Interaction of silver and gold nanoparticles in mammalian cancer: as real topical bullet for wound healing— A comparative study

  • Allur Subramaniyan Sivakumar
  • Chandran Krishnaraj
  • Sunirmal Sheet
  • Dileep Reddy Rampa
  • Da Rae Kang
  • Shah Ahmed Belal
  • Abhay Kumar
  • In Ho Hwang
  • Soon-Il Yun
  • Yang Soo Lee
  • Kwan Seob ShimEmail author


The present study evaluates in vitro cytotoxic effects and the mode of interaction of biologically synthesized Ag and Au nanoparticles (NPs) using Brassica oleracea L. var. capitata f. rubra (BOL) against HT-1080 cancer cells and bacterial cells as well as their wound healing efficacy using a mouse model. UV–visible spectroscopy, scanning electron microscopy, high-resolution transmission electron microscopy, and energy-dispersive X-ray analysis have ascertained the formation of nano-sized Ag and Au particles. Fourier transform infrared analysis has confirmed that polyphenol and amide groups in BOL act as capping as well as reducing agents. The free radical scavenging activity under in vitro conditions is found to be higher for the Ag NPs when compared to the Au NPs. Acridine orange–ethidium bromide dual staining and comet assay have indicated that the cytotoxic effects are mediated through nuclear morphological changes and DNA damage. The intracellular localization of Ag and Au NPs in HT-1080 cells and their subsequent effect on apoptosis and necrosis were analyzed by flow cytometry while the mode of interaction was established by scanning electron microscopy under field emission mode and by bio-transmission electron microscopy. These methods of analysis clearly revealed that the Ag and Au NPs have easily entered and accumulated into the cytosol and nucleus, resulting in activation of inflammatory and apoptosis pathways, which in turn cause damage in DNA. Further, mRNA and protein expression of caspase-3 and caspase-7, TNF-α, and NF-κB have provided sufficient clues for induction of intrinsic and extrinsic apoptosis and inflammatory pathways in Ag NP- and Au NP-treated cells. Evaluation of wound healing properties of Ag and Au NPs using a mouse model indicates rapid healing of wounds. In addition, no clear toxic effects and no nuclear abnormalities in peripheral blood cells are observed. Ag NPs appear to be a better anticancer therapeutic agent than Au NPs. Nonetheless, both Ag NPs and Au NPs show potential for promoting topical wound healing without any toxic effects.

Graphical abstract

Schematic representation of biological synthesis of Ag and Au NPs and its application on cancer and wound healing:


Silver Gold Nanoparticles Apoptosis Cancer Wound healing Inflammation Peripheral blood 



The TEM and SEM samples were analyzed using the JEM-2010 (JEOL) installed at the Center for University-Wide Research Facilities (CURF) at Chonbuk National University. We thank Mrs. Eun-Jin Choi at the CURF at Chonbuk National University.

Compliance with ethical standards

All procedures relating to the animals and their care conformed to the international guideline “Principles of Laboratory Animals Care” (NIH Publication No. 85-23, revised 1985).

Supplementary material

11626_2017_150_MOESM1_ESM.jpg (86 kb)
Fig. S1 UV–visible spectra of the biosynthesized Ag NPs and Au NPs observed at 430 nm. Digital photographs showing the (A) leaf extract alone, (B) reddish dark brown color formation of Ag NPs after the addition of 1 mM AgNO3 into the aqueous Brassica oleracea L. var. capitata f. rubra extract, and (C) light brownish color formation of Au NPs after the addition of 1 mM HAuCl4 into the aqueous plant extract. (JPEG 85 kb)
11626_2017_150_MOESM2_ESM.jpg (173 kb)
Fig. S2 FE-SEM images of the Ag and Au NPs observed after the addition of 1 mM AgNO3 and 1 mM HAuCl4 into the aqueous BOL extract: (a) Ag NPs and (b) Au NPs. (JPEG 173 kb)
11626_2017_150_MOESM3_ESM.jpg (42 kb)
Fig. S3 Energy-dispersive X-ray analysis performed on the biosynthesized Ag and Au NPs: (a) Au NPs and (b) Ag NPs. (JPEG 42 kb)


  1. Ahmed MF, Rao A (2014) Simultaneous determination of phenolic compounds in Brassica oleracea L. var capitata. by high-performance liquid chromatography. Int J Pharm Pharm Sci 6:534Google Scholar
  2. Brand-Williams W, Cuvelier ME, Berset CLWT (1995) Use of a free radical method to evaluate antioxidant activity. LWT-Food Sci Tech 28:25CrossRefGoogle Scholar
  3. Foldbjerg R, Olesen P, Hougaard M, Dang DA, Hoffmann HJ, Autrup H (2009) PVP-coated silver nanoparticles and silver ions induce reactive oxygen species, apoptosis and necrosis in THP-1 monocytes. Toxicol Lett 190:156CrossRefPubMedGoogle Scholar
  4. Guo H, Qian H, Idris NM, Zhang Y (2010) Singlet oxygen-induced apoptosis of cancer cells using upconversion fluorescent nanoparticles as a carrier of photosensitizer. Nanomed Nanotech 6:486CrossRefGoogle Scholar
  5. Hu W, Lee SK, Jung MJ, Heo SI, Hur JH, Wang MH (2010) Induction of cell cycle arrest and apoptosis by the ethyl acetate fraction of Kalopanax pictus leaves in human colon cancer cells. Bioresour Technol 101:9366CrossRefPubMedGoogle Scholar
  6. Jeyaraj M, Rajesh M, Arun R, MubarakAli D, Sathishkumar G, Sivanandhan G, Dev GK, Manickavasagam M, Premkumar K, Thajuddin N, Ganapathi A (2013a) An investigation on the cytotoxicity and caspase-mediated apoptotic effect of biologically synthesized silver nanoparticles using Podophyllum hexandrum on human cervical carcinoma cells. Colloids Surf B Biointerfaces 102:708CrossRefPubMedGoogle Scholar
  7. Jeyaraj M, Sathishkumar G, Sivanandhan G, MubarakAli D, Rajesh M, Arun R, Kapildev G, Manickavasagam M, Thajuddin N, Premkumar K, Ganapathi A (2013b) Biogenic silver nanoparticles for cancer treatment: an experimental report. Colloids Surf B Biointerfaces 106:86CrossRefPubMedGoogle Scholar
  8. Kulandaivelu B, Gothandam KM (2016) Cytotoxic effect on cancerous cell lines by biologically synthesized silver nanoparticles. Braz Arch Boil technol 59:1Google Scholar
  9. Kumar CG, Mamidyala SK (2011) Extracellular synthesis of silver nanoparticles using culture supernatant of Pseudomonas aeruginosa. Colloids Surf B 84:462CrossRefGoogle Scholar
  10. Lafon-Hughes L, Di Tomaso MV, Mendez-Acuna L, Martınez-Lopez W (2008) Chromatin-remodelling mechanisms in cancer. Mutat Res 658:191CrossRefPubMedGoogle Scholar
  11. Lakshmi S, Dhanya GS, Joy B, Padmaja G, Remani P (2008) Inhibitory effect of an extract of Curcuma zedoariae on human cervical carcinoma cells. Med Chem Res 17:335CrossRefGoogle Scholar
  12. Liu H, Xu Y, Wen S, Zhu J, Zheng L, Shen M, Zhao J, Zhang G, Shi X (2013) Facile hydrothermal synthesis of low generation dendrimer-stabilized gold nanoparticles for in vivo computed tomography imaging applications. Polym Chem 4:1788CrossRefGoogle Scholar
  13. Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2−ΔΔCT method. Methods 25:402CrossRefPubMedGoogle Scholar
  14. Nasongkla N, Bey E, Ren J, Ai H, Khemtong G, Guthi JS, Chin SF, Sherry AD, Boothman DA, Gao J (2006) Multifunctional polymeric micelles as cancer-targeted, MRI-ultrasensitive drug delivery systems. Nano Lett 6:2427CrossRefPubMedGoogle Scholar
  15. Pardee AB, Stein GS (2011) The biology and treatment of cancer: understanding cancer. Wiley, HobokenGoogle Scholar
  16. Percival SL, Bowler PG, Russell D (2005) Bacterial resistance to silver in wound care. J Hosp Infect 60:1CrossRefPubMedGoogle Scholar
  17. Pradeep T (2009) Noble metal nanoparticles for water purification: a critical review. Thin Solid Films 517:6441CrossRefGoogle Scholar
  18. Ramar M, Manikandan B, Marimuthu PN, Raman T, Mahalingam A, Subramanian P, Karthick S, Munusamy A (2015) Synthesis of silver nanoparticles using Solanum trilobatum fruits extract and its antibacterial, cytotoxic activity against human breast cancer cell line MCF 7. Spectro chimica Acta A 140:223CrossRefGoogle Scholar
  19. Saha K, Agasti SS, Kim C, Li X, Rotello VM (2012) Gold nanoparticles in chemical and biological sensing. Chem Rev 112:2739CrossRefPubMedPubMedCentralGoogle Scholar
  20. Salunke GR, Ghosh S, Kumar RS, Khade S, Vashisth P, Kale T, Chopade S, Pruthi V, Kundu G, Bellare JR, Chopade BA (2014) Rapid efficient synthesis and characterization of silver, gold, and bimetallic nanoparticles from the medicinal plant Plumbago zeylanica and their application in biofilm control. Int J Nanomedicine 9:2635PubMedPubMedCentralGoogle Scholar
  21. Sharma G, Sharma AR, Kurian M, Bhavesh R, Nam JS, Lee SS (2014) Green synthesis of silver nanoparticle using Myristica fragrans (nutmeg) seed extract and its biological activity. Dig J Nanomater Biostruct 9:325Google Scholar
  22. Singh NP, McCoy MT, Tice RR, Schneider EL (1988) A simple technique for quantitation of low levels of DNA damage in individual cells. Exp Cell Res 175:184CrossRefPubMedGoogle Scholar
  23. Singh J, Upadhyay AK, Bahadur A, Singh B, Singh KP, Rai M (2006) Antioxidant phytochemicals in cabbage (Brassica oleracea L. var. capitata). Sci Hortic-Amsterdam 108:233CrossRefGoogle Scholar
  24. Singh S, Jain DVS, Singla ML (2013) Sol–gel based composite of gold nanoparticles as matix for tyrosinase for amperometric catechol biosensor. Sensor Actuat B-Chem 182:161CrossRefGoogle Scholar
  25. Sivakumar AS, Anuradha CV (2011) Effect of galangin supplementation on oxidative damage and inflammatory changes in fructose-fed rat liver. Chem Biol Interact 193:141CrossRefPubMedGoogle Scholar
  26. Sivakumar AS, Ochirbat C, Cho SH, Yang J, Hwang I (2014) Antiapoptotic effect of a novel synthetic peptide from bovine muscle and MPG peptide on H2O2-induced C2C12 cells. In vitro cell dev biol anim 50:630CrossRefPubMedGoogle Scholar
  27. Vivek R, Thangam R, Muthuchelian K, Gunasekaran P, Kaveri K, Kannan S (2012) Green biosynthesis of silver nanoparticles from Annona squamosa leaf extract and its in vitro cytotoxic effect on MCF-7 cells. Process Biochem 47:2405CrossRefGoogle Scholar
  28. Woolfenden BS, Wilson RL (1982) Radical cations as reference chromogens in studies of one-electron transfer reactions: pulse radiolysis studies of 2, 2-azinobis-(3-ethylbenzthiazoline-6-sulfonate). J Chem Soc Perkin Trans:805–812Google Scholar

Copyright information

© The Society for In Vitro Biology 2017

Authors and Affiliations

  • Allur Subramaniyan Sivakumar
    • 1
  • Chandran Krishnaraj
    • 2
  • Sunirmal Sheet
    • 3
  • Dileep Reddy Rampa
    • 4
  • Da Rae Kang
    • 1
  • Shah Ahmed Belal
    • 1
  • Abhay Kumar
    • 5
  • In Ho Hwang
    • 6
  • Soon-Il Yun
    • 2
  • Yang Soo Lee
    • 3
  • Kwan Seob Shim
    • 1
    Email author
  1. 1.Department of Animal BiotechnologyChonbuk National UniversityJeonjuRepublic of Korea
  2. 2.Department of Food Science and TechnologyChonbuk National UniversityJeonjuRepublic of Korea
  3. 3.Department of Wood Science and Technology, College of Agriculture and Life SciencesChonbuk National UniversityJeonjuRepublic of Korea
  4. 4.Department of BIN Convergence Technology, College of EngineeringChonbuk National UniversityJeonjuRepublic of Korea
  5. 5.R&D CentreM/s. Eureka Forbes LimitedBangaloreIndia
  6. 6.Department of Animal ScienceChonbuk National UniversityJeonjuRepublic of Korea

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