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

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 7^th–30^th 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.

Keywords

limbal stem cell deficiency / limb / epithelium of the oral mucosa / cornea / fibrovascular pannus / rabbits

References

[1] Davanger M, Evensen A. Role of the Pericorneal Papillary Structure in Renewal of Corneal Epithelium. Nature. 1971;229(5286): 560–561. DOI: 10.1038/229560a0

[2] Davanger M., Evensen A. Role of the Pericorneal Papillary Structure in Renewal of Corneal Epithelium // Nature. 1971. Vol. 229, No. 5286. P. 560–561. DOI: 10.1038/229560a0

[3] Popandopulo AG, Kavelina AS, Ivanova ON, Drozhzhina GI. Rol limbalnykh kletok v regeneratsii rogovitsy. Scientific journal Tavricheskiy Mediko-Biologicheskiy Vestnik. 2013;16(1–2):158–160. (In Russ.)

[4] Попандопуло А.Г., Кавелина А.С., Иванова О.Н., Дрожжина Г.И. Роль лимбальных клеток в регенерации роговицы // Таврический медико-биологический вестник. 2013. T. 16, № 1–2. С. 158–160.

[5] Daniels JT, Dart JK, Tuft SJ, Khaw PT. Corneal stem cells in review. Wound Repair and Regen. 2001;9(6):483–494. DOI: 10.1046/j.1524-475x.2001.00483.x

[6] Daniels J.T., Dart J.K., Tuft S.J., Khaw P.T. Corneal stem cells in review // Wound Repair and Regen. 2001. Vol. 9, No. 6. P. 483–494. DOI: 10.1046/j.1524-475x.2001.00483.x

[7] Shevlyuk NN, Radchenko AV, Stadnikov AA. The Structural and Functional Basis of Physiological and Reparative Regeneration of Corneal Tissues. Journal of Anatomy and Histopathology. 2019;8(2): 82–90. (In Russ.) DOI: 10.18499/2225-7357-2019-8-2-82-90

[8] Шевлюк Н.Н., Радченко А.В., Стадников А.А. Структурно-функциональные основы физиологической и репаративной регенерации тканей роговицы // Журнал анатомии и гистопатологии. 2019. T. 8, № 2. С. 82–90. DOI: 10.18499/2225-7357-2019-8-2-82-90

[9] Nikolaeva LR, Chentsova EV. Limbic cellular insufficiency. The Russian annals of ophthalmology. 2006;122(3):43–47. (In Russ.)

[10] Николаева Л.Р., Ченцова Е.В. Лимбальная клеточная недостаточность // Вестник офтальмологии. 2006. T. 122, № 3. С. 43–47.

[11] Li W, Hayashida Y, Chen Y-T, Tseng SC. Niche regulation of corneal epithelial stem cells at the limbus. Cell Res. 2007;17(1):26–36. DOI: 10.1038/sj.cr.7310137

[12] Li W., Hayashida Y., Chen Y.-T., Tseng S.C. Niche regulation of corneal epithelial stem cells at the limbus // Cell Res. 2007. Vol. 17, No. 1. P. 26–36. DOI: 10.1038/sj.cr.7310137

[13] Ramos T, Deborah S, Ahmad S. An update on ocular surface epithelial stem cells: cornea and conjunctiva. Stem Cells Int. 2015;2015:601731. DOI: 10.1155/2015/601731

[14] Ramos T., Deborah S., Ahmad S. An update on ocular surface epithelial stem cells: cornea and conjunctiva // Stem Cells Int. 2015. Vol. 2015. ID601731. DOI: 10.1155/2015/601731

[15] Dua HS, Saini JS, Azuara-Blanco A, Gupta P. Limbal stem cell deficiency: Concept, aetiology, clinical presentation, diagnosis and management. Indian J Ophthalmol. 2000;48(2):83–92.

[16] Dua H.S., Saini J.S., Azuara-Blanco A., Gupta P. Limbal stem cell deficiency: Concept, aetiology, clinical presentation, diagnosis and management // Indian J Ophthalmol. 2000. Vol. 48, No. 2. P. 83–92.

[17] Shapiro MS, Friend J, Thoft RA. Corneal re-epithelialization from the conjunctiva. Investig Ophthalmol Vis Sci. 1981;21(1–1):135–142.

[18] Shapiro M.S., Friend J., Thoft R.A. Corneal re-epithelialization from the conjunctiva // Investig Ophthalmol Vis Sci. 1981. Vol. 21, No. 1–1. P. 135–142.

[19] Deng SX, Sejpal K, Bakhtiari P. Presentation, diagnosis and management of limbal stem cell deficiency. Middle East Afr J Ophthalmol. 2013;20(1):5–10. DOI: 10.4103/0974-9233.106381.

[20] Deng S.X., Sejpal K., Bakhtiari P. Presentation, diagnosis and management of limbal stem cell deficiency // Middle East Afr J Ophthalmol. 2013. Vol. 20, No. 1. P. 5–10. DOI: 10.4103/0974-9233.106381

[21] Kenyon KR, Tseng SC. Limbal asutograft transplantation for ocular surface disorders. Ophthalmology. 1989;96(5):709–723. DOI: 10.1016/S0161-6420(89)32833-8

[22] Kenyon K.R., Tseng S.C. Limbal asutograft transplantation for ocular surface disorders // Ophthalmology. 1989. Vol. 96, No. 5. P. 709–723. DOI: 10.1016/S0161-6420(89)32833-8

[23] Tsai RJ-F, Tseng SCG. Human allograft limbal transplantation for corneal surface reconstruction. Cornea. 1994;13(5):389–400. DOI: 10.1097/00003226-199409000-00003

[24] Tsai R.J.-F., Tseng S.C.G. Human allograft limbal transplantation for corneal surface reconstruction // Cornea. 1994. Vol. 13, No. 5. P. 389–400. DOI: 10.1097/00003226-199409000-00003

[25] Kwitko S, Marinho D, Barcaro S, et al. Allograft conjunctival transplantation for bilateral ocular surface disorders. Ophthalmology. 1995;102(7):1020–1025. DOI: 10.1016/S0161-6420(95)30918-9

[26] Kwitko S., Marinho D., Barcaro S., et al. Allograft conjunctival transplantation for bilateral ocular surface disorders // Ophthalmology. 1995. Vol. 102, No. 7. P. 1020–1025. DOI: 10.1016/S0161-6420(95)30918-9

[27] Nakamura T, Inatomi T, Sotozono C, et al. Ocular surface reconstruction using stem cell and tissue engineering. Prog Retin Eye Res. 2016;51:187–207. DOI: 10.1016/j.preteyeres.2015.07.003

[28] Nakamura T., Inatomi T., Sotozono C., et al. Ocular surface reconstruction using stem cell and tissue engineering // Prog Retin Eye Res. 2016. Vol. 51. P. 187–207. DOI: 10.1016/j.preteyeres.2015.07.003

[29] Nakamura T, Inatomi T, Sotozono C, et al. Successful primary culture and autologous transplantation of corneal limbal epithelial cells from minimal biopsy for unilateral severe ocular surface disease. Acta Ophthalmologica Scandinavica. 2004;82(4):468–471. DOI: 10.1111/j.1395-3907.2004.00285.x

[30] Nakamura T., Inatomi T., Sotozono C., et al. Successful primary culture and autologous transplantation of corneal limbal epithelial cells from minimal biopsy for unilateral severe ocular surface disease // Acta Ophthalmologica Scandinavica. 2004. Vol. 82, No. 4. P. 468–471. DOI: 10.1111/j.1395-3907.2004.00285.x

[31] Rama P, Matuska S, Paganoni G, et al. Limbal stem-cell therapy and long-term corneal regeneration. N Engl J Med. 2010;363(2): 147–155. DOI: 10.1056/NEJMoa0905955

[32] Rama P., Matuska S., Paganoni G., et al. Limbal stem-cell therapy and long-term corneal regeneration // N Engl J Med. 2010. Vol. 363, No. 2. P. 147–155. DOI: 10.1056/NEJMoa0905955

[33] Schwab IR, Reyes M, Isseroff RR. Successful transplantation of bioengineered tissue replacements in patients with ocular surface disease. Cornea. 2000;19(4):421–426. DOI: 10.1097/00003226-200007000-00003

[34] Schwab I.R., Reyes M., Isseroff R.R. Successful transplantation of bioengineered tissue replacements in patients with ocular surface disease // Cornea. 2000. Vol. 19, No. 4. P. 421–426. DOI: 10.1097/00003226-200007000-00003

[35] Khlopin NG. Obshchebiologicheskie i ehksperimental’nye osnovy gistologii. Leningrad: AN SSSR, 1946. 491 p. (In Russ.)

[36] Хлопин Н.Г. Общебиологические и экспериментальные основы гистологии. Ленинград: АН СССР, 1946. 491 с.

[37] Kinoshita S, Nakamura T. Development of cultivated mucosal epithelial sheet transplantation for ocular surface reconstruction. Artificial Organs. 2004;28(1):22–27. DOI: 10.1111/j.1525-1594.2004.07319.x

[38] Kinoshita S., Nakamura T. Development of cultivated mucosal epithelial sheet transplantation for ocular surface reconstruction // Artificial Organs. 2004. Vol. 28, No. 1. P. 22–27. DOI: 10.1111/j.1525-1594.2004.07319.x

[39] Nakamura T, Endo K-I, Cooper LJ, et al. The successful culture and autologous transplantation of rabbit oral mucosal epithelial cells on amniotic membrane. Investig Ophthalmol Vis Sci. 2003;44(1): 106–116. DOI: 10.1167/iovs.02-0195

[40] Nakamura T., Endo K.-I., Cooper L.J., et al. The successful culture and autologous transplantation of rabbit oral mucosal epithelial cells on amniotic membrane // Investig Ophthalmol Vis Sci. 2003. Vol. 44, No. 1. P. 106–116. DOI: 10.1167/iovs.02-0195

[41] 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

[42] 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. Vol. 351, No. 12. P. 1187–1196. DOI: 10.1056/NEJMoa040455

[43] Inatomi T, Nakamura T, Koizumi N, et al. Midterm results on ocular surface reconstruction using cultivated autologous oral mucosal epithelial transplantation. Am J Ophthalmol. 2006;141(2):267–275. DOI: 10.1016/j.ajo.2005.09.003

[44] Inatomi T., Nakamura T., Koizumi N., et al. Midterm results on ocular surface reconstruction using cultivated autologous oral mucosal epithelial transplantation // Am J Ophthalmol. 2006. Vol. 141, No. 2. P. 267–275. DOI: 10.1016/j.ajo.2005.09.003

[45] Bezushko AV, Dubovikov AS, Kulikov AN, et al. Мodification of Mechanical Limbal Stem Cell Deficiency Model. Ophthalmology in Russia. 2018;15(1):51–57. (In Russ.) DOI: 10.18008/1816-5095-2018-1-51-57

[46] Безушко А.В., Дубовиков А.С., Куликов А.Н., и др. Модификация механической модели лимбальной недостаточности // Офтальмология. 2018. T. 15, № 1. С. 51–57. DOI: 10.18008/1816-5095-2018-1-51-57

[47] Voino-Yasenetskii VV. Razrastanie i izmenchivost’ tkanei glaza pri ego zabolevaniyakh i travmakh. Kiеv: Vishcha shkola,1979. 165 p. (In Russ.)

[48] Войно-Ясенецкий В.В. Разрастание и изменчивость тканей глаза при его заболеваниях и травмах. Киев: Вища школа,1979. 165 с.

[49] Inamochi A, Tomioka A, Kitamoto K, et al. Simple oral mucosal epithelial transplantation in a rabbit model. Sci Rep. 2019;9(1):18088. DOI: 10.1038/s41598-019-54571-7

[50] Inamochi A., Tomioka A., Kitamoto K., et al. Simple oral mucosal epithelial transplantation in a rabbit model // Sci Rep. 2019. Vol. 9, No. 1. ID 18088. DOI: 10.1038/s41598-019-54571-7

RIGHTS & PERMISSIONS

Сhentsova E.V., Borovkova N.V., Tselaya T.V., Storozheva M.V., Ponomarev I.N., Makarov M.S.