Maternal Intake of Either Fructose or the Artificial Sweetener Acesulfame-K Results in Differential and Sex-Specific Alterations in Markers of Skin Inflammation and Wound Healing Responsiveness in Mouse Offspring: A Pilot Study

Abstract

Growing evidence has demonstrated that maternal artificial sweetener (AS) consumption may not be a beneficial alternative when compared to sugar-sweetened beverages and potentially leads to metabolic dysfunction in adult offspring. Compromised skin integrity and wound healing associated with type 2 diabetes can lead to complications such as diabetic pressure injury (PI). In this context, the skin plays an important role in the maintenance of metabolic homeostasis, yet there is limited information on the influence of sugar- or AS-sweetened beverages during pregnancy on developmental programming and offspring skin homeostasis. This study examined the impact of maternal fructose or acesulfame-k consumption on offspring wound healing. Female C57Bl/6 mice received a chow diet ad libitum with either water (CD), fructose (FR; 34.7 mM fructose), or AS (AS; 12.5 mM Acesulfame-K) throughout pregnancy and lactation. PIs were induced in offspring at 9 weeks of age (n = 6/sex/diet). PIs and healthy skin biopsies were collected for later analysis. Maternal AS intake increased skin inflammatory markers in healthy biopsies while an FR diet increased Tgfb expression, and both diets induced subtle changes in inflammatory markers post-wound inducement in a sex-specific manner. Furthermore, a maternal FR diet had a significant effect on pressure wound severity and early wound healing delay, while AS maternal diet had a sex-specific effect on the course of the healing process. This study demonstrates the need for a better understanding of developmental programming as a mediator of later-life skin integrity and wound responsiveness.

Keywords: DOHaD; adipose tissue; animal model; artificial sweetener; developmental programming; fructose; inflammation; maternal nutrition; pressure injury; skin; wound healing.

Figure 1

Pressure injury timeline and placement. (A) timeline of magnet placement, (B) Placement of magnets on mouse to induce pressure injury, (C) Method to characterise skin injury in male and female mice offspring. Protocol modified from Stadler et al. [25].

Figure 2

Gene expression in healthy skin. The impact of Ace-K (AS) and fructose (FR) intake compared to controls (CD) at D0 on skin gene expression in female and male offspring. (A) Tnfα, (B) IL1b, (C) Nlrp3, (D) Vegfa, (E) Tgfb, (F) Ppargc1, (G) Pparg. Data were analysed using one-way ANOVA. Data are expressed as mean ± SEM. * p < 0.05 w.r.t CD. + p < 0.05 w.r.t AS. n = 3/group. NS: Not significant.

Figure 3

Wound characterisation at D3. The impact of Ace-K (AS) or fructose (FR) intake compared to Controls (CD) at D3 on wound characterisation in female and male C57BL/6 offspring. (A) Percentage of pressure injury occurrence (B) Representative images of wounds taken at D3 in male offspring (top) and female offspring (below). (C) Total surface area of pressure injury in female and male offspring; n = 12 wounds per group (2 wounds per mouse). * p < 0.05 vs. CD and AS.

Figure 4

Characterisation of surface area of inflamed lesions, skin breakdown, or ischemic crowns in female and male offspring at D3 and D5. * p < 0.05 vs. CD.

Figure 5

Sex-specific effects of the maternal diet on the wound healing process in offspring. * p < 0.05 vs. female.

Figure 6

Maternal diet effect on the wound healing process in female and male offspring. * p < 0.05 vs. CD and vs. AS; # p < 0.05 vs. respective D3, red FR and green AS.

Figure 7

D5 and D9 pressure injury gene expression. The impact of Ace-K (AS) and fructose (FR) intake compared to D5 Controls (CD) at D5 and D9 on skin pressure injury gene expression in female and male C57BL/6 offspring mice. (A) Tnfα, (B) I11b, (C) Nlrp3, (D) Vegfa, (E) Tgfb, (F) Ppargc1, (G) Pparg. Data were analysed using two-way ANOVA. Data are expressed as mean ± SEM. * p < 0.05 w.r.t CD D5. + p < 0.05 w.r.t AS D5. ^ p < 0.05 w.r.t FR D5. # p < 0.05 w.r.t CD D9. ~ p < 0.05 w.r.t AS D9. n = 4–5/group. NS = Not significant.