Cell and Tissue Banking

, Volume 19, Issue 4, pp 629–636 | Cite as

Optimal number of chemical extraction treatments for maintaining the biological properties of an allogeneic tendon

  • Peng Chen
  • Changqing Jiang
  • Li Shen
  • Wentao Zhang
  • Lixin ZhuEmail author


The aim of this study was to explore the biological effects of the amount of chemical extraction treatments performed on an allogeneic tendon through histomorphology, biological mechanics testing, and an immunogenicity assay. Sixteen New Zealand rabbits (body weight 2.5–3.0 kg) were randomly divided into four groups: group A (chemical extraction once), group B (chemical extraction twice), group C (chemical extraction three times), and group D (blank control group), with four rabbits in each group. The Achilles tendons of each rabbit were separated and subjected to a chemical extraction process with Triton X-100 and sodium deoxycholate, followed by hematoxylin and eosin staining, electron microscopy observation, biomechanical testing, and mixed lymphocyte culture. There were no significant differences in the surface color and fiber bundles between groups A and B and the blank control group, whereas group C showed clear differences from the blank control group with a rough surface, loose fibers, and poor tension. There were no significant differences in the biomechanics among the four groups. The four groups showed significant differences in the lymphocyte conversion ratio, with reduced rates of lymphocyte conversion along with increasing treatment numbers. Two chemical extractions of the tendon allowed for retaining most of the integrity of the original tendon fiber while removing immunogenicity with good biological properties. These findings lay a foundation for application of this method to human tendons so as to provide a good tissue source for tendon transplantation.


Allogeneic tendon Chemical extraction Biological properties Tissue engineering 



We are grateful to the valuable comments of the anonymous reviewers. This work is partly supported by Health and Family Science Foundation of Shenzhen under Grant Numbers. 201606007 and Sanming Project of Medicine in Shenzhen (No. SZSM201612078).

Compliance with ethical standards

Conflict of interest

The author’s declare that they have no conflicts of interest.


  1. Arai T, Kanje M, Lundborg G, Sondell M, Liu XL, Dahlin LB (2000) Axonal outgrowth in muscle grafs made acellular by chemical extraction. Restor Neurol Neurosci 17:165–174PubMedGoogle Scholar
  2. Brockbank KG, Chen ZZ, Song YC (2010) Vitrification of porcine articular cartilage. Cryobiology 60:217–221CrossRefPubMedGoogle Scholar
  3. Brown BN, Valentin JE, Stewart-Akers AM, McCabe GP, Badylak SF (2009) Macrophage phenotype and remodeling outcomes in response to biologic scaffolds with and without a cellular component. Biomaterials 30:1482–1491CrossRefPubMedPubMedCentralGoogle Scholar
  4. Cartmell JS, Dunn MG (2000) Effect of chemical treatments on tendon cellularity and mechanical properties. J Biomed Mater Res 49(1):134–140CrossRefPubMedGoogle Scholar
  5. Fan WX, Ma XH, Ge D, Liu TQ, Cui ZF (2009) Cryoprotectants for the vitrification of corneal endothelial cells. Cryobiology 58:28–36CrossRefPubMedGoogle Scholar
  6. Huang Q, Ingham E, Rooney P, Kearney JN (2013) Production of a sterilised decellularised tendon allograft for clinical use. Cell Tissue Bank 14(4):645–654CrossRefPubMedGoogle Scholar
  7. Isachenko V, Lapidus I, Isachenko E, Krivokharchenko A, Kreienberg R, Woriedh M, Bader M, Weiss JM (2009) Human ovarian tissue vitrification versus conventional freezing: morphological, endocrinological, and molecular biological evaluation. Reproduction 138(2):319–327CrossRefPubMedGoogle Scholar
  8. Jiang Chang-qing, Jun Hu, Xiang Jian-ping, Zhu Jia-kai, Liu Xiao-lin, Luo Peng (2016) Tissue-engineered rhesus monkey nerve grafs for the repair of long ulnar nerve defects: similar outcomes to autologous nerve grafs. Neural Regener Res 11(11):1845–1850CrossRefGoogle Scholar
  9. Kim HW, Yeo IJ, Hwang KE, Song DH, Kim YJ, Ham YK, Jeong TJ, Choi YS, Kim CJ (2016) Isolation and characterization of pepsin-soluble collagens from bones, skins, and tendons in duck feet. Korean J Food Sci Anim Resour 36(5):665–670CrossRefPubMedPubMedCentralGoogle Scholar
  10. Kowalski JB, Mosley GA, Merritt K, Osborne J (2012) Assessment of bioburden on human and animal tissues: part 1–results of method development and validation studies. Cell Tissue Bank 13(1):129–138CrossRefPubMedGoogle Scholar
  11. Kuleshova LL, Lopata A (2002) Vitrification can be more favorable than slow cooling. Fertile Sterile 78:449–454CrossRefGoogle Scholar
  12. Lee KI, Lee JS, Kang KT, Shim YB, Kim YS, Jang JW, Moon SH, D’Lima DD (2018) In vitro and in vivo performance of tissue-engineered tendons for anterior cruciate ligament reconstruction. Am J Sports Med 46(7):1641–1649CrossRefPubMedGoogle Scholar
  13. Li T, Mai Q, Gao J, Zhou C (2010) Cryopreservation of human embryonic stem cells with a new bulk vitrification method. Biol Reprod 82(5):848–853CrossRefPubMedGoogle Scholar
  14. Mohamad NA, Mustafa S, El Sheikha AF, Khairil Mokhtar NF, Ismail A, Ali ME (2016) Modification of gelatin-DNA interaction for optimised DNA extraction from gelatin and gelatin capsule. J Sci Food Agric 96(7):2344–2351CrossRefPubMedGoogle Scholar
  15. Mokrejs P, Sukop S, Svoboda P (2012) Three-stage extraction of gelatines from tendons of abattoir cattle: 2–properties of gelatines. Appl Biochem Biotechnol 168(2):434–445CrossRefPubMedGoogle Scholar
  16. Nakano T, Betti M, Pietrasik Z (2010) Extraction, isolation and analysis of chondroitin sulfate glycosaminoglycans. Recent Pat Food Nutr Agric 2(1):61–74CrossRefPubMedGoogle Scholar
  17. Omae H, Sun YL, An KN, Amadio PC, Zhao C (2012) Engineered tendon with decellularized xenotendon slices and bone marrow stromal cells: an in vivo animal study. J Tissue Eng Regener Med 6(3):238–244CrossRefGoogle Scholar
  18. Raghavan SS, Woon CY, Kraus A, Megerle K, Choi MS, Pridgen BC, Pham H, Chang J (2012) Human flexor tendon tissue engineering: decellularization of human flexor tendons reduces immunogenicity in vivo. Tissue Eng Part A 18:796–805CrossRefPubMedGoogle Scholar
  19. Srinivasan A, Sehgal PK (2010) Characterization of biocompatible collagen fibers: a promising candidate for cardiac patch. Tissue Eng Part C Methods 16(5):895–903CrossRefPubMedGoogle Scholar
  20. Sripunya N, Somfai T, Inaba Y, Nagai T, Imai K, Parnpai R (2010) A comparison of cryotop and solid msurface vitrification methods for the cryopreservation of in vitro matured bovine oocytes. J Reprod Dev 56:176–181CrossRefPubMedGoogle Scholar
  21. Wildemann B, Kadow-Romacker A, Pruss A, Haas NP, Schmidmaier G (2007) Quantification of growth factors in allogenic bone grafts extracted with three different methods. Cell Tissue Bank. 8(2):107–114CrossRefPubMedGoogle Scholar
  22. Yang JL, Yao X, Qing Q, Zhang Y, Jiang YL, Ning LJ, Luo JC, Qin TW (2018) An engineered tendon/ligament bioscaffold derived from decellularized and demineralized cortical bone matrix. J Biomed Mater Res A 106(2):468–478CrossRefPubMedGoogle Scholar

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© Springer Nature B.V. 2018

Authors and Affiliations

  1. 1.ZhuJiang Hospital of Southern Medical UniversityGuangzhouChina
  2. 2.Peking University Shenzhen HospitalShenzhenChina
  3. 3.Pingshan Women and Children’s HospitalShenzhenChina

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