June 2013
Volume 54, Issue 15
Free
ARVO Annual Meeting Abstract  |   June 2013
Biological Differences between Cryopreserved and Dehydrated Amniotic Membrane Tissue Grafts
Author Affiliations & Notes
  • Lorraine Chua
    Ocular Surface Research & Education Foundation, Miami, FL
    Research & Development, TissueTech, Inc., Miami, FL
  • Marissa Cooke
    Wallace H. Coulter Department of Biomedical Engineering, Georgia Institute of Technology and Emory University, Atlanta, GA
  • Christian Mandrycky
    Wallace H. Coulter Department of Biomedical Engineering, Georgia Institute of Technology and Emory University, Atlanta, GA
  • Ek Kia Tan
    Ocular Surface Research & Education Foundation, Miami, FL
    Research & Development, TissueTech, Inc., Miami, FL
  • Julie O'Connell
    Amniox Medical, Marietta, GA
  • Scheffer Tseng
    Ocular Surface Research & Education Foundation, Miami, FL
    Research & Development, TissueTech, Inc., Miami, FL
  • Todd McDevitt
    Wallace H. Coulter Department of Biomedical Engineering, Georgia Institute of Technology and Emory University, Atlanta, GA
    Parker H. Petit Institute for Bioengineering and Bioscience, Georgia Institute of Technology, Atlanta, GA
  • Footnotes
    Commercial Relationships Lorraine Chua, TissueTech Inc. (E); Marissa Cooke, None; Christian Mandrycky, None; Ek Kia Tan, TissueTech, Inc. (E); Julie O'Connell, None; Scheffer Tseng, NIH, NEI (F), TissueTech, Inc. (F), TissueTech, Inc. (E), TissueTech, Inc. (P); Todd McDevitt, Amniox Medical (C)
  • Footnotes
    Support None
Investigative Ophthalmology & Visual Science June 2013, Vol.54, 3894. doi:https://doi.org/
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      Lorraine Chua, Marissa Cooke, Christian Mandrycky, Ek Kia Tan, Julie O'Connell, Scheffer Tseng, Todd McDevitt; Biological Differences between Cryopreserved and Dehydrated Amniotic Membrane Tissue Grafts. Invest. Ophthalmol. Vis. Sci. 2013;54(15):3894. doi: https://doi.org/.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

Purpose: Amniotic membrane (AM) processing methods can dramatically impair both the structural integrity, and biological activity, of critical matrix cell signaling factors essential for the intended use of the product. To analyze the effect of different AM available commercially for the ophthalmology market on conserving the therapeutic potential of the tissue, we compared cryopreserved (CryoTek™) and dehydrated (Purion®) processed tissue grafts in physical and biochemical assays.

Methods: Cryopreserved thin (CT-Thin) and thick (CT-Thick) tissues were compared to dehydrated, (EF) and (AF) AM tissue grafts. Structural properties of cryopreserved and dehydrated AM were assessed by histological staining while biochemical properties were measured in soluble tissue extracts by comparing hyaluronan (HA) content and molecular weight (MW) spectrum; and critical proteoglycan (HC-HA) and protein (PTX3) signaling factors.

Results: Histochemical staining demonstrated dehydrated tissues having a more compact extracellular matrix compared to the cryopreserved tissues even after the prescribed hydration duration. HA quantity was highest in CT-Thick, and although content was relatively similar in CT-Thin and EF/AF, the MW analysis revealed all cryopreserved samples contained high MW HA, while the dehydrated samples contained low MW HA. Essential signaling proteins, HC-HA and PTX3 detected by western blots were present in cryopreserved samples but were either compromised, or completely absent, in dehydrated tissues.

Conclusions: The cryopreservation process better preserves the structural integrity and biochemistry of AM tissue grafts in comparison to dehydrated grafts, and suggests the therapeutic benefit of dehydrated AM may be compromised as a result.

Keywords: 765 wound healing • 687 regeneration • 490 cytokines/chemokines  
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