title: Reconstruction of limbal stem cell deficient corneal surface with induced human bone marrow mesenchymal stem cells on amniotic membrane.
abstract: The cornea can be damaged by a variety of clinical disorders or chemical, mechanical, and thermal injuries. The objectives of this study were to induce bone marrow mesenchymal stem cells (BMSCs) to corneal lineage, to form a tissue engineered corneal substitute (TEC) using BMSCs, and to treat corneal surface defects in a limbal stem cell deficiency model. BMSCs were induced to corneal lineage using limbal medium for 10 days. Induced BMSCs demonstrated upregulation of corneal stem cell markers; β1-integrin, C/EBPδ, ABCG2, and p63, increased protein expression of CK3 and p63 significantly compared with the uninduced ones. For TEC formation, passage 1 BMSCs were trypsinized and seeded on amniotic membrane in a transwell co-culture system and were grown in limbal medium. Limbal stem cell deficiency models were induced by alkaline injury, and the TEC was implanted for 8 weeks. Serial slit lamp evaluation revealed remarkable improvement in corneal regeneration in terms of corneal clarity and reduced vascularization. Histologic and optical coherence tomography analyses demonstrated comparable corneal thickness and achieved stratified epithelium with a compact stromal layer resembling that of normal cornea. CK3 and p63 were expressed in the newly regenerated cornea. In conclusion, BMSCs can be induced into corneal epithelial lineage, and these cells are viable for the formation of TEC, to be used for the reconstruction of the corneal surface in the limbal stem cell deficient model.
authors.affiliation: Tissue Engineering Center, Universiti Kebangsaan Malaysia Medical Center, Kuala Lumpur, Malaysia.
authors.forename: Che Man
authors.initials: CM
authors.lastname: Rohaina
authors.affiliation: Department of Ophthalmology, Faculty of Medicine, Universiti Kebangsaan Malaysia, Kuala Lumpur, Malaysia.
authors.forename: Kong Yong
authors.initials: KY
authors.lastname: Then
authors.affiliation: Tissue Engineering Center, Universiti Kebangsaan Malaysia Medical Center, Kuala Lumpur, Malaysia.
authors.forename: Angela Min Hwei
authors.initials: AM
authors.lastname: Ng
authors.affiliation: Department of Ophthalmology, Faculty of Medicine, Universiti Kebangsaan Malaysia, Kuala Lumpur, Malaysia.
authors.forename: Wan Haslina
authors.initials: WH
authors.lastname: Wan Abdul Halim
authors.affiliation: Tissue Engineering Center, Universiti Kebangsaan Malaysia Medical Center, Kuala Lumpur, Malaysia.
authors.forename: Aida Zairani Mohd
authors.initials: AZ
authors.lastname: Zahidin
authors.affiliation: Tissue Engineering Center, Universiti Kebangsaan Malaysia Medical Center, Kuala Lumpur, Malaysia; Ear, Nose, and Throat Consultant Clinic, Ampang Puteri Specialist Hospital, Selangor, Malaysia.
authors.forename: Aminuddin
authors.initials: A
authors.lastname: Saim
authors.affiliation: Tissue Engineering Center, Universiti Kebangsaan Malaysia Medical Center, Kuala Lumpur, Malaysia; Department of Physiology, Faculty of Medicine, Universiti Kebangsaan Malaysia, Kuala Lumpur, Malaysia. Electronic address: ruszy@medic.ukm.my.
authors.forename: Ruszymah B H
authors.initials: RB
authors.lastname: Idrus
mesh_terms: D000650:Amnion / Q000166:cytology*; D000818:Animals; D015415:Biomarkers / Q000378:metabolism; D001854:Bone Marrow Cells / Q000166:cytology* / Q000378:metabolism; D002478:Cells, Cultured; D003315:Cornea / Q000166:cytology*; D004195:Disease Models, Animal; D005786:Gene Expression Regulation; D006801:Humans; D053558:Keratin-3 / Q000235:genetics; D008565:Membrane Proteins / Q000235:genetics; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000378:metabolism; D051379:Mice; D051381:Rats; D011923:Rats, Nude; D060888:Real-Time Polymerase Chain Reaction; D033581:Stem Cell Transplantation; D023822:Tissue Engineering
chemical_list: D015415:Biomarkers; C081876:CKAP4 protein, human; C479360:KRT3 protein, human; D053558:Keratin-3; D008565:Membrane Proteins