mersin univ saglık bilim derg Mersin Üniversitesi Sağlık Bilimleri Dergisi 1308-0830 Mersin University 10.26559/mersinsbd.594531 Health Care Administration Sağlık Kurumları Yönetimi The effect of modified gold nanoparticles on the function of neurons of mice hipocampal brain slices Modifiye altın nanoparçacıkların fare hipokampal kesitlerindeki nöronal fonksiyonlar üzerine etkileri https://orcid.org/0000-0003-1348-1336 Tuna Bilge Güvenç YEDITEPE UNIVERSITY https://orcid.org/0000-0002-2211-9665 Yavuz Yavuz YEDİTEPE ÜNİVERSİTESİ https://orcid.org/0000-0002-3375-8956 Kuku Gamze YEDİTEPE ÜNİVERSİTESİ https://orcid.org/0000-0002-1212-1550 Maharramov Akif YEDİTEPE ÜNİVERSİTESİ https://orcid.org/0000-0002-2674-6535 Yilmaz Bayram YEDİTEPE ÜNİVERSİTESİ https://orcid.org/0000-0002-1952-9495 Saricam Melike YEDİTEPE ÜNİVERSİTESİ https://orcid.org/0000-0001-5231-4659 Ercan Merve YEDITEPE UNIVERSITY https://orcid.org/0000-0002-3844-5190 Culha Mustafa YEDİTEPE ÜNİVERSİTESİ https://orcid.org/0000-0002-7762-8109 Dogan Soner YEDİTEPE ÜNİVERSİTESİ 08 30 2019 12 2 328 340 07 22 2019 08 09 2019 Copyright © 2008, Mersin Üniversitesi Sağlık Bilimleri Dergisi 2008 Mersin Üniversitesi Sağlık Bilimleri Dergisi

Aim: Itis still unclear whether modifications of gold nanoparticles (AuNp) affect thefunction of the nervous system. Aim of this study was to investigate the bioelectricaleffect of glucose (GlucoseAuNp), oligonükleotite (d(A)15AuNP) or peptide(AspAuNp) modified AuNp, on hippocampal mouse neurons after ex vivoadministration. Methods: AuNPmodifications were characterized by UV/Vis spectroscopy, Dynamic LightScattering (DLS) and Transmission Electron Microscopy (TEM). Subsequently, AuNpmodifications were added ex vivo to sections of hippocampal regions obtainedfrom the brains of 4-6 week old male Balb-c mice, and then spontaneous andstimulated action potentials (AP) were recorded by patch clamp with whole-celltechnique. The Peak amplitude, rising slope, half with and firing rate of Apswere analyzed. Results:Characterization results showed that AuNp modifications have 5-7 nm size andcolloidal shape. The firing rate of the APs measured in the GlucoseAuNp (n=9)group was higher compared to the AspAuNp (n=7), d(A)15AuNP (n=9), and control(n=9) groups (respectively 10.1±5.5 9.5±3.8, 6.2±3.0 pic/s, and 4.4±3.2, pic/s,p>0.05). In addition, current voltage (I-V) analysis showed that themembrane potential in response to current stimuli ((-150)–(150 pA)) of theGlucoseAuNp (n=8) shifted negative for negative current stimulations andpositive for positive current stimulations compared to the control (n=8) andd(A)15AuNP (n=9) groups. Conclusion:The surface AuNp modifications will be used in medical applications mightincrease the firing rate of Aps and stimulate neurological patholojies inhippocampal neurons and it may be suggested that more biocompatible AuNp formsshould be identified.

Amaç: Altın nanoparçacık (AuNP) modifikasyonlarının sinir sistemifonksiyonunu nasıl etkileyebileceği tam olarak bilinmemektedir. Bu çalışmada, AuNp’leringlikoz (GlikozAuNp), oligonükleotid (15 baz, d(A)15AuNP) veya peptid (negatif amino asitiçeren, AspAuNp) ile işlevselleştirilmiş formlarının fare hipokampalnöronları üzerindeki biyoelektriksel etkisinin ex vivo olarak araştırılması amaçlanmıştır. Yöntem: AuNP modifikasyonları UV/Vis spektroskopi, Dinamik IşıkSaçılması (DLS) ve Geçirimli Elektron Mikroskobu (TEM) ile karakterize edildive ex vivo olarak 4-6 haftalık Balb-cerkek farelerin beyin hipokampüs bölgelerinden alınan kesitlere eklendi,spontan ve uyarılmış aksiyon potansiyelleri (AP) yama kıskacı, tüm hücretekniği ile kaydedildi. Spontan AP’lerinin en büyük genlik değeri, yükseleneğimi, yarı genişliği, iki pik arasında geçen süre ve ateşleme hızı analizedildi. Bulgular: Karakterizasyonsonuçları AuNP’lerin yüzey modifikasyonlarının 5-7 nm çapında ve koloidalolduğunu gösterdi. Spontan AP’lerin ateşleme hızı GlikozAuNp (n=9) gurubunda,kontrol (n=9), AspAuNp (n=6) ve d(A)15AuNP (n=9) eklenen gruplara göre en fazlaolarak ölçüldü (sırasıyla 10.1±5.5 4.4±3.2 9.5±3.8 ve 6.2±3.0 pik/s, p>0.05).Ek olarak, uyarılmış AP’lerin özellikleriniyansıtan akım voltaj (I-V) analizinde, GlikozAuNp (n=8) gurubunun akımuyarılarına ((-150)–(+150 pA)) cevaben oluşturdukları membran potansiyellerikontrol (n=8) ve d(A)15AuNP (n=9) gruplarının cevabına göre negatif akımbölgesinde negatife, pozitif akım uyarısı bölgesinde pozitife kaydı. Sonuç: Medikal uygulamalardakullanılacak AuNP’lerin yüzey modifikasyonlarının hipokampal nöronlarınAP’lerinin ateşleme hızını arttırarak, patolojik koşulları tetikleyebileceklerive daha biyouyumlu formların belirlenmesi gerektiği önerilebilir.

 Altın nanoparçacıklar aksiyon potansiyeli Yüzey modifikasyonları Gold nanoparticles Action potential Surface Modifications TUBITAK 215S052 1) Jung S, Bang M, Kim BS, Lee S, Kotov NA, Kim B, Jeon D.”Intracellular gold nanoparticles increase neuronal excitability and aggravate seizure activity in the mouse brain”, PLoS One. 2014.13;9(3):e91360. 2) Salinas K, Kereselidze Z, DeLuna F, Peralta XG, Santamaria F. “Transient extracellular application of gold nanostars increases hippocampal neuronal activity”, J Nanobiotechnology. 2014. 20;12(1):31. 3) Ghosh P, Han G, De M, Kim CK, Rotello VM. “Gold nanoparticles in delivery applications”, Adv Drug Deliv Rev. 2008.;60:1307e15. 4) Feng G, Kong B, Xing J, Chen J. “Enhancing multimodality functional and molecular imaging using glucose-coated gold nanoparticles”, Clin Radiol. 2014. 69(11):1105-11. 5) Sperling, R. A., Gil, P. R., Zhang, F., Zanella, M. ve Parak, W. J. “Biological Applications of Gold Nanoparticles”, Chemical Society. 24 Nisan 2008. 6) Polak P, Shefi O. “Nanometric agents in the service of neuroscience: Manipulation of neuronal growth and activity using nanoparticles” Nanomedicine. 2015 Aug;11(6):1467-79. 7) Chithrani BD, Chan WC.”Elucidating the mechanism of cellular uptake and removal of protein-coated gold nanoparticles of different sizes and shapes”, Nano Lett. 2007.Jun;7(6):1542-50. 8) Conner SD, Schmid SL. “Regulated portals of entry into the cell”, Nature. 2003. 422:37–44. 9) Shukla R, Bansal V, Chaudhary M, Basu A, Bhonde RR. “Biocompatibility of gold nanoparticles and their endocytotic fate inside the cellular compartment: A microscopic overview”, Langmuir . 2005. 21: 10644–10654. 10) Boisselier E, Astruc D. “Gold nanoparticles in nanomedicine: preparations, imaging, diagnostics, therapies and toxicity”, Chem Soc Rev. 2009. 38: 1759–1782. 11) Sonavane G, Tomoda K, Makino K.”Biodistribution of colloidal gold nanoparticles after intravenous administration: Effect of particle size”, Colloids Surf B Biointerfaces. 2008. 66:274–280. 12) Chen YS, Hung YC, Liau I, Huang GS. “Assessment of the In Vivo Toxicity of Gold Nanoparticles”, Nanoscale Res Lett. 2009. 8;4(8):858-864. 13) Chen YS, Hung YC, Lin LW, Liau I, Hong MY, Huang GS. “Size-dependent impairment of cognition in mice caused by the injection of gold nanoparticles”, Nanotechnology. 2010.3;21(48):485102. 14) Kim JH, Kim KW, Kim MH, Yu YS. “Intravenously administered gold nanoparticles pass through the blood-retinal barrier depending on the particle size, and induce no retinal toxicity”, Nanotechnology. 2009. 20:505101. 15) Sur, I., Cam, D., Kahraman, M., Baysal, A., Culha, M. "Interaction of multi-functional silver nanoparticles with living cells", Nanotechnology. 2010. 21, 175104. 16) Chen J, Hessler JA, Putchakayala K, Panama BK, Khan DP, Hong S, Mullen DG, Dimaggio SC, Som A, Tew GN. “Cationic nanoparticles induce nanoscale disruption in living cell plasma membranes”, The Journal of Physical Chemistry B..2009. 113:11179–11185. 17) Goodman CM, McCusker CD, Yilmaz T, Rotello VM. ”Toxicity of gold nanoparticles functionalized with cationic and anionic side chains”, Bioconjug Chem. 2004. ; 15:897e900. 18) Gromnicova R, Davies HA, Sreekanthreddy P, Romero IA, Lund T, Roitt IM, Phillips JB, Male DK. “Glucose-coated gold nanoparticles transfer across human brain endothelium and enter astrocytes in vitro.” PLoS One. 2013 Dec 5;8(12):e81043. 19) Stampfl A, Maier M, Radykewicz R, Reitmeir P, Göttlicher M, Niessner R. Langendorff. “Heart: a model system to study cardiovascular effects of engineered nanoparticles”, ACS Nano. 2011.;5:5345–5353. 20) Piella J,Bastus NG,Puntes V. Size-Controlled Synthesis of Sub-10-nanometer Citrate-Stabilized Gold Nanoparticles and Related Optical Properties. Chem. Mater. January 20, 2016. 20162841066-1075. 21) Bernardes, G.J.L., Davis, B.G. “Direct thionation of reducing sugars”, Protocol Exchange. 2007. 22) Hurst SJ, Hill HD, Mirkin CA. "Three-dimensional hybridization" with polyvalent DNA-gold nanoparticle conjugates. J Am Chem Soc. 2008 Sep 10;130(36):12192-200. 23) Spadavecchia, J., Movia, D., Moore, C., Maguire, C.M., Moustaoui, H., Casale, S.. “Targeted polyethylene glycol gold nanoparticles for the treatment of pancreatic cancer: from synthesis to proof-of-concept in vitro studies”, Int J Nanomedicine. 2016. 11, 791–822. 24) Haiss W1, Thanh NT, Aveyard J, Fernig DG. Determination of size and concentration of gold nanoparticles from UV-vis spectra. Anal Chem. 2007 Jun 1;79(11):4215-21. 25) Guo, F., Yu, N., Cai, J. Q., Quinn, T., Zong, Z. H., Zeng, Y. J. & Hao, L. Y. “Voltage-gated sodium channels Nav1.1, Nav1.3 and b1 subunit were up-regulated in the hippocampus of spontaneously epileptic rat” Brain Res. Bull. 2008. 75, 179–187. 26) Meisler, M. H. ve Kearney, J. A. “Sodium channel mutations in epilepsy and other neurological disorders”, J. Clin. Invest. 2005.115, 2010–2017.