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[Preliminary study of cryopreservation of tissue engineered tendon]
Zheng Zhou1, Zhi-ming Yang, Hui-qi Xie
1Key Laboratory of Transplant Engineering and Immunology, Ministry of Public Health: Institute of Reparative and Reconstructive Surgery Laboratory of Tissue Engineering, West China Hospital, Sichuan University, Chengdu, Sichuan, P. R. China 610041.
Objective:
To investigate a cryophylactic agent (CPA) to protect tissue engineered tendon (TET) in deep low temperature.
Methods:
Sixty-four BALB/C inbred nude mice were chosen, which included 4 as blank control group, left sides of 60 as experimental group and their right sides as control group. Transformed human embryonic tendon cells of the 54th passage and artificial materials of carbon fiber (CF) and polyglycolic acid (PGA) were co-cultured in vitro to construct TET. TET was frozen in liquid nitrogen with four kinds of CPA (groups A, B, C, and D) for 2 months. They were thawed quickly and transplanted into hind limbs of nude mice to repair the defects of Achilles tendon, which was 5 mm in length and 65.7% of total Achilles tendon. In control group, no cryopreservation treatment was taken. The morphological, histological, ultrastructure, and immunohistochemistry examinations were made and short tandem repeat loci were detected 2, 4, 6, 8, and 12 weeks later.
Results:
In the experimental group, the morphological properties of tendon cells resumed gradually and the capability of synthesizing collagen enhanced by degrees. Tendon cells survived and could secret type I collagen and there was less difference between experimental and control groups 12 weeks after transplantation. In group A, vacuole in mitochondrion of tendon cell decreased, tendon cell arranged in order and abundant collagen fibers were found and linked.
Conclusion:
The cryopreservation agent in group A can protect TET in deep low temperature.