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The expression of inflammatory cytokines in chemical or microkeratomeassisted corneal epithelial fla

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The expression of inflammatory cytokines in chemical- or microkeratome-assisted ... 2,4, Hyuk- Jin Choi MD1,2, Hyun Joo Lee1,2, Mee Kum Kim, MD1,2, Won Ryang Wee, ... – PowerPoint PPT presentation

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Title: The expression of inflammatory cytokines in chemical or microkeratomeassisted corneal epithelial fla


1
The expression of inflammatory cytokines in
chemical- or microkeratome-assisted corneal
epithelial flaps in rabbits
  • Taehyung Lim, MD1,2,4, Hyuk- Jin Choi MD1,2, Hyun
    Joo Lee1,2, Mee Kum Kim, MD1,2, Won Ryang Wee,
    MD1,2, Jin Hak Lee,2,3
  • Department of Ophthalmology, Seoul National
    University Hospital1, Seoul, Korea
  • Seoul Artificial Eye Center, Seoul National
    University Hospital Clinical Research Institute2,
    Seoul, Korea
  • Department of Ophthalmology, Bundang Seoul
    National University Hospital3, Seoul, Korea
  • HanGil Eye Hospital4, Incheon, Korea
  • Authors have not any financial support

2
Purpose
  • To compare the expression of inflammatory
    cytokines in chemical- or microkeratome-assisted
    corneal epithelial flaps in rabbit animal models.

3
Subject Methods
  • 40 eyes of 20 New Zealand White rabbits
  • Bilateral study
  • Corneal epithelial flap(diameter of 5.0 mm) was
    made using the following ways
  • Group A
  • 1. Application of 20 alcohol for 30 seconds,
    2 or 3 times
  • 2. Corneal epithelial flap was made using a
    micro hoe (Katena, U.S.)
  • 3. The flap was replaced
  • Group B
  • 1.The flap was made using epimicrokeratome(Ama
    deus, ?? Hz)
  • 2. The flap was replaced

4
  • All the eyes wore therapeutic contact lenses and
    received tarsorrhaphies
  • Eyes were enucleated at 3 days after the
    procedure
  • Tissue sections were stained with HE and
    immunohistochemistry (MMP-9, TNF-alpha)
  • (hamster anti mouse TNF-alpha Santa Cruz,
    sc-12744, mouse anti MMP-9, Chemicon, MAB3309)
  • IL-6, TNF-alpha were evaluated by RT-PCR and were
    compared between those two groups.
  • (Rabbit TNF-a Forward PRIMER
    atggtcaccctcagatcagc Reverse PRIMER
    ttgaccgctgaagagaacct,
  • Rabbit IL-6 Forward PRIMER
    tcctggagaccatcaaggagReverse PRIMER
    gggtggcttcttcattcaaa )

5
Results
  • HE stain showed more irregular arrangement of
    epithelial basal cells in Group A than in Group B

Normal
Group A
Group B
6
  • RT-PCR
  • - The expression level of mRNA of IL-6,
    MMP-9, TNF-alpha showed no significant difference
    between those two groups

Mean SD
7
Immunohistochemistry
  • MMP-9

Negative Control, x 200
Positive Control, x 200
8
  • MMP-9 (x 200)

Group A
Group B
9
Immunohistochemistry
  • TNF-alpha

Negative Control, x 200
Positive Control, x 200
10
  • TNF-alpha
  • (x400)

Group A
Group B
11
  • MMP-9 and TNF-alpha were expressed and were more
    prominent than negative control tissue in all
    groups
  • There was no significant difference between two
    groups

12
Conclusion
  • The expression of inflammatory cytokines in
    epimicrokeratome-assisted flap might be no
    different from alcohol-assisted flap, suggesting
    that the inflammatory response of the cornea
    might be similar between Epi-LASIK and LASEK.
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