Even sinusoidal tissue distant from the Affi-Gel bead vehicles showed abundant apoptosis after SHH inhibitor treatment. Apoptosis was not induced in control rats (Fig. 7) or in SHH protein-treated rats (SHH protein-treated = 1.6 6 0.7).
SHH Protein Treatment at the Time of CN Injury Prevents Post-CN Injury-induced Apoptosis
Affi-Gel beads soaked in SHH protein were injected into the corpora cavernosa at the time of bilateral CN injury in adult Sprague Dawley rats.
After 2 days, apoptosis was abundant and there was no difference in the number of apoptotic cells in the presence or absence of SHH protein (ratio of apoptotic cells to all cells within ~330 im of a bead in SHH protein CN2 = 0.85 6 0.11 and in PBS CN2 = 0.82 6 0.06 treated penes). At 4 days post-CN injury, the amount of apoptosis was decreased in the presence of SHH protein (Fig. 7, ratio of apoptotic cells to all cells within ~330 im of a bead in SHH protein-treated CN4 = 0.64 6 0.05 and pBs control = 0.8 6 0.06 penes, P-value = 0.02). When double the concentration of SHH protein was applied, apoptosis was suppressed in a larger region surrounding the bead vehicles and apoptosis was further reduced by 2.5-fold (ratio of apoptotic cells to all cells within ~330 im of a bead in 2X SHH protein-treated CN4 = 0.33 6 0.09 and control heat-inactivated 2X SHH protein-treated CN4 = 0.82 6 0.03 penes, P-value = 3.39E-05).