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Page 1: Endothelial viability of precut pre-Descemetic grafts ... · Endothelial viability of precut pre-Descemetic grafts prepared using an innovative microkeratome technology Rudy M.M.A.

Endothelial viability of precut pre-Descemetic grafts prepared using an innovative microkeratome technology

Rudy M.M.A. Nuijts, Friso W.F. van Marion, Jurriaan Brekelmans, Petra Steijger-Vermaat, Teja Wesseling, Mor M. Dickman

EuCornea, Amsterdam 5 October 2013

Page 2: Endothelial viability of precut pre-Descemetic grafts ... · Endothelial viability of precut pre-Descemetic grafts prepared using an innovative microkeratome technology Rudy M.M.A.

Financial disclosure

• R.M.M.A. Nuijts Acufocus (S), Alcon (C,L,S), Asico (C,P),

Bausch & Lomb(C), Gebauer (S),

HumanOptic (S), Ophtec (S), TheaPharma (C)

• J. Brekelmans None

• P. Steijger-Vermaat None

• T. Wesseling None

• F. W.F. van Marion None

• M.M. Dickman None

This study was financially supported by:

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Eye Bank Association of America. Eye Banking Statistical Report, 2012.

DMEK outcomes and embracement

Time point Pre-op 1 month 3 months 6 months 12 months 24 months

DSAEK n=401 - n=137 n=395 n=188 n=23

DMEK n=398 n=189 n=189 n=307 n=156 n=11

UT-DSAEK n=188 n=188 n=179 n=160 n=119 n=41

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Why not DMEK?

• Donor preparation

• Donor positioning and adherence

– Total detachment 0% - 6.7%

– Partial detachment 4.7 - 6.7%

– Re-bubbling

• Early post-operative 1% - 52%

• Total (up to 6 months) 1% - 82%

– Primary graft failure 0% - 8.1%

• Re-operations

– Secondary DSAEK 0% - 6.7%

– Secondary DMEK 0% - 4.7%

– Secondary PKP 0% - 7.1%

Author N Study type

Van Dijk, K., et al. 2013

300 Prospective

Dapena, I., et al. 2013

178 Retrospective

Parker, J., et al. 2013

50 Prospective

Yoeruek, E.,et al. 2012

70 Retrospective

Tourtas, T.,et al. 2012

38 Retrospective

Heindl, L.M.,et al. 2011

50Prospectivecase series

Guerra, F.P.,et al. 2011

136Prospective

interventionalseries

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DMEK donor preparation

- SCUBA

- Pneumatic dissection

- DMEK-S

- Partial trephination

- Pre-peeled DMEK

A B

C

D

A. Krabcova, I., 2011B. Price Vision Group, 2012C. McKee, H.D., 2012D. Muraine, M., 2013

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Standardized automatic donor preparation?!

• Thickness predictability

• Graft thickness profile

• Endothelial cell viability

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Gebauer SLc Expert Microkeratome

Adjustable cavity Porous rigid reference member (RMM)

Vacuum based corneal applanation

Dissection of any desired thickness(30-950 micrometer)

Patent US 5368604

Patent US 5521657A

Corneal profiling Set of RRM curvatures

Correction for peripheral to centralcorneal thickness differences

Patent US 5521657A

Technical specifications 10.000 rpm

1.0 mm/sec

9.5 mm trephination diameter

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PDAEK (Pre-Descemetic Automated Endothelial Keratoplasty)

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Graft thickness predictability

Post-cut – without profile correction

Pre-cut

n=21Expected SD±6.0µmPost-cut SD±7.7µm

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Corneal profiling

Corrected (with profile) Uncorrected (without profile)

Pre-cut

Post-cut

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DAPI/Hoechst CY3/Ethidium

FITC/Calcein Merge

HEC – Endothelial Viability

Hoechst- All cells- DNA- Blue

Ethidium- Dead cells- DNA- Red

Calcein- Living cells- Cytoplasm- Green

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Total endothelial viability (calcein) staining

75%

71% 67%

71%

65%

3

1

4 5

2

CCT 80 µm

Viable ECD 1769CCT 80 µm

Viable ECD 1527

CCT 30 µm

Viable ECD 1407

CCT 50 µm

Viable ECD 1611CCT 50 µm

Viable ECD 1477

75%

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Trypan blue staining

1 2 3 4 5

% Viability 75 71 71 67 65

H-ECD 2375 2154 1988 2419 2290

vECD 1769 1527 1407 1611 1477

1 2 3 4 5

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Histology

Bowman graft (~ 25 µm thick)

Pre-Descemetic graft (~ 25 µm thick)

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Take home messages

• We established a POC for standardized harvesting of PDAEK lamellae:

– Highly accurate thickness predictability (± 7.7µm)

– Corneal profiling allows harvesting planar grafts, potentially minimizing postoperative hyperopic shift

– Satisfactory central endothelial density and morphology

– Clinical outcomes remain to be investigated

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Thank you for your kind attention!

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