![Exposure to cold induced painful VOC in nonhyperalgesic HbSS mice. (A) Mechanical hyperalgesia appeared in HbSS(nh), but not in HbAA mice, 1 hour after exposure to cold and persisted for at least 24 hours. ∗Different from BL at P = .002 (F[3,40] = 3916) and #different from HbAA mice at P = .002 (F[1,40] = 18.1), 2-way repeated-measures ANOVA with Bonferroni t test, n = 5 to 7 mice/group. (B) Unlike HbAA mice, HbSS(nh) mice exhibited heat hyperalgesia, which began 1 hour after exposure to cold and disappeared by 24 hours. ∗Different from BL at P = .004 (F[4,44] = 4.57) and #different from HbAA mice at P = .002 (F[1,44] = 15.5, 2-way repeated-measures ANOVA with Bonferroni t test, n = 6 to 7 mice/group). (C) Exposure to cold caused prolonged deep tissue hyperalgesia in both sexes of HbSS(nh) mice. Because means of grip force were lower in female mice compared with male mice in both groups (†different from female of the same genotype at P < .05, Kruskal–Wallis ANOVA on ranks test with Dunn's method), the data for each sex were analyzed separately. ∗Different from BL in HbSS(nh) mice of both sexes at P < .001 (F[4,132] = 15.2) and #different from HbAA mice in both sexes at P < .001 (F[3,132] = 18.2), 2-way repeated-measures ANOVA with Bonferroni t test, n = 6 to 12 mice/group. (D) Cold-induced hyperalgesia was accompanied by an increase in heart rate in HbSS mice (F[1,44] = 10.5, P = .008, 2-way repeated-measures ANOVA). HbAA mice had no changes in heart rate (P = 1.0). ∗Different from BL at P < .01 and #different from HbAA mice at P < .01, Bonferroni t test, n = 6 to 7 mice/group. (E) Exposure to cold produced microvascular stasis that was greater in HbSS(nh) mice. ∗Different from HbAA mice at P < .001 (F[1,24] = 240.4), 2-way repeated-measures ANOVA with Bonferroni t test, n = 4 mice/group. (F) Exposure to cold caused transient hypoxia in HbSS(nh), but not HbAA, mice. Hypoxia (hypoxemia) was defined as a decrease in SpO2 in blood. ∗Different from BL at P < .001 (F[4,44] = 9.39) and #different from HbAA mice at P = .016 (F[1,44] = 8.00), 2-way repeated-measures ANOVA with Bonferroni t test, n = 6 to 7 mice/group. (G) Exposure to cold did not produce hypothermia in HbSS or HbAA mice at any time (F[4,44] = 0.177, P = .949, 2-way repeated-measures ANOVA with Bonferroni t test, n = 6 to 7 mice/group).](https://ash.silverchair-cdn.com/ash/content_public/journal/blood/140/16/10.1182_blood.2022017309/13/blood_bld-2022-017309-gr1.jpeg?Expires=1735514542&Signature=UH8g4WMB6H9qW04rj3n0QUmJsPVxPjl0xv6-E9~83nSWVsh5qmx3-rSyeciW5FIWua3CGPZXkTE9CHg3Jm4wuB5hgMu8JOU6MBRbPjc0uCkjuQmAAnTJO-C9rfePjmPkVxcyE2wWj7bZPEuCk3WJGtRhMivKQsTLLc-OKWQcKMYmH3GlataTSoLOdMLarW9MDP8G-Q8XNIvBkpb5uo01A2bp0Wqhr5S3QbzcjAB~Wf2iSntdrvSOlVbf20zdS~brHTUTP4eYpZ7DegSCpm4Az7F9soi1CvbUI~v7JwZTPHfG3E~3DfK4o2wjhFwPzqIS~Bl0CWvMjeUh03pgi8IpTg__&Key-Pair-Id=APKAIE5G5CRDK6RD3PGA)
Exposure to cold induced painful VOC in nonhyperalgesic HbSS mice. (A) Mechanical hyperalgesia appeared in HbSS(nh), but not in HbAA mice, 1 hour after exposure to cold and persisted for at least 24 hours. ∗Different from BL at P = .002 (F[3,40] = 3916) and #different from HbAA mice at P = .002 (F[1,40] = 18.1), 2-way repeated-measures ANOVA with Bonferroni t test, n = 5 to 7 mice/group. (B) Unlike HbAA mice, HbSS(nh) mice exhibited heat hyperalgesia, which began 1 hour after exposure to cold and disappeared by 24 hours. ∗Different from BL at P = .004 (F[4,44] = 4.57) and #different from HbAA mice at P = .002 (F[1,44] = 15.5, 2-way repeated-measures ANOVA with Bonferroni t test, n = 6 to 7 mice/group). (C) Exposure to cold caused prolonged deep tissue hyperalgesia in both sexes of HbSS(nh) mice. Because means of grip force were lower in female mice compared with male mice in both groups (†different from female of the same genotype at P < .05, Kruskal–Wallis ANOVA on ranks test with Dunn's method), the data for each sex were analyzed separately. ∗Different from BL in HbSS(nh) mice of both sexes at P < .001 (F[4,132] = 15.2) and #different from HbAA mice in both sexes at P < .001 (F[3,132] = 18.2), 2-way repeated-measures ANOVA with Bonferroni t test, n = 6 to 12 mice/group. (D) Cold-induced hyperalgesia was accompanied by an increase in heart rate in HbSS mice (F[1,44] = 10.5, P = .008, 2-way repeated-measures ANOVA). HbAA mice had no changes in heart rate (P = 1.0). ∗Different from BL at P < .01 and #different from HbAA mice at P < .01, Bonferroni t test, n = 6 to 7 mice/group. (E) Exposure to cold produced microvascular stasis that was greater in HbSS(nh) mice. ∗Different from HbAA mice at P < .001 (F[1,24] = 240.4), 2-way repeated-measures ANOVA with Bonferroni t test, n = 4 mice/group. (F) Exposure to cold caused transient hypoxia in HbSS(nh), but not HbAA, mice. Hypoxia (hypoxemia) was defined as a decrease in SpO2 in blood. ∗Different from BL at P < .001 (F[4,44] = 9.39) and #different from HbAA mice at P = .016 (F[1,44] = 8.00), 2-way repeated-measures ANOVA with Bonferroni t test, n = 6 to 7 mice/group. (G) Exposure to cold did not produce hypothermia in HbSS or HbAA mice at any time (F[4,44] = 0.177, P = .949, 2-way repeated-measures ANOVA with Bonferroni t test, n = 6 to 7 mice/group).
