1. Volume 2, Issue 3, Pages 144-153 (July 2018)
Implantation of a heterologous dermo-epidermal skin substitute in a patient with deep dermal burn that enhances biomechanical and functional recovery: Case report 
M.C. Ornelas-Flores, J. García-López, Y. Melgarejo-Ramírez, R. Sánchez-Sánchez, G. Leyva-Gómez, N. Zacaula-Juárez, O. González- Mendoza, H.A. Manzo-Castrejón, F.E. Ferreira-Aparicio, E. Márquez-Gutiérrez, M.E. Martínez-Pardo, M.C. Velasquillo-Martínez, J.C. Ibarra-Ponce de León, A.M. Brena-Molina 
Burns Open 
Volume 2, Issue 3, Pages (July 2018)
DOI: /j.burnso
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2. Fig. 1
Development of hDE-SS. RHA was cultured with keratinocytes and fibroblasts for separation to obtain two layers (scale model: 1 cm2) for prior analyses (A). Before application on patient, epidermal layer (keratinocytes-RHA) and dermal layer (fibroblasts-RHA) (size: 25 cm2 each) in vitro (B). Pretibial region of the right leg with a deep dermal degree burn prior to excision (C). Burn after excision (D).
Burns Open 2018 2, DOI: ( /j.burnso )
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3. Fig. 2
Radio-sterilized Human Amnion (RHA). Before use. The air dried and radio-sterilized human amnion is a thin and transparent tissue (A). The RHA has a squared texture on its surface, which results from its direct contact with the nylon mesh sheet (B). Presence and organization of RHA epithelial-cell nuclei stained in fluorescent red (C). Scale bars: 0.01 cm (A and B); 100 µm (C).
Burns Open 2018 2, DOI: ( /j.burnso )
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4. Fig. 3
Viability of keratinocytes and fibroblast in vitro and onto RHA. Keratinocytes colonies (white arrow) are onto feeder layer (yellow arrow), maintained a rounded morphology (A). Fibroblasts in primary culture (B) have an elongated morphology and are spindle-shaped (blue arrow). The viability percentage graph shows that cell viability is high in both dermal populations in vitro, while the percentage of mortality is less in fibroblasts than in keratinocytes (C). Keratinocytes and fibroblasts cultured onto RHA are viable, maintaining their morphology as in in vitro, as well as the nuclei of epithelial cells of RHA (D, E) that are present in the RHA control, before culture (F, pink arrow). Live cells are fluorescent green in color, and dead cells are fluorescent red. Scale bars: 100 µm. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)
Burns Open 2018 2, DOI: ( /j.burnso )
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5. Fig. 4
Phenotype maintenance of keratinocytes and fibroblasts. Basal keratinocytes expressed Cytokeratin 5 (CK5) stained in fluorescent red and differentiated keratinocytes expressed Cytokeratin 10 (CK10) stained in fluorescent green, in vitro (A and B) and onto RHA (E and F), respectively. Fibroblasts express 1B10 (the fibroblast surface marker) stained in fluorescent green, in vitro (C) and onto RHA (G). Expression of α-sma is present only in fibroblasts in vitro (D) and, onto RHA, it is negative (H). The nuclei were stained in fluorescent blue (DAPI). Scale bars: 100 µ and 200 µm.
Burns Open 2018 2, DOI: ( /j.burnso )
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6. Fig. 5
Implant of human Dermo-Epidermal Skin Substitute (hDE-SS). Application of hDE-SS that appears as a transparent shiny layer over the burn (A, green arrows). Application of meshed autografts (white arrow) surrounding the hDE-SS (yellow arrow) for the treatment of the complete deep dermal burn (B). (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)
Burns Open 2018 2, DOI: ( /j.burnso )
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7. Fig. 6
Clinical evolution of repair of a deep dermal burn treated with hDE-SS. Evaluation of macroscopic repair. Postoperative Day 7, the burned area presents complete integration of the amnion, erythematous areas with presence of granulation tissue (A, Blue box: hDE-SS, yellow box: meshed skin autograft); for day 14 without erythema at the surgical edges (B). At day 21, there is no apparent contraction or scarring by secondary intention (C). At day 35 with a homogeneous appearance that respects pigmentation (D). At day 92, the physical appearance of the area implanted with the hDE-SS is similar to that of the grafted areas. (E) Magnification of the implanted area: Close-up evolution of the hDE-SS adhered to the burned skin, with the apparent decrease in erythema at day 21 (F-H) and an increase at day 35 (I, black arrow). Day 92 with the presence of hair follicles and a decrease in erythema (J). Magnification of Autograft: Erythema evolution is similar in all cases; pigmentation is also similar, and only with a clear difference at day 35 (K-N); but at day 92, the tones again matched the hDE-SS (O).
Burns Open 2018 2, DOI: ( /j.burnso )
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8. Fig. 7
Cutometer®. Maximal extension of skin (R0) (A), net elasticity (R5) (B), and viscoelastic amount of elasticity (R6) (C) determined by the suction method demonstrate a continuous recovery of skin functionality after the burn injury. Amnion hydration by Corneometer® represents an increase in the amount of water in the stratum corneum as an indication of the recovery of the quality of the skin’s architecture (D). Pigmentation levels by Mexameter® indicate an increase in melanin levels (E); however, there is no decrease in erythema levels (F). Green line: wound area without treatment, reference; blue line: amnion; red line: grafts, and orange line region of the foot without injury, control. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)
Burns Open 2018 2, DOI: ( /j.burnso )
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9. Supplementary Figure 1
Abdominoplasty and foreskin fibroblasts in vitro and onto RHA. Foreskin fibroblasts. Proliferation in vitro is higher than onto RHA; however, the expression of α-sma is present in both (A and C). Abdominoplasty fibroblasts. Expression of α-sma is present only in in vitro, while it is absent onto RHA, but both maintain their proliferation (B and D). Nuclear protein, Ki67 (proliferation) in fluorescent green. α-sma, in fluorescent red and nuclei in fluorescent blue. Scale bars: 100 µm.
Burns Open 2018 2, DOI: ( /j.burnso )
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10. Supplementary Figure 2
Cutometer® follow-up of patient. Macroscopic figures show the areas analyzed with the Cutometer in patient: Reference, is the area near the deep dermal degree burn (green square). Amnion, treated area with hDE-SS (blue square). Graft, treated area with autograft (red square). Control, distant area (foot) without burn injury, not shown in macroscopic images. The microscopic images were taken with a digital microscope on a 40× scale. Reference present a repaired area and presence of follicles. Amnion has erythema presence and few follicles. In graft, the mesh and a contraction are evident. Control, healthy skin without changes.
Burns Open 2018 2, DOI: ( /j.burnso )
Copyright © 2018 The Authors Terms and Conditions