title: Angiogenic properties of dehydrated human amnion/chorion allografts: therapeutic potential for soft tissue repair and regeneration.
abstract: BACKGROUND
  Chronic wounds are associated with a number of deficiencies in critical wound healing processes, including growth factor signaling and neovascularization. Human-derived placental tissues are rich in regenerative cytokines and have been shown in randomized clinical trials to be effective for healing chronic wounds. In this study, PURION® Processed (MiMedx Group, Marietta, GA) dehydrated human amnion/chorion membrane tissue allografts (dHACM, EpiFix®, MiMedx) were evaluated for properties to support wound angiogenesis.
  
  
  METHODS
  Angiogenic growth factors were identified in dHACM tissues using enzyme-linked immunosorbent assays (ELISAs), and the effects of dHACM extract on human microvascular endothelial cell (HMVEC) proliferation and production of angiogenic growth factors was determined in vitro. Chemotactic migration of human umbilical vein endothelial cells (HUVECs) toward pieces of dHACM tissue was determined using a standard in vitro transwell assay. Neovascularization of dHACM in vivo was determined utilizing a murine subcutaneous implant model.
  
  
  RESULTS
  Quantifiable levels of the angiogenic cytokines angiogenin, angiopoietin-2 (ANG-2), epidermal growth factor (EGF), basic fibroblast growth factor (bFGF), heparin binding epidermal growth factor (HB-EGF), hepatocyte growth factor (HGF), platelet derived growth factor BB (PDGF-BB), placental growth factor (PlGF), and vascular endothelial growth factor (VEGF) were measured in dHACM. Soluble cues promoted HMVEC proliferation in vitro and increased endogenous production of over 30 angiogenic factors by HMVECs, including granulocyte macrophage colony-stimulating factor (GM-CSF), angiogenin, transforming growth factor β3 (TGF-β3), and HB-EGF. 6.0 mm disks of dHACM tissue were also found to recruit migration of HUVECs in vitro. Moreover, subcutaneous dHACM implants displayed a steady increase in microvessels over a period of 4 weeks, indicative of a dynamic intra-implant neovascular process.
  
  
  CONCLUSIONS
  TAKEN TOGETHER, THESE RESULTS DEMONSTRATE THAT DHACM GRAFTS: 1) contain angiogenic growth factors retaining biological activity; 2) promote amplification of angiogenic cues by inducing endothelial cell proliferation and migration and by upregulating production of endogenous angiogenic growth factors by endothelial cells; and 3) support the formation of blood vessels in vivo. dHACM grafts are a promising wound care therapy with the potential to promote revascularization and tissue healing within poorly vascularized, non-healing wounds.
authors.affiliation: MiMedx Group, Inc., 1775 West Oak Commons Ct., Marietta, GA, USA.
authors.forename: Thomas J
authors.initials: TJ
authors.lastname: Koob
authors.affiliation: MiMedx Group, Inc., 1775 West Oak Commons Ct., Marietta, GA, USA.
authors.forename: Jeremy J
authors.initials: JJ
authors.lastname: Lim
authors.affiliation: MiMedx Group, Inc., 1775 West Oak Commons Ct., Marietta, GA, USA.
authors.forename: Michelle
authors.initials: M
authors.lastname: Massee
authors.affiliation: MiMedx Group, Inc., 1775 West Oak Commons Ct., Marietta, GA, USA.
authors.forename: Nicole
authors.initials: N
authors.lastname: Zabek
authors.affiliation: Division of Plastic and Reconstructive Surgery, Department of Surgery, Stanford University School of Medicine, Palo Alto, CA, USA.
authors.forename: Robert
authors.initials: R
authors.lastname: Rennert
authors.affiliation: Division of Plastic and Reconstructive Surgery, Department of Surgery, Stanford University School of Medicine, Palo Alto, CA, USA.
authors.forename: Geoffrey
authors.initials: G
authors.lastname: Gurtner
authors.affiliation: The Angiogenesis Foundation, Cambridge, MA, USA.
authors.forename: William W
authors.initials: WW
authors.lastname: Li
keywords: Amnion; Amnion/chorion grafts; Angiogenesis; Chorion; Chronic wounds; Endothelial cells; Growth factors; Soft tissue regeneration; VEGF; Wound healing; dHACM