Biological properties of dehydrated human amnion/chorion composite graft: implications for chronic wound healing

Thomas J Koob, Robert Rennert, Nicole Zabek, Michelle Massee, Jeremy J Lim, Johnna S Temenoff, William W Li, Geoffrey Gurtner, Thomas J Koob, Robert Rennert, Nicole Zabek, Michelle Massee, Jeremy J Lim, Johnna S Temenoff, William W Li, Geoffrey Gurtner

Abstract

Human amnion/chorion tissue derived from the placenta is rich in cytokines and growth factors known to promote wound healing; however, preservation of the biological activities of therapeutic allografts during processing remains a challenge. In this study, PURION® (MiMedx, Marietta, GA) processed dehydrated human amnion/chorion tissue allografts (dHACM, EpiFix®, MiMedx) were evaluated for the presence of growth factors, interleukins (ILs) and tissue inhibitors of metalloproteinases (TIMPs). Enzyme-linked immunosorbent assays (ELISA) were performed on samples of dHACM and showed quantifiable levels of the following growth factors: platelet-derived growth factor-AA (PDGF-AA), PDGF-BB, transforming growth factor α (TGFα), TGFβ1, basic fibroblast growth factor (bFGF), epidermal growth factor (EGF), placental growth factor (PLGF) and granulocyte colony-stimulating factor (GCSF). The ELISA assays also confirmed the presence of IL-4, 6, 8 and 10, and TIMP 1, 2 and 4. Moreover, the relative elution of growth factors into saline from the allograft ranged from 4% to 62%, indicating that there are bound and unbound fractions of these compounds within the allograft. dHACM retained biological activities that cause human dermal fibroblast proliferation and migration of human mesenchymal stem cells (MSCs) in vitro. An in vivo mouse model showed that dHACM when tested in a skin flap model caused mesenchymal progenitor cell recruitment to the site of implantation. The results from both the in vitro and in vivo experiments clearly established that dHACM contains one or more soluble factors capable of stimulating MSC migration and recruitment. In summary, PURION® processed dHACM retains its biological activities related to wound healing, including the potential to positively affect four distinct and pivotal physiological processes intimately involved in wound healing: cell proliferation, inflammation, metalloproteinase activity and recruitment of progenitor cells. This suggests a paracrine mechanism of action for dHACM when used for wound healing applications.

Keywords: Amnion; Chorion; Chronic ulcers; Dermal fibroblasts; Growth factors; Mesenchymal stem cells; Wound healing; dHACM.

©2013 The Authors. International Wound Journal published by John Wiley & Sons Ltd and Medicalhelplines.com Inc.

Figures

Figure 1
Figure 1
Relative amount of growth factors eluting from dehydrated human amnion/chorion tissue allografts (dHACM) after 24 hours in normal saline. Each bar represents the average ±standard deviation of five samples tested.
Figure 2
Figure 2
Effects of extracts of dehydrated human amnion/chorion tissue allografts (dHACM) on adult human dermal fibroblast proliferation. The control wells contained Dulbecco's modified Eagle's medium (DMEM) only. The positive control contained DMEM supplemented with calf serum and established the capacity of adult human dermal fibroblast (HDFa) cells to proliferate in response to active factors. dHACM extract concentration in the DMEM is shown on the x‐axis. Each value represents the average ± standard deviation of five wells.
Figure 3
Figure 3
Cell density of migrated mesenchymal stem cells (MSCs) in response to dehydrated human amnion/chorion tissue allografts (dHACM), as well as negative and positive controls, for three MSC donors. (A) Representative micrographs and (B) cell density measurements indicated that greater migration was observed in response to larger samples, relative to their smaller counterparts. MSC migration in positive controls was significantly greater than all other samples (P≤0·05). * Indicates significantly greater migration than negative controls (serum‐free) in same MSC donor (P≤0·05). # Indicates significantly greater migration than 1·5 mm group in same MSC donor (P≤0·05).
Figure 4
Figure 4
Effects of dehydrated human amnion/chorion tissue allografts (dHACM) on mesenchymal progenitor cell recruitment in an in vivo mouse model. Upper panels demonstrate the fluorescence‐activated cell sorting (FACS) gating scheme and identification of progenitor cells. Bottom chart shows the relative number of mesenchymal stem cells in specimens and dHACM and sham implant site on days 3, 7, 14 and 28 post implant. Values shown are means ± standard deviations, n = 4 specimens. ** Indicates P < 0·05 when comparing dHACM to normal skin and sham implant via one‐way analysis of variance (ANOVA).

References

    1. John T. Human amniotic membrane transplantation: past, present and future. Ophthalmol Clin North Am 2003;16:43–65.
    1. Cornwell KG, Landsman A, James KS. Extracellular matrix biomaterials for soft tissue repair. Clin Podiatr Med Surg 2009;26:507–23.
    1. Gruss JS, Jirsch DW. Human amniotic membrane: a versatile wound dressing. Can Med Assoc J 1978;118:1237–46.
    1. Sawhney CP. Amniotic membrane as a biological dressing in the management of burns. Burns 1989;15:339–42.
    1. Forbes J, Fetterolf D. Dehydrated amniotic membrane allografts for the treatment of chronic wounds: a case study. J Wound Care 2012;21:290–6.
    1. Sheikh ES, Sheikh ES, Fetterolf DE. Use of dehydrated human amniotic membrane allografts to promote healing in patients with refractory non healing wounds. Int Wound J 2013. DOI: 10.1111/iwj.12035.
    1. Zelen C, Serena TE, Denoziere G, Fetterolf DE. A prospective randomized comparative parallel study of amniotic membrane wound graft in the management of diabetic foot ulcers. Int Wound J 2013. DOI: 10.1111/iwj.12097.
    1. Serena T, Fetterolf D. Clinical Research: Dehydrated human amniotic membrane (dHAM) treatment of lower extremity venous ulceration (CR23). Poster presented at SAWC Annual Spring Meeting in Atlanta, GA. April 2012.
    1. Ennis W, Sui A, Papineau E, Plummer M, Altman I, Meneses P. Clinical experience with a novel regenerative template for hard to heal wounds. SAWC Annual Spring Meeting in Atlanta, GA. April 2012.
    1. Lopez‐Valladares MJ, Rodriguez‐Ares MT, Tourino R, Gude F, Teresa Silva M, Couceiro J. Donor age and gestational age influence on growth factor levels in human amniotic membrane. Acta Opththalmol 2010;88:e211–6.
    1. Russo A, Bonci P, Bonci P. The effects of different preservation processes on the total protein and growth factor content in a new biological product developed from human amniotic membrane. Cell Tissue Bank 2012;13:353–61.
    1. US Patent 8,357,403– Placenta Tissue Grafts.
    1. US Patent 8,372,437‐Placenta Tissue Grafts.
    1. US Patent 8,409,626‐Placenta Tissue Grafts.
    1. Box GEP, Cox DR. An analysis of transformation. J R Stat Soc (B) 1964;26:211–52.
    1. Schultz GS, Davidson JM, Kirsner RS, Bornstein P, Herman IM. Dynamic reciprocity in the wound microenvironment. Wound Repair Regen 2011;19:134–48.
    1. Weber RL, Iacono VJ. The cytokines: a review of interleukins. Periodontal Clin Investig 1997;19:17–22.
    1. Hocking AM, Gibran NS. Mesenchymal stem cells: paracrine signaling and differentiation during cutaneous wound repair. Exp Cell Res 2010;316:2213–9.
    1. Rodriguez‐Menocal L, Salgado M, Ford D, Van Badiavas E. Stimulation of skin and wound fibroblast migration by mesenchymal stem cells derived from normal donors and chronic wound patients. Stem Cells Transl Med 2012;1:221–9.
    1. Wu Y, Chen L, Scott PG, Tredget EE. Mesenchymal stem cells enhance wound healing through differentiation and angiogenesis. Stem Cells 2007;25:2648–59.
    1. Chen JS, Wong VW, Gurtner GC. Therapeutic potential of bone marrow‐derived mesenchymal; stem cells for cutaneous wound healing. Front Immunol 2012;3:192.
    1. Lawrence WT, Diegelmann RF. Growth factors in wound healing. Clin Dermatol 1994;12:157–69.
    1. Barrientos S, Stojadinovic O, Golinko MS, Brem H, Tomic‐Canic M. Growth factors and cytokines in wound healing. Wound Repair Regen 2008;16:585–601.
    1. Cooper DM, Yu EZ, Hennessey P, Ko F, Robson MC. Determination of endogenous cytokines in chronic wounds. Ann Surg 1994;219:688–92.
    1. Schultz G, Rotatori DS, Clark W. EGF and TGF‐α in wound healing and repair. J Cell Biochem 1991;45:346–52.
    1. Pierce GF, Tarpley JE, Tseng J, Bready J, Chang D, Kenney WC, Rudolph R, Robson MC, Vande Berg J, Reid P. Detection of platelet‐derived growth factor (PDGF)‐AA in actively healing human wounds treated with recombinant PDGF‐BB and absence of PDGF in chronic nonhealing wounds. J Clin Invest 1995;96:1336–50.
    1. Robson MC, Phillips LG, Thomason A, Robson LE, Pierce GF. Platelet‐derived growth factor BB for the treatment of chronic pressure ulcers. Lancet 1992;339:23–5.
    1. Brown GL, Curtsinger L, Jurkiewicz MJ, Nahai F, Schultz G. Stimulation of healing of chronic wounds by epidermal growth factor. Plast Reconstr Surg 1991;88:189–94.
    1. Robson MC, Phillips LG, Lawrence WT, Bishop JB, Youngerman JS, Hayward PG, Broemeling LD, Heggers JP. The safety and effect of topically applied recombinant basic fibroblast growth factor on the healing of chronic pressure sores. Ann Surg 1992;216:401–8.
    1. Steed DL. Clinical evaluation of recombinant human platelet—derived growth factor for the treatment of lower extremity diabetic ulcers. J Vasc Surg 1995;21:71–81.
    1. Brown RL, Breeden MP, Greenhalgh DG. PDGF and TGF‐α act synergistically to improve wound healing in the genetically diabetic mouse. J Surg Res 1994;56:562–70.
    1. Lynch SE, Colvin RB, Antoniades HN. Growth factors in wound healing. Single and synergistic effects on partial thickness porcine skin wounds. J Clin Invest 1989;84:640–6.
    1. Hennessey PJ, Black CT, Andrassy RJ. Epidermal growth factor and insulin act synergistically during diabetic healing. Arch Surg 1990;125:926–9.
    1. Robson MC. The role of growth factors in the healing of chronic wounds. Wound Repair Regen 1997;5:12–7.

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