title: Decellularization of human stromal refractive lenticules for corneal tissue engineering.
abstract: Small incision lenticule extraction (SMILE) becomes a procedure to correct myopia. The extracted lenticule can be used for other clinical scenarios. To prepare for allogeneic implantation, lenticule decellularization with preserved optical property, stromal architecture and chemistry would be necessary. We evaluated different methods to decellularize thin human corneal stromal lenticules created by femtosecond laser. Treatment with 0.1% sodium dodecylsulfate (SDS) followed by extensive washes was the most efficient protocol to remove cellular and nuclear materials. Empty cell space was found inside the stroma, which displayed aligned collagen fibril architecture similar to native stroma. The SDS-based method was superior to other treatments with hyperosmotic 1.5 M sodium chloride, 0.1% Triton X-100 and nucleases (from 2 to 10 U/ml DNase and RNase) in preserving extracellular matrix content (collagens, glycoproteins and glycosaminoglycans). The stromal transparency and light transmittance was indifferent to untreated lenticules. In vitro recellularization showed that the SDS-treated lenticules supported corneal stromal fibroblast growth. In vivo re-implantation into a rabbit stromal pocket further revealed the safety and biocompatibility of SDS-decellularized lenticules without short- and long-term rejection risk. Our results concluded that femtosecond laser-derived human stromal lenticules decellularized by 0.1% SDS could generate a transplantable bioscaffold with native-like stromal architecture and chemistry.
authors.affiliation: Tissue Engineering and Stem Cell Group, Singapore Eye Research Institute, Singapore.|Eye-ACP, Duke-NUS Graduate Medical School, Singapore.
authors.forename: Gary Hin-Fai
authors.initials: GH
authors.lastname: Yam
authors.affiliation: Tissue Engineering and Stem Cell Group, Singapore Eye Research Institute, Singapore.
authors.forename: Nur Zahirah Binte M
authors.initials: NZ
authors.lastname: Yusoff
authors.affiliation: Tissue Engineering and Stem Cell Group, Singapore Eye Research Institute, Singapore.
authors.forename: Tze-Wei
authors.initials: TW
authors.lastname: Goh
authors.affiliation: Tissue Engineering and Stem Cell Group, Singapore Eye Research Institute, Singapore.
authors.forename: Melina
authors.initials: M
authors.lastname: Setiawan
authors.affiliation: Tissue Engineering and Stem Cell Group, Singapore Eye Research Institute, Singapore.
authors.forename: Xiao-Wen
authors.initials: XW
authors.lastname: Lee
authors.affiliation: Tissue Engineering and Stem Cell Group, Singapore Eye Research Institute, Singapore.|Singapore National Eye Center, Singapore.
authors.forename: Yu-Chi
authors.initials: YC
authors.lastname: Liu
authors.affiliation: Tissue Engineering and Stem Cell Group, Singapore Eye Research Institute, Singapore.|Eye-ACP, Duke-NUS Graduate Medical School, Singapore.|Singapore National Eye Center, Singapore.|School of Materials Science and Engineering, Nanyang Technological University, Singapore.
authors.forename: Jodhbir S
authors.initials: JS
authors.lastname: Mehta
mesh_terms: D000818:Animals; D003315:Cornea / Q000601:surgery*; D003319:Corneal Stroma / Q000737:chemistry / Q000166:cytology* / Q000187:drug effects; D048988:Corneal Surgery, Laser; D016039:Corneal Transplantation; D004195:Disease Models, Animal; D006801:Humans; D011817:Rabbits; D012967:Sodium Dodecyl Sulfate / Q000494:pharmacology; D023822:Tissue Engineering / Q000379:methods*; D020858:Tissue and Organ Harvesting; D014184:Transplantation, Homologous
chemical_list: D012967:Sodium Dodecyl Sulfate