Reconstruction of rabbits' corneal epithelium with human epidermal stem cells delivery by amniotic membrane contact lens

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Author:
WAN Peng-xia(State Key Laboratory ofOphthalmology, Zhongshan Ophthalmic Center, Sun Yat -sen University, Guangzhou 510060, China)
WANG Zhi-chong(State Key Laboratory ofOphthalmology, Zhongshan Ophthalmic Center, Sun Yat -sen University, Guangzhou 510060, China)
MA Ping(State Key Laboratory ofOphthalmology, Zhongshan Ophthalmic Center, Sun Yat -sen University, Guangzhou 510060, China)
GAO Nan(State Key Laboratory ofOphthalmology, Zhongshan Ophthalmic Center, Sun Yat -sen University, Guangzhou 510060, China)
Journal Title:
CHINESE JOURNAL OF BIOMEDICAL ENGINEERING
Issue:
Volume 15, Issue 01, 2009
DOI:
10.3760/cma.j.issn.1674-1927.2009.01.004
Key Word:
Amnion;Contact lenses;Stem cells transplantation;Limbal stem celldeficiency;Epithelium, corneal

Abstract: Objective To introduce contact lens made up of bio-membrane fixing device (BMFD) and amniotic membrane (AM) as a cell delivery system on ocular surface and evaluate its efficiency in transplanting human epidermal stem cells (Epi-SCs) to the limbal stem cell deficiency (LSCD) rabbits' ocular surface for corneal epithelium reconstruction. Methods The female LSCD rabbit models with the limbal stem cells and corneal epithelium removed were established, and were divided into 3 groups. The concentrated suspension of male human Epi-SCs was injected into the space between the LSCD rabbits' ocular surface and the BMFD/AM contact lens (AM contact lens for short) in AM + Epi-SCs group (n=20), wearing the AM contact lens was performed in AM patching group (n=20), and only drug treatment was performed in control group (n=20). All the rabbits were followed up with slit-lamp examination and corneal sodium fluorescent staining, and the corneal restoration was evaluated. Pathological and immunohistochemical stained tissue sections were examined till completely re-epithelization. Human Y-STR gene was detected to trace the implanted Epi-SCs by PCR. Results The complete re-epithelization time of cornea in AM + Epi-SCs group was (5.60±0.46) d, which was much shorter than that of the other two groups [AM patching group: (9.25±0.51) d; control group: (12.45 ± 0.65) d, all P<0.05]. The reconstructed epithelium in AM + Epi-SCs group showed K3/K12 (+), Mucin 5AC (-), K4 (-) detected by fluorescent immunohistochemistry, which indicated a normal phenotype of corneal epithelium. And human Y-STR gene was found too. By contrast, the corneal epithelium showed conjunctiva phenotype in control group with K4 (+), Mucin 5AC(+), K3/K12(-). Conclusion AM contact lens can serve as a cell delivery system and help corneal epithelium reconstruction in LSCD rabbits by combined with the human Epi-SCs suspension injection.

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