González-Andrades, MiguelCarriel, VictorRivera-Izquierdo, MarioGarzón, IngridGonzález-Andrades, ElenaMedialdea, SantiagoAlaminos, MiguelCampos, Antonio2016-08-082016-08-082015-04-10González-Andrades M, Carriel V, Rivera-Izquierdo M, Garzón I, González-Andrades E, Medialdea S, et al. Effects of Detergent-Based Protocols on Decellularization of Corneas With Sclerocorneal Limbus. Evaluation of Regional Differences. Transl Vis Sci Technol. 2015 ; 4(2):13http://hdl.handle.net/10668/2308JOURNAL ARTICLE;PURPOSE 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. TRANSLATIONAL RELEVANCE Decellularized whole corneas may be potential therapeutic agents for lamelar keratoplasty.enCornea regionsDecellularization protocolsTissue engineeringSclerocorneal limbusExtracellular matrixAnimalesCompuestos de benzalconioLámina limitante anteriorColágenoCórneaTrasplante de córneaLámina limitante posteriorDetergentesLimbo de la córneaOctoxinolProteoglicanosSodioDodecil sulfato de sodioPorcinosMedical Subject Headings::Organisms::Eukaryota::AnimalsMedical Subject Headings::Chemicals and Drugs::Organic Chemicals::Amines::Quaternary Ammonium Compounds::Benzylammonium CompoundsMedical Subject Headings::Anatomy::Sense Organs::Eye::Anterior Eye Segment::Cornea::Bowman MembraneMedical Subject Headings::Chemicals and Drugs::Macromolecular Substances::Polymers::Biopolymers::CollagenMedical Subject Headings::Anatomy::Sense Organs::Eye::Anterior Eye Segment::CorneaMedical Subject Headings::Analytical, Diagnostic and Therapeutic Techniques and Equipment::Surgical Procedures, Operative::Ophthalmologic Surgical Procedures::Refractive Surgical Procedures::Corneal TransplantationMedical Subject Headings::Anatomy::Sense Organs::Eye::Anterior Eye Segment::Cornea::Descemet MembraneMedical Subject Headings::Chemicals and Drugs::Chemical Actions and Uses::Specialty Uses of Chemicals::Surface-Active Agents::DetergentsMedical Subject Headings::Anatomy::Sense Organs::Eye::Anterior Eye Segment::Cornea::Limbus CorneaeMedical Subject Headings::Chemicals and Drugs::Organic Chemicals::Alcohols::Glycols::Ethylene Glycols::Polyethylene Glycols::OctoxynolMedical Subject Headings::Chemicals and Drugs::Carbohydrates::Polysaccharides::ProteoglycansMedical Subject Headings::Chemicals and Drugs::Inorganic Chemicals::Elements::Metals, Alkali::SodiumMedical Subject Headings::Organisms::Eukaryota::Animals::Chordata::Vertebrates::Mammals::Artiodactyla::SwineMedical Subject Headings::Chemicals and Drugs::Organic Chemicals::Alcohols::Fatty Alcohols::Dodecanol::Sodium Dodecyl Sulfateresearch article25909036open access10.1167/tvst.4.2.132164-2591PMC4404970