Figure 4. Genetic depletion or pharmacological inhibition of TLR4 improves diabetic wound healing.

A: Wounds were created in DIO Tlr4^−/− and DIO Tlr4^+/+ mice. Representative photographs of the wounds of DIO Tlr4^−/− and DIO Tlr4^+/+ on days 0 and 4 post injury are shown. The change in wound area was recorded daily by blinded observer and analyzed with ImageJ software (n = 5). *p<0.05 by Mann-Whitney U test. B: Wound myeloid cells CD11b⁺[CD3⁻CD19⁻Ly6G⁻] were isolated on day 5 in DIO Tlr4^−/−, ND Tlr4^−/−, DIO Tlr4^+/+, and ND Tlr4^+/+ and Il1b and Tnfa expression was quantified using qPCR (n=5 per group, replicated in triplicate). **p<0.01 by ANOVA followed by Newman–Keuls post hoc. C: Wounds were created in DIO mice and treated with daily injections of TAK-242 (3 mg/kg) or PBS control injection (n = 5). Wound size was measured by blinded observed in Image J NIH. Representative photographs of the wounds of DIO mice with PBS injection and DIO mice with TAK-242 injection on days 0 and 4 post injury are shown (n=4 per group). The arrow denotes the first day of pharmaceutical injection prior to wound creation. *p<0.05; **p<0.01 by Mann-Whitney U test. D: Wound myeloid cells CD11b⁺[CD3⁻CD19⁻Ly6G⁻] were isolated on day 3 in DIO+PBS and DIO+ TAK-242 and Il1b and Tnfa expression was quantified using qPCR (n=3 per group, replicated in triplicate). **p<0.01 by Mann-Whitney U test. E: Wounds were created in DIO Tlr4^f/fLyz2^Cre+ and DIO Tlr4^f/fLyz2^Cre- mice. Representative photographs of the wounds of DIO Tlr4^f/fLyz2^Cre+ and DIO Tlr4^f/fLyz2^Cre- on days 0 and 6 post injury are shown. The change in wound area was recorded daily by blinded observer and analyzed with ImageJ software (n = 5 per group). All data are representative of 2–3 independent experiments. Data are presented as the mean±SEM.