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Cell Biochemistry and Biophysics

, Volume 73, Issue 1, pp 27–33 | Cite as

The Preparation of Acellular Dermal Matrices by Freeze–Thawing and Ultrasonication Process and the Evaluation of Its Antigenicity

  • Yazhen Xu
  • Guoan ZhangEmail author
  • Yang Chang
  • Yu-xuan Qiu
  • Cheng Wang
Original Paper

Abstract

Antigenicity is the biggest obstacle of xenogeneic acellular dermal matrices (ADM) as dermal scaffold in treatment of large-area skin defect. We prepared ADM by repeated freezing and thawing and ultrasonic process, and then injected the ADM homogenate and degradation product into porcine skin to evaluate the effectiveness of the decellularized method and the antigenicity of porcine ADM. In this work, chinese miniature pigs (n = 10) were sensitized by subcutaneous injection with human ADM degradation products on days 0, 7, and 14. After 21 days, their abdominal skin was divided into five regions for intradermal injection of porcine ADM homogenate (PADM), PADM degradation products, human ADM homogenate (HADM), HADM degradation products, and physiological saline (negative control). Positive controls (n = 2) were processed with fresh human skin homogenate by the same method. The skin manifestations in related areas were observed at 24 and 48 h and then the skin was subjected to histopathological and immunohistochemical analysis. The results showed that skin erythema and hydroderma were not observed in all groups but in positive control group. The histopathological and immunohistochemical results confirmed that no inflammatory cell infiltration, irregular extracellular matrix, IL-2, and IFN-γ expression were observed in all four test groups. Our results suggest that the combination with repeated freeze–thawing and ultrasonication can be an effective method to prepare ADM, which has great potential in clinical application.

Keywords

Porcine Skin Collagen Antigenic determinant ADM 

Notes

Acknowledgments

We thank the faculty and staff in Jishuitan hospital Burns laboratory for their assistance to prepare ADM, Professor Gu Ming’ group of Chinese Academy of Sciences for their assistance to analyze amino acid sequence; the faculty and staff in Peking University Health Science Center Animal Lab for their assistance in animal experiment and animal management.

Supplementary material

12013_2015_569_MOESM1_ESM.docx (163 kb)
Supplementary material 1 (DOCX 163 kb)
12013_2015_569_MOESM2_ESM.docx (155 kb)
Supplementary material 2 (DOCX 155 kb)

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Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  • Yazhen Xu
    • 1
  • Guoan Zhang
    • 1
    Email author
  • Yang Chang
    • 1
  • Yu-xuan Qiu
    • 1
  • Cheng Wang
    • 2
  1. 1.Department of Burns and Plastic SurgeryPeking University Fourth School of Clinical MedicineBeijingPeople’s Republic of China
  2. 2.Beijing Jishuitan HospitalBeijingPeople’s Republic of China

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