Transplantation of autologous cultivated oral mucosal epithelial sheets for limbal stem cell deficiency at Siriraj Hospital: a case series

Wipawee Booranapong, Panida Kosrirukvongs, Sunisa Duangsa-Ard, Kanda Kasetsinsombat, Khanit Sa-Ngiamsuntorn, Adisak Wongkajornsilp, Wipawee Booranapong, Panida Kosrirukvongs, Sunisa Duangsa-Ard, Kanda Kasetsinsombat, Khanit Sa-Ngiamsuntorn, Adisak Wongkajornsilp

Abstract

Background: The loss of limbal stem cells owing to either corneal burn or inflammation leads to the repopulation of opaque skin over the raw surface of the cornea. It has been proposed that reconstitution of oral mucosal stem cells over this raw surface will mimic the limbal stem cells and restore vision. The efficacy and safety of applying a sheet of cultivated oral mucosal cells as an autologous graft for corneal replacement were evaluated.

Case presentation: The study was conducted during 2014-2015 and involved a total of six patients, of whom three had suffered a chemical burn and three had Stevens-Johnson Syndrome (SJS). Oral mucosal tissue was dissected from each patient, seeded onto irradiated J2 fibroblast feeder cells for 14 days, and analyzed for quality and safety 1 day before being transplanted onto the cornea of the affected eyes. After transplantation, topical antibiotic and anti-inflammatory eye drops were instilled four times daily, and the patients wore contact lenses. Subjects were clinically followed for visual acuities and adverse effects at 2, 4, and 6 weeks, 3 and 6 months, and 1 year post-transplantation. Data were presented descriptively. Visual acuities in patients improved at 2 weeks post-surgery. However, two patients with SJS had corneal ulcer at 2 weeks postoperatively. At the 1-year postoperative examination, the eyes of two patients were in good condition with decreased vascularization and epithelial defect.

Conclusions: Cultivated oral mucosal epithelial sheet transplantation in limbal stem cell deficiency had a favorable efficacy. In this study, patients with chemical burn had more clinical benefit than those with SJS. Trial registration ClinicalTrials.gov: NCT02415218. Registered retrospectively 4 Apr 2015 ( https://ichgcp.net/clinical-trials-registry/NCT02415218 ).

Keywords: Corneal replacement; Limbal deficiency; Oral mucosal epithelial sheet.

Conflict of interest statement

The authors declared that they have no competing interest.

© 2022. The Author(s).

Figures

Fig. 1
Fig. 1
Hematoxylin and eosin (H&E) staining (objective lens: 40×) and immunofluorescence staining (objective lens: 20×) for the markers of epithelial stem/progenitor cells (tumor protein p63 [p63]), corneal differentiation (cytokeratin 3 [AE5]), and barrier function (membrane-anchored mucin-16 [MUC16] and tight junction protein-1 [ZO-1]) in the epithelial cell sheets prepared from the oral mucosal cells from subject #1 (A) and #3 (B)
Fig. 2
Fig. 2
The colony-forming assay of the oral mucosal epithelial cells from subjects #1 (A) and #3 (B)
Fig. 3
Fig. 3
Ophthalmic examinations on the inflicted eyes from successful responders (A, subjects #1 and #3) and less successful responders (B, subjects #2, #5, and #6) were taken before the operation and after the operation at the designated time points

References

    1. Ilari L, Daya SM. Long-term outcomes of keratolimbal allograft for the treatment of severe ocular surface disorders. Ophthalmology. 2002;109(7):1278–1284. doi: 10.1016/S0161-6420(02)01081-3.
    1. Gomes JA, Santos MS, Ventura AS, Donato WB, Cunha MC, Hofling-Lima AL. Amniotic membrane with living related corneal limbal/conjunctival allograft for ocular surface reconstruction in Stevens-Johnson syndrome. Arch Ophthalmol. 2003;121(10):1369–1374. doi: 10.1001/archopht.121.10.1369.
    1. Samson CM, Nduaguba C, Baltatzis S, Foster CS. Limbal stem cell transplantation in chronic inflammatory eye disease. Ophthalmology. 2002;109(5):862–868. doi: 10.1016/S0161-6420(02)00994-6.
    1. Tsubota K, Satake Y, Kaido M, et al. Treatment of severe ocular-surface disorders with corneal epithelial stem-cell transplantation. N Engl J Med. 1999;340(22):1697–1703. doi: 10.1056/NEJM199906033402201.
    1. Williams KA, Brereton HM, Aggarwal R, et al. Use of DNA polymorphisms and the polymerase chain reaction to examine the survival of a human limbal stem cell allograft. Am J Ophthalmol. 1995;120(3):342–350. doi: 10.1016/S0002-9394(14)72164-6.
    1. Tsai RJ, Tseng SC. Effect of stromal inflammation on the outcome of limbal transplantation for corneal surface reconstruction. Cornea. 1995;14(5):439–449. doi: 10.1097/00003226-199509000-00001.
    1. Tseng SC, Prabhasawat P, Barton K, Gray T, Meller D. Amniotic membrane transplantation with or without limbal allografts for corneal surface reconstruction in patients with limbal stem cell deficiency. Arch Ophthalmol. 1998;116(4):431–441. doi: 10.1001/archopht.116.4.431.
    1. Yamato M, Konno C, Utsumi M, Kikuchi A, Okano T. Thermally responsive polymer-grafted surfaces facilitate patterned cell seeding and co-culture. Biomaterials. 2002;23(2):561–567. doi: 10.1016/S0142-9612(01)00138-7.
    1. Shimizu T, Yamato M, Isoi Y, et al. Fabrication of pulsatile cardiac tissue grafts using a novel 3-dimensional cell sheet manipulation technique and temperature-responsive cell culture surfaces. Circ Res. 2002;90(3):e40. doi: 10.1161/hh0302.105722.
    1. Hayashida Y, Nishida K, Yamato M, et al. Ocular surface reconstruction using autologous rabbit oral mucosal epithelial sheets fabricated ex vivo on a temperature-responsive culture surface. Invest Ophthalmol Vis Sci. 2005;46(5):1632–1639. doi: 10.1167/iovs.04-0813.
    1. Hayashi R, Yamato M, Takayanagi H, et al. Validation system of tissue-engineered epithelial cell sheets for corneal regenerative medicine. Tissue Eng Part C Methods. 2010;16(4):553–560. doi: 10.1089/ten.tec.2009.0277.
    1. Hori Y, Sugiyama H, Soma T, Nishida K. Expression of membrane-associated mucins in cultivated human oral mucosal epithelial cells. Cornea. 2007;26(9 Suppl 1):S65–S69. doi: 10.1097/ICO.0b013e31812f6c95.
    1. Sumide T, Nishida K, Yamato M, et al. Functional human corneal endothelial cell sheets harvested from temperature-responsive culture surfaces. FASEB J. 2006;20(2):392–394. doi: 10.1096/fj.04-3035fje.
    1. Nishida K, Yamato M, Hayashida Y, et al. Corneal reconstruction with tissue-engineered cell sheets composed of autologous oral mucosal epithelium. N Engl J Med. 2004;351(12):1187–1196. doi: 10.1056/NEJMoa040455.
    1. Ikuno Y, Ikeda T, Sato Y, Tano Y. Tractional retinal detachment after branch retinal vein occlusion. Influence of disc neovascularization on the outcome of vitreous surgery. Ophthalmology. 1998;105(3):417–423. doi: 10.1016/S0161-6420(98)93021-4.
    1. Sotozono C, Ang LP, Koizumi N, et al. New grading system for the evaluation of chronic ocular manifestations in patients with Stevens-Johnson syndrome. Ophthalmology. 2007;114(7):1294–1302. doi: 10.1016/j.ophtha.2006.10.029.
    1. Yoshikawa Y, Ueta M, Nishigaki H, Kinoshita S, Ikeda T, Sotozono C. Predictive biomarkers for the progression of ocular complications in chronic Stevens-Johnson syndrome and toxic Eeidermal necrolysis. Sci Rep. 2020;10(1):18922. doi: 10.1038/s41598-020-76064-8.

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