title: In vitro human skin irritation test for evaluation of medical device extracts.
abstract: The aim of this study was to determine if the EpiDerm™ reconstructed human skin model (MatTek Corp.) could be an acceptable alternative to the ISO 10993-required rabbit skin irritation test for assessing medical device biocompatibility. Eleven medical device polymers were tested. Four extracts were prepared per polymer, two each with saline and sesame oil; half were spiked with two R-38 irritants, lactic acid for saline extracts and heptanoic acid for the sesame oil extracts. Tissue viability was assessed by MTT reduction and the proinflammatory response was assessed by IL-1α release. LOAELs of 2% for lactic acid in saline and 0.7% for heptanoic acid in sesame oil were determined. A cell viability reduction of >50% was indicative of skin irritation. Cells exposed to saline extracts spiked with 3.25% lactic acid had significantly reduced mean cell viabilities (12.6-17.2%). Cells exposed to sesame oil extracts spiked with 1.25% heptanoic acid also exhibited reduced mean cell viabilities (25.5%-41.7%). All spiked cells released substantial amounts of IL-1α (253.5-387.4pg/ml) signifying a proinflammatory response. These results indicate that the EpiDerm™ model may be a suitable in vitro replacement for the assessment of the irritation potential of medical device extracts.
authors.affiliation: Medtronic, Inc., 11520 Yellow Pine St. N. W., C202, Minneapolis, MN 55448, USA. Electronic address: jesus.w.casas@medtronic.com.
authors.forename: J W
authors.initials: JW
authors.lastname: Casas
authors.forename: G M
authors.initials: GM
authors.lastname: Lewerenz
authors.forename: E A
authors.initials: EA
authors.lastname: Rankin
authors.forename: J A
authors.initials: JA
authors.lastname: Willoughby
authors.forename: L C
authors.initials: LC
authors.lastname: Blakeman
authors.forename: J M
authors.initials: JM
authors.lastname: McKim
authors.forename: K P
authors.initials: KP
authors.lastname: Coleman
mesh_terms: D000269:Adhesives / Q000737:chemistry*; D000826:Animal Testing Alternatives; D002470:Cell Survival / Q000187:drug effects; D045424:Complex Mixtures / Q000633:toxicity*; D004864:Equipment and Supplies; D006244:Hardness; D006801:Humans; D066298:In Vitro Techniques; D053582:Interleukin-1alpha / Q000378:metabolism; D011108:Polymers / Q000737:chemistry*; D012715:Sesame Oil / Q000737:chemistry; D023422:Skin Irritancy Tests; D012965:Sodium Chloride / Q000737:chemistry
chemical_list: D000269:Adhesives; D045424:Complex Mixtures; D053582:Interleukin-1alpha; D011108:Polymers; D012965:Sodium Chloride; D012715:Sesame Oil
keywords: 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide; AST; EC(50); ECVAM; ECVAM’s Science Advisory Committee; EIA; ELISA; ESAC; European Centre for Validation of Alternative Methods; German National Centre for the Documentation and Evaluation of Alternatives to Animal Experiments; ISO; In vitro; International Organization for Standardization; LDH; LOAEL; MTT; Medical device; NHEK; NOAEL; National Institute of Public Health and the Environment, Netherlands; PBS; REACH; RIVM; Registration, Evaluation, Authorisation and Restriction of Chemicals; RhE; Skin irritation; ZEBET; alternative skin test; effective concentration 50; enzyme immunoassay; enzyme-linked immunosorbent assay; lactate dehydrogenase; lowest observable adverse effect level; no observed adverse effect level; normal, human-derived epidermal keratinocytes; phosphate buffered saline; reconstructed human epidermis