[Growth Factors and Interleukins in Amniotic Membrane Tissue Homogenate].

PURPOSE Application of amniotic membrane homogenate eye drops may be a potential treatment alternative for therapy resistant corneal epithelial defects. The purpose of this study was to determine the concentrations of epidermal growth factor (EGF), fibroblast growth factor basic (bFGF), hepatocyte growth factor (HGF), keratinocyte growth factor (KGF), interleukin-6 (IL-6) and interleukin-8 (IL-8) in amniotic membrane homogenates. METHODS Amniotic membranes of 8 placentas were prepared and thereafter stored at - 80 °C using the standard methods of the LIONS Cornea Bank Saar-Lor-Lux, Trier/Westpfalz. Following defreezing, amniotic membranes were cut in two pieces and homogenized in liquid nitrogen. One part of the homogenate was prepared in cell-lysis buffer, the other part was prepared in PBS. The tissue homogenates were stored at - 20 °C until enzyme-linked immunosorbent assay (ELISA) analysis for EGF, bFGF, HGF, KGF, IL-6 and IL-8 concentrations. RESULTS Concentrations of KGF, IL-6 and IL-8 were below the detection limit using both preparation techniques. The EGF concentration in tissue homogenates treated with cell-lysis buffer (2412 pg/g tissue) was not significantly different compared to that of tissue homogenates treated with PBS (1586 pg/g tissue, p = 0.72). bFGF release was also not significantly different using cell-lysis buffer (3606 pg/g tissue) or PBS treated tissue homogenates (4649 pg/g tissue, p = 0.35). HGF release was significantly lower using cell-lysis buffer (23,555 pg/g tissue), compared to PBS treated tissue (47,766 pg/g tissue, p = 0.007). CONCLUSION Containing EGF, bFGF and HGF, and lacking IL-6 and IL-8, the application of amniotic membrane homogenate eye drops may be a potential treatment alternative for therapy-resistant corneal epithelial defects.

[1]  B. Seitz,et al.  Growth factor and interleukin concentration in amniotic membrane suspension (AMS) and amniotic tissue homogenates (AMTH) , 2014 .

[2]  A. Murakami,et al.  Role of the IL-6 classic- and trans-signaling pathways in corneal sterile inflammation and wound healing. , 2011, Investigative ophthalmology & visual science.

[3]  N. Bazan,et al.  EGF stimulates lipoxin A4 synthesis and modulates repair in corneal epithelial cells through ERK and p38 activation. , 2011, Investigative ophthalmology & visual science.

[4]  S. Kozma,et al.  Differences in Wound Healing in Mice with Deficiency of IL-6 versus IL-6 Receptor , 2010, The Journal of Immunology.

[5]  Jia Yin,et al.  Growth factors and corneal epithelial wound healing , 2010, Brain Research Bulletin.

[6]  Jin A. Choi,et al.  Effects of amniotic membrane suspension in human corneal wound healing in vitro , 2009, Molecular vision.

[7]  A. Behrens,et al.  Corneal wound healing is modulated by topical application of amniotic fluid in an ex vivo organ culture model. , 2008, Experimental eye research.

[8]  S. Qi,et al.  Effects of basic fibroblast growth factor on the expression of extracellular matrix and matrix metalloproteinase‐1 in wound healing , 2008, Clinical and experimental dermatology.

[9]  Jonathan Chupka,et al.  Detection of cytokine protein expression in mouse lung homogenates using suspension bead array , 2006, Journal of Inflammation.

[10]  M. Beckmann,et al.  Integration patterns of cryopreserved amniotic membranes into the human cornea. , 2006, Ophthalmology.

[11]  B. Seitz,et al.  Histopathology and ultrastructure of human corneas after amniotic membrane transplantation. , 2006, Archives of ophthalmology.

[12]  H. Tanihara,et al.  Role of Cytokines and Chemokines in Pseudomonal Keratitis , 2005, Cornea.

[13]  M. Boulton,et al.  Hepatocyte growth factor and keratinocyte growth factor regulation of epithelial and stromal corneal wound healing , 2005, Journal of cataract and refractive surgery.

[14]  A. Tarkkanen,et al.  Epidermal growth factor is a constant component of normal human tear fluid , 2005, Graefe's Archive for Clinical and Experimental Ophthalmology.

[15]  S. Werner,et al.  Regulation of wound healing by growth factors and cytokines. , 2003, Physiological reviews.

[16]  B. Seitz,et al.  Corneal calcification after amniotic membrane transplantation , 2003, The British journal of ophthalmology.

[17]  G. A. Limb,et al.  Human corneal epithelial cells require MMP-1 for HGF-mediated migration on collagen I. , 2003, Investigative ophthalmology & visual science.

[18]  H. Bazan,et al.  HGF- and KGF-induced activation of PI-3K/p70 s6 kinase pathway in corneal epithelial cells: its relevance in wound healing. , 2001, Experimental eye research.

[19]  S. Shimmura,et al.  Antiinflammatory Effects of Amniotic Membrane Transplantation in Ocular Surface Disorders , 2001, Cornea.

[20]  S. Tseng,et al.  Suppression of interleukin 1α and interleukin 1β in human limbal epithelial cells cultured on the amniotic membrane stromal matrix , 2001 .

[21]  N. Koizumi,et al.  Growth factor mRNA and protein in preserved human amniotic membrane , 2000 .

[22]  R. Mohan,et al.  Expression of HGF, KGF, EGF and receptor messenger RNAs following corneal epithelial wounding. , 1999, Experimental eye research.

[23]  H. Sheardown,et al.  Tear EGF concentration following corneal epithelial wound creation. , 1996, Journal of ocular pharmacology and therapeutics : the official journal of the Association for Ocular Pharmacology and Therapeutics.

[24]  T. Inatomi,et al.  Keratinocyte growth factor accelerates corneal epithelial wound healing in vivo. , 1995, Investigative ophthalmology & visual science.

[25]  K. Matsumoto,et al.  Roles of HGF as a pleiotropic factor in organ regeneration. , 1993, EXS.

[26]  J. Rubin,et al.  Human KGF is FGF-related with properties of a paracrine effector of epithelial cell growth. , 1989, Science.