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Abstract
Purpose
To investigate the characteristics and the results of long-term follow-up of limbal epithelial cells cultivated in vivo on amniotic membranes (LIVAM) in corneal limbal deficiency.
Methods
Twenty-two eyes of twenty-two patients diagnosed with corneal limbal deficiency underwent transplantation of in vivo cultivated corneal limbal epithelial cells on the amniotic membrane. Biopsy and immunohistochemical staining (AE5, MUC5AC) of the amniotic membrane cultivated for one week were performed to verify that the cultivated epithelial cells on the amniotic membrane were corneal epithelial cells. Impression cytology was performed to evaluate the characteristics of the transplanted corneal limbal epithelial cells at postoperative 1 week, 3 months, 6 months, and 1 year.
Results
Successful epithelial growth was observed on the amniotic membrane at one week. The epithelial cells were confirmed to be corneal epithelial cells by immunohistochemical staining. Transplanted in vivo cultivated corneal epithelial cells were confirmed to have corneal specificity by impression cytology and immunohistochemical staining at postoperative 1 week, 3 months, 6 months, and 1 year.
Conclusions
In vivo cultured corneal epithelial cells showed morphological and immunohistochemical findings similar to those of normal corneal epithelial cells. Transplanted in vivo cultivated corneal epithelial cells were maintained and showed the characteristics of corneal epithelial cells. Transplantation of in vivo cultivated corneal limbal epithelial cells can be performed to reconstruct the corneal limbus in treating corneal limbal deficiency.
References
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Figure 1.
Schematic drawing of the orientation of each cellulose acetate paper applied on the ocular surface.
Figure 2.
(A) In vivo cultivated corneal epithelial cells on human amniotic membrane for a week. (Hematoxylin-Eosin stain, Original magnification ×100), (B) Periodic Acid-Schiff stain (original magnification ×100), (C) Anti-AE5 immunohistochemical stain (original magnification ×100). Light microscopy demonstrates well stratified and cuboidal corneal epithelial cells firmly attached to basement membrane of human amniotic membrane. There is no goblet cell in the PAS staining. In immunohistochemical staining, cultivated epithelial cells show positive staining for AE5.
Figure 3.
Impression cytology from the cornea, transplanted amniotic membrane and conjunctiva at postoperative a week (original magnification ×100). PAS stain: abundant periodic acid-schiff negative epithelial cells on corneal and amniotic membrane side, but periodic acid-Schiff positive on the conjunctival side (arrow); Anti-AE5 immunohistochemical stain: positive staining on the corneal and amniotic membrane side; Anti-MUC5AC immunohistochemical stain: positive staining only on the conjunctival side.
Figure 4.
Impression cytology from the cornea, transplanted amniotic membrane and conjunctiva at postoperative 3 months (original magnification ×100). PAS stain: abundant periodic acid-Schiff negative epithelial cells on the corneal and amniotic membrane side, but periodic acid-Schiff positive on the conjunctival side (arrow); Anti-AE5 immunohistochemical stain: positive staining on the corneal and amniotic membrane side; Anti-MUC5AC immunohistochemical stain: positive staining only on the conjunctival side.
Figure 5.
Impression cytology from the the cornea, transplanted amniotic membrane and conjunctiva at postoperative 6 months (original magnification ×100). PAS stain: abundant periodic acid-Schiff negative epithelial cells on the corneal and amniotic membrane side, but periodic acid-Schiff positive on the conjunctival side (arrow); Anti-AE5 immunohistochemical stain: positive staining on the corneal and amniotic membrane side; Anti-MUC5AC immunohistochemical stain: positive staining only on the conjunctival side.
Figure 6.
Impression cytology from the cornea, transplanted amniotic membrane and conjunctiva of patient with scleral recurrence at postoperative 6 months (original magnification ×100). Anti-AE5 immunohistochemical stain: positive staining on the corneal and amniotic membrane side; Anti-MUC5AC immunohistochemical stain: positive staining on the transplanted amniotic membrane and conjunctival side.
Figure 7.
Impression cytology from the cornea, transplanted amniotic membrane and conjunctiva at postoperative 1 year (original magnification ×100). PAS stain: abundant periodic acid-Schiff negative epithelial cells on the corneal and amniotic membrane side, but periodic acid-Schiff positive on the conjunctival side (arrow); Anti-AE5 immunohistochemical stain: positive staining on the corneal and amniotic membrane side; Anti-MUC5AC immunohistochemical stain: positive staining only on the conjunctival side.
Table 1.
The results of immunohistochemistry of transplanted epithelial cells cultivated in vivo on the amniotic membrane
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