Cell and Tissue Banking

, Volume 20, Issue 1, pp 95–108 | Cite as

Epithelial analysis of simple limbal epithelial transplantation in limbal stem cell deficiency by in vivo confocal microscopy and impression cytology

  • Pinnita Prabhasawat
  • Angkoon Luangaram
  • Pattama Ekpo
  • Kaevalin Lekhanont
  • Wimolwan Tangpagasit
  • Chawikan Boonwong
  • Naharuthai Inthasin
  • Chareenun ChirapapaisanEmail author


Simple limbal epithelial transplantation (SLET) is a relatively new treatment for severe limbal stem cell deficiency. Outcomes of treatment are typically determined based on clinical manifestations. In this prospective-multicenter study, we aimed to analyze the epithelial phenotypes of the corneas after SLET using IVCM and IC, and correlated them with clinical findings. Ten eyes of nine patients, who underwent SLET (five autologous SLET and five living-related SLET) were recruited. A set of examinations included slit-lamp biomicroscopy, corneal in vivo confocal microscopy (IVCM), and impression cytology (IC) was performed in all eyes at least twice (≥ 3-month interval) postoperatively. Then, a correlation between findings of the three examinations was analyzed. There were seven eyes with clinical success (no central neovascularization) showed pure corneal epithelial phenotype or mixed corneal-conjunctival phenotypes (mostly cornea) in either IVCM or IC. Three eyes with clinical failure, presented with peripheral and central neovascularization, showed total or predominant conjunctival phenotype in IVCM and sole conjunctival phenotype in IC. From a total of 22 sets of examinations, there was a high correlation between clinical manifestation vs. IC (κ = 0.844, observed agreement = 81.82%) and a substantial correlation between clinical manifestation vs. IVCM (κ = 0.727, observed agreement = 76.19%) and between IVCM versus IC (κ = 0.729, observed agreement = 76.19%). In conclusion, IVCM and IC facilitate determination of epithelial phenotype of the cornea after SLET. There was a substantial to high correlation between IVCM, IC and clinical presentations. Findings observed by IVCM and IC may allow early detection of epithelial alterations in eyes underwent SLET before clinical recognition.


Limbal stem cell deficiency (LSCD) In vivo confocal microscopy (IVCM) Impression cytology (IC) Cornea Epithelial phenotype 



The authors gratefully acknowledge contributors from Faculty of Medicine Siriraj Hospital, Mahidol University: Wiwit Tantibhedhyangkul, Ph.D and Patimaporn Wongprompitak, Ph.D (Department of Immunology) for encouragement and valuable comments; Anupong Veeraburinon (Research Division) for assistance with manuscript development and data collection; Pratuangsri Chonpimai (Department of Ophthalmology) for assistance with in vivo confocal microscopy; Mathuwan Srikong and Kritphol Rattanawarinchai (Medical Education Technology Centre) for preparing the figures; and Nuttawat Saenyasiri, MD (Department of Immunology) for providing technical support of immunofluorescence imaging. Furthermore, we would like to acknowledge all patients for their kind cooperation and engagement throughout the study.


This work was supported by Research Development Fund from Faculty of Medicine Siriraj Hospital, Mahidol University (Grant Number R015932015). The funder was not involved in the study design, collection, analysis, and interpretation of data; writing of the report; or the decision to submit the paper for publication.

Compliance with ethical standards

Conflict of interest

The authors declare that there is no conflict of interest regarding the publication of this paper.

Ethical approval

Ethical review boards from Siriraj Hospital, Ramathibodi Hospital, and Thammasat University Hospital have been approved this study before enrolling the cases (Approval Number, Si051/2016: Siriraj Hospital; 2559/255: Ramathibodi Hospital; and 047/2559: Thammasat University Hospital).

Informed consent

Informed consent was obtained from all subjects recruited in this study.

Supplementary material

10561_2018_9746_MOESM1_ESM.tif (137.9 mb)
Supplemental Figure 1: Preoperative and postoperative slit lamp images of all eyes underwent SLET (TIFF 141203 kb)
10561_2018_9746_MOESM2_ESM.tif (7.4 mb)
Supplemental Figure 2: Positive and negative controls of immunofluorescence staining (TIFF 7570 kb)
10561_2018_9746_MOESM3_ESM.docx (19 kb)
Supplementary material 3 (DOCX 18 kb)


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Copyright information

© Springer Nature B.V. 2019

Authors and Affiliations

  • Pinnita Prabhasawat
    • 1
  • Angkoon Luangaram
    • 1
  • Pattama Ekpo
    • 2
  • Kaevalin Lekhanont
    • 3
  • Wimolwan Tangpagasit
    • 4
  • Chawikan Boonwong
    • 5
  • Naharuthai Inthasin
    • 2
  • Chareenun Chirapapaisan
    • 1
    Email author
  1. 1.Department of Ophthalmology, Faculty of Medicine Siriraj HospitalMahidol UniversityBangkokThailand
  2. 2.Department of Immunology, Faculty of Medicine Siriraj HospitalMahidol UniversityBangkokThailand
  3. 3.Department of Ophthalmology, Faculty of Medicine Ramathibodi HospitalMahidol UniversityBangkokThailand
  4. 4.Department of Ophthalmology, Faculty of MedicineThammasat UniversityPathumthaniThailand
  5. 5.Research Division, Faculty of Medicine Siriraj HospitalMahidol UniversityBangkokThailand

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