Fig 5. Immunohistochemical reactions showing histological features of the non-cellular component of reticular dermal matrices at different incubation times (20X magnification).

In control samples (T0) vimentin (A), laminin (D) and collagen IV (G) immunohistochemical reactions showed the presence of an intact basement membrane (stained in brown), which was progressively degraded during the decellularization process; in DMEM treated samples, the immunostaining was focal and weak after 5 weeks (T5) of treatment (B: vimentin; E: laminin; H: collagen IV). Basement membrane degradations appeared more evident in T5 samples treated with NaOH, in which vimentin (C) and laminin (F) reactions were negative, and only collagen IV staining was still positive (I). Vimentin immunostaining also showed that both DMEM and NaOH were able to completely eliminate fibroblasts at T5.