Short-term modulation of pain-related behaviors in wild-type mice and female HbSS-BERK mice. (A) Experimental design for the single-session treatment with behavioral study. (B-C) Short-term inhibition of heat pain sensitivity in wild-type mice. The impact of tFUS on heat pain-associated behaviors was assessed in both (B) male (n = 8) and (C) female (n = 8) wild-type mice. Single-session tFUS with a PRF of 40 Hz at left S1HL resulted in a significant increase in latency difference between contralateral (right) and ipsilateral (left) hind paw compared with baseline and sham-treated mice. ns, not significant, ∗∗P < .01 using t test with Wilcoxon matched–pairs signed rank test; #P < .05 using t test with Mann-Whitney test. (D-F) Short-term modulation of thermal and mechanical hyperalgesia in female HbSS-BERK mice. Quantification of averaged Δ hPWL from hot-plate test, Δ hPWF from cold-plate test, and Δ hPWT from von Frey test was conducted before and after tFUS with control groups (negative control and sham treatment) in HbSS-BERK mice. (D) Single-session tFUS with a PRF of 40 Hz at left S1HL (n = 10) led to a significant increase in the averaged Δ hPWL from hot-plate test compared with baseline, sham treatment (n = 10), and negative control (n = 10). Control groups did not show apparent treatment effect compared with pretreatment baseline. A negative value of Δ hPWL was observed with a PRF of 3 kHz at left S1HL (n = 10), which was significantly different from the baseline and response with the PRF of 40 Hz. (E) Single-session tFUS with PRF of 40 Hz at left S1HL (n = 10) and insula (n = 10) resulted in a significant change in the Δ hPWF from cold-plate test at 8 minutes after treatment compared to the sham treatment and baseline. (F) tFUS with PRF of 40 Hz at left S1HL (n = 10) and insula (n = 10) led to a prominent elevation of Δ hPWT from von Frey test compared with sham-treated (n = 10) and negative control mice (n = 10). A lower withdrawal threshold of the contralateral (right) hind paw was observed with tFUS at PRF of 3 kHz at left S1HL (n = 10) compared with PRF of 40 Hz at S1HL and insula. ∗P < .05, ∗∗P < .01 using t test with Wilcoxon matched–pairs signed rank test; #P < .05, ##P < .01, ###P < .001, ####P < .0001 using t test with Mann-Whitney test. ns, not significant.

Short-term modulation of pain-related behaviors in wild-type mice and female HbSS-BERK mice. (A) Experimental design for the single-session treatment with behavioral study. (B-C) Short-term inhibition of heat pain sensitivity in wild-type mice. The impact of tFUS on heat pain-associated behaviors was assessed in both (B) male (n = 8) and (C) female (n = 8) wild-type mice. Single-session tFUS with a PRF of 40 Hz at left S1HL resulted in a significant increase in latency difference between contralateral (right) and ipsilateral (left) hind paw compared with baseline and sham-treated mice. ns, not significant, ∗∗P < .01 using t test with Wilcoxon matched–pairs signed rank test; #P < .05 using t test with Mann-Whitney test. (D-F) Short-term modulation of thermal and mechanical hyperalgesia in female HbSS-BERK mice. Quantification of averaged Δ hPWL from hot-plate test, Δ hPWF from cold-plate test, and Δ hPWT from von Frey test was conducted before and after tFUS with control groups (negative control and sham treatment) in HbSS-BERK mice. (D) Single-session tFUS with a PRF of 40 Hz at left S1HL (n = 10) led to a significant increase in the averaged Δ hPWL from hot-plate test compared with baseline, sham treatment (n = 10), and negative control (n = 10). Control groups did not show apparent treatment effect compared with pretreatment baseline. A negative value of Δ hPWL was observed with a PRF of 3 kHz at left S1HL (n = 10), which was significantly different from the baseline and response with the PRF of 40 Hz. (E) Single-session tFUS with PRF of 40 Hz at left S1HL (n = 10) and insula (n = 10) resulted in a significant change in the Δ hPWF from cold-plate test at 8 minutes after treatment compared to the sham treatment and baseline. (F) tFUS with PRF of 40 Hz at left S1HL (n = 10) and insula (n = 10) led to a prominent elevation of Δ hPWT from von Frey test compared with sham-treated (n = 10) and negative control mice (n = 10). A lower withdrawal threshold of the contralateral (right) hind paw was observed with tFUS at PRF of 3 kHz at left S1HL (n = 10) compared with PRF of 40 Hz at S1HL and insula. ∗P < .05, ∗∗P < .01 using t test with Wilcoxon matched–pairs signed rank test; #P < .05, ##P < .01, ###P < .001, ####P < .0001 using t test with Mann-Whitney test. ns, not significant.

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