Nanotechnology. 2017 Sep 8;28(36):365101.

Investigation of novel superparamagnetic Ni0.5Zn0.5Fe2O4@albumen nanoparticles for controlled delivery of anti-cancer drug

Mohd Qasim1, Khushnuma Asghar1, Gangappa Dharmapuri2, and D. Das1*

1School of Engineering Sciences and Technology, University of Hyderabad, Hyderabad 500046, India

2Department of Animal Biology, School of Life Sciences, University of Hyderabad, Hyderabad-500046, India.

*Corresponding author: Tel: +91 40 23134454 / Fax: +91-40-23011087



In spite of intense research effort, cancer is still one of the most deadly diseases in the world. Several types of anticancer drugs have been commercialized but these conventional molecular drugs have several limitations including rapid clearance and high toxicity toward normal cells [1]. Loading the drug in a nanocarrier and then parcelling it to the tumour tissues offer several benefits over the conventional drug delivery approach [2]. Several types of materials, depending on its biocompatibility, biodegradability and hydrophilicity, have been explored to develop variety of drug nanocarriers. Natural biopolymers, especially albumin proteins, are most promising materials for nanocarrier development due to their inherent biocompatibility, biodegradability and hydrophilicity. Among various proteins, water soluble, biocompatible and highly amenable (because of the presence of large no of functional groups) Egg albumen is expected to have enormous potential to be used as nanocarrier material. Also, it is inexpensive alternative to commonly used human serum albumin (HSA) and bovin serum albumin (BSA). Incorporation of magnetic nanoparticles (MNP) in a nanocarrier give them excellent magnetic targeting ability, which is not possible in their intrinsic form. Excellent magnetic properties and potential anticancer activity of NZF NPs motivated us to use them as magnetic component in this novel nano-formulation. In our earlier work we have reported egg albumen assisted coating of Zn0.95Ni0.05O on NZF nanoparticles (NP) [3] and recently, we have reported NZF@mSiO2 core shell nanocarrier for biomedical applications [4].

In the present investigation we have amalgamated the properties of superparamagnetic Ni0.5Zn0.5Fe2O4 and albumen nanoparticle together to produce multifunctional nanosystem for tumour diagnosis and therapy. We, for the first time, have formulated novel, multifunctional Ni0.5Zn0.5Fe2O4@Egg albumen (NZF@Alb) and doxorubicin loaded Ni0.5Zn0.5Fe2O4@albumen (NZF@Alb-Dox) core-shell nanoparticles by facile environmental friendly synthesis process using inexpensive chicken egg albumen and characterized them for different physiochemical properties by various techniques. NZF@Alb-Dox and NZF@Alb NPs were obtained following the procedure schematically shown in figure 1. Detailed formation mechanism of NZF@Alb-Dox NPs and NZF@Alb NPs have been discussed in our published article [1]. It has been noted that development of NZF@Alb-Dox NPs was probably driven by electrostatic and /or hydrophobic interaction among NZF, Dox, and albumen. XRD analysis has shown successful formation of spinel cubic NZF (crystalline size ~21 nm), presence of crystalline NZF in the amorphous albumen matrix and also suggested coating of albumen does not alter crystalline nature of NZF.



Figure 1. Schematic representation of formation of NZF@Alb-Dox and NZF@Alb NPs.


It has been observed from FESEM (figure 2(a)) and TEM (figure 2(b)) analysis of NZF NPs that particles are nearly spherical in shape with mean particle sizes of ~ 22 nm. The larger particle sizes of NZF@Alb NPs compared to NZF NPs was attributed to the coating of albumen as shown in Figure 2(c). Mean particle size of nanocomposite was found to be ~100 nm. Layer of amorphous albumen (grey color) on crystalline NZF NPs (black) has clearly been observed in low and high magnification TEM micrographs of NZF@Alb NPs (figure 2c-d). Detailed HRTEM analysis using inverse fast Fourier Transformation (IFFT) technique of albumen coating and NZF region of both NZF@Alb and NZF@Alb-Dox NPs have been carried out and discussed in our published work (figure 4 and 5 of ref [1]). NZF and NZF@Alb NPs were found to be super-paramagnetic in nature with high Ms values, which are essential requirement of MNPs for biomedical applications. Further, the obtained Ms value (36 emu/gm) of NZF@Alb NPs is very good for magnetically guided drug delivery application. FTIR analysis was used to further support the coating of albumen on NZF NPs and loading of Dox in NZF@Alb-Dox.



Figure 2. FESEM (a) and TEM (b) micrographs of NZF NP. (c) TEM micrograph of NZF@Alb NP. Figure (d) shows enlarge image of red framed region in (c). Coating of albumen (grey) on black NZF NP can be seen in (d).


Reported nano-formulation showed a very good Dox loading ability with the Dox loading efficiency and loading capacity of ~90 % and ~5 % respectively, which was attributed to the good electrostatic interaction between oppositely charged Dox and albumen molecules. Dox release profiles from NZF@Alb-Dox NPs have been studied in two pH environments i.e. pH of cancer cells (especially for microenvironment in endosomes and lysosomes) and blood plasma. Slow and desired sustained Dox release values i.e. 41.5% (at pH 5) and 32.5% (at pH 7.4) were observed even after 50 hours of soaking. A sustained, controlled and pH dependent release of Dox from NZF@Alb-Dox NPs was observed, which is very promising and desirable behavior for anticancer drug-delivery applications (figure 3). These pH-responsive nature of NZF@Alb-Dox NPs may diminish the unwanted release of Dox in the blood circulation system and thus can improve the efficacy of Dox by releasing it preferably at the cancer site. Release kinetics of Dox were also investigated using different mathematical models including zero order, first order, Higuchi and Korsmeyer–Peppas models and described in our published work [1].


Figure 3. Schematic illustrating pH-dependent Dox release behavior from NZF@Alb-Dox NPs.


NZF and NZF@Alb NPs did not show much destruction of normal cells (RAW 264.7 cells) and are biocompatible in nature. Slight increase in biocompatibility was observed for NZF@Alb.  MTT assay against Hela cells has shown a dose dependent promising anticancer activity (IC50 ~20 µg/ml) of magnetically guidable NZF@Alb-Dox NPs. Further, prepared nano-formulation was found to have excellent long-term chemical and physical stability (good dispersibility and colloidal stability), which is also a primary requirement of nanocarriers for biomedical applications. Detailed physicochemical characteristics of NZF@Alb-Dox NPs have been demonstrated in figure 4. The result of our present investigation reveals that this inexpensive, scalable and pH responsive nanoformulation could be very much useful for biomedical applications including targeted and controlled delivery of anticancer drugs for cancer treatment.


Figure 4. Summarized characteristics of NZF@Alb-Dox NPs.


In summary, for the first time, novel pH responsive, water dispersible, biocompatible, inexpensive and magnetically targetable multifunctional NZF@Alb and Dox loaded NZF@Alb-Dox core-shell nanoparticles have been prepared by a facile environmental friendly method using inexpensive chicken egg albumen, NZF NP and Dox. An extensive characterization of the synthesized nanocarriers shows suitable physiochemical properties for magnetically targeted and controlled drug delivery applications. It is demonstrated that easily available egg albumen could be a profitable alternative than commercially available expensive BSA and HSA. NZF@Alb-Dox nanoformulation have shown ideal drug loading/release behavior and excellent anticancer activity against Hela cells. Thus, the present study could be extremely useful for the advancement of albumin based nanocarriers design and development for biomedical applications. The reported results, presented herein, are significant because it has a great potential to pave a new way for the formulation of variety of novel nanocarriers for cancer diagnosis and therapy using inexpensive and biocompatible biopolymers-based sources.

The present work was carried out at the Laboratory of Functional Ceramic Materials ( headed by Prof Dibakar Das in the School of Engineering Sciences and Technology, University of Hyderabad, India. Prof Das’s research group is actively engaged in the research activities of novel functional materials, magnetic materials, magnetic nanomaterials and nanocomposites.



Photographs of authors of the present investigation.


About the authors:

Mohd Qasim

Mohd Qasim did M.Sc. in Physical Chemistry and MTech in Nanotechnology from the Aligarh Muslim University, India. Later, he joined for PhD in Nanoscience and Technology in the School of Engineering Sciences and Technology (SEST), University of Hyderabad, India under the supervision of Prof. Dibakar Das. He has been working on synthesis and characterization of advanced multifuntional magnetic nanoparticles for biomedical applications. Recently, he has submitted his PhD thesis in the same area. His research interest mainly covers development of multifunctional nanomaterials, magnetic nanoparticles, nanocarriers, nanocomposites and their biomedical applications. He published around 14 research papers in refereed Journals and conference proceedings and has presented his research work in various national and international conferences. Mohd Qasim can be reached at

Khushnuma Asghar

Khushnuma Asghar obtained M.Sc. in Physical Chemistry, PG Diploma in Nanotechnology and MTech in Nanotechnology from the Aligarh Muslim University, India. Currently, she is pursuing PhD in Material Engineering at the School of Engineering Sciences and Technology (SEST), University of Hyderabad, India under the supervision of Prof. Dibakar Das. Her research interest mainly covers development of functional nanomaterials and its applications. She has published several research papers in refereed Journals and conference proceedings and has presented her research work in various national and international conferences. Khushnuma Asghar can be reached at

Dr. Gangappa Dharmapuri

Dr. D. Gangappa, has completed his Ph.D from the Department of Biotechnology, Sri Krishnadevaraya University, Anantapuram in 2015. Currently he is working as Post-Doctoral fellow (DST-SERB- NPDF) in the Department of Animal Biology at the School of Life Sciences, University of Hyderabad, Hyderabad. He can be reached at

Prof. Dibakar Das

Dibakar Das is currently a Professor in the School of Engineering Sciences and Technology (SEST) at the University of Hyderabad, India. He obtained Ph.D. from IIT Bombay in 2004. Dr. Das carried out a significant part of his thesis work at the University of Colorado at Boulder, and National Institute of Standard and Technology (NIST) Boulder, USA. Following Ph. D., he joined the University of Cincinnati, Ohio, USA as a Postdoctoral Research Assistant in 2004. After three and half years of postdoctoral research at the University of Cincinnati, he joined Sinmat Inc. in Gainesville, Florida, USA, as a Senior Scientist, in 2007. Serving Sinmat for about 2 years he joined the School of Engineering Sciences and Technology (SEST) at the University of Hyderabad in 2009 August. Dr. Das is elected fellows of the Indian Institute of Ceramics (IIC) and Telangana Academy of Sciences. He has published more than 90 peer-reviewed articles in journals and conference proceedings of international repute. His research interest includes CMP of WBG semiconductors, lead-free piezoelectric ceramics, ferrite ceramics for magnetoelastic, magnetoelectric, multifuntional magnetic nanomaterials, drug nanocarriers, nanocomposite and their biological applications. More details about Prof. Das’s research activities could be found at his research group web page Prof. Das can be reached at or



[1] Mohd Qasim, Khushnuma Asghar, Gangappa Dharmapuri and D Das, Nanotechnology 28 (2017) 365101 (18pp)

[2] Tian Y, Jiang X, Chen X, Shao Z and Yang W 2014 Adv. Mater. 26 7393–8

[3] Qasim M, Asghar K, Singh B R, Prathapani S, Khan W, Naqvi A H and Das D 2015 Spectrochim. Acta Part A: Mol. Biomol. Spectrosc. 137 1348–56

[4] Qasim M, Asghar K and Das D 2017 chapter 5 Proceedings of the 3rd Pan American Materials Congress. The Minerals, Metals & Materials Series ed M Meyers et al (Springer Int. Publishing) pp 47–54