Phenotypic Evaluation of Severely Damaged Ocular Surface after Reconstruction by Cultured Limbal Epithelial Cell Transplantation

Abstract

Objective: To report the clinical outcome of autologous cultured limbal epithelial cell transplantation (CLECT) followed by deep anterior lamellar keratoplasty (DALK) in paediatric eyes and to correlate the clinical outcome with the phenotype of rejuvenated corneal epithelium. Methods: Four patients with total limbal stem cell deficiency (LSCD) underwent autologous CLECT. Cultivated cell sheets were transplanted onto the damaged ocular surface followed by DALK surgery. Excised corneal buttons were subjected to histopathological analysis. Data recorded included age, sex, laterality, nature of injury, follow-up period, severity of stem cell deficiency, visual acuity, Schirmer's test and impression cytology. Results: At a mean follow-up period of 19.5 ± 7.4 (range 9-26) months after CLECT, all 4 eyes showed epithelialized and clinically stabilized ocular surface. Manual DALK was performed in all 4 eyes, with a mean follow-up of 9.75 ± 4.5 (range 5-15) months. All eyes exhibited smooth and clear corneal epithelium with improved visual acuity. Excised corneal buttons demonstrated organized corneal epithelial morphology and showed expression of cornea-specific CK3/12 marker. Conclusion: Restoration of severely damaged ocular surface following chemical injury by using 2-stage meticulous approaches offers a new modality for the treatment of severe LSCD. Transplantation of cultivated autologous limbal epithelial cell sheet followed by DALK surgery can efficiently restore the corneal phenotype with improved vision.

Systematic Review and Meta-analysis on Transplantation of Ex Vivo Cultivated Limbal Epithelial Stem Cell on Amniotic Membrane in Limbal Stem Cell Deficiency

“…As for the ex vivo culture, we specifically focus on the following 4 aspects: culture system, the type of AM, and 30,39,49 20,34,46,47 30,39,49 Y 37,42 …”

“…may be used as a cell substrate in the ex vivo culture. But human amniotic membrane (HAM), which promotes epithelialization, inhibits fibrosis, has anti-inflammatory and antiangiogenic properties, and possesses antimicrobial and antiviral properties and a high hydraulic conductivity, 22,23 with low or no immunogenicity, 24 has become the widely used material [25][26][27][28][29][30][31][32][33][34][35][36][37][38][39][40][41][42] acting as a surrogate environmental stem cell niche. 43 The use of ex vivo cultured LESCs to treat corneal LSCD in humans was first described by Pellegrini et al 44 in 1997. Subsequently, plenty of case series and reviews have reported the efficiency and evidence of this treatment.…”

“…43 The use of ex vivo cultured LESCs to treat corneal LSCD in humans was first described by Pellegrini et al 44 in 1997. Subsequently, plenty of case series and reviews have reported the efficiency and evidence of this treatment. 12,[25][26][27][28][29][30][31][32][33][34][35][36][37][38][39][40][41][42][43][45][46][47] However, with the lack of sufficient cases or valid measurements, they failed to provide a convincing assessment of this surgery. Moreover, there is controversy regarding the clinical outcome between different sources of limbal stem cells, such as autograft and allograft.…”

Limbal Epithelial Stem Cells in Corneal Regeneration

“…The successful restoration of damaged ocular surface may be due to the regeneration of demolished LESC niche, which may in turn repopulate the stem cells crucial for maintaining the epithelial integrity. In one of our report 34 , we further described our experience using a meticulous approach to reconstruct the severely damaged ocular surface by combining our previously reported LSCT method with the deep anterior lamellar corneal transplantation using donor corneas to restore corneal clarity and vision by reducing corneal vascularisation. Further more histopathology of excised host lamellar corneal buttons demonstrated organised corneal epithelial morphology with the expression of CK3/12 in rejuvenated corneal.…”

“…Further more histopathology of excised host lamellar corneal buttons demonstrated organised corneal epithelial morphology with the expression of CK3/12 in rejuvenated corneal. 34 …”