br Introduction br Nanotechnology is the technology applied
Nanotechnology is the technology applied at the “nano” level which means short or dwarf . Nanotechnology deals with nanoparticles which are either metallic or non-metallic nanoparticles. Metallic na-noparticles especially silver(Ag), gold(Au) and platinum(Pt) have at-tracted the interest to the researchers because of their application in biomedical, pharmaceutical  and medical field like biosensing for medical imaging . Among various metallic nanoparticles silver na-noparticles (AgNPs) have unique physical, chemical, optical, electrical and biomedical properties which makes them potential applications in medicine for catalysis, biosensing, imaging, drug delivery and
fabrication of nanodevices [4–6]. Engineered AgNPs have been de-signed in the intention to improve the physical properties of nano-particles in order to improve the targeted drug delivery to various or-gans . Nanoparticles whose size is below 10 nm have extreme application because there chemical and physical effect arising from quantum size effect which confers them with great potential in elec-tronics, drugs, pigments, and biosensors .
Nanoparticles can be synthesized by chemical and physical methods which are expensive, time-consuming and sometimes hazardous che-mical is used for the synthesis . However using a green approach (various biological sources) for the biosynthesis of nanoparticles like fungi, plant, algae, and bacteria are cost-effective, less expensive, and
E-mail addresses: [email protected] (S. Majeed), [email protected] (M. Danish).
yield of nanoparticles is good [10,11]. The green approach depends upon (1) selection of solvent medium, (2) chosen of eco-friendly re-ducing agent, and (3) selection/identification of toxic free substances for the stability of AgNPs. Therefore, biological nanofactories for the production of nanoparticles is preferred over the physical and chemical methods. Cancer of various organs like liver, lung, colon, breast, and bones have the leading mortality rate in the world wide are expected to reach up to 22 million in the next coming two decades . Generally cancers can be treated through chemotherapy, radiation therapy, and surgery. The chemotherapy involves the various cytotoxic drugs to damage the cancer SB 203580 apart from this these drugs also damages the normal cells . The side effect of chemotherapy (anticancer agents) could be minimized by using controlled targeted drug delivery which can kill only cancer cells without harming the normal cell by using nano-particles. The AgNPs can play an important role in engineering with anticancer drugs with maximum therapeutic effect .
Bovine Serum albumin (BSA) is a serum protein having the prop-erties of plasma proteins which works well under the pH 7 to 9 having so many hydrophobic binding sites. The BSA is natural transporter of various drugs fatty acid hence could be used as transport of various therapeutic drugs that can bind covalently as well as non –covalently having the property to release the drugs in a controlled manner . The chrysin BSA nanoparticles enhanced the cytotoxic effect against HEF-2 cancer cell with controlled and sustained release showing the BSA are biocompatible [16,17].
This study aims to synthesize the cAgNPs from Bacillus cereus (ATCC 14579) and capped them with bovine serum albumin (BSA). These AgNPS and cAgNPS were characterized by UV Visible spectroscopy, FTIR, FESEM, EDX and TEM analysis. Proteins degradation was de-termined by thermogravimetric analysis (TGA) to confirm the protein loss. The cytotoxicity AgNPs and cAgNPs were analyzed against MCF-7, HCT-116, and MG-63 cells lines and alongside compared with normal 3T3 fibroblast cell line on dose-dependent manner. Further LDH assay was analyzed from LDH kit, apoptotic activity, and DNA damage was analyzed by Ethidium bromide/Acridine orange (EB/AO) dual staining method and DNA fragmentation by using agarose gel electrophoresis.
2. Material and methods
2.1. Cell culture cell line and reagents
All the four cell lines, breast cancer MCF-7, intestinal colon cancer HCT-116, bone cancer osteosarcoma MG-63 and normal fibroblast 3T3 cell lines were procured from national center of cell science studies Poona (NCCS) India. All the four cell lines were grown on Dulbecco's modified eagle's medium (DMEM) supplied with 1% (v/v) glutamine,100 units mL−1 streptomycin, fetal bovine serum (FBS) 10% (v/v), penicillin 100 units mL−1 under carbon dioxide(5%) environ-ment at 37 °C.
2.2. Culture collection
The Bacillus cereus was procured from American type culture col-lection (ATCC-) with strain number −14,579 was cultured on the nu-trient agar plate. The plate was incubated at 37 °C for 24 h. Peptone broth was prepared in a conical flask containing 150 mL of water. The flask was swirled around to ensure perfect mixing of peptone powder with water. The B. cereus was then cultured in peptone broth by in-oculating a few colonies of the bacteria from the cultured plate. The flask was incubated at 37 °C for 24 h to let complete growth of bacteria. The turbid bacterial solution was centrifuged at 10,000 rpm for 15 min. The upper layer supernatant was transferred into the separate flask and was later subjected for AgNPs synthesis.