J Korean Med Sci.  2008 Jun;23(3):502-508. 10.3346/jkms.2008.23.3.502.

The Effect of In Vivo Grown Corneal Epithelium Transplantation on Persistent Epithelial Defects with Limbal Stem Cell Deficiency

• Kim JT
• Chun YS
• Song KY
• Kim JC

• KMID: 1786891
• DOI: http://doi.org/10.3346/jkms.2008.23.3.502

Abstract

We report our experience with corneal epithelium, grown in vivo, transplantation in three patients with persistent epithelial defect (PED). The three patients had ocular surface disease unresponsive to standard treatments and were therefore chosen for transplantation. They underwent transplantation of epithelial sheets, grown in vivo, to the most affected eye. In vivo cultivation was carried out in the cornea of a living related donor. After epithelialization was completed, the epithelium grown on an amniotic membrane was harvested gently; it was then transplanted into the patient's eye after debridement of fibrovascular tissue. The cultivated epithelium was completely epithelialized by 2 weeks; it was well-differentiated with well-formed hemidesmosome. On immunohistochemical staining, p63, connexin 43, and Integrin beta4 were expressed in the cells on the epithelial sheet. The PED was covered completely and maintained for 4 weeks in all cases. However, corneal erosion recurred after 5 weeks in two cases. This novel technique demonstrates the corneal epithelial cells can be expanded in vivo successfully on denuded amniotic membrane of a healthy cornea and harvested safely. A corneal epithelial sheet, grown in vivo, can be transplanted to treat eye with a severe ocular surface disease, such as total limbal deficiency.

Keyword

Amnion; Epithelium, Corneal; Limbus Corneae; Tissue Expansion

MeSH Terms

Adult
Cell Culture Techniques
Cells, Cultured
Corneal Diseases/etiology/pathology/*surgery
Corneal Transplantation/*methods
Epithelial Cells/cytology/*transplantation
Epithelium, Corneal/cytology/*transplantation
Eye Burns/complications
Humans
Limbus Corneae/*pathology
Male
Middle Aged
Stem Cells/*pathology
Stevens-Johnson Syndrome/complications

Figure

• 

  Fig. 1 Slit lamp findings of donor cornea and structural findings of in vivo expanded corneal epithelium on donor cornea at 2 weeks of cultivation (case 1). (A) Denuded AM was transplanted onto the cornea using a continuous suture with #10-0 Nylon after epithelial peeling. Epithelialization on the AM was complete by 2 weeks after the AM transplantation. (B) HE staining demonstrated well differentiated multilayer epithelium in the in vivo expanded epithelial sheet under the light microscopy (×200). (C) EM showed a well developed hemidesmosome (arrows).

• 

  Fig. 2 Immunohistochemical staining of p63 (A), Connexin 43 (B) and Integrin β4 (C) from the corneal epithelial culture of the donor's cornea at 2 weeks of cultivation (case 1) (×400). (A) p63 was expressed by the epithelial basal cells. (B) Connexin 43 was expressed in the intercellular space. (C) Integrin β4 was expressed in the intercellular space, predominantly the basal cell layer.

• 

  Fig. 3 A 44-yr-old patient with Steven Johnson syndrome (case 1) had PED (8 mm diameter), present for more than 5 months (A, C). However, the epithelial defect was completely covered with the in vivo expanded epithelial sheet 2 weeks after surgery (B, D).

• 

  Fig. 4 A 37-yr-old patient with alkali burns (case 3) had PED, presented for more than 5 months (A, C). The epithelial defect was completely covered, but erosion in another area developed 2 weeks after surgery (B, D).

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