The EBAI CME 2013 - 21st and 22nd of September 2013, Golden Valley Resort, Ghodbunder Road, Thane West, Mumbai.

1. Dr Abhijit Wadekar
M.S., D.N.B, (Ophthal)
Cornea Fellow L.V.P.E.I, Hyderabad
Leela Eye Institute
Kalyan

2.  Surrounds the developing fetus
 Derived from fetal tissue
 Amnion is translucent structure adjacent to amniotic
fluid and provides nutrients
 Chorion is opaque membrane attached to the decidua

3. Histology
 Single layer of cuboidal epithelial cells
 A thick Basement Membrane
 Avascular stromal matrix , loosely attached to the
chorion

4. Introduction
 Innermost layer forming the fetal membrane.
 First used by Davis in 1910 in skin transplant.
 First documented ophthalmic application for
conjunctival surface reconstruction in 1940 by De
Roth.
 Sorsby et al in 1946 reported its use in acute ocular
burns.
 Kim and Tseng successfully reintroduced usage in 1995

5. Properties of Amniotic membrane
Facilitates migration of epithelial cells
Reinforcement of basal cell adhesion
Induction of epithelial cell differentiation and promotes goblet cell
differentiation
Anti-inflammatory action reduces scarring and vascularization
bacteriostatic properties
Lack of Immunogenicity

6.  Cytokines, growth factors and protease inhibitors
(IL4,6) EPG, FGF, TGF, HGF and 2- microglobulin.
 Exclusion of inflammatory cells with anti-protease
activities
 Suppression of TGF- signalling system and
myofibroblast differentiation of normal fibroblasts.
 Prolongation of life span and clonogenicity of
epithelial progenitor cells.
 Promotion of non-goblet cells epithelial
differentiation
 Promotion of goblet cells differentiation when
combined with epithelial fibroblast

7. Collection
 Consenting Sero negative (Hepatitis B and C virus,
Syphilis and HIV) maternal donors determined pre
surgery
 elective caesarian section
 Under sterile conditions
 An antibiotic cocktail covering most of Gram –positive
and Negative bacteria and fungi in Balanced Salt
Solution for 24 hours
 Second wash of BSS
 Amnion separated from chorion

8. Harvesting Amniotic membrane

9. Preservation
 BSS containing Ampicilin, Streptomycin and
Amphotericin B used.
 Scrapped manually to remove debris
 Amnion is clear and Transparent
 Nitrocellulose membranes of required size
 Epithelial/basement layer up.

10.  Preserved in solution containing Modified
Eagle’s Medium (MEM) and glycerol in 1:1 ratio
with 3.3% l-glutamine, 25 ug/ml Gentamicin, 50
units/ml penicillin, 100ug/ml ciprofloxacin and
0.5 mg/ml Amphotericin B
 Stored in 50 ML wide mouthed screw capped
irradiated transparent bottles at -80 Degrees

11. Indications
Used either as a ‘substrate’ to replace
damaged ocular tissue or
as a ‘Patch’ (Biological dressing) or
 Combination of both

12. Corneal diseases
Symptomatic Bullous keratopathy

13. Chemicalinjury
Limbal stem cell
deficiency (Partial
or total): combined
with stem cell graft

14. Persistent epithelial defects
Corneal ulceration

15. Conjunctival diseases
Stevens-Johnson Syndrome
Symblepharon lysis
Conjunctival surface reconstruction
Pterygium surgery
Post Trabeculectomy:bleb leakage or
revision

16. Conjunctival cicatrisation/scar

17. Limbal stem cell transplant on
Amniotic membrane

18. Surgical techniques
Thawed at room temperature
Rinsed three times in BSS
Gently separated from the
nitrocellulose paper
Epithelium-basement membrane side
up

19. Stromal side up, away from the eye.
The stromal side of the membrane is
sticky
Trimmed to the appropriate shape
10-0 nylon interrupted sutures
Intercepted 10-0 nylon sutures

20. A large therapeutic contact lens
Occasionally, a tarsorrhaphy
Sutures and contact lens are often
removed after 2 to 4 weeks
Post-operative topical treatment

22. Tissue-cultured limbal stem cells and
amniotic membrane
Disintegrates within 2 to 4 weeks
Storage medium should be light pink

23. Complications
Suture granuloma
Persist ant inflammation
Hematoma dehiscence
Shrinkage of the graft
Failure to achieve the intended effect
Infection
Dislodged or loose AM
Hemorrhage
Early disintegration

24. Thank You