AFRL-HE-WP-TP-2006-0019Air Force Research Laboratory

Effect of Silver Nanoparticles on SRCActivity in Male Germ-Line Stem Cells

Laura K. Braydich-Stolle

University of DaytonDayton OH 45437-0009

Saber HussainJohn J. Schlager

Air Force Research LaboratoryHuman Effectiveness DirectorateApplied Biotechnology Branch

Wright-Patterson AFB OH 45433-5707

Marie-Claude Hofmann

University of DaytonDayton OH 45437-0009

March 2006

FINAL REPORT -JUNE 2003 TO SEPTEMBER 2005

20060403510Approved for public release; Air Force Research Laboratory

distribution unlimited Human Effectiveness Directorate

Biosciences and Protection DivisionApplied Biotechnology BranchWright-Patterson AFB, OH 45433-5707

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March 2006 Technical Paper June 2003 - September 20054. TITLE AND SUBTITLE 5a. CONTRACT NUMBEREffect of Silver Nanoparticles on SRC Activity in Male Germ- F33615-03-C-6325

Line Stem Cells 5b. GRANT NUMBER

5c. PROGRAM ELEMENT NUMBER

6. AUTHOR(S) 5d. PROJECT NUMBER*L.K. Braydich-Stolle; **Saber Hussain, **John J. Schlager, *Marie-Claude 2312

Hofmann 5e. TASK NUMBER

5f. WORK UNIT NUMBER

7. PERFORMING ORGANIZATION NAME(S) AND ADDRESS(ES) 8. PERFORMING ORGANIZATION REPORTAND ADDRESS(ES) NUMBER*Air Force Materiel Command **University of

Air Force Research Laboratory DaytonHuman Effectiveness Directorate Daytotn OH AFRL-HE-WP-TP-2006-0019Wright-Patterson AFB OH 45433-7022

9. SPONSORING / MONITORING AGENCY NAME(S) AND ADDRESS(ES) 10. SPONSOR/MONITOR'S ACRONYM(S)Air Force Materiel Command AFRL/HEPB

Air Force Research Laboratory

Human Effectiveness Directorate 11. SPONSORIMONITOR'S REPORT

Biosciences and Protection Division NUMBER(S)

Wright-Patterson AFB OH 45433-5707

12. DISTRIBUTION / AVAILABILITY STATEMENTApproved for public release; distribution is unlimited.

13. SUPPLEMENTARY NOTESThese are briefing charts. Clearance: AFRL/WS-06-0428, 15 Feb 06.

14. ABSTRACTGametogenesis is a complex biological process that is particularly sensitive to environmental insults such as chemicals and physicalstressors. Exposure to specific chemicals has been shown to inhibit fertility through a negative impact on germ cell proliferation anddifferentiation that can lower sperm count. In addition, toxicants might produce mutations that could have negative consequences on thedevelopment of the offspring. A previous study showed that a spermatogonial stem cell line, called C 18-4, provides a sensitive model toassess the cytotoxicity of nanoparticles in the male germ line. We also found that Ag-15nm was more toxic than Al-30nm and MoO3-30nmfor spermatogonial stem cell proliferation. The purpose of the present study was to determine the effects of various sizes of Agnanoparticles on the C18-4 cell line using standard cytotoxic assays. We also assessed the impact of Ag nanoparticles on Src signaling,since the activation of this kinase allows normal spermatogonial stem cells to proliferate. Mitochondrial function (MTS) data showed thatAg-130nm was not toxic to the cells, but smaller particles showed toxicity. Membrane leakage was increased by treatment with the Ag-25nm and Ag-30nm particles, even at low concentrations (5 pg/ml). However, no membrane leakage was apparent when the cells weretreated with Ag- 130nm. Further, we have found that the activity of Fyn (a member of the Src family of cytoplasmic kinases) issignificantly reduced when the cells are treated with Ag nanoparticles. This inhibition increases with the size of the nanoparticles, with noactivity detected above a size of 30 nm..15. SUBJECT TERMSUninhabited Systems16. SECURITY CLASSIFICATION OF: 17. LIMITATION 18. NUMBER 19a. NAME OF RESPONSIBLE PERSON

Unclassified OF ABSTRACT OF PAGES Saber Hussain

a. REPORT b. ABSTRACT c. THIS PAGE UL 30 19b. TELEPHONE NUMBER (include area

UNC UNC UNC code)(937) 904-9517

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