Cartilage tissue engineering for auricular reconstruction: in vitro evaluation of potential genotoxic and cytotoxic effects of scaffold materials.
Tissue engineering of autologous cartilage transplants is suggested as a new approach in reconstruction of external auricular deformities. 1.6-Hexanediol (HD), 1.8-diazabicyclo[5.4.0]undec-7-ene (DBU) and 6-hydroxyhexanoic acid (HHA) are matrices of the open-pored polyurethane three-dimensional scaffold. Since these bioresorbable materials may interact with the human organism, cytotoxic effects on human chondrocytes and lymphocytes and genotoxic effects on human lymphocytes were monitored. Staining with propidium iodide and fluorescence diacetate as well as the EZ4U proliferation assay served for the detection of cytotoxic effects of the materials on human chondrocytes. Trypan blue staining was used to monitor cytotoxicity on lymphocytes. Genotoxic effects on lymphocytes in terms of strand breaks, alkali labile sites and incomplete excision repair were determined by the alkaline single cell microgel electrophoresis (Comet) assay. Cytotoxic effects in chondrocytes and lymphocytes as well as genotoxic effects in lymphocytes were dose-dependent with threshold values of 5mg/mL HD, 0.5mg/mL DBU and 0.03 mg/mL HHA showing no effects. These data suggest that these matrices could be safely used for scaffolds made of polyurethane unless these compounds are not released at a rate giving higher concentrations at the site of implantation or in body fluids, respectively.
Havla
Joachim B
JB
Department of Toxicology, Walther Straub Institute, Ludwig-Maximilians University, Munich, Germany.
Lotz
Amelie S
AS
Richter
Elmar
E
Froelich
Katrin
K
Hagen
Rudolf
R
Staudenmaier
Rainer
R
Kleinsasser
Norbert H
NH
D019086:Bridged Bicyclo Compounds, Heterocyclic / Q000633:toxicity*; D002208:Caproates / Q000633:toxicity*; D002358:Cartilage, Articular; D002470:Cell Survival; D019902:Chondrocytes / Q000187:drug effects; D020552:Comet Assay; D004305:Dose-Response Relationship, Drug; D005260:Female; D006018:Glycols / Q000633:toxicity*; D006801:Humans; D006880:Hydroxy Acids; D008214:Lymphocytes / Q000187:drug effects; D008297:Male; D009153:Mutagens; D023822:Tissue Engineering / Q000009:adverse effects*; D054457:Tissue Scaffolds / Q000009:adverse effects*
D019086:Bridged Bicyclo Compounds, Heterocyclic; D002208:Caproates; D006018:Glycols; D006880:Hydroxy Acids; D009153:Mutagens; C039565:6-hydroxyhexanoic acid; C031033:1,8-diazabicyclo(5.4.0)undec-7-ene; C027765:hexamethylene glycol