title: Effects of Detergent-Based Protocols on Decellularization of Corneas With Sclerocorneal Limbus. Evaluation of Regional Differences.
abstract: OBJECTIVE
  In this work, we decellularized whole porcine corneas including the sclerocorneal limbus (SCL) and we evaluated regional differences in order to identify an efficient method to decellularize whole corneas for future clinical use.
  
  
  METHODS
  We analyzed the efficiency of four decellularization protocols based on benzalkonium chloride (BAK), Igepal, sodium dodecyl sulfate (SDS), and Triton X-100 detergents on whole porcine corneas.
  
  
  RESULTS
  Results showed that the decellularization efficiency of most protocols was low, with the specific protocol resulting in more efficient levels of decellularization being 0.1% SDS for 48 hours, especially in the medium and posterior cornea regions. A significant correlation was found between the decellularization efficiency and the concentration of agent used (P= 0.0174;r= 0.1540), but not for the incubation time (P> 0.05). The analysis of cornea components preservation demonstrated that all protocols were able to preserve the integrity of the Bowman's layer and Descemet's membrane. Although the collagen structure was partially altered, the global decellularization groups showing highest preservation of the ECM collagen contents and orientation were Igepal and SDS, with differences among the three regions of the cornea. All global groups showed high levels of proteoglycan and glycoprotein preservation after decellularization, with the best results were found in the SDS group followed by the Igepal group.
  
  
  CONCLUSIONS
  These results suggest that very powerful protocols are necessary for whole-cornea decellularization. For the generation of lamelar corneas for clinical use, decellularization regional differences should be taken into account.
  
  
  CONCLUSIONS
  Decellularized whole corneas may be potential therapeutic agents for lamelar keratoplasty.
authors.affiliation: Tissue Engineering Group, Department of Histology, University of Granada, Spain and Instituto de Investigación Biosanitaria ibs.GRANADA, Spain ; Division of Ophthalmology, University Hospital San Cecilio, Granada, Spain.
authors.forename: Miguel
authors.initials: M
authors.lastname: González-Andrades
authors.affiliation: Tissue Engineering Group, Department of Histology, University of Granada, Spain and Instituto de Investigación Biosanitaria ibs.GRANADA, Spain.
authors.forename: Victor
authors.initials: V
authors.lastname: Carriel
authors.affiliation: Tissue Engineering Group, Department of Histology, University of Granada, Spain and Instituto de Investigación Biosanitaria ibs.GRANADA, Spain.
authors.forename: Mario
authors.initials: M
authors.lastname: Rivera-Izquierdo
authors.affiliation: Tissue Engineering Group, Department of Histology, University of Granada, Spain and Instituto de Investigación Biosanitaria ibs.GRANADA, Spain.
authors.forename: Ingrid
authors.initials: I
authors.lastname: Garzón
authors.affiliation: Tissue Engineering Group, Department of Histology, University of Granada, Spain and Instituto de Investigación Biosanitaria ibs.GRANADA, Spain.
authors.forename: Elena
authors.initials: E
authors.lastname: González-Andrades
authors.affiliation: Division of Ophthalmology, University Hospital Virgen de las Nieves, Granada, Spain.
authors.forename: Santiago
authors.initials: S
authors.lastname: Medialdea
authors.affiliation: Tissue Engineering Group, Department of Histology, University of Granada, Spain and Instituto de Investigación Biosanitaria ibs.GRANADA, Spain.
authors.forename: Miguel
authors.initials: M
authors.lastname: Alaminos
authors.affiliation: Tissue Engineering Group, Department of Histology, University of Granada, Spain and Instituto de Investigación Biosanitaria ibs.GRANADA, Spain.
authors.forename: Antonio
authors.initials: A
authors.lastname: Campos
keywords: cornea regions; decellularization protocols; extracellular matrix; sclerocorneal limbus; tissue engineering