The lethal heat dose for 50% primary human fibroblast cell death is 48 °C.
Burn
Fibroblasts
Hyperthermia
Pathogenesis
Thermal
Journal
Archives of dermatological research
ISSN: 1432-069X
Titre abrégé: Arch Dermatol Res
Pays: Germany
ID NLM: 8000462
Informations de publication
Date de publication:
Oct 2022
Oct 2022
Historique:
received:
17
09
2019
accepted:
18
03
2021
revised:
11
02
2021
pubmed:
29
3
2021
medline:
26
8
2022
entrez:
28
3
2021
Statut:
ppublish
Résumé
Understanding the effect of heat on skin cells is important for the prevention of burn injury. Knowledge of the heat dose required to kill cells can be used to study the cellular mechanisms involved in thermal injury cell death, to assist with the development of novel burn treatments. In this study, primary human skin dermal fibroblasts were exposed to temperatures from 37 to 54 °C for 1 h and the relative cell viability of heat-treated and control cells was assessed. Cell damage and viability were assessed by light microscopy, MTT assay and live/dead staining. The LD50 for 1 h of heat exposure was 48 °C for primary fibroblasts; and there was evidence that thermal damage to cells begins to occur at 43 °C. This study presents a reproducible method for examining the effect of heat on primary human cells grown in culture on a cellular level and can be used in the future to study the mechanisms behind heat-induced cell death, to inform burn injury prevention efforts and effective post-burn treatment.
Identifiants
pubmed: 33774732
doi: 10.1007/s00403-021-02217-y
pii: 10.1007/s00403-021-02217-y
doi:
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
809-814Subventions
Organisme : National Health and Medical Research Council
ID : APP1130862
Informations de copyright
© 2021. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.
Références
Ahmed K, Zaidi SF, Mati-ur-Rehman RR, Kondo T (2020) Hyperthermia and protein homeostasis: cytoprotection and cell death. J Therm Biol. https://doi.org/10.1016/j.jtherbio.2020.102615
doi: 10.1016/j.jtherbio.2020.102615
pubmed: 33454039
Birch HL, Wilson AM, Goodship AE (1997) The effect of exercise-induced localised hyperthermia on tendon cell survival. J Exp Biol 200:1703–1708
doi: 10.1242/jeb.200.11.1703
Boonkaew B, Kempf M, Kimble R, Cuttle L (2014) Cytotoxicity testing of silver-containing burn treatments using primary and immortal skin cells. Burns 40:1562–1569. https://doi.org/10.1016/j.burns.2014.02.009
doi: 10.1016/j.burns.2014.02.009
pubmed: 24767717
Chaabane W, User SD, El-Gazzah M, Jaksik R, Sajjadi E, Rzeszowska-Wolny J, Los MJ (2013) Autophagy, apoptosis, mitoptosis and necrosis: interdependence between those pathways and effects on cancer. Arch Immunol Ther Exp 61:43–58. https://doi.org/10.1007/s00005-012-0205-y
doi: 10.1007/s00005-012-0205-y
Dokladny K, Myers OB, Moseley PL (2015) Heat shock response and autophagy-cooperation and control. Autophagy 11:200–213. https://doi.org/10.1080/15548627.2015.1009776
doi: 10.1080/15548627.2015.1009776
pubmed: 25714619
pmcid: 4502786
Fajardo LF (1984) Pathological effects of hyperthermia in normal tissues. Cancer Res 44:4826s–4835s
pubmed: 6467235
Fernandes ACM, de Franca JP, Gaiba S, Aloise AC, de Oliveira AF, Moraes AADS, de Franca LP, Ferreira LM (2014) Development of experimental in vitro burn model. Acta Cir Bras 29:15–20. https://doi.org/10.1590/S0102-86502014001400004
doi: 10.1590/S0102-86502014001400004
pubmed: 25229509
Gerweck LE (1985) Hyperthermia in cancer therapy: the biological basis and unresolved questions. Cancer Res 45:3408–3414
pubmed: 3893686
Gurtner GC, Werner S, Barrandon Y, Longaker MT (2008) Wound repair and regeneration. Nature 453:314. https://doi.org/10.1038/nature07039
doi: 10.1038/nature07039
pubmed: 18480812
Harmon BV, Corder AM, Collins RJ, Gobe GC, Allen J, Allan DJ, Kerr JF (1990) Cell death induced in a murine mastocytoma by 42–47 degrees C heating in vitro: evidence that the form of death changes from apoptosis to necrosis above a critical heat load. Int J Radiat Biol 58:845–858. https://doi.org/10.1080/09553009014552221
doi: 10.1080/09553009014552221
pubmed: 1977828
Harmon BV, Takano YS, Winterford CM, Gobe GC (1991) The role of apoptosis in the response of cells and tumours to mild hyperthermia. Int J Radiat Biol 59:489–501. https://doi.org/10.1080/09553009114550441
doi: 10.1080/09553009114550441
pubmed: 1671698
Ibtisham F, Zhao Y, Nawab A, Liguang H, Wu J, Xiao M, Zhao Z, An L (2018) The effect of high temperature on viability, proliferation, apoptosis and anti-oxidant status of chicken embryonic fibroblast cells. Braz J Poult Sci 20:463–470. https://doi.org/10.1590/1806-9061-2017-0685
doi: 10.1590/1806-9061-2017-0685
Lepock JR (2005) How do cells respond to their thermal environment? Int J Hyperth 21:681–687. https://doi.org/10.1080/02656730500307298
doi: 10.1080/02656730500307298
Lin PS, Butterfield CE (1977) Hyperthermic treatment (43 degrees C) rapidly impedes attachment of fibroblasts to culture substrates. Cell Biol Int Rep 1:57–61
doi: 10.1016/0309-1651(77)90010-8
Lu L, Zhang L, Wai MS, Yew DT, Xu J (2012) Exocytosis of MTT formazan could exacerbate cell injury. Toxicol Vitro 26:636–644. https://doi.org/10.1016/j.tiv.2012.02.006
doi: 10.1016/j.tiv.2012.02.006
Matsuura Y, Noda K, Suzuki S, Kawai K (2019) Glucocorticoids suppress fibroblast apoptosis in an in vitro thermal injury model. Burns 45:173–179. https://doi.org/10.1016/j.burns.2018.08.002
doi: 10.1016/j.burns.2018.08.002
pubmed: 30253958
Muschter D, Geyer F, Bauer R, Ettl T, Schreml S, Haubner F (2018) A comparison of cell survival and heat shock protein expression after radiation in normal dermal fibroblasts, microvascular endothelial cells, and different head and neck squamous carcinoma cell lines. Clin Oral Investig 22:2251–2262. https://doi.org/10.1007/s00784-017-2323-8
doi: 10.1007/s00784-017-2323-8
pubmed: 29307045
Nakashima A, Cheng SB, Kusabiraki T, Motomura K, Aoki A, Ushijima A, Ono Y, Tsuda S, Shima T, Yoshino O, Sago H, Matsumoto K, Sharma S, Saito S (2019) Endoplasmic reticulum stress disrupts lysosomal homeostasis and induces blockade of autophagic flux in human trophoblasts. Sci Rep 9:11466. https://doi.org/10.1038/s41598-019-47607-5
doi: 10.1038/s41598-019-47607-5
pubmed: 31391477
pmcid: 6685987
O’Neill KL, Fairbairn DW, Smith MJ, Poe BS (1998) Critical parameters influencing hyperthermia-induced apoptosis in human lymphoid cell lines. Apoptosis 3:369–375
doi: 10.1023/A:1009689407261
Ohnishi K, Scuric Z, Yau D, Schiestl RH, Okamoto N, Takahashi A, Ohnishi T (2006) Heat-induced phosphorylation of NBS1 in human skin fibroblast cells. J Cell Biochem 99:1642–1650. https://doi.org/10.1002/jcb.20995
doi: 10.1002/jcb.20995
pubmed: 16823774
Purschke M, Laubach HJ, Anderson RR, Manstein D (2010) Thermal injury causes DNA damage and lethality in unheated surrounding cells: active thermal bystander effect. J Investig Dermatol 130:86–92. https://doi.org/10.1038/jid.2009.205
doi: 10.1038/jid.2009.205
pubmed: 19587691
Rueden CT, Schindelin J, Hiner MC, DeZonia BE, Walter AE, Arena ET, Eliceiri KW (2017) Image J2: ImageJ for the next generation of scientific image data. BMC Bioinform. https://doi.org/10.1186/s12859-017-1934-z
doi: 10.1186/s12859-017-1934-z
Shellman YG, Howe WR, Miller LA, Goldstein NB, Pacheco TR, Mahajan RL, LaRue SM, Norris DA (2008) Hyperthermia induces endoplasmic reticulum-mediated apoptosis in melanoma and non-melanoma skin cancer cells. J Investig Dermatol 128:949–956. https://doi.org/10.1038/sj.jid.5701114
doi: 10.1038/sj.jid.5701114
pubmed: 17989736
Siddiqui SH, Subramaniyan SA, Kang DR, Park J, Khan M, Choi HW, Shim K (2020) Direct exposure to mild heat stress stimulates cell viability and heat shock protein expression in primary cultured broiler fibroblasts. Cell Stress Chaperon. https://doi.org/10.1007/s12192-020-01140-x
doi: 10.1007/s12192-020-01140-x
Takano YS, Harmon BV, Kerr JFR (1991) Apoptosis induced by mild hyperthermia in human and murine tumor-cell lines - a study using electron-microscopy and DNA gel-electrophoresis. J Pathol 163:329–336. https://doi.org/10.1002/path.1711630410
doi: 10.1002/path.1711630410
pubmed: 1903443
Ter Horst B, Chouhan G, Moiemen NS, Grover LM (2018) Advances in keratinocyte delivery in burn wound care. Adv Drug Deliv Rev 123:18–32. https://doi.org/10.1016/j.addr.2017.06.012
doi: 10.1016/j.addr.2017.06.012
pubmed: 28668483
pmcid: 5764224
van Meerloo J, Kaspers GJ, Cloos J (2011) Cell sensitivity assays: the MTT assay. Methods Mol Biol 731:237–245. https://doi.org/10.1007/978-1-61779-080-5_20
doi: 10.1007/978-1-61779-080-5_20
pubmed: 21516412
Wang JF, Jiao H, Stewart TL, Shankowsky HA, Scott PG, Tredget EE (2007) Fibrocytes from burn patients regulate the activities of fibroblasts. Wound Repair Regen 15:113–121. https://doi.org/10.1111/j.1524-475X.2006.00192.x
doi: 10.1111/j.1524-475X.2006.00192.x
pubmed: 17244327
Zhang T, Hu HQ, Tao Z, Niu B, Jiao SS, Zhang J, Li YY, Cao BZ (2016) A novel method for primary neuronal culture and characterization under different high temperature. Vitro Cell Dev Biol Anim 52:823–828. https://doi.org/10.1007/s11626-016-0047-8
doi: 10.1007/s11626-016-0047-8