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FGF-2 DAPI DMEM IL-1 FGF-2 Actin DMEM IL-1 25 KDa 20 KDa

Published byErnest Dalton Modified over 6 years ago

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Presentation on theme: "FGF-2 DAPI DMEM IL-1 FGF-2 Actin DMEM IL-1 25 KDa 20 KDa"— Presentation transcript:

1 FGF-2 DAPI DMEM IL-1 FGF-2 Actin DMEM IL-1 25 KDa 20 KDa
Ex vivo incubation of whole cornea with IL-1b (24h) FGF-2 DAPI DMEM IL-1 FGF-2 Actin DMEM IL-1 25 KDa 20 KDa

2 Actin Normal Freezing DAPI 20m
Morphologic change after freezing injury Actin Normal Freezing DAPI 20m

3 A proposed signal pathway in vitro
LY294002 SB203580

4 IL-1b concentration in aqueous humor after freezing FGF-2 expression in endothelium

5 LY294002 SB203580

6 Evaluation of Corneal Endothelial Damage after Argon Laser Iridotomy in Pigmented Rabbit Eyes
Junho You MD, Chulmin Yun MD, Jongyun Lee MD, Yongyun Kim MD, PhD, Hyomyung Kim, MD, PhD, Jongsuk Song MD, PhD Department of Ophthalmology, Korea University College of Medicine, Seoul, Korea

7 Introduction Corneal endothelial cell loss Argon laser iridotomy(ALI)
Lens extraction and IOL implantation Argon laser iridotomy Systemic amantadine administration Argon laser iridotomy(ALI) Corneal complications (during or immediately following procedure) corneal opacities and edema epithelial and endothelial burns Bullous keratopathy after ALI 2nd most common cause of corneal transplantation in Japan Jun Shimazaki et al. Cornea. 2007

8 2 hours after LI (OS)

9 Introduction The studies of corneal endothelial cell apoptosis
after argon laser iridotomy Not many so far

10 Purpose To evaluate the corneal endothelial damage after argon laser iridotomy through several stain methods of corneal endothelium and suggest an animal-model for the study of corneal endothelial cell apoptosis.

11 Methods 12 Pigmented rabbits
OD : argon laser iridotomy OS : normal eye (control) One argon laser iridotomy was performed on each quadrant of iris Power : 500 mW Duration : 200 ms Interval : 300 ms Counts : 400 times

12 Methods Measuring the corneal thickness Pre-laser iridotomy(OU)
Post-laser iridotomy(OD) (immediately, 1day, 2days, 3days, 4days) by using ultrasound corneal pachymetry The BVI Pocket Pachymeter (BV International, Clermont-Ferrand, France)

13 Methods Identification of Corneal endothelial cell damage H & E stain
Alizarin red/Trypan blue stain TUNEL stain Live/Dead cell assay Endothelial cell count (in 0.04mm2 sized square)

14 Results The change of Corneal thickness

15 Results : H&E stain Post-LI control Post-LI #1day Post-LI #4day

16 Results : TUNEL stain Post-LI control

17 Results : Alizarin red/trypan blue stain
Post-LI #1day Post-LI #4day X 10 X 20 X 20 Post-LI X 40 X 20 control X 40

18 Results Corneal endothelial cell count (#4day) P-value : 0.02 Rabbit
Cell count( /mm2) Normal eye Post-LI 1 5675 4050 2 5300 4725 3 5325 4350 4 5250 4375 mean 5387.5 P-value : 0.02

19

20 Results : Live/Dead cell assay
Post-LI #1day X20 X40 X20 X40 Post-LI Post-LI #4day normal X10 X20 X10 X20

21

22

23 IL-1b IHC Laser Tx Post-LI #1day Control

24 Conclusion H & E stain and Alizarin red/Trypan blue stain are less sensitive methods to evaluate corneal endothelial damage after argon laser iridotomy than TUNEL stain or live/dead cell assay. Evaluation of corneal endothelial damage after argon laser iridotomy in pigmented rabbit eyes is thought to be a good animal-model for the study of corneal endothelial cell apoptosis.

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