TY - JOUR
T1 - Identification of cornifelin and early growth response-1 gene as novel biomarkers for in vitro eye irritation using a 3D reconstructed human cornea model MCTT HCE™
AU - Choi, Seunghye
AU - Lee, Miri
AU - Lee, Su Hyon
AU - Jung, Haeng Sun
AU - Kim, Seol Yeong
AU - Chung, Tae Young
AU - Choe, Tae boo
AU - Chun, Young Jin
AU - Lim, Kyung Min
N1 - Funding Information:
This work is supported by Ministry of Food and Drug Safety of Korea, Grant No. 13172MFDS987 and the National Research Foundation (NRF) Grant of the Korea government (MEST), No. NRF-2013R1A1A1007072.
Publisher Copyright:
© 2014, Springer-Verlag Berlin Heidelberg.
PY - 2015/10/1
Y1 - 2015/10/1
N2 - Evaluation of the eye irritation is essential in the development of new cosmetic products. Draize rabbit eye irritation test has been widely used in which chemicals are directly applied to rabbit eye, and the symptoms and signs of eyes are scored. However, due to the invasive procedure, it causes substantial pain and discomfort to animals. Recently, we reported in vitro eye irritation test method using a 3D human corneal epithelial model (MCTT HCE™) which is reconstructed from remaining human tissues after a corneal transplantation. This model exhibited an excellent predictive capacity for 25 reference chemicals (sensitivity 100 %, specificity 77 % and accuracy 88 % vs. GHS). To improve the test performance, we explored new biomarkers for the eye irritation through transcriptomic approach. Three surfactants were selected as model eye irritants that include sodium lauryl sulfate, benzalkonium chloride and triton X-100. After test chemicals were treated, we investigated differentially expressed genes through a whole-gene microarray (Affymetrix GeneChip® Human Gene 2.0 ST Array, 48,000 probes). As a result, we identified that mRNAs of cornifelin (CNFN), a constituent of the insoluble cornified cell envelope of stratified squamous epithelia, and early growth response-1 (EGR1), a nuclear transcriptional regulator, were significantly up-regulated by all three irritants. Up-regulation of CNFN and EGR1 was further confirmed by Q-RT-PCR, and immunohistochemistry revealed increased level of CNFN in irritant-treated tissues, supporting the relevance of CNFN and EGR1 as new biomarkers for eye irritation.
AB - Evaluation of the eye irritation is essential in the development of new cosmetic products. Draize rabbit eye irritation test has been widely used in which chemicals are directly applied to rabbit eye, and the symptoms and signs of eyes are scored. However, due to the invasive procedure, it causes substantial pain and discomfort to animals. Recently, we reported in vitro eye irritation test method using a 3D human corneal epithelial model (MCTT HCE™) which is reconstructed from remaining human tissues after a corneal transplantation. This model exhibited an excellent predictive capacity for 25 reference chemicals (sensitivity 100 %, specificity 77 % and accuracy 88 % vs. GHS). To improve the test performance, we explored new biomarkers for the eye irritation through transcriptomic approach. Three surfactants were selected as model eye irritants that include sodium lauryl sulfate, benzalkonium chloride and triton X-100. After test chemicals were treated, we investigated differentially expressed genes through a whole-gene microarray (Affymetrix GeneChip® Human Gene 2.0 ST Array, 48,000 probes). As a result, we identified that mRNAs of cornifelin (CNFN), a constituent of the insoluble cornified cell envelope of stratified squamous epithelia, and early growth response-1 (EGR1), a nuclear transcriptional regulator, were significantly up-regulated by all three irritants. Up-regulation of CNFN and EGR1 was further confirmed by Q-RT-PCR, and immunohistochemistry revealed increased level of CNFN in irritant-treated tissues, supporting the relevance of CNFN and EGR1 as new biomarkers for eye irritation.
KW - Alternative to animal test methods
KW - Cornifelin
KW - Early growth response-1 gene
KW - Eye irritation
KW - Transcriptomics
UR - http://www.scopus.com/inward/record.url?scp=84940607710&partnerID=8YFLogxK
U2 - 10.1007/s00204-014-1390-8
DO - 10.1007/s00204-014-1390-8
M3 - Article
C2 - 25377654
AN - SCOPUS:84940607710
VL - 89
SP - 1589
EP - 1598
JO - Archives of Toxicology
JF - Archives of Toxicology
SN - 0340-5761
IS - 9
ER -