Article Figures & Data
Figures
- FIGURE 1.
(A) pO2 values in arthritic and healthy ankles (left box plot) and the contralateral muscle issues (right box plot) on day 6 after GPI (n = 5) or control serum (n = 4) injection. (B) We correlated ankle swelling (mm) and pO2 values from arthritic and healthy ankles.
- FIGURE 2.
Representative 18F-FMISO (A; first column) and 18F-FAZA PET (B; first column) images from arthritic (black column) and healthy control (white column) mice on day 6 after GPI or control serum injection (representative slices of 90-min datasets). Ninety-minute 18F-FMISO (GPI, n = 2; control, n = 1) (A; second column) and 18F-FAZA (GPI, n = 2; control, n = 1) (B; second column) time–activity curves on day 6 after GPI or control serum application. Quantification of pooled static 18F-FMISO (GPI, n = 8; control, n = 5) (A; third column) and 18F-FAZA PET (GPI, n = 8; control, n = 4) (B; third column) of arthritic and healthy ankles presented as mean of %ID/g ± SD. (C) Quantification of pooled static 18F-FMISO PET scans presented as mean of %ID/g in arthritic and healthy ankles on day 1 (GPI, n = 3; control, n = 2), day 3 (GPI, n = 2; control, n = 2), and day 6 (GPI, n = 8; control, n = 4) after serum transfer (left bars). We correlated ankle swelling (mm) with %ID/g values from arthritic ankles on days 1, 3, and 6 after serum transfer (right). Mean ± SD; **P < 0.001.
- FIGURE 3.
Localization of exact 18F-FMISO uptake sites in arthritic and control ankles, using PET/MRI. Representative healthy mouse after control serum injection exhibited no signs of hypoxia.
- FIGURE 4.
(A) Representative 18F-FAZA autoradiography analysis of slices from arthritic ankles on day 6 after GPI serum transfer (left) and healthy ankles after control serum application (right) confirming in vivo 18F-FAZA PET results. (B) Pimonidazole immunohistochemistry from arthritic ankles 6 d after GPI serum transfer (right), indicating areas of enhanced hypoxia compared with healthy ankles (left, *).
- FIGURE 5.
(A) Representative hematoxylin and eosin–stained slides from GPI arthritic ankles (n = 2) on day 6 after GPI serum transfer are shown (upper images). HIF-1α (630×) staining (n = 2) demonstrated increased number of hypoxic cells comprising predominantly neutrophils and synoviocytes (lower images). Control ankles (n = 2) exhibited no positive staining. (B) HIF-1α and HIF-2α WB analyses of arthritic (n = 2) and healthy ankles (n = 2) 6 d after GPI or control serum transfer revealed strongly enhanced HIF-1α and HIF-2α protein expressions. Control β-actin staining was performed as indicated at 40 kDa. (C) RT-PCR was performed with ankle tissue from mice 6 (black column) and 12 h (white column) after single GPI or control serum injection and with mouse ankle tissue from day 2 after first and 6 and 12 h after second GPI or control serum injection. We detected highest relative expressions of HIF-1α, HIF-2α, IL-1β, and TNF, 6 h after second GPI serum transfer. c = cytosolic fraction; h = hours; n = nuclear fraction; p.i. = after injection.
- FIGURE 6.
(A) L-012 chemiluminescence imaging in mice after GPI or control serum injection using OI. Arthritic ankles exhibited high levels of L-012 expression compared with control ankles (left). Quantitative analysis confirmed a significantly greater L-012 signal on days 3 (80,544 p/s/cm2/sr, n = 8) and 6 (165,639 p/s/cm2/sr, n = 8) after GPI serum injection in arthritic compared with healthy ankles (2,904 p/s/cm2/sr) (n = 9) (right). (B) We correlated ankle swelling (mm) with average (avg) radiance (p/s/cm2/sr) of L-012 in arthritic and healthy ankles at days 3 and 6 after GPI or control serum injection. (C) DHR-FACS analyses of cells gained from lavage of arthritic and healthy ankles revealed significantly greater ROS levels in arthritic ankles than control ankles. Mean value ± SD; **P < 0.0001.
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