunogenicity of heart valves

GONG Yanjie1, WANG Long2, DONG Laidong3, LI Dong1, SONG Guangmin2, JU Xiuli1   

  1. 1. Department of Paediatrics;
    2. Department of Cardiac Surgery;
    3. Body Donation Receiving Station, Qilu Hospital of Shandong University, Jinan 250012, Shandong, China
  • Received:2015-11-27 Online:2016-08-10 Published:2016-08-10

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Abstract: Objective To explore the proper cryoprotectants and cooling methods for storing heart valves in liquid nitrogen. Methods Each heart valve from 6 healthy donors were divided into 7 specimens: 1 was observed and tested directly as control group, and other 6 were divided into 3 cryoprotectant groups(Group A, B and C), with 2 specimens in each groups. Two cooling methods were applied, including -80 ℃ direct cooling(G1 groups)and programmed cooling(G2 groups). The histological changes were observed with hematoxylin-eosin staining, Weigert-Van Gieson(ET+VG)staining, and Masson Trichrome staining. The ultrastructural changes were observed with transmission electron microscopy(TEM). The expression of human leukocyte antigen(HLA)was detected with Western blotting and immunohistochemistry. The immunogenicity of each group was compared. Results Cryopreservation injured the organizational structure of valves, while valves stored in DMSO and chondroitin sulfate sodium showed the least lesions and obviously down-regulated expression of HLA-I. Collagen fibrils were less stained, while the orientation was more disorderly in the programmed cooling groups than in the -80 ℃direct cooling groups, but the expression of HLA-I was lower. Conclusion DMSO in combination with chondroitin sulfate sodium can best preserve the heart valves, with the lowest immunogenicity. The method of -80 ℃direct cooling is conducive for the preservation of collagen fibrils, but it produces higher immunogenicity than programmed cooling.

Key words: Transmission electron microscopy, Heart valves, Cryopreservation, Histology, Human leukocyte antigen

CLC Number: 

  • R654.2
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