1 / 28

CORNEAL PRESERVATION & STORAGE

CORNEAL PRESERVATION & STORAGE. DR.REKHA. Lions Eye hospital BANGALORE. IMPORTANCE OF STORAGE. SUCCESS OF PENENTRATING KERATOPLASTY DEPEND UPON CORNEAL ENDOTHELIAL INTEGRITY. CORNEAL STORAGE SYSTEM. AIM

curtislanier
Télécharger la présentation

CORNEAL PRESERVATION & STORAGE

An Image/Link below is provided (as is) to download presentation Download Policy: Content on the Website is provided to you AS IS for your information and personal use and may not be sold / licensed / shared on other websites without getting consent from its author. Content is provided to you AS IS for your information and personal use only. Download presentation by click this link. While downloading, if for some reason you are not able to download a presentation, the publisher may have deleted the file from their server. During download, if you can't get a presentation, the file might be deleted by the publisher.

E N D

Presentation Transcript

  1. CORNEAL PRESERVATION & STORAGE DR.REKHA. Lions Eye hospital BANGALORE

  2. IMPORTANCE OF STORAGE SUCCESS OF PENENTRATING KERATOPLASTY DEPEND UPON CORNEAL ENDOTHELIAL INTEGRITY.

  3. CORNEAL STORAGE SYSTEM AIM • To maintain the endothelial viability & integrity. • Prevent corneal autolysis during storage • Long term corneal storage.

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

  5. 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)

  6. 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.

  7. McCAREY&KUFMAN STORAGE Corneo-scleral excision is necessary& immersed with endothelial side up. • Contains Tc199,dextran-40 1%, PH7.0-7.5,Osmolality 295=355 & gentamycin sulphate75-150 micro gm/ml ,HEPES as buffer. Phenol red as indicator. • Scheduling of surgery

  8. STORAGE MEDIA K-SOL:Tc 199,MEM&Earles media, HEPES,Gentamicin,Chondroitin sulphate2.5% . 1988 proprionibacteria contamination DEXOL:MEM, 1.35%Chondrotin Sulphate, 1mM Sodium pyruvate, 1mM non- essential aminoacids, Antioxidants, 1%dextran40. .

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

  10. 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.

  11. FUNCTIONS OF CONTENTS BASE MEDIA a)Tissue-culture199-aminoacids,salts,buffer&energy to support life. b)Minimum Essential Media(MEM) with Earles salt- matches natural salts &buffers of human tissue,electrolites similar to aqueous

  12. FUNCTIONS OF CONTENTS BUFFER HEPES(N-2-hydroxyethyl-piperazine-N-2ethane-sulfonic acid) -used for cultivation of sensitive mammalian cells -provides optimal physiological Ph in the range7-7.2

  13. 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.

  14. COMMON FEATURES OF MEDIA CHONDROTIN SULFATE Polysaccharide,negatively charged Endothelial integrity&acts as a osmotic agent.Superior to dextran.Longer storage timeENRICHED Various salts,aminoacids essential &non- essential,vit.,phosphates &antioxidents to enhance cell & tissue viability & health during storage.

  15. COMMON MEDIA FEATURES ATP precursors--> Provide energy for pumping function. Anti-Oxidents--->Neutralize metabolic waste,maintain DNS synthesis Vitamins-->Provides additional nutritional cell supplements. Sodium Piruvate&Glucose--> Energy supply Colour Phenol Red-->Visual aid for pH indication.

  16. CHECK MEDIA The cornea which is stored in MKmedia,Dexol,optisolGS.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

  17. Ph COLOUR REFERENCE CHART • 7.00 7.25 7.50 contimanation

  18. PROCELL MEM,1.35%chondrotin 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)

  19. 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%

  20. ORGAN CULTURE contents Eagles media,Earles salt with out L-glutamine,L-glutamine1%final conc.,Decomlemented calf serum10%final conc.Penicilllin 100units/ml,, Gentamicin100microgram/ml. AmphotericianB 0.25% microgram/ml.

  21. TISSUE CULTURE MEDIA EUROSOL New Duration of storage 28days. Temp 31deg.C. Better then the European culture media. Maintains intact endothelium, epithelium & Keratocyte.

  22. ORGAN CULTURE ADVANTAGES:Endothelial integrity 120days,Epithelial integrity35days,Culture report prior to transplantation,stored at 4deg,centi. DISADVANTAGES:Cornea thickens with time,Microbial contamination, Complicated expensive techniques, Greater loss of endothelial afterPK.

  23. 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.

  24. CRYOPRESERVATION ADVANTAGES:Long-term storage at- 80deg.C. DISADVANTAGES:Technically 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

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

  26. NOTE • HYPOTHERMIC (CS+Hyperkalaemic) • 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

  27. 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.

  28. Thank you

More Related