Changes in signal intensity reflective of vascular leakage were followed for 32 min (Fig. 6C). The accumulation of contrast agent was restricted to the graft and was not observed in the surrounding muscle (Fig. 6D). Histological staining of the biotinylated contrast agent was performed using avidin-FITC. Consistent with the MRI data, FITC staining was restricted to blood vessels within the muscle, whereas it leaked into the graft (Fig. 6E). The contrast agent was also detected in the follicular fluid. This leakage of the contrast agent in the ovarian grafts was similar to that observed in intact rats that were injected with the contrast agent intravenously 45 min prior to retrieval of the ovaries. Accordingly, the vascular properties of the ovarian grafts in the intramuscular transplantation appeared to be similar to those of the intact ovary.
The distribution of the contrast agent in the ovarian grafts was significantly higher than in the adjacent muscle according to both the MRI and the histology intensity quantification (P < 0.05; two-tailed, unpaired Mest; Fig. 7, A and B; n = 7). The correlation between the MRI and the histology was highly significant (Fig. 7C; P = 0.008; r2 = 0.46). buy glucotrol
FIG. 7. Distribution and quantification of vascular permeability in ovarian grafts and in the adjacent muscle retrieved after 28 d (n = 3) and 35 d (n = 4). A) Vascular permeability (APS) was mapped by MRI from the rate of accumulation of albumin-based contrast agent during the first 10 min pos-tintravenous administration (APSMRI [^M/ min]). B) Quantification of vascular permeability by fluorescence microscopy using avidin-FITC staining of the biotinylated Mr contrast material. Permeability was assessed from fluorescence intensity in the graft and the adjacent muscle relative to the intensity in the blood vessels (APShistolo-gy). In both A and B, values from seven animals are expressed as mean ± SEM. Asterisks denote a significant difference between the ovarian grafts and the muscle both in the APSmri and the APShiStology (P < 0.05; two-tailed unpaired t-test). C) Correlation between MRI and histological measurements of vascular permeability for ovarian grafts (closed circles) and the adjacent muscle (open circles).