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Photochemical tissue bonding: A potential strategy for treating limbal stem cell deficiency
Author(s) -
Gu Chuan,
Ni Tao,
Verter Erol E.,
Redmond Robert W.,
Kochevar Irene E.,
Yao Min
Publication year - 2011
Publication title -
lasers in surgery and medicine
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.888
H-Index - 112
eISSN - 1096-9101
pISSN - 0196-8092
DOI - 10.1002/lsm.21066
Subject(s) - stem cell , chemistry , biophysics , microbiology and biotechnology , biology
Background and Objective To determine the feasibility of attaching human amniotic membrane (HAM), pre‐cultured with limbal stem cells (LSCs), to cornea using a novel, light‐activated tissue bonding method. Materials and Methods LSCs were isolated from rabbit eyes, and then cultured on de‐epithelialized HAM to create grafts (HAM/LSC). These were then transplanted onto rabbit eyes with surgically created limbal stem cell deficiency (LSCD). The grafts were secured either by sutures or by a light‐activated method called photochemical tissue bonding (PTB). Outcomes included corneal opacity, inflammation, neovascularization, and collagen alignment. Results The isolated and cultured cells were verified to be LSCs based on their K19+/intergrin β1+/P63+/K3 profile. Securing the HAM/LSC graft with PTB provided better outcomes. At 28 days post‐surgery, the corneal opacity scores were significantly lower after securing the graft with PTB compared with suture attachment (0.8 ± 0.5 vs. 1.8 ± 0.5, P < 0.01). Similarly, neovascularization scores were lower after PTB (0.8 ± 0.5 vs. 1.5 ± 0.6, P < 0.01). Quantification of MPO and CD31 levels from immunofluorecent staining indicated that PTB stimulated less neutrophil infiltration (5.3 ± 2.2 vs. 13.3 ± 3.1, P < 0.01) and less new blood vessels formation (2.0 ± 0.8 vs. 6.3 ± 1.3, P < 0.01) at the wound site. The collagen alignment in PTB‐treated corneas, as shown by immunofluorescence and second harmonic generation image, was better organized in the PTB‐treated group than in the suture group. Conclusion Bonding LSC grafts with PTB produced improved outcomes compared to suture attachment. This light‐activated method is a promising modality for treating patients with LSCD. Lasers Surg. Med. 43:433–442, 2011. © 2011 Wiley‐Liss, Inc.