title: The safety of photochemical tissue bonding for treating damaged corneal epithelium using limbal stem cells pre-cultured on human amniotic membrane.
abstract: We previously demonstrated the feasibility of treating limbal stem cell deficiency (LSCD) with limbal stem cells (LSCs) pre-cultured on human amniotic membrane (HAM), using a suture-free technique called photochemical tissue bonding (PTB). However, important issues regarding the safety and the influence of PTB on LSCs have not been elucidated. In this study, LSCs, isolated from rabbit eyes and identified by cell markers, were labeled with BrdU prior to cultivation on de-epithelialized HAM to fabricate grafts. Rabbit LSCD models were created and randomly divided into groups for transplantation of fabricated grafts using sutures or PTB (n=10). Possible phototoxicity of PTB to LSCs was analyzed in vitro and in vivo. Restoration of corneal epithelium was evaluated at 28 days after grafting. Our results showed that phototoxicity did not occur in the LSCs cultured on HAM after PTB in vitro. Transplantation of grafts with PTB restored the damaged cornea epithelium effectively and no significant influences on LSC characteristics were found in both sutured and PTB groups. BrdU positive cells were tracked at 28 days post grafting suggesting that the restored epithelium was derived from the in vitro fabricated HAM/LSC graft. These data suggest that PTB is a safe and potential strategy for securing LSC/HAM grafts that produces with better outcomes than sutured attachment.
authors.affiliation: Department of Burn and Plastic Surgery, No. 3 People's Hospital, and Institute of Traumatic Medicine, School of Medicine, Shanghai Jiao Tong University, Shanghai 201900, China; Wellman Center for Photomedicine, Massachusetts General Hospital, Harvard Medical School, Boston, MA 02114, USA; Department of Plastic and Reconstructive Surgery, Ninth People's Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai 200011, China.
authors.forename: Chuan
authors.initials: C
authors.lastname: Gu
authors.affiliation: Department of Plastic and Reconstructive Surgery, Ninth People's Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai 200011, China.
authors.forename: Jun
authors.initials: J
authors.lastname: Yang
authors.affiliation: Department of Ophthalmology, Chengyang People's Hospital, Qingdao City, Shandong Province 266109, China.
authors.forename: Ying
authors.initials: Y
authors.lastname: Yuan
authors.affiliation: Department of Burn and Plastic Surgery, No. 3 People's Hospital, and Institute of Traumatic Medicine, School of Medicine, Shanghai Jiao Tong University, Shanghai 201900, China; Wellman Center for Photomedicine, Massachusetts General Hospital, Harvard Medical School, Boston, MA 02114, USA.
authors.forename: Min
authors.initials: M
authors.lastname: Yao
authors.affiliation: Department of Burn and Plastic Surgery, No. 3 People's Hospital, and Institute of Traumatic Medicine, School of Medicine, Shanghai Jiao Tong University, Shanghai 201900, China. Electronic address: xiong@medmail.co.
authors.forename: Xiong
authors.initials: X
authors.lastname: Zhang
mesh_terms: D000650:Amnion / Q000737:chemistry / Q000378:metabolism*; D000818:Animals; D002470:Cell Survival; D002478:Cells, Cultured; D003316:Corneal Diseases / Q000628:therapy; D004195:Disease Models, Animal; D019573:Epithelium, Corneal / Q000166:cytology* / Q000473:pathology; D006801:Humans; D008856:Microscopy, Fluorescence; D011817:Rabbits; D033581:Stem Cell Transplantation; D013234:Stem Cells / Q000166:cytology* / Q000378:metabolism; D054457:Tissue Scaffolds