CORNEAL PRESERVATION

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CORNEAL PRESERVATION STORAGE

• DR.REKHA.
• Lions Eye hospital
• BANGALORE

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IMPORTANCE OF STORAGE

• SUCCESS OF PENENTRATING
• KERATOPLASTY
• DEPEND UPON CORNEAL
• ENDOTHELIAL INTEGRITY.

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CORNEAL STORAGE SYSTEM

• AIM
• To maintain the endothelial viability
• integrity.
• Prevent corneal autolysis during storage
• Long term corneal storage.

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HISTORY

• First corneal preservation.
• 1911 Mgitot in J.A.M.A.
• Stored cornea in Hemolyzed blood serum at
  5-7deg.centigrade.
• Cornea viable for2days

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STORAGE TYPES

• SHORT-4deg.C(days)
• Moist chamber(1day),M.K.Media(2-4days)
• INTERMEDIATE-14days- 4deg.C(wks)
• K-Sol, Dexsol,OptisolGS,Procell, Eusol-C
• LONG(months)
• EuropeanOrgan culture-120days(37deg.C)
• Eurosol-31deg.C-28days(New)
• Cryoprev.-Unlimited-(-80degC)

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MOIST CHAMBER METHOD

• Filatov Castroviejo.
• Whole globe is stored in a sterile jar.
• Jar has saline Humidification.
• Temp of 4degrees C.
• Popular untill 1970.
• Simple in- expensive.
• Grafting on emergency basis.

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McCAREYKUFMAN
STORAGE

• Corneo-scleral excision is necessary
  
• immersed with endothelial side up.
• Contains
• Tc199,dextran-40 1, PH7.0-7.5,Osmolality
  
• 295355 gentamycin sulphate75-150
• micro gm/ml ,HEPES as buffer.
• Phenol red as indicator.
• Scheduling of surgery

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STORAGE MEDIA

• K-SOL Tc 199,MEMEarles media,
• HEPES,Gentamicin,Chondroitin
• sulphate2.5 .
• 1988 proprionibacteria contamination
• DEXOL MEM, 1.35Chondrotin
• Sulphate, 1mM Sodium pyruvate,
• 1mM non- essential aminoacids,
  
• Antioxidants, 1dextran40.
• .

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OPTISOL.G.S.

• Introduced in 1991
• Contents
• MEM, 1.35Chondrotin
• Sulphate, 1mMSodiumpyruvate,1mM
• non-essential aminoacids, Antioxidants,
  
• 1dextran40,ATP,Iron,cholestrol,L-
• hydroxyproline,Vitamine,2antibiotics-
• Genticyn,Stepomycin. Expensive

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EUSOL-C

• Store at 4deg.C
• Storage time 14days, SIMILAR TO DEXOL
• CONTENTS
• Dextran,Sodium Piruvate,Glucose,Essential
  non-essential aminoacids, mineral salts,
  Vitamins, Gentamin, hepesbuffer, Bicarbonate,
  Phenol Red.

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FUNCTIONS OF CONTENTS

• BASE MEDIA
• a)Tissue-culture199-aminoacids,salts,bufferenergy
  to support life.
• b)Minimum Essential Media(MEM) with Earles salt-
  matches natural salts buffers of human
  tissue,electrolites similar to aqueous

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FUNCTIONS OF CONTENTS

• BUFFER
• HEPES(N-2-hydroxyethyl-piperazine-N-2ethane-sulfo
  nic acid)
• -used for cultivation of sensitive mammalian
  cells
• -provides optimal physiological Ph in the
  range7-7.2

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COMMON FEATURES
OF MEDIA

• ANTIBIOTIC
• Gentamicin only /Streptomycin
• DEXTRAN
• Polysaccharide,negatively charged.
• Prevents tissue swelling.
• Used alone does not provide sufficient tissue
  stability viability for extended storage.

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COMMON FEATURES
OF MEDIA

• CHONDROTIN SULFATE
• Polysaccharide,negatively charged
• Endothelial integrityacts as a osmotic
• agent.Superior to dextran.Longer storage
  
• time ENRICHED
• Various salts,aminoacids essential non-
• essential,vit.,phosphates antioxidents to
  enhance cell tissue viability health during
  storage.

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COMMON MEDIA FEATURES

• ATP precursors--gt Provide energy for pumping
  function.
• Anti-Oxidents---gtNeutralize metabolic
  waste,maintain DNS synthesis
• Vitamins--gtProvides additional nutritional cell
  supplements.
• Sodium PiruvateGlucose--gt Energy supply
• Colour Phenol Red--gtVisual aid for pH
• indication.

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CHECK MEDIA

• The cornea which is stored in MKmedia,Dexol,optis
  olGS.Thefollowing parameters to be checked before
  using
• Intact seal,Expiry date,Turbity, Colour (rose
  red) ,precipitates FB.
• COLOUR CHANGE INDICATES
• Yellow-- Bacterial contamination.
• Red--- Unacceptable Ph.
• Cloudy-- Contamination

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Ph COLOUR REFERENCE
CHART

• 7.00
• 7.25
• 7.50
• contimanation

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PROCELL

• MEM,1.35chondrotin sulphate,1mM sodium
  pyruvate1mM,Non-essential aminoacids,Antioxidants,
  Dextran40,
• Humen insulin10mic.g/ml Human epidermal
  growth factor(hEGF10ng/ml) to improve long term
  endothelial survival after PKP.(experiments have
  not proved it, opth1994 feb101(2)252-9 LassJH et
  all)

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ORGAN CULTURE

• 1936,by Archer Trevor-Roper.
• Being used since1974.
• Refrigeration not required.
• Complicated,expensive well trained
  
• microbiologist.
• EUROPEAN ORGAN CULTURE
• 31-37deg.C, EMEM, MEM, IMDM ,
• HEPES, NaHCO3,Peni,Strep,Ampo-B,
• BOVINE SERUM 2-10ML,Dextran4-10

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ORGAN CULTURE

• contents
• Eagles media,Earles salt with out
  L-glutamine,L-glutamine1final conc.,Decomlemented
  calf serum10final conc.Penicilllin
  100units/ml,, Gentamicin100microgram/ml.
  AmphotericianB 0.25 microgram/ml.

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TISSUE CULTURE MEDIA
EUROSOL

• New
• Duration of storage 28days.
• Temp 31deg.C.
• Better then the European culture media.
• Maintains intact endothelium, epithelium
• Keratocyte.

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ORGAN CULTURE

• ADVANTAGESEndothelial integrity
  120days,Epithelial integrity35days,Culture report
  prior to transplantation,stored at 4deg,centi.
• DISADVANTAGESCornea thickens with time,Microbial
  contamination, Complicated expensive techniques,
  Greater loss of endothelial afterPK.

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CRYOPRESERVATION

• Kaufman Capella.1954
• Cornea is stored in Liquid Nitrogen with
  DimethylSulfoxide(DMSO).
• DMSO,Prevents intracellular damage by ice
  crystals.
• Not a procedure used.Holds a lot of research
  interest.

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CRYOPRESERVATION

• ADVANTAGESLong-term storage at-
• 80deg.C.
• DISADVANTAGESTechnically
• difficult,Limited tissue tolerance to freez-
• thaw. Rapid thaw at 37 deg.C.
• 1964
• 18months
• Tissue strored in Liquid nitrogen at --197deg C
• At the time of thawing the cial containing cornea
  is placed in water bath at temp of 60dec C at
  1min.
• Trained techiques for Freezing thawing

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GLYCERINE PRESERVATION

• Patch graft or Lamellar Keraoplasty.
• Corneo-scleral button.
• 100glycerine.
• Enothelium is nonviable
• Use with in 1 year.
• Stored at room temp.

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NOTE

• HYPOTHERMIC (CSHyperkalaemic)
• Storage 2wks,epithelial not well preserved,not
  used
• ORGAN CULTURE
• At one time predominating in Europe
• Organ culture storage-4wks
• CRYOPRESERVATION
• Not used, tissure strored for emergency use

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FUTURE

• Improve endothelial cell layer viability
• compact stroma.
• Maintain Keratocyte viability.
• Reduce corneal autolysis during storage.
• Thin cornea in spite of long time storage.
• No contamenation
• Easy to use transport
• Inexpensive.

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Thank you