Effects of fetal exposure to carbon nanoparticles on reproductive function in male offspring  Seiichi Yoshida, Ph.D., Kyoko Hiyoshi, Ph.D., Shigeru Oshio, Ph.D., Hirohisa Takano, M.D., Ph.D., Ken Takeda, Ph.D., Takamichi Ichinose, Ph.D.  Fertility and Sterility  Volume 93, Issue 5, Pages 1695-1699 (March 2010) DOI: 10.1016/j.fertnstert.2009.03.094 Copyright © 2010 American Society for Reproductive Medicine Terms and Conditions

Figure 1 Testicular tissue of male offspring exposed to carbon nanoparticles (CB) as fetuses. (1) 5-week-old mouse; (2) 10-week-old mouse; (3) 15-week-old mouse; (a) control; (b) CB group. ×200 magnification. At all three ages, seminiferous epithelial damage (arrows) was seen in the CB group. Vacuolation was seen with some seminiferous tubules (asterisks). Fertility and Sterility 2010 93, 1695-1699DOI: (10.1016/j.fertnstert.2009.03.094) Copyright © 2010 American Society for Reproductive Medicine Terms and Conditions

Figure 2 Percentage of degenerated seminiferous tubules in cross-sections of carbon nanoparticles (CB)–treated and control mice. Estimation of testicular damage was conducted by counting the number of tubular cross-sections and determining the percentage of total degenerated tubules in three cross-sections per testis. Mean±SD (n=8). ∗∗∗p<.001 vs. control. Fertility and Sterility 2010 93, 1695-1699DOI: (10.1016/j.fertnstert.2009.03.094) Copyright © 2010 American Society for Reproductive Medicine Terms and Conditions

Figure 3 Effects of fetal carbon nanoparticle (CB) exposure on spermatogenesis. Right testis sample was weighed and seminiferous tubules released from the tunica albuginea into a saline solution in a homogenizer tube. Tissue was homogenized and diluted to the desired volume, and a sample of the diluted suspension was taken to count sperm heads or sperm cells in a hemocytometer chamber. Daily sperm production (DSP) was calculated as described in Materials and Methods. Mean±SD (n=16). ∗P<.05; ∗∗∗P<.001; vs. control. Fertility and Sterility 2010 93, 1695-1699DOI: (10.1016/j.fertnstert.2009.03.094) Copyright © 2010 American Society for Reproductive Medicine Terms and Conditions

Figure 4 Effects of fetal carbon nanoparticle (CB) exposure on serum testosterone. Testosterone levels in serum were determined by ELISA. Mean±SD (n=15–16). Fertility and Sterility 2010 93, 1695-1699DOI: (10.1016/j.fertnstert.2009.03.094) Copyright © 2010 American Society for Reproductive Medicine Terms and Conditions