Abstract
We have investigated the wound-healing effects of mesenchymal stem cells (MSCs) in combination with human amniotic membrane (HAM) when grafted into full-thickness skin defects of rabbits. Five defects in each of four groups were respectively treated with HAM loaded with autologous MSCs (group A), HAM loaded with allologous MSCs (group B), HAM with injected autologous MSCs (group C), and HAM with injected allologous MSCs (group D). The size of the wounds was calculated for each group at 7, 12, and 15 days after grafting. The wounds were subsequently harvested at 25 days after grafting. Sections stained with hematoxylin and eosin were used to determine the quality of wound healing, as based on the characteristics and amount of granulated tissue in the epidermal and dermal layers. Groups A and B showed the most pronounced effect on wound closure, with statistically significant improvement in wound healing being seen on post-operative days 7, 12, and 15. Although a slight trend toward improved wound healing was seen in group A compared with group B, no statistically significant difference was found at any time point between the two groups. Histological examination of healed wounds from groups A and B showed a thin epidermis with mature differentiation and collagen bundle deposition plus recovered skin appendages in the dermal layer. In contrast, groups C and D showed thickened epidermis with immature epithelial cells and increased fibroblast proliferation with only partially recovered skin appendages in the dermal layer. Thus, the graft of HAM loaded with MSCs played an effective role during the healing of skin defects in rabbits, with no significant difference being observed in wound healing between autologous and allologous MSC transplantation.
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Adinolfi M, Akle CA, McColl I, Fensom AH, Tansley L, Connolly P, et al (1982) Expression of HLA antigens, beta 2-microglobulin and enzymes by human amniotic epithelial cells. Nature 295:325–327
Akashi T, Miyagi T, Ando N, Suzuki Y, Nemoto T, Eishi Y, et al (1999) Synthesis of basement membrane by gastrointestinal cancer cell lines. J Pathol 187:223–228
Akle CA, Adinolfi M, Welsh KI, Leibowitz S, McColl I (1981) Immunogenicity of human amniotic epithelial cells after transplantation into volunteers. Lancet II:1003–1005
Badiavas EV, Abedi M, Butmarc J, Falanga V, Quesenberry P (2003) Participation of bone marrow derived cells in cutaneous wound healing. J Cell Physiol 196:245–250
Bartholomew A, Sturgeon C, Siatskas M, Ferrer K, Mcintosh K, Patil S, et al (2002) Mesenchymal stem cells suppress lymphocyte proliferation in vitro and prolong skin graft survival in vivo. Exp Hematol 30:42–48
Benirschke K, Kaufman P (2000) Pathology of the human placenta. Springer, New York, pp 273–281
Deng W, Han Q, Liao L, Li C, Ge W, Zhao Z, et al (2005) Engrafted bone marrow-derived FIK-11 mesenchymal stem cells regenerate skin tissue. Tissue Eng 11:110–119
Devine SM, Peter S, Martin BJ, Barry F, Mcintosh KR (2001) Mesenchymal stem cells: stealth and suppression. Cancer J 2:76–82
Dinicola M, Carlo-Stella C, Magni M, Milanesi IM, Longoni PD, Matteucci P, et al (2002) Human bone marrow stromal cells suppress T-lymphocyte proliferation induced by cellular or nonspecific mitogenic stimuli. Blood 99:3838–3843
Fathke C, Wilson L, Hutter J, Kapoor V, Smith A, Hocking A, et al (2004) Contribution of bone marrow-derived cells to skin: collagen deposition and wound repair. Stem Cells 22:821–822
Fu X, Fang L, Li X, Cheng B, Sheng Z (2006) Enhanced wound-healing quality with bone marrow mesenchymal stem cells autografting after skin injury. Wound Repair Regen 14:325–335
Hammer A, Hutter H, Blaschitz A, Mahnert W, Hartmann M, Uchanska-Ziegler B, et al (1997) Amnion epithelial cells, in contrast to trophoblast cells, express all classical HLA class I molecules together with HLA-G. Am J Reprod Immunol 37:161–171
Herendael B van, Oberti C, Brosens I (1978) Microanatomy of the human amniotic membranes. A light microscopic, transmission, and scanning electron microscopic study. Am J Obstet Gynecol 131:872–880
Kataoka K, Medina RJ, Kageyama T, Miyazaki M, Yoshino T, Makino T, et al (2003) Participation of adult mouse bone marrow cells in reconstitution of skin. Am J Pathol 163:1227–1231
Kim CH, Kim SS, Sohn SK, Kim DH, Song CG, Kim HJ (2008) The effect of human amniotic membrane, epidermal cells and marrow mesenchymal stem cells in healing a skin defect. J Korean Orthop Assoc 43:276–286
Koizumi NJ, Inatomi TJ, Sotozono CJ, Fullwood NJ, Quantock AJ, Kinoshita S (2000) Growth factor mRNA and protein in preserved human amniotic membrane. Curr Eye Res 20:173–177
Korbling M, Katz RL, Khanna A, Ruifrok AC, Rondon G, Albitar M, et al (2002) Hepatocytes and epithelial cells of donor origin in recipients of peripheral-blood stem cells. N Engl J Med 346:738–746
Kubo M, Sonoda Y, Muramatsu R, Usui M (2001) Immunogenicity of human amniotic membrane in experimental xenotransplantation. Invest Ophthalmol Vis Sci 42:1539–1546
Lee JH, Kosinski PA, Kemp DM (2005) Contribution of human bone marrow stem cells to the individual skeletal myotubes followed by myogenic gene activation. Exp Cell Res 307:174–182
Linden PJ van der, Groetj AF de, Dunselman GA (1996) Endometrial cell adhesion in an in vitro model using intact amniotic membranes. Fertil Steril 65:76–80
Lwebuga-Mukasa JS, Thulin G, Madri JA, Barrett C, Warshaw JB (1984) An acellular human amnionic membrane model for in vitro culture of type II pneumocytes: the role of the basement membrane in cell morphology and function. J Cell Physiol 121:215–225
Mangi AA, Noiseux N, Kong D, He H, Rezvani M, Ingwall JS, et al (2003) Mesenchymal stem cells modified with Akt provent remodeling and restore preference of infarcted hearts. Nat Med 9:1195–1201
McIntosh K, Bartholomew A (2000) Stromal cell modulation of the immune system. A potential role for mesenchymal stem cells. Graft 3:324–328
Modesti A, Scarpa S, D’Orazi G, Simonelli L, Caramia FG (1989) Localization of type IV and V collagens in the stroma of human amnion. Prog Clin Biol Res 296:459–463
Mohamad H (2001) Anatomy and embryology of human placenta. Amnion and chorion. World Scientific, London
Nakagawa H, Akita S, Fukui M, Fujii T, Akino K (2005) Human mesenchymal stem cells successfully improve skin-substitute wound healing. Cutan Biol 153:2–36
Okamoto R, Yajima T, Yamazki M, Kanai T, Mukai M, Okamoto S, et al (2002) Damaged epithelia regenerated by bone marrow-derived cells in the human gastrointestinal tract. Nat Med 8:1011–1017
Orlic D, Kajstura J, Chimenti S, Jakoniuk I, Anderson SM, Li B, et al (2001) Bone marrow cells regenerate infarcted myocardium. Nature 410:701–705
Ortiz-Urda S, Lin Q, Green CL, Keene DR, Marinkovich MP, Khavari PA (2003) Injection of genetically engineered fibroblasts corrects regenerated human epidermolysis bullosa skin tissue. J Clin Invest 111:251–255
Pittenger MF, Mackay AM, Beck SC, Jaiswal RK, Douglas R, Mosca JD, et al (1999) Multilineage potential of adult human mesenchymal stem cells. Science 284:143–147
Prasad J, Feller I, Thomson P (1986) Use of amnion for the treatment of Stevens-Johnson syndrome. J Trauma 26:945–946
Satoh H, Kishi K, Tanaka T, Kubota Y, Nakajima T, Akasaka Y, et al (2004) Transplanted mesenchymal stem cells are effective for skin regeneration in acute cutaneous wounds. Cell Transplant 13:405–412
Sheridan RL, Tompkins RG (1990) Skin substitutes in burns. Burns 25:97–103
Stamm C, Westphal B, Kleine HD, Petzsch M, Kittner C, Klinge H, et al (2003) Autologous bone-marrow stem-cell transplantation for myocardial regeneration. Lancet 361:45–46
Subrahmanyam M (1995) Amniotic membrane as a cover for microskin grafts. Br J Plast Surg 48:477–478
Suzuki S, Matsuda K, Isshiki N, Tamada Y, Ikada Y (1990) Experimental study of a newly developed bilayer artificial skin. Biomaterials 11:356–360
Talmi YP, Sigler L, Inge E, Finkelstein Y, Zohar Y (1991) Antibacterial properties of human amniotic membranes. Placenta 12:285–288
Toda A, Okabe M, Yoshida T, Nikaido T (2007) The potential of amniotic membrane/amnion-derived cells for regeneration of various tissues. J Pharmacol Sci 105:215–228
Trelford J, Trelford-Sauder M (1979) The amnion in surgery, past and present. Am J Obstet Gynecol 134:833–845
Ueta M, Kweon MN, Sano Y, Sotozono C, Yamada J, Koizumi N, et al (2002) Immunosuppressive properties of human amniotic membrane for mixed lymphocyte reaction. Clin Exp Immunol 129:464–470
Wang M, Yoshida A, Kawashima H, Ishizaki M, Takahashi H, Hori J (2006) Immunogenicity and antigenicity of allogeneic amniotic epithelial transplants grafted to the cornea, conjunctiva, and anterior chamber. Invest Ophthalmol Vis Sci 47:1522–1532
Wilshaw SP, Kearney JN, Fisher J, Ingham E (2006) Production of an acellular amniotic membrane matrix for use in tissue engineering. Tissue Eng 12:2117–2129
Wolf HJ, Schmidt W, Drenckhahn D (1991) Immunocytochemical analysis of the cytoskeleton of the human amniotic epithelium. Cell Tissue Res 266:385–389
Yan G, Su Y, Ai G (2004) Study on human amniotic membrane loaded with marrow mesenchymal stem cells and epidermis cells in promoting healing of wound combined with radiation injury. Zhongguo Xiu Fu Chong Jian Wai Ke Za Zhi 18:497–501
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This study was supported by research funds from Dong-A University.
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Kim, S.S., Song, C.K., Shon, S.K. et al. Effects of human amniotic membrane grafts combined with marrow mesenchymal stem cells on healing of full-thickness skin defects in rabbits. Cell Tissue Res 336, 59–66 (2009). https://doi.org/10.1007/s00441-009-0766-1
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DOI: https://doi.org/10.1007/s00441-009-0766-1