Table 1.
Demographics of the subjects. Chronic ocular surface complications score (COCS; range: 0–15) was adapted from previous report [14].
Fig 1.
Comparison of the stem cell properties of Stevens—Johnson syndrome (SJS) and non-SJS cultured oral mucosal epithelial cells.
(A) A representative independent colony-forming efficiency (CFE) experiment. (B) The CFE of SJS cells is comparable to the CFE of non-SJS cells (p>0.05, t-test). (C) High expression of proliferative and stem cell markers (Ki-67 and p63) in the non-SJS and SJS cells (X100). SJS indicates mucosal epithelial cells from SJS subjects; non-SJS indicates mucosal epithelial cells from non-SJS subjects.
Fig 2.
Proliferative and migratory capabilities of cultured oral mucosal epithelial cells from Stevens—Johnson syndrome (SJS) and non-SJS subjects.
(A) Cells were counted on days 2, 3, and 5 and did not differ between SJS and non-SJS cells (p>0.05, two-way ANOVA). (B) Migratory potential was significantly delayed in SJS cells when compared to non-SJS cells (*p<0.05, ***p<0.001, RM two-way ANOVA). (C) Image showing cell migration from the edge (black line) (X100). The black line indicates the area that cells migrated each day. SJS indicates mucosal epithelial cells from SJS subjects; non-SJS indicates mucosal epithelial cells from non-SJS subjects.
Fig 3.
Expression of cytokeratins in oral mucosal epithelial cells and limbal epithelial (LE) cells.
Cytokeratin K1 was not expressed in Stevens—Johnson syndrome (SJS) or non-SJS mucosal epithelial cells or limbal epithelial cells (LEs). K3, K4, K13 and K19 were expressed in SJS and non-SJS mucosal epithelial cells as well as LEs (X100). SJS indicates mucosal epithelial cells from SJS subjects; non-SJS indicates mucosal epithelial cells from non-SJS subjects.
Fig 4.
Cytokine profiles of the oral mucosal epithelial cells from Stevens—Johnson syndrome (SJS) and non-SJS subjects.
ELISA assay showed that the level of VEGF was higher, and the level of EGF was lower in the cells from SJS oral mucosa (SJS) than those in the cells from normal oral mucosa (non-SJS). When compared to the expression of cytokine/growth factors in limbal epithelial cells (LEs), the level of VEGF and TFG-α were higher, while the level of MMP2 was lower in mucosal epithelial cells regardless of SJS status. The levels of FGF, EGF, and IL-1α were higher in non-SJS mucosal cells than in LEs. All data represent the mean ± standard deviation. (*p<0.05, **p<0.01, ***p<0.001, ****p<0.0001, One-way ANOVA)
Fig 5.
In vivo re-epithelialization by Stevens—Johnson syndrome (SJS) and non-SJS cultured oral mucosal epithelial cell sheets (COMECs) in a limbal stem cell-deficient model.
(A) Digital measurement of the epithelial defect area (black arrow) and calculation of re-epithelialized corneal area. (B) The re-epithelialized area was smaller in SJS-COMECs than in non-SJS-COMECs at postoperative day (POD) 3 (**p<0.01, RM two-way ANOVA) but was not statistically different between SJS and non-SJS-COMECs on POD 7. SJS indicates cells from SJS subjects; non-SJS indicates cells from non-SJS subjects.
Fig 6.
Epithelial characteristics of the ocular surface transplanted with Stevens—Johnson syndrome (SJS) and non-SJS COMECs on day 7 post-transplantation.
Corneas with COMECs from both SJS and non-SJS subjects expressed K3, K4 and K13 (X200). K1 and K8 were not expressed in both groups. K19 was expressed in cornea transplanted with non-SJS COMECs, and it was barely expressed in cornea transplanted with SJS COMECs. The control indicates a normal center cornea.
Fig 7.
Characteristics of the progenitor cell expression with Stevens—Johnson syndrome (SJS) and non-SJS COMECs on day 7 post-transplantation.
(A) Cornea with COMECs from both SJS and non-SJS subjects highly expressed p63, ABCG2, and Ki-67 (X400). (B) Percentage of p63- or Ki-67-positive cells in the corneas transplanted with SJS-COMECs was not different from that in the corneas transplanted with non-SJS-COMECs (p>0.05, Mann-Whitney U test). The percentage of p63 was lower in both COMECs-transplanted corneas from SJS and from non-SJS subjects than that in normal human limbal tissues as control (p<0.05, Mann-Whitney U test). The control indicates normal limbal tissues of human.