title: Rocking media over ex vivo corneas improves this model and allows the study of the effect of proinflammatory cytokines on wound healing.
abstract: OBJECTIVE
  The aim of this work was to develop an in vitro cornea model to study the effect of proinflammatory cytokines on wound healing.
  
  
  METHODS
  Initial studies investigated how to maintain the ex vivo models for up to 4 weeks without loss of epithelium. To study the effect of cytokines, corneas were cultured with the interleukins IL-17A, IL-22, or a combination of IL-17A and IL-22, or lipopolysaccharide (LPS). The effect of IL-17A on wound healing was then examined.
  
  
  RESULTS
  With static culture conditions, organ cultures deteriorated within 2 weeks. With gentle rocking of media over the corneas and carbon dioxide perfusion, the ex vivo models survived for up to 4 weeks without loss of epithelium. The cytokine that caused the most damage to the cornea was IL-17A. Under static conditions, wound healing of the central corneal epithelium occurred within 9 days, but only a single-layered epithelium formed whether the cornea was exposed to IL-17A or not. With rocking of media gently over the corneas, a multilayered epithelium was achieved 9 days after wounding. In the presence of IL-17A, however, there was no wound healing evident. Characterization of the cells showed that wherever epithelium was present, both differentiated cells and highly proliferative cells were present.
  
  
  CONCLUSIONS
  We propose that introducing rocking to extend the effective working life of this model and the introduction of IL-17A to this model to induce aspects of inflammation extend its usefulness to study the effects of agents that influence corneal regeneration under normal and inflamed conditions.
authors: Department of Materials Science and Engineering, Kroto Research Institute, University of Sheffield, Sheffield, United Kingdom.
authors: Pallavi
authors: P
authors: Deshpande
authors: Department of Materials Science and Engineering, Kroto Research Institute, University of Sheffield, Sheffield, United Kingdom.
authors: Ílida
authors: Í
authors: Ortega
authors: Department of Materials Science and Engineering, Kroto Research Institute, University of Sheffield, Sheffield, United Kingdom.
authors: Farshid
authors: F
authors: Sefat
authors: Sudhakar and Sreekanth Ravi Stem Cell Laboratory, LV Prasad Eye Institute, Kallam Anji Reddy Campus, LV Prasad Marg, Hyderabad, Telangana, India.
authors: Virender S
authors: VS
authors: Sangwan
authors: Department of Materials Science and Engineering, Kroto Research Institute, University of Sheffield, Sheffield, United Kingdom.
authors: Nicola
authors: N
authors: Green
authors: Department of Materials Science and Engineering, Kroto Research Institute, University of Sheffield, Sheffield, United Kingdom.
authors: Frederik
authors: F
authors: Claeyssens
authors: Department of Materials Science and Engineering, Kroto Research Institute, University of Sheffield, Sheffield, United Kingdom.
authors: Sheila
authors: S
authors: MacNeil
mesh_terms: D000818:Animals; D003315:Cornea / Q000187:drug effects*; D003470:Culture Media; D046508:Culture Techniques / Q000295:instrumentation; D019573:Epithelium, Corneal / Q000187:drug effects; D020381:Interleukin-17 / Q000494:pharmacology*; D007378:Interleukins / Q000494:pharmacology*; D008070:Lipopolysaccharides / Q000494:pharmacology*; D009924:Organ Culture Techniques / Q000295:instrumentation*; D011817:Rabbits; D012038:Regeneration / Q000187:drug effects; D014945:Wound Healing / Q000187:drug effects*; D000097043:Interleukin-22
chemical_list: D003470:Culture Media; D020381:Interleukin-17; D007378:Interleukins; D008070:Lipopolysaccharides; D000097043:Interleukin-22
keywords: cornea organ culture; inflammation; wound healing
references: Invest Ophthalmol Vis Sci. 2006 May;47(5):1947-55
references: 16639002
references: Ann N Y Acad Sci. 2010 Jan;1183:211-21
references: 20146717
references: Prog Retin Eye Res. 2001 Sep;20(5):639-73
references: 11470454
references: Arch Ophthalmol. 2008 Jan;126(1):23-8
references: 18195214
references: Eur Urol. 2008 Jun;53(6):1263-9
references: 18262717
references: J Cell Biol. 1986 Jul;103(1):49-62
references: 2424919
references: Cytokine. 2014 Jul;68(1):1-8
references: 24787050
references: Clin Diagn Lab Immunol. 2005 Jan;12(1):60-7
references: 15642986
references: Curr Eye Res. 1991 Aug;10(8):739-49
references: 1914506
references: J Clin Invest. 2009 Dec;119(12):3573-85
references: 19920355
references: Exp Eye Res. 1968 Jan;7(1):103-12
references: 5640357
references: Indian J Ophthalmol. 2006 Mar;54(1):29-34
references: 16531667
references: Invest Ophthalmol Vis Sci. 2006 Nov;47(11):4794-802
references: 17065490
references: Nat Med. 2007 Jun;13(6):711-8
references: 17496900
references: Emerg Microbes Infect. 2013 Sep;2(9):e60
references: 26038490
references: Proc Natl Acad Sci U S A. 1971 Jan;68(1):229-33
references: 4322262
references: Biomaterials. 2013 Jul;34(21):5088-106
references: 23591389
references: Invest Ophthalmol Vis Sci. 1999 Mar;40(3):575-81
references: 10067960
references: Exp Eye Res. 1996 May;62(5):555-64
references: 8759523
references: J Immunol. 2002 Nov 15;169(10):5897-903
references: 12421973
references: Mucosal Immunol. 2009 May;2(3):243-53
references: 19242409
references: Eur Respir J. 2005 May;25(5):789-96
references: 15863634
references: Int J Inflam. 2012;2012:819467
references: 22229105
references: Exp Eye Res. 1996 Apr;62(4):325-7
references: 8795451
references: J Biomech. 1974 May;7(3):225-38
references: 4846263
references: J Leukoc Biol. 1994 Dec;56(6):672-86
references: 7996043
references: Tissue Eng Part A. 2009 Oct;15(10):2889-902
references: 19265461
references: Invest Ophthalmol Vis Sci. 2006 Jul;47(7):2840-6
references: 16799023
references: J Immunol. 2012 Mar 1;188(5):2410-8
references: 22266282
references: BMC Neurosci. 2010;11:72
references: 20537148
references: Arch Ophthalmol. 1998 Nov;116(11):1470-4
references: 9823348
references: Clin Exp Immunol. 2007 Dec;150(3):407-15
references: 17900301
references: Cornea. 2000 Jul;19(4):421-6
references: 10928750
references: Int Immunol. 2012 Mar;24(3):147-58
references: 22207130
references: Br J Ophthalmol. 1994 May;78(5):401-8
references: 8025077
references: PLoS One. 2011;6(6):e21180
references: 21695050
references: Int J Exp Pathol. 2014 Jun;95(3):216-28
references: 24713057
references: Infect Immun. 2000 Mar;68(3):1731-4
references: 10678999
references: Prog Retin Eye Res. 2001 Sep;20(5):625-37
references: 11470453
references: Invest Ophthalmol Vis Sci. 1998 Dec;39(13):2626-36
references: 9856772
references: Burns. 2013 Sep;39(6):1170-7
references: 23597846
references: Gastroenterology. 2005 Sep;129(3):969-84
references: 16143135
references: Immunol Res. 2008;41(2):87-102
references: 18172584
references: FASEB J. 2011 Aug;25(8):2659-68
references: 21518851
references: Exp Eye Res. 2003 Jun;76(6):735-43
references: 12742356
references: Proc Natl Acad Sci U S A. 2001 Mar 13;98(6):3156-61
references: 11248048
references: Arthritis Rheum. 2005 Apr;52(4):1037-46
references: 15818686
references: Proc Natl Acad Sci U S A. 1981 Jul;78(7):4334-8
references: 6945587
references: J Immunol. 2005 Mar 15;174(6):3695-702
references: 15749908
references: Semin Cell Dev Biol. 2008 Apr;19(2):125-33
references: 17920963
references: J Neurosci Methods. 2001 Sep 30;110(1-2):17-24
references: 11564520
references: Stem Cells. 2010 Dec;28(12):2172-81
references: 20936708
references: Tissue Eng Part A. 2011 Mar;17(5-6):741-50
references: 20929285