title: Cytotoxicity of HBD3 for dendritic cells, normal human epidermal keratinocytes, hTERT keratinocytes, and primary oral gingival epithelial keratinocytes in cell culture conditions.
abstract: Human β-defensin 3 (HBD3) is a prominent host defense peptide. In our recent work, we observed that HBD3 modulates pro-inflammatory agonist-induced chemokine and cytokine responses in human myeloid dendritic cells (DCs), often at 20.0 μM concentrations. Since HBD3 can be cytotoxic in some circumstances, it is necessary to assess its cytotoxicity for DCs, normal human epidermal keratinocytes (NHEKs), human telomerase reverse transcriptase (hTERT) keratinocytes, and primary oral gingival epithelial (GE) keratinocytes in different cell culture conditions. Cells, in serum free media with resazurin and in complete media with 10% fetal bovine serum and resazurin, were incubated with 5, 10, 20, and 40 μM HBD3. Cytotoxicity was determined by measuring metabolic conversion of resazurin to resorufin. The lethal dose 50 (LD50, mean μM±Std Err) values were determined from the median fluorescent intensities of test concentrations compared to live and killed cell controls. The LD50 value range of HBD3 was 18.2-35.9 μM in serum-free media for DCs, NHEKs, hTERT keratinocytes, and GE keratinocytes, and >40.0 μM in complete media. Thus, HBD3 was cytotoxic at higher concentrations, which must be considered in future studies of HBD3-modulated chemokine and cytokine responses in vitro.
authors.affiliation: Division of Pharmaceutics and Translational Therapeutics, Department of Pharmaceutical Sciences and Experimental Therapeutics, College of Pharmacy, The University of Iowa, Iowa City, IA 52242, USA. Electronic address: nattawut-leelakanok@uiowa.edu.
authors.forename: Nattawut
authors.initials: N
authors.lastname: Leelakanok
authors.affiliation: Dows Institute for Dental Research, College of Dentistry, The University of Iowa, Iowa City, IA 52242, USA. Electronic address: carol-bratt@uiowa.edu.
authors.forename: Carol L
authors.initials: CL
authors.lastname: Fischer
authors.affiliation: Dows Institute for Dental Research, College of Dentistry, The University of Iowa, Iowa City, IA 52242, USA. Electronic address: amber-bates@uiowa.edu.
authors.forename: Amber M
authors.initials: AM
authors.lastname: Bates
authors.affiliation: College of Dentistry, The University of Nebraska Medical Center, Lincoln, NE 68583, USA. Electronic address: janet.guthmiller@unmc.edu.
authors.forename: Janet M
authors.initials: JM
authors.lastname: Guthmiller
authors.affiliation: Department of Periodontics, College of Dentistry, The University of Iowa, Iowa City, IA 52242, USA. Electronic address: georgia-johnson@uiowa.edu.
authors.forename: Georgia K
authors.initials: GK
authors.lastname: Johnson
authors.affiliation: Division of Pharmaceutics and Translational Therapeutics, Department of Pharmaceutical Sciences and Experimental Therapeutics, College of Pharmacy, The University of Iowa, Iowa City, IA 52242, USA. Electronic address: aliasger-salem@uiowa.edu.
authors.forename: Aliasger K
authors.initials: AK
authors.lastname: Salem
authors.affiliation: Dows Institute for Dental Research, College of Dentistry, The University of Iowa, Iowa City, IA 52242, USA; Department of Periodontics, College of Dentistry, The University of Iowa, Iowa City, IA 52242, USA. Electronic address: kim-brogden@uiowa.edu.
authors.forename: Kim A
authors.initials: KA
authors.lastname: Brogden
authors.affiliation: Division of Pharmaceutics and Translational Therapeutics, Department of Pharmaceutical Sciences and Experimental Therapeutics, College of Pharmacy, The University of Iowa, Iowa City, IA 52242, USA. Electronic address: nicole-brogden@uiowa.edu.
authors.forename: Nicole K
authors.initials: NK
authors.lastname: Brogden
mesh_terms: D002460:Cell Line; D002470:Cell Survival; D002478:Cells, Cultured; D016895:Culture Media, Serum-Free; D003713:Dendritic Cells / Q000187:drug effects*; D004305:Dose-Response Relationship, Drug; D004847:Epithelial Cells / Q000187:drug effects*; D005881:Gingiva / Q000166:cytology; D006801:Humans; D015603:Keratinocytes / Q000187:drug effects*; D007928:Lethal Dose 50; D013997:Time Factors; D023083:beta-Defensins / Q000633:toxicity*
chemical_list: D016895:Culture Media, Serum-Free; C422950:DEFB103A protein, human; D023083:beta-Defensins
keywords: Cytotoxicity; Defensins; Dendritic cells; Epidermal Keratinocytes; Gingival epithelial GE keratinocytes; HBD3; hTERT Keratinocytes