Transplantation of the oral mucosa epithelial layer in the treatment of corneal defects with limbal stem cell deficiency

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Abstract

BACKGROUND: Corneal epithelialization occurs due to proliferation and differentiation of limbal stem epithelial cells. Death of these cells or damage of its microenvironment leads to limbal stem cell deficiency. In bilateral total limbal damage (both eyes), autologues limbal stem cells transplantation becomes impossible. So, it is revalent to find new sources of autologous progenitor cells. One of such sources are buccal cells from cheek mucosa.

AIM: To study the reparative effect of buccal cells in oral mucosa autologous epithelial layer in a mechanical limbal stem cell deficiency model.

MATERIALS AND METHODS: The study was conducted on 7 Chinchilla rabbits (14 eyes). At the first stage, rabbits underwent bilateral total limbectomy and mechanical de-epithelialization of the cornea resulted in fibrovascular pannus development. Then, a full-layer flap of the cheek mucosa measuring 5 × 5 mm was taken, and epithelial layer was separated by 0.5% dispase solution. After superficial keratectomy to transparent layers, a layer of buccal epithelium was placed to the cornea and covered with a soft contact lens. In controls soft contact lens, was placed on the cornea. Temporary tarsorrhaphy was performed for 5 days. In the postoperative period, the area of the deepithelized surface, neovascularization and corneal transparency were evaluated.

RESULTS: On the 7th–30th day, a reduction of erosion was noted in experimental and control eyes, but the dynamics of recovery processes did not significantly differ. On day 60, the area of erosion in the experimental eyes was significantly less than in the control (p = 0.038). Recurrence of erosion was noted in 4 control and 3 experimental eyes.

CONCLUSIONS: In our model of limbal stem cell deficiency, the use of a buccal epithelium layer did not reveal a pronounced reparative effect.

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About the authors

Ekaterina V. Сhentsova

Helmholtz National Medical Research Center for Eye Diseases

Email: chentsova27@yandex.ru
ORCID iD: 0000-0002-8394-1038
SPIN-code: 8191-8338

Dr. Sci. (Med.), Professor, Head of the traumatology and reconstructive surgery department

Russian Federation, Moscow

Natalia V. Borovkova

N.V. Sklifosovsky Research Institute for Emergency Medicine

Email: borovkovanv@yandex.ru
ORCID iD: 0000-0002-8897-7523
SPIN-code: 9339-2800

Dr. Sci. (Med.), Head of the biotechnology and transfusiology department

Russian Federation, Moscow

Tatyana V. Tselaya

Helmholtz National Medical Research Center for Eye Diseases

Author for correspondence.
Email: tatyana.tselaya@yandex.ru
ORCID iD: 0000-0003-3013-685X

Postgraduate Student

Russian Federation, Moscow

Maya V. Storozheva

N.V. Sklifosovsky Research Institute for Emergency Medicine

Email: mayya.storozheva@yandex.ru
ORCID iD: 0000-0003-1927-2404
SPIN-code: 7789-3277

Researcher

Russian Federation, Moscow

Ivan N. Ponomarev

N.V. Sklifosovsky Research Institute for Emergency Medicine

Email: rzam@yandex.ru
ORCID iD: 0000-0002-2523-6939
SPIN-code: 4705-9314

Cand. Sci. (Med.), Senior Researcher

Russian Federation, Moscow

Maxim S. Makarov

N.V. Sklifosovsky Research Institute for Emergency Medicine

Email: mcsimmc@yandex.ru
SPIN-code: 3543-5800

Cand. Sci. (Med.), Senior Researcher

Russian Federation, Moscow

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Supplementary files

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2. Fig. 1. Slit lamp photograph of the eye immediately after total deepithelization and limbectomy

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3. Fig. 2. Slit lamp photograph of the eye on the day 30 after deepithelization and limbectomy

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4. Fig. 3. Anterior segment of the eye the after keratectomy with an epithelial layer and soft contact lens

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5. Fig. 4. Buccal cells in the epithelial layer. Vital staining with tripaflavin acridine-orange. a — the epithelial layer from the basal layer; b — the epithelial layer from the outer layer of the epithelium; c — the underlying connective tissue of the cheek biopsy. The cells are absent on the basement membrane

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6. Fig. 5. Slit lamp photographs of the eyes after soft contact lens removal: a — control; b — experiment. With fluorescein staining

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7. Fig. 6. Slit lamp photographs of the eyes on the day 7 after soft contact lens removal: a — experiment; a — control. With fluorescein staining

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8. Fig. 7. Slit lamp photographs of the eyes on the day 14 after removal of soft contact lens: a — experiment; b — control. With fluorescein staining

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9. Fig. 8. Slit lamp photographs of the eyes on the day 30 after removal of soft contact lens: a — experiment; b — control. With fluorescein staining

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10. Fig. 9. Slit lamp photographs of the eyes on the day 60 after removal of soft contact lens: a — experiment; b — control. With fluorescein staining

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11. Fig. 10. Slit lamp photographs of the eyes on the day 90 after removal of soft contact lenssoft contact lens: a — experiment; b — control. With fluorescein staining

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Copyright (c) 2022 Сhentsova E.V., Borovkova N.V., Tselaya T.V., Storozheva M.V., Ponomarev I.N., Makarov M.S.

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