Effects of temperature and doxorubicin exposure on keratinocyte damage in vitro

Abstract

Cancer chemotherapy treatment often leads to hair loss, which may be prevented by cooling the scalp during drug administration. The current hypothesis for the hair preservative effect of scalp cooling is that cooling of the scalp skin reduces blood flow (perfusion) and chemical reaction rates. Reduced perfusion leads to less drugs available for uptake, whereas the reduced temperature decreases uptake of and damage by chemotherapy. Altogether, less damage is exerted to the hair cells, and the hair is preserved. However, the two mechanisms in the hypothesis have not been quantified yet. To quantify the effect of reduced drug damage caused by falling temperatures, we investigated the effect of local drug concentration and local tissue temperature on hair cell damage using in vitro experiments on keratinocytes. Cells were exposed for 4 h to a wide range of doxorubicin concentrations. During exposure, cells were kept at different temperatures. Cell viability was determined after 3 d using a viability test. Control samples were used to establish a concentrationviability curve. Results show that cell survival is significantly higher in cooled cells (T<22°C) than in non-cooled cells (T=37°C), but no significant differences are visible between T=10°C and T=22°C. Based on this result and previous work, we can conclude that there is an optimal temperature in scalp cooling. Further cooling will only result in unnecessary discomfort for the patient and should therefore be avoided.

Use of in vitro human keratinocyte models to study the effect of cooling on chemotherapy drug-induced cytotoxicity

“…1, upper panels), with the observation for doxorubicin being in agreement with previous studies (Janssen et al, 2008). The ability of cooling to protect from cytotoxicity was striking for several, mid-range drug concentrations, particularly for doxorubicin and 4-OH-CP; in fact for some drug doses cell biomass returned from below 20% to nearly 100% of the control value (Fig.…”

“…This was primarily because it has been suggested by various studies that a scalp temperature of 22 °C or less is required for hair preservation (Komen et al, 2013) and also it was the temperature previously tested in vitro by Janssen et al (2008). As shown in Fig.…”

“…It has previously been reported that cooling below 22 °C did not provide any further protection against doxorubicin-mediated keratinocyte cytotoxicity, even when the culture temperature was reduced to 10 °C during drug treatment (Janssen et al, 2008). Using HaCaTa cells, we tested whether temperature values below 22 °C could provide further protection against doxorubicin, 4-OH-CP and docetaxel.…”

“…Only a limited number of studies have previously addressed the effect of chemotherapy drugs on normal human epidermal keratinocytes (NHEKs) and a single study assessed the effect of cooling on doxorubicin-mediated cytotoxicity on NHEKs (Janssen et al, 2008). Using NHEKs cultured in low calcium (0.09 mM) serum-free medium, conditions under which they adopt a highly-proliferative basal cell phenotype, we assessed the cytotoxic effect of a panel of three routinely used chemotherapy drugs (Grevelman and Breed, 2005), in particular the taxane docetaxel, the anthracycline doxorubicin and the alkylating agent cyclophosphamide, all of which are associated with CIA.…”

“…Ex vivo models, such as those by Paus and colleagues that are based on isolation and culture of human hair follicles, represent an elegant model for studying cyclophosphamide-induced CIA (Bodo et al, 2007). A more reductive culture model to study chemotherapy drug-induced cytotoxicity involves the use of human neonatal epidermal keratinocytes and a previous report has provided some, though limited, evidence for an effect of culture temperature on the cytotoxicity of doxorubicin on such cells (Janssen et al, 2008). The principle behind the use of human epidermal keratinocytes is that they are maintained under culture conditions that render them highly-proliferative, thus resembling the rapidly-dividing population of native matrix keratinocytes.…”