Air Force Research Laboratory

Transcription

1 AFRL-HE-WP-TP Air Force Research Laboratory Effect of Silver Nanoparticles on SRC Activity in Male Germ-Line Stem Cells Laura K. Braydich-Stolle University of Dayton Dayton OH Saber Hussain John J. Schlager Air Force Research Laboratory Human Effectiveness Directorate Applied Biotechnology Branch Wright-Patterson AFB OH Marie-Claude Hofmann University of Dayton Dayton OH March 2006 FINAL REPORT -JUNE 2003 TO SEPTEMBER Approved for public release; distribution unlimited Air Force Research Laboratory Human Effectiveness Directorate Biosciences and Protection Division Applied Biotechnology Branch Wright-Patterson AFB, OH

2 Form Approved REPORT DOCUMENTATION PAGE OMB No Public reporting burden for this collection of information is estimated to average 1 hour per response, including the time for reviewing instructions, searching existing data sources, gathering and maintaining the data needed, and completing and reviewing this collection of information. Send comments regarding this burden estimate or any other aspect of this collection of information, including suggestions for reducing this burden to Department of Defense, Washington Headquarters Services, Directorate for Information Operations and Reports ( ), 1215 Jefferson Davis Highway, Suite 1204, Arlington, VA Respondents should be aware that notwithstanding any other provision of law, no person shall be subject to any penalty for failing to comply with a collection of information if it does not display a currently valid OMB control number. PLEASE DO NOT RETURN YOUR FORM TO THE ABOVE ADDRESS. 1. REPORT DATE (DD-MM-YYYY) 2. REPORT TYPE 3. DATES COVERED (From - To) March 2006 Technical Paper June September TITLE AND SUBTITLE 5a. CONTRACT NUMBER Effect of Silver Nanoparticles on SRC Activity in Male Germ- F 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 REPORT AND ADDRESS(ES) NUMBER *Air Force Materiel Command **University of Air Force Research Laboratory Dayton Human Effectiveness Directorate Daytotn OH AFRL-HE-WP-TP Wright-Patterson AFB OH 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 Biosciences and Protection Division Wright-Patterson AFB OH DISTRIBUTION / AVAILABILITY STATEMENT Approved for public release; distribution is unlimited. 11. SPONSORIMONITOR'S REPORT NUMBER(S) 13. SUPPLEMENTARY NOTES These are briefing charts. Clearance: AFRL/WS , 15 Feb ABSTRACT Gametogenesis is a complex biological process that is particularly sensitive to environmental insults such as chemicals and physical stressors. Exposure to specific chemicals has been shown to inhibit fertility through a negative impact on germ cell proliferation and differentiation that can lower sperm count. In addition, toxicants might produce mutations that could have negative consequences on the development of the offspring. A previous study showed that a spermatogonial stem cell line, called C 18-4, provides a sensitive model to assess the cytotoxicity of nanoparticles in the male germ line. We also found that Ag-15nm was more toxic than Al-30nm and MoO3-30nm for spermatogonial stem cell proliferation. The purpose of the present study was to determine the effects of various sizes of Ag nanoparticles 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 that Ag-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 were treated with Ag- 130nm. Further, we have found that the activity of Fyn (a member of the Src family of cytoplasmic kinases) is significantly reduced when the cells are treated with Ag nanoparticles. This inhibition increases with the size of the nanoparticles, with no activity detected above a size of 30 nm SUBJECT TERMS Uninhabited Systems 16. 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) Standard Form 298 (Rev. 8-98) Prescribed by ANSI Std

3 >14 0 IC LL Cf.) o *2 0 w ca co ) 4m E 7* 00 U)) 4-4 ma U) ci CL t4-.4 LiI U- E < Lia * U

4 4. to%~ 0 0 F~I C) r *4 0 4"

5 All U) C4 a 0) ' C 0 L.L

6 00 U U Im S 4-a U U I L U)

7 IIl I/I 7411" CIO o2,, > rool ( -t a(d AN ~- 4W~p *O E~ o cri CU a 9-0) qc 0 0~C c-ca- CD

8 144-7:4...

9 I. 1~C a -f-i

10 co 0 4-a- CtC) -- V - 0 t 3 t* 4-4, 4-4-a

11 4-a 20 4-a

12 * ~0- > IK EE SU

13 0 (3) SI0 0 E 0 Q* > 0 (0 4- U) (j. cz (I >4

14 E0 L M (1 (Z 00 92: ~ z0<2ý ~ ~ C.4u-) ~.

15 4* 0 >1 * U) U) x 0 0 C) 4- >I* >1 CM0 - I0

16 - *7 1 ½ 4 4) U Pt 02 S 4! vs * a V "I S o * ii> 1' - 79< I. *-.w

17 If) co m (al U) U)r E E 75~ U) ca. 0 ol o a In ~ I 0 "1 c4 (,ailuoo jo %) uallnpai SILAI Q,.1 a) o -0 0 u) 0 U-) 2 2 =L 0 04i 'ai c0 -~ t al 0 0D D 0D 0 CD C 0 C0 0: CD I0 t 0'4 0: 00 to ýt (N (Ioiluoo jo %) uoilanpa jis~j 04 al CO Ct CN 0 C CD CJ CN C (oioluo3 jo %) uoil~npoi S1ILd

18 E E L E E EE -0- a: --- Co, CL) 00 a- - F) c.-m CD (IOoIUOO10 0" CO Lo C) LO)ý %) OBABOIHHa

19 LL Tt 1 VM M N N jiojuoi 10% I fii

20 *0 E (n 0C 0 1 n -*' a) 0 C: 0 0 m r) E 0x ta a) 00-1o I- U ~10 U cm m S- L) 0

21 0) o E C4 N 00 I C) C: 0 E 4) E4 00 m < co 0 Q4 4-a0 I 1 0 I S 1j (D C

22 0) 0 0 0) 0 LO ) 1 0 LO E a~cm LO 0) o0 IT Cl0 0 D I (MU06V~U

23 0 0) C: 4.' 0 0) L.O 00E CD 0) c3 LA-O 0 C 0 ) 0 tn0w Dt 0 [Thu06~a 1-6ci 6 C;U

24 5 0 UU 0@ 4-aa * - U 0~ 0 UJQ 0 < c 0-0 S E CU 1 U E

25 40 4-a IVol

26 '4ý S- -L 0 a * I cmm ac 0C

27 00 4U _Il z U c 0 D0

28 CDW 0 C o c CD 0) E obco EE _ I 0 C.) pell~oi~sod j B 10sqn

29 >11 * o Q- L) 0) (1 ) 0 (1 Q U)z * 0 0

30 C:s (D C C a U) 0 Of0 I I a) E 0 I a. M -j 2