Topically applied metal chelator reduces thermal injury progression in a rat model of brass comb burn - PubMed (original) (raw)
Topically applied metal chelator reduces thermal injury progression in a rat model of brass comb burn
Cheng Z Wang et al. Burns. 2015 Dec.
Abstract
Oxidative stress may be involved in the cellular damage and tissue destruction as burn wounds continues to progress after abatement of the initial insult. Since iron and calcium ions play key roles in oxidative stress, this study tested whether topical application of Livionex formulation (LF) lotion, that contains disodium EDTA as a metal chelator and methyl sulfonyl methane (MSM) as a permeability enhancer, would prevent or reduce burns.
Methods: We used an established brass comb burn model with some modifications. Topical application of LF lotion was started 5 min post-burn, and repeated every 8 h for 3 consecutive days. Rats were euthanized and skin harvested for histochemistry and immunohistochemistry. Formation of protein adducts of 4-hydroxynonenal (HNE), malonadialdehyde (MDA) and acrolein (ACR) and expression of aldehyde dehydrogenase (ALDH) isozymes, ALDH1 and ALDH2 were assessed.
Results: LF lotion-treated burn sites and interspaces showed mild morphological improvement compared to untreated burn sites. Furthermore, the lotion significantly decreased the immunostaining of lipid aldehyde-protein adducts including protein -HNE, -MDA and -ACR adducts, and restored the expression of aldehyde dehydrogenase isozymes in the unburned interspaces.
Conclusion: This data, for the first time, demonstrates that a topically applied EDTA-containing lotion protects burns progression with a concomitant decrease in the accumulation of reactive lipid aldehydes and protection of aldehyde dehydrogenase isozymes. Present studies are suggestive of therapeutic intervention of burns by this novel lotion.
Keywords: Brass comb burn; Burn progression; Iron chelation; Oxidative stress; Reactive aldehydes; Thermal injury; Wound healing.
Copyright © 2015 Elsevier Ltd and ISBI. All rights reserved.
Figures
Fig 1
A, Brass comb probe consisting of three (3) 10-mm teeth separated by two (2) 10-mm notches modified from the previous Regas and Ehrlich model (1992). B, Diagram of the bottom view of the modified brass comb consisting of three (3)10×19 mm rectangles of burn sites separated by two (2)10×19 mm rectangles of interspaces. The second diagram is of the tissue sampling in each comb burn wound; two tissue blocks (9 × 30 mm each) were harvested.
Fig 2
Representative photographs of the burn wounds: A (5 min after injury) and B (72 hrs after injury). C (5 min after injury) and D (72 hrs after injury) with LF lotion treatment. Note that at 72 hrs the LF lotion-treated wound (D) showed a size of unburned interspace similar to that of the same wound 5 min after burn injury (C), while the untreated burn wound (B) had a significantly smaller interspace when compared to that of the same wound 5 min after burn (A).
Fig 3
Representative H&E stained microphotographs of the burn sites: Middle of burn sites showing microscopic characteristics of burn wound without (A–C) or with (D–F) LF lotion treatment started 5 min post injury. B and C, higher power of the upper and lower insert boxes in A, respectively. E and F, higher power of the upper and lower insert boxes in D, respectively. SkM, skeletal muscle; SG, sebaceous gland; HF, hair follicles; △, dilation and congestion of capillaries, venules and arterioles; ▲, blocked vessels filled with denatured clots; *, inflammatory cell infiltration. ↑, Scale bar = 200 µm in A & D; 50 µm in B, C, E, F
Fig 4
Representative microphotographs of Masson’s trichrome staining of burn sites, 72 hrs post injury: A, Control, without burn; B, the middle of a burn site, burn alone; C, the middle of a burn site, burn plus LF lotion treatment post injury. SkM, skeletal muscle; SG, sebaceous gland; HF, hair follicles; ▲, blocked vessels filled with denatured clots; *, areas with inflammatory cell infiltration. Scale bar = 200 µm in A-C
Fig 5
Representative H&E stained microphotographs of the interspaces: Middle of interspaces showing microscopic characteristics without (A–C) or with (D–F) LF lotion treatment. A–C, 72 hrs post injury; D–F, 72 hrs post injury plus LF lotion treatment. B and C, higher power of the upper and lower insert boxes in A, respectively. E and F, higher power of the upper and lower insert boxes in D, respectively. SkM, skeletal muscle; SG, sebaceous gland; HF, hair follicles; △, dilation and congestion of capillaries, venules or arterioles; *, inflammatory cell infiltration. Scale bar = 200 µm in A & D, 50 µm in B,C, E,F
Fig 6
Representative H&E microphotographs of survived interspace epidermis: A, control without burn; B, 72 hrs post injury; C, 72 hrs post injury plus LF lotion treatment. Yellow arrow line marks the length of survived interspace epidermis. SkM, skeletal muscle; SG, sebaceous gland; HF, hair follicles; ▲, blocked vessels filled with denatured clots; ↓, necrotic epidermis. Scale bar = 500 µm in A-C
Fig 7
Representative Masson’s trichrome staining of the half interspace (4 mm from its middle line). A, control without burn; B, 72 hrs after burn; C, 72 hrs after burn plus LF lotion treatment post injury. SkM, skeletal muscle; SG, sebaceous gland; HF, hair follicles; ▲, blocked vessels filled with denatured clots; ↓, necrotic epidermis. Scale bar = 500 µm in A–C
Fig 8
Representative protein-HNE IHC microphotographs of interspaces : The methyl green counterstained IHC microphotographs show epidermis and dermis (A–C) and hypodermis (D–F). A & D, control, without burn; B & E, 72 hrs post burn; C & F, 72 hrs after a burn plus LF lotion treatment post injury; Ep, epidermis; SG, sebaceous gland; HF, hair follicles; ↓, patent vessels (D, F); △, dilated capillary in dermis (B); ▲, dilated and blocked (E) vessels in hypodermis. Scale bar = 100 µm (A–F)
Fig 9
Representative protein-MDA IHC microphotographs of interspaces: The methyl green counterstained IHC microphotographs show protein-MDA staining in the interspaces of A control, without burn; B, , 72 hrs after burn; C, 72 hrs after burn plus LF lotion treatment post injury. Ep, epidermis; SG, sebaceous gland; HF, hair follicle. Scale bar = 50 µm (A–C)
Fig 10
Representative ALDH1A1 and ALDH2 IHC microphotographs of interspaces: The methyl green counterstained IHC microphotographs show immunostaining of ALDH1A1 (A-C) and ALDH2 (D–F) in sebaceous gland (SG) and hair follicle (HF) epithelial cells. A & D, Control, without burn; B & E, 72 hrs after burn; C & F, 72 hrs after burn plus LF lotion treatment post injury. Ep, epidermis; SG, sebaceous gland; HF, hair follicle. Scale bar, 100 µm (A–F)
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