FIG. 1.

Macrophages isolated from chronic wounds in diabetic patients exhibit a proinflammatory phenotype that may be induced by the wound environment. Macrophages were isolated from chronic wound biopsy specimens and expression of proinflammatory markers IL-1β, MMP-9, TNF-α, and IL-6 (A–D) and healing-associated/anti-inflammatory markers CD206, IGF-1, TGF-β, and IL-10 (E–H) were assessed by real-time PCR. For comparison, blood-derived macrophages from healthy volunteers were left nonstimulated (Non), stimulated with TNF-α and IFN-γ (classically activated [CA]), or stimulated with chronic wound–conditioned medium (WCM). For the in vitro experiments, release of IL-1β, TNF-α, IGF-1, and TGF-β (I–L) into cell culture medium was measured by ELISA. In addition, chronic wound biopsy cryosections were immunostained for IL-1β and the macrophage marker CD68, with images taken at 20× (M–O; scale bar = 100 μm) and at 40× (P–R; scale bar = 50 μm); location of 40× images shown in box on 20× images. Nuclei stained with DAPI. For all graphs, bars = mean ± SD and n = 5 for both in vivo and in vitro experiments. *Mean value significantly different from that for nonstimulated controls, P < 0.05. diff, difference.