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FOXO1 has a Dual Function to Promote Normal but Inhibit Diabetic Wound Healing

  • Dana T. GravesEmail author
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Part of the Recent Clinical Techniques, Results, and Research in Wounds book series (RCTRRW, volume 3)

Abstract

FOXO1 is a transcription factor that regulates cellular events, several of which are important to wound healing. When FOXO1 is specifically deleted in keratinocytes of normal wounds, the closure of the wound is impaired. In normal wounds, FOXO1 drives expression of TGFβ1 and antioxidants to facilitate keratinocyte migration. Furthermore, TGFβ1 induced by FOXO1 in keratinocytes plays an important role in promoting connective tissue wound healing. Surprisingly, when FOXO1 is deleted in diabetic wounds, the opposite occurs, and healing is accelerated. In diabetic wounds, FOXO1 is unable to induce TGFβ1 expression and instead enhances production of CCL-10, SerpinB2, and IFN36γ. Expression of the latter factors impedes keratinocyte migration at high levels. Thus, FOXO1 plays a critical role in regulating wound healing behavior of keratinocytes and has opposite functions in normal and diabetic wounds. This is due to the fact that the genes induced by FOXO1 under normal conditions differ than those induced under diabetic conditions.

References

  1. 1.
    Kaul K, Tarr JM, Ahmad SI, Kohner EM, Chibber R (2012) Introduction to diabetes mellitus. Adv Exp Med Biol 771:1–11PubMedGoogle Scholar
  2. 2.
    Graves DT, Kayal RA (2008) Diabetic complications and dysregulated innate immunity. Front Biosci 13:1227–1239PubMedPubMedCentralGoogle Scholar
  3. 3.
    Hameedaldeen A, Liu J, Batres A, Graves GS, Graves DT (2014) FOXO1, TGF-beta regulation and wound healing. Int J Mol Sci 15:16257–16269PubMedPubMedCentralGoogle Scholar
  4. 4.
    Ochoa O, Torres FM, Shireman PK (2007) Chemokines and diabetic wound healing. Vascular 15:350–355PubMedGoogle Scholar
  5. 5.
    Chatzigeorgiou A, Harokopos V, Mylona-Karagianni C, Tsouvalas E, Aidinis V, Kamper EF (2010) The pattern of inflammatory/anti-inflammatory cytokines and chemokines in type 1 diabetic patients over time. Ann Med 42:426–438PubMedGoogle Scholar
  6. 6.
    Khanna S, Biswas S, Shang Y, Collard E, Azad A, Kauh C, Bhasker V, Gordillo GM, Sen CK, Roy S (2010) Macrophage dysfunction impairs resolution of inflammation in the wounds of diabetic mice. PLoS One 5:e9539PubMedPubMedCentralGoogle Scholar
  7. 7.
    Nwomeh BC, Yager DR, Cohen IK (1998) Physiology of the chronic wound. Clin Plast Surg 25:341–356PubMedGoogle Scholar
  8. 8.
    Wu Y, Chen L, Scott PG, Tredget EE (2007) Mesenchymal stem cells enhance wound healing through differentiation and angiogenesis. Stem Cells 25:2648–2659PubMedGoogle Scholar
  9. 9.
    Chen L, Tredget EE, Wu PY, Wu Y (2008) Paracrine factors of mesenchymal stem cells recruit macrophages and endothelial lineage cells and enhance wound healing. PLoS One 3:e1886PubMedPubMedCentralGoogle Scholar
  10. 10.
    Singer NG, Caplan AI (2011) Mesenchymal stem cells: mechanisms of inflammation. Ann Rev Pathol 6:457–478Google Scholar
  11. 11.
    Vojtassak J, Danisovic L, Kubes M, Bakos D, Jarabek L, Ulicna M, Blasko M (2006) Autologous biograft and mesenchymal stem cells in treatment of the diabetic foot. Neuro Endocrinol Lett 27(Suppl 2):134–137PubMedGoogle Scholar
  12. 12.
    Kuo YR, Wang CT, Cheng JT, Wang FS, Chiang YC, Wang CJ (2011) Bone marrow-derived mesenchymal stem cells enhanced diabetic wound healing through recruitment of tissue regeneration in a rat model of streptozotocin-induced diabetes. Plast Reconstr Surg 128:872–880PubMedGoogle Scholar
  13. 13.
    Roh C, Lyle S (2006) Cutaneous stem cells and wound healing. Pediatric Res 59:100R–103RGoogle Scholar
  14. 14.
    Lau K, Paus R, Tiede S, Day P, Bayat A (2009) Exploring the role of stem cells in cutaneous wound healing. Exp Dermatol 18:921–933PubMedGoogle Scholar
  15. 15.
    Werner S, Grose R (2003) Regulation of wound healing by growth factors and cytokines. Physiol Rev 83:835–870PubMedGoogle Scholar
  16. 16.
    Martin P (1997) Wound healing—aiming for perfect skin regeneration. Science 276:75–81PubMedGoogle Scholar
  17. 17.
    Singer AJ, Clark RA (1999) Cutaneous wound healing. N Engl J Med 341:738–746PubMedGoogle Scholar
  18. 18.
    Galkowska H, Wojewodzka U, Olszewski WL (2006) Chemokines, cytokines, and growth factors in keratinocytes and dermal endothelial cells in the margin of chronic diabetic foot ulcers. Wound Repair Regen 14:558–565PubMedGoogle Scholar
  19. 19.
    Lan CC, Liu IH, Fang AH, Wen CH, Wu CS (2008) Hyperglycaemic conditions decrease cultured keratinocyte mobility: implications for impaired wound healing in patients with diabetes. Br J Dermatol 159:1103–1115PubMedGoogle Scholar
  20. 20.
    Lucas T, Waisman A, Ranjan R, Roes J, Krieg T, Muller W, Roers A, Eming SA (2010) Differential roles of macrophages in diverse phases of skin repair. J Immunol 184:3964–3977PubMedGoogle Scholar
  21. 21.
    Martinez FO, Sica A, Mantovani A, Locati M (2008) Macrophage activation and polarization. Front Biosci 13:453–461PubMedGoogle Scholar
  22. 22.
    Al-Mulla F, Leibovich SJ, Francis IM, Bitar MS (2011) Impaired TGF-beta signaling and a defect in resolution of inflammation contribute to delayed wound healing in a female rat model of type 2 diabetes. Mol Biosyst 7:3006–3020PubMedGoogle Scholar
  23. 23.
    Wen Y, Gu J, Li SL, Reddy MA, Natarajan R, Nadler JL (2006) Elevated glucose and diabetes promote interleukin-12 cytokine gene expression in mouse macrophages. Endocrinology 147:2518–2525PubMedGoogle Scholar
  24. 24.
    Han YP, Tuan TL, Wu H, Hughes M, Garner WL (2001) TNF-alpha stimulates activation of pro-MMP2 in human skin through NF-(kappa)B mediated induction of MT1-MMP. J Cell Sci 114:131–139PubMedPubMedCentralGoogle Scholar
  25. 25.
    Hubner G, Brauchle M, Smola H, Madlener M, Fassler R, Werner S (1996) Differential regulation of pro-inflammatory cytokines during wound healing in normal and glucocorticoid-treated mice. Cytokine 8:548–556PubMedGoogle Scholar
  26. 26.
    Ponugoti B, Dong G, Graves DT (2012) Role of forkhead transcription factors in diabetes-induced oxidative stress. Exp Diabetes Res 2012:939751PubMedPubMedCentralGoogle Scholar
  27. 27.
    Siqueira MF, Li J, Chehab L, Desta T, Chino T, Krothpali N, Behl Y, Alikhani M, Yang J, Braasch C, Graves DT (2010) Impaired wound healing in mouse models of diabetes is mediated by TNF-alpha dysregulation and associated with enhanced activation of forkhead box O1 (FOXO1). Diabetologia 53:378–388PubMedGoogle Scholar
  28. 28.
    Wallace HJ, Stacey MC (1998) Levels of tumor necrosis factor-alpha (TNF-alpha) and soluble TNF receptors in chronic venous leg ulcers—correlations to healing status. J Inv Dermatol 110:292–296Google Scholar
  29. 29.
    Kaiser GC, Polk DB (1997) Tumor necrosis factor alpha regulates proliferation in a mouse intestinal cell line. Gastroenterology 112:1231–1240PubMedGoogle Scholar
  30. 30.
    Liu R, Bal HS, Desta T, Behl Y, Graves DT (2006) Tumor necrosis factor-alpha mediates diabetes-enhanced apoptosis of matrix-producing cells and impairs diabetic healing. Am J Pathol 168:757–764PubMedPubMedCentralGoogle Scholar
  31. 31.
    Hasnan J, Yusof MI, Damitri TD, Faridah AR, Adenan AS, Norbaini TH (2010) Relationship between apoptotic markers (Bax and Bcl-2) and biochemical markers in type 2 diabetes mellitus. Singapore Med J 51:50–55PubMedGoogle Scholar
  32. 32.
    Chan YC, Roy S, Khanna S, Sen CK (2012) Downregulation of endothelial microRNA-200b supports cutaneous wound angiogenesis by desilencing GATA binding protein 2 and vascular endothelial growth factor receptor 2. Arterioscler Thromb Vasc Biol 32:1372–1382PubMedPubMedCentralGoogle Scholar
  33. 33.
    Goren I, Muller E, Pfeilschifter J, Frank S (2006) Severely impaired insulin signaling in chronic wounds of diabetic ob/ob mice: a potential role of tumor necrosis factor-alpha. Am J Pathol 168:765–777PubMedPubMedCentralGoogle Scholar
  34. 34.
    Alikhani M, Roy S, Graves DT (2010) FOXO1 plays an essential role in apoptosis of retinal pericytes. Mol Vis 16:408–415PubMedPubMedCentralGoogle Scholar
  35. 35.
    Behl Y, Krothapalli P, Desta T, Roy S, Graves DT (2009) FOXO1 plays an important role in enhanced microvascular cell apoptosis and microvascular cell loss in type 1 and type 2 diabetic rats. Diabetes 58:917–925PubMedPubMedCentralGoogle Scholar
  36. 36.
    Alblowi J, Kayal RA, Siqueria M, McKenzie E, Krothapalli N, McLean J, Conn J, Nikolajczyk B, Einhorn TA, Gerstenfeld L, Graves DT (2009) High levels of tumor necrosis factor-alpha contribute to accelerated loss of cartilage in diabetic fracture healing. Am J Pathol 175:1574–1585PubMedPubMedCentralGoogle Scholar
  37. 37.
    Desta T, Li J, Chino T, Graves DT (2010) Altered fibroblast proliferation and apoptosis in diabetic gingival wounds. J Dental Res 89:609–614Google Scholar
  38. 38.
    Alikhani M, Maclellan CM, Raptis M, Vora S, Trackman PC, Graves DT (2007) Advanced glycation end products induce apoptosis in fibroblasts through activation of ROS, MAP kinases, and the FOXO1 transcription factor. Am J Physiol Cell Physiol 292:C850–C856PubMedGoogle Scholar
  39. 39.
    Weigel D, Jurgens G, Kuttner F, Seifert E, Jackle H (1989) The homeotic gene fork head encodes a nuclear protein and is expressed in the terminal regions of the Drosophila embryo. Cell 57:645–658PubMedGoogle Scholar
  40. 40.
    Golson ML, Kaestner KH (2016) Fox transcription factors: from development to disease. Development 143:4558–4570PubMedPubMedCentralGoogle Scholar
  41. 41.
    Jean D, Harbison M, McConkey DJ, Ronai Z, Bar-Eli M (1998) CREB and its associated proteins act as survival factors for human melanoma cells. J Biol Chem 273:24884–24890PubMedGoogle Scholar
  42. 42.
    Jacobs FM, van der Heide LP, Wijchers PJ, Burbach JP, Hoekman MF, Smidt MP (2003) FoxO6, a novel member of the FoxO class of transcription factors with distinct shuttling dynamics. J Biol Chem 278:35959–35967PubMedGoogle Scholar
  43. 43.
    Adamopoulos IE, Sabokbar A, Wordsworth BP, Carr A, Ferguson DJ, Athanasou NA (2006) Synovial fluid macrophages are capable of osteoclast formation and resorption. J Pathol 208:35–43PubMedGoogle Scholar
  44. 44.
    Birkenkamp K, Coffer P (2003) FOXO transcription factors as regulators of immune homeostasis: molecules to die for? J Immunol 171:1623–1629PubMedGoogle Scholar
  45. 45.
    Coomans de Brachene A, Demoulin JB (2016) FOXO transcription factors in cancer development and therapy. Cell Mol Life Sci 73:1159–1172PubMedGoogle Scholar
  46. 46.
    Barthel A, Schmoll D, Unterman TG (2005) FoxO proteins in insulin action and metabolism. Trends Endocrinol Metab 16:183–189PubMedGoogle Scholar
  47. 47.
    Urbanek P, Klotz LO (2017) Posttranscriptional regulation of FOXO expression: microRNAs and beyond. Br J Pharmacol 174(12):1514–1532PubMedGoogle Scholar
  48. 48.
    Battiprolu PK, Hojayev B, Jiang N, Wang ZV, Luo X, Iglewski M, Shelton JM, Gerard RD, Rothermel BA, Gillette TG, Lavandero S, Hill JA (2012) Metabolic stress-induced activation of FoxO1 triggers diabetic cardiomyopathy in mice. J Clin Inv 122:1109–1118Google Scholar
  49. 49.
    Brunet A, Kanai F, Stehn J, Xu J, Sarbassova D, Frangioni JV, Dalal SN, DeCaprio JA, Greenberg ME, Yaffe MB (2002) 14-3-3 transits to the nucleus and participates in dynamic nucleocytoplasmic transport. J Cell Biol 156:817–828PubMedPubMedCentralGoogle Scholar
  50. 50.
    Lalmansingh AS, Karmakar S, Jin Y, Nagaich AK (2012) Multiple modes of chromatin remodeling by Forkhead box proteins. Biochim Biophys Acta 1819:707–715PubMedGoogle Scholar
  51. 51.
    Tikhanovich I, Cox J, Weinman SA (2013) Forkhead box class O transcription factors in liver function and disease. J Gastroenterol Hepatol 28(Suppl 1):125–131PubMedPubMedCentralGoogle Scholar
  52. 52.
    Wang Y, Zhou Y, Graves DT (2014) FOXO transcription factors: their clinical significance and regulation. Biomed Res Int 2014:925350PubMedPubMedCentralGoogle Scholar
  53. 53.
    Dong G, Wang Y, Xiao W, Pujado S, Xu F, Tian C, Xiao E, Choi Y, Graves DT (2015) FOXO1 regulates dendritic cell activity through ICAM-1 and CCR7. J Immunol 194(8):3745–3755PubMedPubMedCentralGoogle Scholar
  54. 54.
    Chung S, Ranjan R, Lee YG, Park GY, Karpurapu M, Deng J, Xiao L, Kim JY, Unterman TG, Christman JW (2015) Distinct role of FoxO1 in M-CSF- and GM-CSF-differentiated macrophages contributes LPS-mediated IL-10: implication in hyperglycemia. J Leukoc Biol 97:327–339PubMedGoogle Scholar
  55. 55.
    Chen Z, Wang Y, Shi C (2015) Therapeutic implications of newly identified stem cell populations from the skin dermis. Cell Transplant 24:1405–1422PubMedGoogle Scholar
  56. 56.
    Ma H, Yin C, Zhang Y, Qian L, Liu J (2016) ErbB2 is required for cardiomyocyte proliferation in murine neonatal hearts. Gene 592:325–330PubMedPubMedCentralGoogle Scholar
  57. 57.
    Limon JJ, So L, Jellbauer S, Chiu H, Corado J, Sykes SM, Raffatellu M, Fruman DA (2014) mTOR kinase inhibitors promote antibody class switching via mTORC2 inhibition. Proc Natl Acad Sci USA 111:E5076–E5085PubMedGoogle Scholar
  58. 58.
    Cheng Z, White MF (2011) Targeting Forkhead box O1 from the concept to metabolic diseases: lessons from mouse models. Antioxid Redox Signal 14:649–661PubMedPubMedCentralGoogle Scholar
  59. 59.
    Zhang C, Ponugoti B, Tian C, Xu F, Tarapore R, Batres A, Alsadun S, Lim J, Dong G, Graves DT (2015) FOXO1 differentially regulates both normal and diabetic wound healing. J Cell Biol 209:289–303PubMedPubMedCentralGoogle Scholar
  60. 60.
    Reinke JM, Sorg H (2012) Wound repair and regeneration. Eur Surg Res 49:35–43PubMedGoogle Scholar
  61. 61.
    Grice EA, Segre JA (2012) Interaction of the microbiome with the innate immune response in chronic wounds. Adv Exp Med Biol 946:55–68PubMedPubMedCentralGoogle Scholar
  62. 62.
    Xu F, Zhang C, Graves DT (2013) Abnormal cell responses and role of TNF-alpha in impaired diabetic wound healing. Biomed Res Int 2013:754802PubMedPubMedCentralGoogle Scholar
  63. 63.
    Goova MT, Li J, Kislinger T, Qu W, Lu Y, Bucciarelli LG, Nowygrod S, Wolf BM, Caliste X, Yan SF, Stern DM, Schmidt AM (2001) Blockade of receptor for advanced glycation end-products restores effective wound healing in diabetic mice. Am J Pathol 159:513–525PubMedPubMedCentralGoogle Scholar
  64. 64.
    Coulombe PA (2003) Wound epithelialization: accelerating the pace of discovery. J Invest Dermatol 121:219–230PubMedGoogle Scholar
  65. 65.
    Raja SK, Garcia MS, Isseroff RR (2007) Wound re-epithelialization: modulating keratinocyte migration in wound healing. Front Biosci 12:2849–2868PubMedGoogle Scholar
  66. 66.
    Alikhani M, Alikhani Z, Boyd C, MacLellan CM, Raptis M, Liu R, Pischon N, Trackman PC, Gerstenfeld L, Graves DT (2007) Advanced glycation end products stimulate osteoblast apoptosis via the MAP kinase and cytosolic apoptotic pathways. Bone 40:345–353PubMedGoogle Scholar
  67. 67.
    Lim JC, Kl K, Mattos M, Fang M, Zhang C, Feinberg D, Sindi H, Li S, Alblowi J, Kayal RA, Einhorn TA, Gerstenfeld LC, Graves DT (2017) TNFα contributes to diabetes impaired angiogenesis in fracture healing. Bone 99:26–38PubMedPubMedCentralGoogle Scholar
  68. 68.
    Kayal RA, Siqueira M, Alblowi J, McLean J, Krothapalli N, Faibish D, Einhorn TA, Gerstenfeld LC, Graves DT (2010) TNF-α mediates diabetes-enhanced chondrocyte apoptosis during fracture healing and stimulates chondrocyte apoptosis Through FOXO1. J Bone Mineral Res 25:1604–1615Google Scholar
  69. 69.
    Ko KI, Coimbra LS, Tian C, Alblowi J, Kayal RA, Einhorn TA, Gerstenfeld LC, Pignolo RJ, Graves DT (2015) Diabetes reduces mesenchymal stem cells in fracture healing through a TNFalpha-mediated mechanism. Diabetologia 58:633–642PubMedPubMedCentralGoogle Scholar
  70. 70.
    Li S, Dong G, Moschidis A, Ortiz J, Benakanakere MR, Kinane DF, Graves DT (2013) P. Gingivalis modulates keratinocytes through FOXO transcription factors. PLoS One 8:e78541PubMedPubMedCentralGoogle Scholar
  71. 71.
    Ponugoti B, Xu F, Zhang C, Tian C, Pacios S, Graves DT (2013) FOXO1 promotes wound healing through the up-regulation of TGF-beta1 and prevention of oxidative stress. J Cell Biol 203:327–343PubMedPubMedCentralGoogle Scholar
  72. 72.
    Xu F, Othman B, Lim J, Batres A, Ponugoti B, Zhang C, Yi L, Liu J, Tian C, Hameedaldeen A, Alsadun S, Tarapore R, Graves DT (2015) Foxo1 inhibits diabetic mucosal wound healing but enhances healing of normoglycemic wounds. Diabetes 64:243–256PubMedGoogle Scholar
  73. 73.
    Gailit J, Welch MP, Clark RA (1994) TGF-beta 1 stimulates expression of keratinocyte integrins during re-epithelialization of cutaneous wounds. J Invest Dermatol 103:221–227PubMedGoogle Scholar
  74. 74.
    Xiao E, Graves DT (2015) Impact of diabetes on the protective role of FOXO1 in wound healing. J Dent Res 94:1025–1026PubMedPubMedCentralGoogle Scholar
  75. 75.
    Bos DC, de Ranitz-Greven WL, de Valk HW (2011) Advanced glycation end products, measured as skin autofluorescence and diabetes complications: a systematic review. Diabetes Technol Ther 13:773–779PubMedGoogle Scholar
  76. 76.
    Gkogkolou P, Bohm M (2012) Advanced glycation end products: Key players in skin aging? Dermatoendocrinology 4:259–270Google Scholar
  77. 77.
    Altunbas A, Lee SJ, Rajasekaran SA, Schneider JP, Pochan DJ (2011) Encapsulation of curcumin in self-assembling peptide hydrogels as injectable drug delivery vehicles. Biomaterials 32:5906–5914PubMedPubMedCentralGoogle Scholar
  78. 78.
    Lindhurst MJ, Sapp JC, Teer JK, Johnston JJ, Finn EM, Peters K, Turner J, Cannons JL, Bick D, Blakemore L, Blumhorst C, Brockmann K, Calder P, Cherman N et al (2011) A mosaic activating mutation in AKT1 associated with the Proteus syndrome. N Engl J Med 365:611–619PubMedPubMedCentralGoogle Scholar
  79. 79.
    Zambruno G, Marchisio PC, Marconi A, Vaschieri C, Melchiori A, Giannetti A, De Luca M (1995) Transforming growth factor-beta 1 modulates beta 1 and beta 5 integrin receptors and induces the de novo expression of the alpha v beta 6 heterodimer in normal human keratinocytes: implications for wound healing. J Cell Biol 129:853–865PubMedGoogle Scholar
  80. 80.
    Hebda PA (1988) Stimulatory effects of transforming growth factor-beta and epidermal growth factor on epidermal cell outgrowth from porcine skin explant cultures. J Invest Dermatol 91(5):440PubMedGoogle Scholar
  81. 81.
    Deveci M, Gilmont RR, Dunham WR, Mudge BP, Smith DJ, Marcelo CL (2005) Glutathione enhances fibroblast collagen contraction and protects keratinocytes from apoptosis in hyperglycaemic culture. Br J Dermatol 152:217–224PubMedGoogle Scholar
  82. 82.
    Zhang C, Lim J, Liu J, Ponugoti B, Alsadun S, Tian C, Vafa R, Graves DT (2017) FOXO1 expression in keratinocytes promotes connective tissue healing. Sci Rep 7:42834PubMedPubMedCentralGoogle Scholar
  83. 83.
    Forbes SJ, Rosenthal N (2014) Preparing the ground for tissue regeneration: from mechanism to therapy. Nat Med 20:857–869PubMedGoogle Scholar
  84. 84.
    Koh TJ, DiPietro LA (2011) Inflammation and wound healing: the role of the macrophage. Expert Rev Molec Med 13:e23Google Scholar
  85. 85.
    Martins-Green M, Petreaca M, Wang L (2013) Chemokines and their receptors are key players in the orchestra that regulates wound healing. Adv Wound Care (New Rochelle) 2:327–347Google Scholar
  86. 86.
    Lacroix M, Bovy T, Nusgens BV, Lapiere CM (1995) Keratinocytes modulate the biosynthetic phenotype of dermal fibroblasts at a pretranslational level in a human skin equivalent. Arch Dermatol Res 287:659–664PubMedGoogle Scholar
  87. 87.
    Walter MN, Wright KT, Fuller HR, MacNeil S, Johnson WE (2010) Mesenchymal stem cell-conditioned medium accelerates skin wound healing: an in vitro study of fibroblast and keratinocyte scratch assays. Exp Cell Res 316:1271–1281PubMedGoogle Scholar
  88. 88.
    Leask A, Holmes A, Black CM, Abraham DJ (2003) Connective tissue growth factor gene regulation. Requirements for its induction by transforming growth factor-beta 2 in fibroblasts. J Biol Chem 278:13008–13015PubMedGoogle Scholar
  89. 89.
    Tong Z, Sant S, Khademhosseini A, Jia X (2011) Controlling the fibroblastic differentiation of mesenchymal stem cells via the combination of fibrous scaffolds and connective tissue growth factor. Tissue Eng Part A 17:2773–2785PubMedPubMedCentralGoogle Scholar
  90. 90.
    Yang CY, Jeon HH, Alshabab A, Chung CH, Graves DT (2017) RANKL deletion in periodontal ligament cells blocks orthodontic tooth movement. IJOS In PressGoogle Scholar

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Authors and Affiliations

  1. 1.Department of Periodontics, School of Dental MedicineUniversity of PennsylvaniaPhiladelphiaUSA

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