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Fig 1.

An External Tissue Expansion Device (ETED).

It contains (a) a suction cup 3.5 cm in diameter and 5 cm in height that connected to (b) the pump system. The pump system contains the pump that supplies negative pressure for 4 branches that were linked to 4 suction cups. The pressure meter is used to monitor the pressure. (c) The application of the ETED on porcine model.

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Fig 2.

The effects of the ETED on skin tissue.

(a) Wrinkle-like skin was observed after ETED treatment, particularly in the 3H-20 group. H&E stains of skin tissue on (b) control group, (c)1H-10 group, (d)1H-20 group, (e) 3H-10 group and (f) 3H-20 group demonstrating that longer ETED treatment results in a more severe effect on the epidermis morphology. C: Control. H: Hour.

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Fig 3.

Images of H&E staining on the epidermis and dermis.

Samples are from (a) control group, (b) 1H-10 group, (c) 1H-20 group, (d) 3H-10 group and (e) 3H-20 group (Scale bar equals to 100 μm). (f) Quantification of subcutaneous thickness under ETED treatment showing increasing thickness with treatment time. * Indicates statistical significance p <0.05.

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Fig 4.

Sections of specimens were stained with Ki67 (brown) to evaluate cell proliferation.

Those specimens are from control group (a), 1H-10 group (b), 1H-20 group (c), 3H-10 group (d) and 3H-20 group (magnification, 50x, Scale bar equals to 100 μm). (e) The percentage of Ki67-positive cells. * Indicates statistical significance p <0.05.

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Fig 5.

The blood vessels were evaluated by staining for CD 31 (Green).

Results are shown for the (a) control group, (b) 1H-10 group, (c) 1H-20 group, (d) 3H-10 group and (e) 3H-20 group (Scale bar equals to 100 μm). Animals treated with the ETED showed an increase in blood vessel density (f) and higher levels of FGF (g), VEGF-A (h) and PDGF-BB (i) compared to the control group. * Indicates statistical significance p <0.05.

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Fig 6.

Hair follicle evaluation analysis.

Keratin-16 stains (brown) of the specimens from (a) control group, (b)1H-10 group, (c)1H-20 group, (d)3H-10 group and (e) 3H-20 group. The presented images were assembled by approximately 20 images, scale bar is approximately 1 mm. (f) Quantification of hair follicle density. * Indicates statistical significance p <0.05. Arrow indicates the location of hair follicles.

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