Fig 1.
Stretching chamber and stretch device.
(A) Over and side view of the chamber. Scale bar = 10 mm. (B) Overhead view of the stretch device. Yellow arrow shows the stretching direction. (C) Schema of the chamber.
Fig 2.
Procedure for establishing HSEs.
Fig 3.
Macroscopic view of the harvested HSEs.
(A) Non-stretch sample. (B) Stretched sample. Stretched sample had a thicker cuticle and reduced transparency compared with the non-stretch sample. Scale bar = 10 mm.
Fig 4.
Histlogic and immunohistologic analysis of HSEs.
(A) Hematoxylin and eosin staining of the non-stretch sample (B) and of the stretched sample. Scale bar = 100 μm. (C and D) The number of basal cells per 100 μm of a dermal—epidermal junction and the thickness of the epidermal keratinized layer showing a significant increase in the ST group. **p < 0.01 and *p < 0.05. (E and F) The expression of involurin was significantly increased in the ST group. Dotted lines indicate basement membrane. Scale bar = 100μm.
Fig 5.
Laminin 5 and collagen IV/VII expression analysis of HSEs by immunofluorescence staining.
(A, B and C) Non-stretch sample. (D, E and F) Stretched sample. Scale bar = 500 μm. (G) Fluorescence intensity of NST group was taken as control and adjusted to the 1 value. Each histogram bar represents the mean value of the normalized and adjusted fluorescence intensity. All three proteins of ST group were significantly greater compared with NST group. **p < 0.01.
Fig 6.
(A) Non-stretch sample. (B) Stretched sample. White arrow: hemidesmosome. Black arrow: lamina densa. Scale bar = 1 μm. (C, D) In the ST group, the length of lamina densa and the number of hemidesmosomes per 100 μm of a dermal—epidermal junction were significantly greater than in the NST group. **p < 0.01.