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Full thickness wound repair by engineered skin grafting in athymic nude mice . ( A ) Representative images of the wounds grafted with PG-HSE, BTM-HSE, or native skin autografts on day of grafting (day 0) and post-grafting (day 14). ( B ) Representative microscopic images of day 14 grafts analysed by immunofluorescence, using a human specific antibody against involucrin. PG-HSE pre-grafting (scale 100 µm); PG-HSE graft (scale 1 mm); BTM-HSE pre-grafting (scale 100 µm); BTM-HSE graft (scale 1 mm); autograft pre-grafting (scale 100 µm; autograft (scale 1 mm); and human native skin (scale 1 mm). Zoomed images scale bar = 50 µm. Orange arrows indicate the graft edge 2 weeks post-grafting and red arrows indicate the initial wound edge. ( C ) Presence of human involucrin in grafts is presented as integrated density (IntDen), that is, sum of pixel value/signal intensity within a selected region of interest calculated using FIJI software version 1.54p. ( D ) Human-specific <t>Ku80</t> marker was detected by immunofluorescence on day 14 post-grafting and % Ku80 positive cells were counted using Nikon NIS-Elements Analysis software (version 5.5). Data were analysed using ordinary one-way ANOVA. Values in ( C , D ) represent mean values +/− SEM in each group (* = p ≤ 0.05, ** = p ≤ 0.01, n = 4–5 per group). PG-HSE, platelet-derived human skin equivalent; BTM-HSE, NovoSorb biodegradable temporising matrix human skin equivalent; autograft, and autologous full thickness skin graft.
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Full thickness wound repair by engineered skin grafting in athymic nude mice . ( A ) Representative images of the wounds grafted with PG-HSE, BTM-HSE, or native skin autografts on day of grafting (day 0) and post-grafting (day 14). ( B ) Representative microscopic images of day 14 grafts analysed by immunofluorescence, using a human specific antibody against involucrin. PG-HSE pre-grafting (scale 100 µm); PG-HSE graft (scale 1 mm); BTM-HSE pre-grafting (scale 100 µm); BTM-HSE graft (scale 1 mm); autograft pre-grafting (scale 100 µm; autograft (scale 1 mm); and human native skin (scale 1 mm). Zoomed images scale bar = 50 µm. Orange arrows indicate the graft edge 2 weeks post-grafting and red arrows indicate the initial wound edge. ( C ) Presence of human involucrin in grafts is presented as integrated density (IntDen), that is, sum of pixel value/signal intensity within a selected region of interest calculated using FIJI software version 1.54p. ( D ) Human-specific <t>Ku80</t> marker was detected by immunofluorescence on day 14 post-grafting and % Ku80 positive cells were counted using Nikon NIS-Elements Analysis software (version 5.5). Data were analysed using ordinary one-way ANOVA. Values in ( C , D ) represent mean values +/− SEM in each group (* = p ≤ 0.05, ** = p ≤ 0.01, n = 4–5 per group). PG-HSE, platelet-derived human skin equivalent; BTM-HSE, NovoSorb biodegradable temporising matrix human skin equivalent; autograft, and autologous full thickness skin graft.
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Full thickness wound repair by engineered skin grafting in athymic nude mice . ( A ) Representative images of the wounds grafted with PG-HSE, BTM-HSE, or native skin autografts on day of grafting (day 0) and post-grafting (day 14). ( B ) Representative microscopic images of day 14 grafts analysed by immunofluorescence, using a human specific antibody against involucrin. PG-HSE pre-grafting (scale 100 µm); PG-HSE graft (scale 1 mm); BTM-HSE pre-grafting (scale 100 µm); BTM-HSE graft (scale 1 mm); autograft pre-grafting (scale 100 µm; autograft (scale 1 mm); and human native skin (scale 1 mm). Zoomed images scale bar = 50 µm. Orange arrows indicate the graft edge 2 weeks post-grafting and red arrows indicate the initial wound edge. ( C ) Presence of human involucrin in grafts is presented as integrated density (IntDen), that is, sum of pixel value/signal intensity within a selected region of interest calculated using FIJI software version 1.54p. ( D ) Human-specific <t>Ku80</t> marker was detected by immunofluorescence on day 14 post-grafting and % Ku80 positive cells were counted using Nikon NIS-Elements Analysis software (version 5.5). Data were analysed using ordinary one-way ANOVA. Values in ( C , D ) represent mean values +/− SEM in each group (* = p ≤ 0.05, ** = p ≤ 0.01, n = 4–5 per group). PG-HSE, platelet-derived human skin equivalent; BTM-HSE, NovoSorb biodegradable temporising matrix human skin equivalent; autograft, and autologous full thickness skin graft.
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Full thickness wound repair by engineered skin grafting in athymic nude mice . ( A ) Representative images of the wounds grafted with PG-HSE, BTM-HSE, or native skin autografts on day of grafting (day 0) and post-grafting (day 14). ( B ) Representative microscopic images of day 14 grafts analysed by immunofluorescence, using a human specific antibody against involucrin. PG-HSE pre-grafting (scale 100 µm); PG-HSE graft (scale 1 mm); BTM-HSE pre-grafting (scale 100 µm); BTM-HSE graft (scale 1 mm); autograft pre-grafting (scale 100 µm; autograft (scale 1 mm); and human native skin (scale 1 mm). Zoomed images scale bar = 50 µm. Orange arrows indicate the graft edge 2 weeks post-grafting and red arrows indicate the initial wound edge. ( C ) Presence of human involucrin in grafts is presented as integrated density (IntDen), that is, sum of pixel value/signal intensity within a selected region of interest calculated using FIJI software version 1.54p. ( D ) Human-specific Ku80 marker was detected by immunofluorescence on day 14 post-grafting and % Ku80 positive cells were counted using Nikon NIS-Elements Analysis software (version 5.5). Data were analysed using ordinary one-way ANOVA. Values in ( C , D ) represent mean values +/− SEM in each group (* = p ≤ 0.05, ** = p ≤ 0.01, n = 4–5 per group). PG-HSE, platelet-derived human skin equivalent; BTM-HSE, NovoSorb biodegradable temporising matrix human skin equivalent; autograft, and autologous full thickness skin graft.

Journal: International Journal of Molecular Sciences

Article Title: Bioengineered Skin from a Platelet-Derived Hydrogel Repairs Full Thickness Wounds in a Pre-Clinical Mouse Model

doi: 10.3390/ijms26209988

Figure Lengend Snippet: Full thickness wound repair by engineered skin grafting in athymic nude mice . ( A ) Representative images of the wounds grafted with PG-HSE, BTM-HSE, or native skin autografts on day of grafting (day 0) and post-grafting (day 14). ( B ) Representative microscopic images of day 14 grafts analysed by immunofluorescence, using a human specific antibody against involucrin. PG-HSE pre-grafting (scale 100 µm); PG-HSE graft (scale 1 mm); BTM-HSE pre-grafting (scale 100 µm); BTM-HSE graft (scale 1 mm); autograft pre-grafting (scale 100 µm; autograft (scale 1 mm); and human native skin (scale 1 mm). Zoomed images scale bar = 50 µm. Orange arrows indicate the graft edge 2 weeks post-grafting and red arrows indicate the initial wound edge. ( C ) Presence of human involucrin in grafts is presented as integrated density (IntDen), that is, sum of pixel value/signal intensity within a selected region of interest calculated using FIJI software version 1.54p. ( D ) Human-specific Ku80 marker was detected by immunofluorescence on day 14 post-grafting and % Ku80 positive cells were counted using Nikon NIS-Elements Analysis software (version 5.5). Data were analysed using ordinary one-way ANOVA. Values in ( C , D ) represent mean values +/− SEM in each group (* = p ≤ 0.05, ** = p ≤ 0.01, n = 4–5 per group). PG-HSE, platelet-derived human skin equivalent; BTM-HSE, NovoSorb biodegradable temporising matrix human skin equivalent; autograft, and autologous full thickness skin graft.

Article Snippet: Slides were blocked as above and incubated with primary antibodies: rabbit anti-human involucrin (1:100, Cat. No. ab234403, Abcam), rabbit anti-human Ku80 (1:50, Cat. No. 214745, Biorbyt, Durham, NC, USA), or rabbit anti-human CK5 (1:250, Cat. No. 905504, BioLegend, San Diego, CA, USA ) overnight.

Techniques: Immunofluorescence, Software, Marker, Derivative Assay