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Series in BioEngineering
Manoranjan Arakha
Suman Jha
Interfacial
Phenomena
on Biological
Membranes
The Series in Bioengineering serves as an information source for a professional
audience in science and technology as well as for advanced students. It covers all
applications of the physical sciences and technology to medicine and the life
sciences. Its scope ranges from bioengineering, biomedical and clinical engineering
to biophysics, biomechanics, biomaterials, and bioinformatics.
More information about this series at http://www.springer.com/series/10358

Manoranjan Arakha Suman Jha
•
Interfacial Phenomena
on Biological Membranes
123
Manoranjan Arakha Suman Jha
National Institute of Technology Rourkela National Institute of Technology Rourkela
Rourkela, Odisha Rourkela, Odisha
India India
ISSN 2196-8861 ISSN 2196-887X (electronic)

ISBN 978-3-319-73325-8 ISBN 978-3-319-73326-5 (eBook)
https://doi.org/10.1007/978-3-319-73326-5
Library of Congress Control Number: 2017963270
© Springer International Publishing AG 2018

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Contents
1 Nanoparticle . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1
1.1 Synthesis of Nanoparticles . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2
1.2 Different Kinds of Nanoparticles . . . . . . . . . . . . . . . . . . . . . . . . . 4
1.2.1 Zinc Oxide Nanoparticle (ZnONP) . . . . . . . . . . . . . . . . . . 5
1.2.2 Iron Oxide Nanoparticle (IONP) . . . . . . . . . . . . . . . . . . . . 5
1.2.3 Silver Nanoparticle (AgNP) . . . . . . . . . . . . . . . . . . . . . . . 5
1.2.4 Liposome . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6
1.2.5 Albumin-Functionalized NPs . . . . . . . . . . . . . . . . . . . . . . 6
1.2.6 Polymeric NPs . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7
1.2.7 Quantum Dot . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7
1.3 Physicochemical Properties of Nanoparticles . . . . . . . . . . . . . . . . 8
1.3.1 Shape, Size, and Curvature . . . . . . . . . . . . . . . . . . . . . . . . 8
1.3.2 Surface Concentration . . . . . . . . . . . . . . . . . . . . . . . . . . . 9
1.3.3 Surface Functionality . . . . . . . . . . . . . . . . . . . . . . . . . . . . 10
1.3.4 Surface Potential . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 11
1.4 Physicochemical Properties of Biological Membranes
and Biomacromolecules . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 11
1.4.1 Cell Membrane . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 11
1.4.2 Bacterial Cell Wall . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 12
1.4.3 Eukaryotic Cell Membrane . . . . . . . . . . . . . . . . . . . . . . . . 13
1.4.4 Nucleic Acid Biomacromolecules . . . . . . . . . . . . . . . . . . . 14
1.4.5 Protein Biomacromolecules . . . . . . . . . . . . . . . . . . . . . . . 15
1.5 Nanoparticle Interfacial Interaction with Biological Membranes
and Biomacromolecules . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 18
1.5.1 Nanoparticle–Biological Membrane Interaction . . . . . . . . . 19
1.5.2 Cellular Internalization of Nanoparticles . . . . . . . . . . . . . . 20
1.5.3 Nanoparticle–Nucleic Acid Interaction . . . . . . . . . . . . . . . 20
1.5.4 Nanoparticle-Protein Interaction . . . . . . . . . . . . . . . . . . . . 21
v
vi Contents
1.6 Applications of Nanoparticle–Biomolecular Interactions

in Biological Sciences . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . ... 23
1.6.1 Nanoparticles as Novel Antibiotics . . . . . . . . . . . . . . . ... 23
1.6.2 Nanoparticle-Mediated Approach for Cancer Diagnosis
and Therapy . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . ... 24
1.6.3 Nanoparticle Acting as a Protein Folding Chaperone . . ... 25
1.6.4 Detection of Protein Aggregation Using Nanoparticles . ... 27
1.6.5 Advantages of Nanoparticles-Based Therapeutics
Over Conventional Therapies for Amyloidoses . . . . . . ... 28
References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . ... 29
2 Synthesis and Characterization of Nanoparticles . . . . . . . . . . ..... 37
2.1 Synthesis of Zinc Oxide Nanoparticle and Its Surface
Modification . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 37
2.1.1 Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 37
2.1.2 Materials and Methods . . . . . . . . . . . . . . . . . . . . . . . . . . . 37
2.1.3 Results and Discussion . . . . . . . . . . . . . . . . . . . . . . . . . . 38
2.1.4 Surface Modification of ZnONP . . . . . . . . . . . . . . . . . . . . 42
2.2 Synthesis and Surface Modification of Iron Oxide
Nanoparticle . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 45
2.2.1 Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 45
2.3 Synthesis of Silver Nanoparticle Using Bacteria from Coal
Mine—A Green Synthesis Approach . . . . . . . . . . . . . . . . . . . . . . 49
2.3.1 Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 49
References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 57
3 Effect of Interfacial Potential on Antimicrobial Propensity
of ZnONPs . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 61
3.1 Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 61
3.2 Materials Methods . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 62
3.2.1 ZnONP-Bacteria Interfacial Potential Measurement . . . . . . 62
3.2.2 Bacterial Cell Viability in Presence of ZnONPs . . . . . . . . . 62
3.2.3 ROS Detection . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 63
3.2.4 Bacterial Morphology on ZnONP Treatment . . . . . . . . . . . 63
3.3 Results and Discussion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 64
3.3.1 ZnONP-Bacteria Interfacial Potential . . . . . . . . . . . . . . . . . 64
3.3.2 Surface Potential Neutralization of B. Subtilis
and E. Coli by ZnONPs . . . . . . . . . . . . . . . . . . . . ...... 68
Contents vii
3.3.3 Enhanced ROS Production in Presence

of ZnONP-Bacteria Interface . . . . . . . ............... 69
3.3.4 Surface Morphology of Bacteria upon
ZnONP Treatment . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 70
3.4 Discussion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 70
3.5 Conclusion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 76
References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 76
4 Effect of Surface Functionality on Antimicrobial Propensity
of Iron Oxide Nanoparticles . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 79
4.1 Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 79
4.2 Materials and Methods . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 80
4.2.1 Growth Kinetic Analysis . . . . . . . . . . . . . . . . . . . . . . . . . 80
4.2.2 CFU Measurement . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 80
4.2.3 ROS Detection . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 81
4.2.4 LIVE/DEAD BacLight Fluorescence Microscopy
Assay . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 81
4.3.1 Effect of the Interfaces upon Bacterial Cell Viability . . . . . 82
4.4 Discussion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 84
4.5 Conclusion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 88
References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 89
5 Effect of ZnONP Surface Defects on Cytotoxic
and Antimicrobial Propensities . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 91
5.1 Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 91
5.2.1 Cell Culture and ZnONP Stock Solution Preparation . . . . . 92
5.2.2 Cytotoxicity of ZnONPs . . . . . . . . . . . . . . . . . . . . . . . . . . 92
5.2.3 ZnONP-Induced ROS Generation . . . . . . . . . . . . . . . . . . . 93
5.2.4 Comet Assay . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 93
5.2.5 Cell Cycle Analysis . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 93
5.2.6 ZnONP-Induced Autophagy . . . . . . . . . . . . . . . . . . . . . . . 94
5.2.7 ZnONP-Induced Apoptosis . . . . . . . . . . . . . . . . . . . . . . . . 94
5.2.8 Morphological Changes . . . . . . . . . . . . . . . . . . . . . . . . . . 95
5.3.1 Cytotoxic Propensity of ZnONPs . . . . . . . . . . . . . . . . . . . 95
5.3.2 Effect of ZnONP Treatment on the Cell Cycle . . . . . . . . . 99
5.3.3 Induction of Autophagy upon ZnONPs Treatment . . . . . . . 100
5.3.4 ZnONPs Treatment Causes Apoptotic Cell Death . . . . . . . 102
5.3.5 HT1080 Morphology upon ZnONPs Treatment . . . . . . . . . 103
5.3.6 Antimicrobial Propensity of ZnONPs . . . . . . . . . . . . . . . . 106
5.3.7 Conclusion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 108
References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 109
viii Contents
6 Effect of Interfacial Assembly of Antimicrobial Peptide

on Conformational and Functional Dynamics of the Peptide . . . . . . 111
6.1 Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 111
6.2.1 Preparation of AgNP–nisin Conjugates . . . . . . . . . . . . . . . 112
6.2.2 Biophysical Characterization of AgNP–nisin
Conjugates . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 113
6.2.3 Antimicrobial Activity of AgNP–nisin Conjugates . . . . . . . 114
6.2.4 Interfacial and Intracellular ROS Detection . . . . . . . . . . . . 114
6.2.5 Membrane Destabilization and Internalization
of AgNP–nisin Conjugates . . . . . . . . . . . . . . . . . . . . . . . . 115
6.3.1 Interfacial Assembly of Nisin at AgNP Interface . . . . . . . . 115
6.3.2 The Interfacial Assembly Enhances the Antimicrobial
Propensity of Nisin . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 122
6.3.3 Oxidative Stress Mediated Antimicrobial Activity
of AgNP–nisin Conjugates . . . . . . . . . . . . . . . . . . . . . . . . 125
6.3.4 Membrane Destabilization by AgNP–nisin Conjugates . . . . 126
6.3.5 Proposed Mechanism of the Assembled Nisin
Antimicrobial Activity . . . . . . . . . . . . . . . . . . . . . . . . . . . 129
6.3.6 Conclusion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 133
References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 133
7 Effect of Globular Protein Interfacial Assembly
on Conformational Dynamics of the Protein . . . . . . . . . . . . . . . . . . . 137
7.1 Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 137
7.2.1 Preparation of ZnONP Solution . . . . . . . . . . . . . . . . . . . . 138
7.2.2 Preparation of Lysozyme–ZnONP Conjugates . . . . . . . . . . 138
7.2.3 Circular Dichroism (CD) Spectropolarimetry . . . . . . . . . . . 138
7.2.4 Intrinsic Tryptophan Fluorescence Spectroscopy . . . . . . . . 139
7.2.5 ANS Fluorescence Studies of Lysozyme . . . . . . . . . . . . . . 139
7.2.6 Lysozyme Tryptophan Fluorescence Quenching Study
Using Acrylamide . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 139
7.2.7 Thioflavin T Assay of Lysozyme Fibrillation Kinetics . . . . 139
7.2.8 Transmission Electron Microscopy Study . . . . . . . . . . . . . 140
7.2.9 Circular Dichroism (CD) Spectropolarimetry . . . . . . . . . . . 140
7.3.1 Interfacial Assembly of Lysozyme at ZnONP Interface . . . 140
7.3.2 Antiamyloidosis Propensity of ZnONP Interface . . . . . . . . 147
7.4 Conclusion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 150
References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 151
Chapter 1
Nanoparticle
Fifty-seven years ago, on the evening of December 29, 1959, best known paper of
nanotechnology entitled ‘There’s Plenty of Room at the Bottom’ was delivered by
physicist professor Richard Feynman to the American Physical Society at the
California Institute of Technology, Pasadena [1]. In the paper, Dr. Richard
Feynman described the possibilities, if we could learn to control single atom and
molecules [1]. The work led the community to the era of nanotechnology. Hence,
the essence of nanotechnology is based on the ability to work at the molecular and
atomic levels to formulate fundamentally new molecular structure with advance
physicochemical properties [1, 2]. The combination of science and technology for
the nature of material at the nanoscale provides a strong foundation for nanotech-
nology [3]. The growth of nanotechnology is a convergence and divergence pro-
cess. For example, the convergence in understanding of nanotechnology reached its
maximum strength by 2000 AD, and one may expect divergence in applications of
nanotechnology in coming decades [3].
Among various nanotechnology fields, nanobiotechnology has been considered
as a most emerging field, all because of its various applications in molecular
diagnostics, material science, and bioengineering. The definition of nanoparticles
has been suggested by National Nanotechnology Initiative (NNI), USA. According
to NNI., particles with size range from 1 to 100 nm in at least one dimension are
called nanoparticles [4]. Moreover, the widely applied term ‘nano’ is adapted from
a Greek word, meaning ‘dwarf.’ ‘Nano’ is also used as a prefix for 10−9 magnitude
[5]. Nanoparticles have drawn great scientific interest as they bridge the physico-
chemical gap developed between bulk (macroscopic) material and atomic or
molecular structure [5]. The unique characteristic properties of nanoparticle are
quite different from macroscopic material. The differences are mainly developed
from high surface-to-volume ratio as well as improved percentage of grain
boundaries of nanoparticles [6]. These unique features of nanoparticles have drawn
the attention of various research groups to employ nanoparticles in various fields of
© Springer International Publishing AG 2018 1

M. Arakha and S. Jha, Interfacial Phenomena on Biological Membranes,
Series in BioEngineering, https://doi.org/10.1007/978-3-319-73326-5_1
2 1 Nanoparticle
science and technology. Hence, to do so, the synthesis of different nanoparticles

with advanced physicochemical properties has become a major interest in the
current era.
1.1 Synthesis of Nanoparticles
Different protocols have been optimized to synthesize different types of nanopar-

ticles that meet the requirements of different nanoparticle-mediated applications.
The synthesis protocols are broadly divided into two categories such as top-down
(physical process) and bottom-up (chemical and biological processes) approaches
[7, 8]. Generally, in top-down approach, the nanoparticles are formulated from their
bulk material or constituents. Examples of some top-down approaches are high
energy ball milling, where the nanoparticles are produced from the milling of their
respective bulk material. Another approach is wire explosion method; conducting
metal nanoparticles are produced from an explosion due to a sudden high current
pulse. In inert-gas condensation method, evaporated atoms from bulk material are
condensed in a matrix, and the respective nanoparticle growth is achieved. Laser
ablation approach is another physical method for nanoparticle synthesis, where
higher energy laser is used to induce evaporation. In the top-down approaches,
shape, size, and composition of nanoparticles can be monitored by controlling
different physicochemical parameters. The methods produce larger quantity of
nanoparticles, however with polydisperse nature [7].
In the latter approach, i.e., in bottom-up (chemical or biological processes)
approach, the elements generated from respective ion reduction assemble into
nanoparticles. Since the process is initiated from atoms, the processes are also
known as bottom-up approaches [7, 8]. For examples, in chemical approaches, the
reducing agent like sodium borohydride (NaBH4) helps in reduction of metal ions
into atoms, and the reduced atoms assemble to form nanosize crystals, hence
nanoparticles. Photochemical synthesis is another method for nanoparticle syn-
thesis, which is assisted by light. Nanoparticles are synthesized with the help of
ultrasound in sonochemical routes. Microemulsion is an approach for the synthesis
of nanoparticles using water in oil or oil in water emulsions. Additionally, in
solvothermal synthesis, the synthesis of nanoparticles happens in a closed system
from solvents at lower temperature, for example the coacervation or emulsion
methods for protein nanoparticle synthesis.
It is reported that decreasing hydrophobic interaction during protein unfolding
triggers the protein nanoparticle formation [9], which is an example of the
bottom-up approach. The protein nanoparticles are formed due to the conforma-
tional changes of protein, which depends upon its composition, cross-linking,
concentration, different chemical conditions like pH, ionic strength, type of solvents
[10]. During unfolding process, protein expresses different interactive groups like
disulfides, thiols, hydroxides. Interestingly, thermal and chemical cross-linking of
1.1 Synthesis of Nanoparticles 3
Fig. 1.1 Protein nanoparticles prepared by a coacervation or phase separation, b emulsion/solvent

extraction method, and c complex coacervation method. Reproduced from Lohcharoenkal et al.
[10]
the groups leads to the formation of protein nanoparticles with entrapped drug
molecule (Fig. 1.1).
Coacervation/desolvation process is one of the two well-known methods for
protein nanoparticle synthesis. The process is based on differential solubility of
proteins in different solvents, which is a function of solvent pH, polarity, ionic
strength, and electrolyte presence. As shown in Fig. 1.1, the process helps in
reduction of protein solubility, which leads to phase separation. The desolvating
agent helps in protein conformational change leading to protein coacervation or
precipitation. Size of the protein nanoparticle, formed in the process, can be con-
trolled by controlling the processing parameters like pH, ionic strength. The formed
nanoparticles are cross-linked by different agents like glutaraldehyde and glyoxal
[11]. In addition to coacervation method, emulsion/solvent extraction is another
method for preparation of protein nanoparticles. In this method, a high-speed
4 1 Nanoparticle
homogenizer/ultrasonic shear is being used to emulsify the aqueous solution of

protein in oil. As shown in Fig. 1.1, the protein nanoparticles are formed at the
interface of water/oil, upon emulsification. Additionally, phosphatidylcholine and/
or Span 80 are generally used as stabilizers for formed protein nanoparticle using
the method [10, 12]. The organic solvents are used to remove the oil phase of the
solution leading to the formation of protein nanoparticles. The method is generally
used to prepare different protein nanoparticles like albumin, whey protein.
Although various physical and chemical methods have been optimized for the
synthesis of nanoparticles, the use of strong and weak chemical reducing agents
along with protective agents like sodium borohydride, sodium citrate, and alcohols
for the synthesis of nanoparticles is not advisable. Most of these chemicals are
expensive, toxic, flammable and possess various environmental issues.
Additionally, the processes show low production rate [8]. In many cases, elevated
temperatures are required for the synthesis of nanoparticles [8, 13]. Hence, in order
to avoid the limitations of physical and chemical methods, biological methods (also
known as green synthesis method) have been adopted for the synthesis of metal and
metal oxide nanoparticles, a step to avoid toxic chemicals and to make the synthesis
process eco-friendly [14, 15]. In addition, the biological methods are less toxic,
safe, and energy efficient than the other two methods. Biological agents, commonly
used for the synthesis of nanoparticles, are plant extracts, extracts from microor-
ganisms and fungi, etc. [14]. It is also reported that the biological agents help in
reduction of metal ions at a faster rate than in other two methods and require
ambient temperature and pressure [8]. Additionally, shape and size of nanoparticles
can be modulated by the change in metal to extract ratios, pH, temperature of the
reduction reaction, agitation, etc. In this approach, metallo-enzyme present in
biological agents reduces metal ions into respective elements or molecule, and the
resultant elements are capped at nanosize by same biomolecules or others present in
the reducing medium. Hence, nanoparticles synthesized by the method have
functionalized surface because of absorption of different cellular moieties as a
capping agent or stabilizer [8].
1.2 Different Kinds of Nanoparticles
According to different applications of nanotechnology, different methods have been

optimized to fabricate different kinds of nanoparticles. Different nanoparticles are
metal/metal oxide nanoparticles for antimicrobial/magnetic/photocatalytic/heavy
metal adsorption applications, liposome nanoparticle for microRNA/drug delivery
application, protein nanoparticle for biocompatible drug delivery application, etc.
Following is the description of different kinds of nanoparticles.
1.2 Different Kinds of Nanoparticles 5
1.2.1 Zinc Oxide Nanoparticle (ZnONP)
Due to the presence of wide band gap (3.37 eV) as well as large excitation binding
energy (60 meV at room temperature), ZnONP possesses unique features like UV
light absorption, semiconducting, catalytic, and antimicrobial properties, etc. [16,
17]. In addition, ZnONP possesses unique features that are completely different
from bulk ZnO material like proportion of atoms on the surface of ZnONP, elec-
tronic band gap, etc. [17]. Hence, ZnONP is widely used in different fields such as
water and air disinfection, anticancer agents, antimicrobial agents, remediation of
hazardous waste, etc. [16]. Along with these applications, these nanoparticles are
also used for solar cells, photocatalytic applications, antivirus agent in coating,
biosensors, biological imaging, etc. [16, 18, 19].
1.2.2 Iron Oxide Nanoparticle (IONP)
Due to supermagnetic nature of iron oxide nanoparticles (IONPs), they are widely
used as passive and active targeted imaging agents [20]. The superparamagnetic
iron oxide nanoparticles (SPIONs) contain iron oxide as the core, and an outer layer
of dextran or other biocompatible compounds to make the core stable in physio-
logical medium [21–23]. The commonly used SPIONs are magnetite (Fe3O4) and
maghemite (cFe2O3). Interestingly, these SPIONs exhibit superparamagnetism in
size-dependent manner, so that in the presence of external magnetic field they
become magnetized and become neutral upon removal of the field. Additionally,
SPIONs are degraded into iron and/or iron oxide molecules, which are metabolized
further and stored in cells in bound form with ferritin, and finally incorporated into
hemoglobin [24]. Since different magnetic nanoparticles, like SPIONs, are bio-
compatible in nature and chemically stable and possess magnetic behavior, they are
widely used in different biomedical applications [25]. These nanoparticles are also
used for delivery of various drugs to their targeted tissues by employing external
magnetic field [25]. Additionally, they are used in analytical chemistry, antigen
diagnosis, tissue repair, pathogen detection, protein separation, etc. [24–26].
1.2.3 Silver Nanoparticle (AgNP)
Even before the evolution of nanotechnology, silver metal had been used in various
applications. Moreover, the understanding of nanotechnology has enhanced the
applications of silver by many folds, but as AgNP [8]. Recently, because of
antimicrobial property, the efforts are being made to incorporate AgNP into dif-
ferent medical devices, surgical instruments, surgical masks, etc. [27]. In addition,
many studies have reported the wound-healing capacities of ionic silver; silver
6 1 Nanoparticle
sulfadiazine is being replaced by AgNP to treat wounds [27–29]. Due to enhanced

physicochemical properties of AgNP, they are widely used in biomedical imaging,
nanomedicine, biosensing, catalysis, drug delivery, nanodevice fabrications, etc.
[27, 30]. Additionally, due to strong antimicrobial activity, the nanoparticle is
widely used in the production of sterile materials [8]. AgNP is synthesized from the
reduction of silver salt with the help of different reducing agents like sodium citrate,
sodium borohydride [27]. However, different stabilizing agents, such as polyvinyl
alcohol, bovine serum albumin (BSA), cellulose, citrate, etc., are used during the
synthesis of AgNPs. Commonly used methods for the nanoparticle synthesis are
chemical reduction in solution, sonochemical method, microwave-assisted method,
microemulsion method, etc. [31]. Along with these synthesis approaches, produc-
tion of AgNPs using green synthesis methods has also drawn the attention of
various research groups.
1.2.4 Liposome
Liposome is considered as one of the first nanoparticles formulation, which was

described in 1965 as a cellular membrane model [20, 32]. These are spherical
vesicles containing a single or multiple bilayers of lipids and self-assemble upon
suspension into aqueous solution [33]. The unique features of liposome, which
increase its use in biological applications, are diverse ranges of available compo-
sitions, able to carry and protect encapsulated/adsorbed molecules, biocompatible,
and biodegradable nature [20, 33, 34]. Hence, the liposomes are used as transfection
agents for different genetic materials like microRNA into a cell. The process is
commonly known as lipofection [35]. The process forms aggregates of cationic
lipids with anionic genetic material. Similarly, the liposomes are also used as
therapeutic drug carriers [32].
1.2.5 Albumin-Functionalized NPs
In biomedical sciences, nanoparticles are widely used as drug delivery agents, since
nanoparticles have the potential to protect the target drug from degradation,
enhance the drug absorption efficacy, improve intracellular penetration and distri-
bution, and modify drug tissue distribution profile [36]. In addition, nanoparticles
are widely accepted agents for drug delivery since they withstand different physi-
ological stresses and improve biological stability, high possibility of oral delivery,
etc. [36]. Among different nanoparticles, protein-based nanoparticles have drawn
the attention of researchers for their better stability during storage as well as
non-cytotoxic nature [36]. Albumin is being used as a drug delivery vehicle for
cancer treatment, since it acts as a natural carrier for different hydrophobic mole-
cules like water-insoluble plasma substances, vitamins, and hormone [37].
1.2 Different Kinds of Nanoparticles 7
Albumin-bound nanoparticles are important class of nanoparticles which carry the

hydrophobic molecules in bloodstream using endogenous albumin pathways [38].
Albumin binds hydrophobic molecules non-covalently and avoids solvent-based
toxicity [39]. Hence, albumin-based nanoparticles are used as drug delivery
vehicles.
1.2.6 Polymeric NPs
Polymeric nanoparticles are another group of nanoparticles those are formed from
the polymers having biocompatible and biodegradable properties and are exten-
sively used as therapeutic drug carriers, for example dendrimer-encapsulated
nanoparticles (DEN), protein nanoparticles (pNP) [40]. These are formulated from
block copolymers having different hydrophobicity. In an aqueous environment,
these copolymers spontaneously self-assembled into core-shell micelle [41]. The
hydrophilic and hydrophobic drugs, proteins, and nucleic acids can be encapsulated
on the polymeric nanoparticles for different biological applications [42]. Polymeric
nanoparticles are very much important because they help in safety and efficacy of
drug they usually carry. Protein nanoparticle, one of the polymeric nanoparticles, is
being treated as potential delivery agent for the anticancer drugs, since pNPs are
relatively safe, easy to prepare, and easy to control size distribution [43]. For
example, albumin-based nanocarrier system has made an impact on cancer treat-
ment [10]. Balancing of attractive and repulsive forces in protein is the key factor
for the formulation of protein nanoparticle. On the other hand, DEN is primarily
used as a catalyst, because of very high surface-to-volume ratio and highly
monodisperse nature [40]. DEN is made of dendrimer, commonly used dendrimer
like poly(aminoamine), which is attached terminally to a metal ion. In the presence
of reducing agents like sodium borohydride, reduction of metal ions into metal
element leads to dendrimer-encapsulated nanoparticle formation. Size of the
nanoparticle can be easily controlled by choosing the degree of aminoamines
polymerization [40].
1.2.7 Quantum Dot
Quantum dots are generally semiconductor particles having size less than 10 nm.
These were first discovered in 1980. Quantum dots are well known for unique
electronic and optical properties [44]. Among different physicochemical properties,
the particles have very broad absorption spectra and possess very narrow emission
spectra. Quantum dots possess long lifetime and emit bright colors, and efficiency
of the quantum dots is high. Hence, the quantum dots are widely used in different
optical applications. Additionally, these are widely used in biological fields for cell
labeling, biomolecule tracking, etc. [45–47]. In addition, QDs have numerous
8 1 Nanoparticle
advantages than other fluorophores like organic dyes and fluorescent proteins [48].
It is important to note that, conventional dyes commonly suffer from narrow
excitation spectra and require specific light wavelength for excitation, which varies
from one dye to another, whereas QDs having broad absorption spectra require
wide range of light wavelengths for excitation. This property of QDs can be
exploited to excite different colored QDs by a single wavelength simultaneously
[48].
1.3 Physicochemical Properties of Nanoparticles
1.3.1 Shape, Size, and Curvature
Shape, size, and curvature of nanomaterials greatly influence the internalization of

nanomaterials inside a cell. It has been reported that kinetic of nanomaterials cel-
lular uptake varies with the shape and size of nanomaterials [49]. When discussing
the internalization of nanomaterials based on their shape, it has been reported that
spherical particle of same size is internalized quickly than the rod shape, since
longer membrane wrapping time is required for rod-shape nanomaterial than
spherical shape. In other ways, size and curvature also play crucial roles for cellular
uptake of nanomaterials, as size and curvature of nanoparticle strongly affect the
binding and activation of membrane receptors, subsequently affect the respective
protein expression, and hence affect the cellular uptake [50]. From various in vitro
and in vivo studies, it was found that nanomaterials inside a biological milieu
display variant outcomes due to aggregation of nanomaterials with various sizes.
However, the shape and size of nanomaterials greatly influence the toxicity/
compatibility of nanomaterials toward mammalian and/or bacterial cells.
The interaction of proteins on nanoparticles surface is also affected by different
physicochemical parameters of nanoparticles. The conjugation of proteins with
colloidal nanoparticles has been widely studied from the development of immune
probes in the 1970s. Some experiments regarding adsorption of proteins or peptides
on different size of gold (Au) or silicon oxide (SiO2) nanoparticles have been
performed. It has been studied that some proteins like lysozyme, catalase, trypsin,
and horseradish peroxidase bind strongly to SiO2NP generally in the size range
from 9 to 40 nm [51]. From these studies, it has been reported that there was partial
loss of protein structure and a significant loss of enzymatic activity [51]. Based on
these ideas, Vertegel et al. have studied different size silica nanoparticles effects on
the activity of adsorbed lysozyme [51]. The work observed that the lysozyme–silica
nanoparticle interaction is strongly influenced by the nanoparticle size (Fig. 1.2)
[51].
Figure 1.2 shows schematic representation for the interaction of lysozyme with
different sizes of silica nanoparticles. Generally, smaller nanoparticles possess high
surface curvature and larger nanoparticles possess smaller surface curvature. Hence,
1.3 Physicochemical Properties of Nanoparticles 9
Fig. 1.2 Schematic diagram of lysozyme interaction with silica nanoparticles of varying sizes.
Reprinted (adapted) with permission from Ref. [51]. Copyright (2017), American Chemical
Society
when a protein interacts with a smaller nanoparticle, the interaction is weak (both
Columbic and hydrophobic), since the edge of the protein molecules is at a greater
distance from nanoparticle surface, and vice versa for larger nanoparticles.
Therefore, for small nanoparticles the structure of protein remains intact, and for
larger it changes depending upon the interaction pattern. Similar results were found
for the interaction of silica nanoparticle with RNase A as studied by Shang et al.
[52]. Hence, these studies concluded that the size and curvature of nanoparticles
significantly influence the structural dynamics of proteins upon interaction.
1.3.2 Surface Concentration
Apart from size and curvature, the surface concentration also plays a major role in
defining conformational dynamics of proteins upon interaction with surface/
interface. Higher surface concentration of nanoparticles facilitates more proteins to
be adsorbed onto the nanoparticles surface, so that a crowded environment is
created which favors the protein–protein interaction. But at lower surface concen-
tration, the interaction between nanoparticles and proteins becomes more prominent
[53]. Figure 1.3 shows the unfolding kinetics of lysozyme upon binding with dif-
ferent concentrations of silica nanoparticles. Wu et al. calculated the unfolded
fraction of adsorbed lysozyme at different concentrations of silica nanoparticles,
keeping the concentration of protein constant [54]. As shown in the figure, lyso-
zyme at low nanoparticle surface concentration was unfolded to a greater amount
than higher surface concentrations in equilibrium state. The result confirmed the
existence of a higher energy barrier in a crowded environment that helps in
unfolding of protein [54].
At a lower nanoparticle surface concentration, the molecular interactions exist
between proteins and hydrophobic surface of silica nanoparticle. The interaction
potential induces the unfolding of protein molecules due to available free space and
absence of energy barrier. But at higher nanoparticle surface concentration, i.e., in
10 1 Nanoparticle
Fig. 1.3 Unfolding kinetics of lysozyme adsorbed onto silica nanoparticle surface at different
surface concentrations at neutral pH and low salt concentration. The normalized tryptophan
fluorescence intensity of lysozyme at 380 nm indicates the unfolded protein fraction with varying
surface concentrations. Reproduced with permission from the Ref. [54]. Copyright © 2008
Elsevier B.V.
crowed environment, the distance between the neighboring protein molecules will
be small. The interaction exists between protein molecules on the nanoparticle
surfaces; hence, the unfolding behavior is limited. The energy barrier raised from
the interaction from protein molecule decreases the extent of unfolding at higher
nanoparticle surface concentration [54].
1.3.3 Surface Functionality
Inside the biological milieu, the interaction of nanomaterials with different bio-
molecules depends on the properties of nanomaterials like shape, size, surface
charge. Here, first we will discuss how the neutral surface charge influences the
interaction of nanoparticles with cell membrane. In addition to shape and size of
nanomaterials, the accessible surface functional groups present over the nanoma-
terial surface dictate many important properties of nanomaterials, like solubility,
nanomaterial cell surface interaction, including the internalization of the nanopar-
ticle. Moreover, upon nanomaterial administration into a biological medium,
nanomaterial absorbs serum proteins which help the nanomaterial for internaliza-
tion by receptor-mediated endocytosis [55].
However, for many biological applications, surface functionality of nanoparti-
cles possessing potential not to interact with the cell membrane is also desirable.
For example, non-specific absorption of protein onto nanoparticle surface can
happen during the in vivo applications, which leads to the aggregation and
1.3 Physicochemical Properties of Nanoparticles 11
clearance from the reticular-endothelial system. Hence, the absorption hinders the
potential of nanoparticle for drugs/genes delivery to target site. The nanoparticle
can also bind to cellular membrane non-specifically, which reduces its efficiency for
targeting. Hence, various research groups have taken attempts to avoid the issue by
coating nanoparticles with neutral ligands, like poly(ethylene glycol), PEG. For
example, Xie, J. et al. have coated Fe3O4 nanoparticles with PEG which resulted in
negligible aggregation of nanoparticles in culture medium and reduction in uptake
of nanoparticles by macrophage cells non-specifically [56].
1.3.4 Surface Potential
In comparison with the charged surfaces, nanomaterial with neutral surface is rel-
atively good delivering agents, since charged surfaces have many non-specific
binding partners in a biological milieu. Hence, most of the charged accessible
functional groups are generally responsible for nanomaterial interaction with cells.
Based on the surface charge of nanomaterials, cationic charged nanoparticles
have shown stronger interaction with anionic cell membrane via electrostatic
interaction and translocate easily into the cell membrane. Due to stronger interac-
tion of cationic nanoparticles with anionic membrane, these nanoparticles are
exploited as synthetic agents for drug and gene delivery [49]. However, various
studies have approved the internalization of negatively charged nanoparticles
through the cell membrane, despite the unfavorable interactions between the
interfaces. In this context, Harush-Frenkel et al. have performed studies on endo-
cytosis of charged nanoparticles through the cell membrane. They observed that the
positively charged particle translocated easily via clathrin-mediated pathway,
whereas the particles with negative charge showed less efficiency for endocytosis
[57]. Surprisingly, the nanoparticles with negative charge bind non-specifically with
some positive particles on the plasma membrane (relatively less than negatively
charged domain) and get internalized through endocytosis [49, 57].
1.4 Physicochemical Properties of Biological Membranes

and Biomacromolecules
1.4.1 Cell Membrane
Generally, the cell membranes of both prokaryotic and eukaryotic systems are
complex as well as dynamic in nature, which are made up of different phospholipid
molecules. In addition, it contains different components such as lipopolysaccha-
rides, extracellular polymeric substances, and protein, embedded and/or superfi-
cially attached to the membrane [58]. Besides providing the buffer against
12 1 Nanoparticle
mechanical stress to cell, cell membrane helps in separating the intracellular

components from extracellular environment. Additionally, the cell membrane
maintains an anisotropic fluid phase which helps in supporting proteins as well as in
regulating molecular transport into and out of the cell [58]. Depending upon
interfacial composition of membrane, the prokaryotic cells have been divided
majorly into two categories, Gram-positive and Gram-negative bacteria. Both kinds
of bacterial interface behave differently to different surfaces like nanoparticle
interface and adherent surfaces.
1.4.2 Bacterial Cell Wall
The primary role of the bacterial cell wall is to provide shape, strength, and rigidity
to the cell. Additionally, it protects the cell from the mechanical damage and
osmotic rupture [59]. Based on their structure and functions, the bacterial cell walls
are categorized as Gram-positive and Gram-negative bacteria (Fig. 1.4). The cell
membrane of Gram-positive bacterium contains a thicker layer (20–50 nm) of
peptidoglycan attached to the teichoic acids. These teichoic acids are generally
embedded in the peptidoglycan layer, whereas the lipoteichoic acids are found to be
extended into the cytoplasmic membrane of the bacteria [59, 60]. However, the cell
membrane of Gram-negative bacterium contains a relatively thinner layer of pep-
tidoglycan, and an additional outer membrane covers the cell membrane. The outer
Fig. 1.4 Structure of

bacterial cell, a Gram-positive
bacterium and
b Gram-negative bacterium.
Reproduced from Hajipour
et al. [59]
1.4 Physicochemical Properties of Biological ... 13
membrane of these bacteria is linked to a thinner peptidoglycan layer by lipoproteins.

The peptidoglycan layer is placed within the periplasmic space formed between the
outer and cytoplasmic membranes. In addition, porins and lipopolysaccharide
molecules are also found in outer membrane. Hence, Gram-negative cell membrane is
more complex both, structurally and chemically, compared to Gram-positive. Due to
the presence of lipopolysaccharide molecules in outer membrane, the surface
potential of Gram-negative bacterium is more negative than Gram-positive bacterium.
However, surface functionality, because of peptidoglycan, is more diverse in
Gram-positive bacterium than Gram-negative bacterium [59].
1.4.3 Eukaryotic Cell Membrane
The basic framework model to understand the structure and function of eukaryotic
cell membrane was the Fluid—Mosaic Membrane Model, first introduced in 1972
[61]. It is composed of phospholipid bilayers with embedded proteins (Fig. 1.5).
Generally, 70% of total lipids present in mammalian cells are phospholipids [62].
Phospholipids contain one polar part and another nonpolar part, hence amphiphilic
in nature [63]. Although plasma membrane is predominantly composed of phos-
pholipids, the lipids have different functional roles, beyond forming the lipid
bilayers.
Plasma membrane, which is selectively permeable membrane, acts as a boundary
for the cell and maintains a viable intracellular environment. When discussing the
permeability of membrane, it comes into focus that membrane allows small and
Fig. 1.5 Structure of eukaryotic membrane, reproduced from Lombard et al. [63]
14 1 Nanoparticle
nonpolar molecules like O2 and CO2 to diffuse through it. Whereas, polar entities
like ions and nanoparticles of relatively larger size are restricted to diffuse across the
lipid bilayer. Generally, proteins of nanoscale size and important ions diffuse across
the lipid bilayer through different passive membrane-transport protein channels.
Other nanoscale molecules and supramolecular assemblies are transported through
a process of endocytosis, where the particles are enclosed in membrane vesicles,
and the cellular uptake happens [49]. It is reported that, in case of some
nanoparticles, the nanoparticles are confined in membrane-bound vesicles (en-
dolysosomes), which restrict their path to reach the cytosol. However, the process is
not limited to all nanoparticles. Additionally, it is reported that many nanoparticles
possess the inherent property to penetrate the cell membrane. In those cases, the
nanoparticles form pores in cell membrane which leads to cellular toxicity, since
they destroy the balance of intracellular and extracellular ions, proteins, and other
vital macromolecular concentrations that are essential for the normal function of a
cell [49]. Additionally, different approaches have been adopted for transporting
nanoparticles into a cell cytoplasm, like (i) endosome disruption and entry of NP
into the cytosol through sponge-effect mechanism [64], or using chloroquine [65]
(ii) direct microinjection of nanomaterials into cells [66, 67], (iii) use of electro-
poration [68], and (iv) conjugation of nanoparticle with natural cell-penetrating/
fusogenic chaperones [49, 69]. However, the interaction of nanoparticles with cell
membrane depends on the chemical functionalities of surface, shape, and size [49].
1.4.4 Nucleic Acid Biomacromolecules
Nucleic acids which are principal constituents of all living organisms are considered
to be possessing significant importance as protein, which are involved in cell
division and growth [70]. Nucleic acids are also considered as biological infor-
mation storage block which are relatively physicochemically stable and possess the
potential of self-organization [71]. The DNA (deoxyribonucleic acid), a long chain
polymer made from nucleotide, is commonly known as hereditary material.
Generally, nucleotide consists of sugar, heterocycle, and phosphate. The ribose or
deoxyribose, and sugar are found in cyclic and furanoside form. These are con-
nected by b-glycosyl linkage and produce four normal nucleosides such as ade-
nosine, guanosine, cytidine, and thymidine. Using the process of hybridization, a
single-stranded DNA binds its complementary strand and forms 1–3-dimensional
structure [71]. From the last decade, the scientists are trying to interact various
nanoparticles with DNA by both covalent and non-covalent interactions for dif-
ferent therapeutic approaches [72]. However, the DNA macromolecule is consid-
ered as suitable material for fabrication of nanostructures, and utilization of DNA
has gained interest for highly specific molecular recognition which is based on the
DNA hybridization [73]. In addition, for specific bindings of DNA with different
nanoparticles, DNA molecules can be modified with different functional groups
[73]. Interestingly, in biomedical sciences, DNA/RNA molecules are the important
molecules which play a key role as carriers for genetic information as well as they
also provide specific interactions with different target molecules like complemen-
tary DNA molecules, cell receptors, and proteins [73]. Hence, nucleic–nanoparticle
interaction plays a vital role in biomedical sciences.
1.4.5 Protein Biomacromolecules
Protein, one of the essential biomolecules to sustain life, consists of long chains of
amino acids. Upon synthesis, nascent protein attains tertiary structure, which
defines the functional attributes of the protein. The tertiary structure of protein is
governed by a network of interaction, predominantly non-covalent interactions like
hydrogen bonds, electrostatic interactions, salt bridge, van der Waals interactions,
hydrophilic/hydrophobic effects, dipole–dipole interactions. The network of inter-
actions, governing tertiary structure, is so fragile that perturbation of even one
non-covalent interaction may lead to conformational rearrangement or complete
loss of the function. Furthermore, conformational changes of proteins are associated
with different properties of proteins like self-assembly, tendency to aggregate,
transportation, and cytotoxicity [74]. Since the biomolecule is most dynamic and
functional entity of a cell, any additives to cell, like nanoparticles and drugs, will
likely to interact and affect the structure and function of the biomolecule. In past
decades, folding has received enormous attention of different researchers to
understand different biological phenomena like from genetic information to
molecular diagnostics [74].
1.4.5.1 Protein Folding
Protein folding is one of the most fundamental and spontaneous phenomena

occurring in a cell, where the entire polypeptide chain efficiently folds into a
physiologically active conformation (called native conformation), and executes
different functions needed for cell survivality [75]. Various studies are being carried
out to understand the mechanism of protein folding along with the kinetic and
thermodynamic analysis of the intermediates [76]. The ‘folding funnel’ or ‘energy
landscape theory’ of protein folding elucidates a statistical description of free
energies of all the molecular configurations (Fig. 1.6).
The unfolded protein starts from the top of the funnel, and as it descends the
funnel, the free energy is lowered due to the formation of intramolecular
16 1 Nanoparticle
Fig. 1.6 Schematic figure showing energy landscape of protein folding and aggregation.
Although the surface shows multiple conformations, with the help of intramolecular contact
formations, conformations ‘funneling’ toward native state or with the help of intermolecular
contact the conformations ‘funneling’ toward amyloid fibrils. Reproduced from Muntau et al. [77]
interactions. The reduced number of possible conformations at each stage simplifies

the folding process and finally reaches the unique natively folded conformations of
the lowest energy level. However, in case of larger proteins and a few small
proteins, the folding funnel does not remain ‘smooth’ but becomes ‘rugged.’ This
happens due to the presence of transiently stable intermediate conformation(s) in
the folding pathway of such proteins [78]. Sometimes during the folding process,
protein gets kinetically trapped in the local energy minima as an intermediate state
and needs to cross a certain energy barrier in order to reach the native state at
bottom of the folding funnel. These transiently populated states, known as inter-
mediate states, are partially folded protein conformations and are required to tra-
verse during the folding into native state. The folding pathway traversed by such
proteins is termed as ‘noncooperative’ or ‘multistate folding.’ In some cases, these
intermediates may lie off the productive folding pathway and are known as ‘off
pathway intermediates.’ Once these intermediates accumulate above a threshold
concentration during the folding process, owing to their exposed hydrophobic core,
the intermediate(s) interact with each other to form aggregates. Thereby, the process
shifts the equilibrium toward protein aggregation, where intermolecular interactions

rule over the intramolecular interactions. The aggregates are generally categorized
as either amorphous or highly ordered amyloid fibril (Fig. 1.6) [76, 77].
1.4.5.2 Protein Misfolding and Diseases
As mentioned above, perturbation of the interaction network is easy for most of the
proteins and can be misarranged/broken by slight change in local chemico-physical
environment of the protein, resulting in partially unfolded to completely unfolded
conformations. In both the conformations, protein loses its physiological function
(s), resulting in degradation by proteostasis network of the cell. However, when
these confirmations go unchecked of proteostasis network in cells, exposed
hydrophobic core drives the conformation into self-assembly of monomers into
pathogenic amyloid fibrils [79]. During the self-assembly into amyloid fibrils, the
process is accompanied by different intermediates with cytotoxic propensity [80,
81]. The cytotoxic nature of intermediate(s) results in several degenerative diseases
like Alzheimer’s disease, Huntington’s disease, Parkinson’s disease, amyloid
polyneuropathy, diabetes type-2, spongiform encephalopathy [79]. The degenera-
tive diseases associated with amyloid fibrils are generally of three types: (1) neu-
rodegenerative diseases like Alzheimer’s diseases: amyloid fibrils mainly
degenerate the brain cells, (2) non-neuropathic localized amyloidosis: the
fibril-mediated cell death occurs to a kind of cells other than neuronal cells, and
(3) non-neuropathic systemic amyloidosis: amyloid fibrils cause cell death to
multiple kinds of cells other than neuronal cells [79].
Since the root of amyloid disease is the formation of misfolded protein con-
formation with amyloidogenic propensity, development of effective drugs against
the conformation has been a key issue for last decade. In spite of many attempts, the
scientific community has failed to develop therapeutic agents to combat/hinder the
amyloid fibrillation. These insoluble deposits (fibrillar assemblies) are irreversible
aggregates of misfolded proteins. Thus, the reversal of protein aggregation would
have been an attractive strategy for the development of therapeutic agents against
the diseases [82]. Accordingly, first vaccine ‘Doblin-based Elan Pharmaceuticals’
AN-1792” was developed against Alzheimer’s diseases. The vaccine helped in
dissolution of visible aggregate and resulted in recovery of dementia in tested mice
models. However, its trial on human proved disaster with meningoencephalitis and
microbleed as the side effects in treated patients [83–85]. Development of
second-generation vaccines with different approaches against amyloid diseases is
still in clinical trials [86]. In last decade, scientists have also looked several other
types of molecules with tendency to induce anti-amyloidogenic conformations/
conditions, like synthetic peptides, heat shock proteins, and chemical compounds
(resveratrol) [87–89]. Apart from the studies, recently nanoparticles have been
suggested as one of a possible inhibitor of amyloid fibrillation [82, 90–92].
18 1 Nanoparticle
1.5 Nanoparticle Interfacial Interaction with Biological

Membranes and Biomacromolecules
In the current era, nanoparticle is considered as promising tool with different

applications in biomedical science, like biosensing, drug delivery, imaging.
However, it is a matter of intensive research to understand the synthetic material
interaction with different biomolecules, cells, and tissues before considering the
applications in biological sciences [93]. In biological medium, nanoparticles come
in myriad shape and size and interact with different biomolecules forming nano–bio
complexes. The physicochemical properties of both the nanoparticle and biomo-
lecule are affected by interaction patterns at nano–bio interface [94]. Additionally,
the interface helps in thermodynamic exchanges between nanomaterial and different
biological components, like membranes, proteins, phospholipids, DNA [94]. As
shown in Fig. 1.7, the different components developed as a result of nanoparticle–
biomolecule interactions are (i) surface of nanoparticle, (ii) solid–liquid interface,
(iii) contact zone of solid–liquid interfaces with biological substrates [94].
However, the most important factors determining the surface properties of
nanoparticle are chemical composition of material, shape and size, angle of cur-
vature, porosity, surface crystallinity, roughness, functionalization, heterogeneity,
hydrophilicity/hydrophobicity, etc. [94]. However, when the nanoparticle is dis-
persed in a medium, the developed quantifiable properties like effective surface
charge (zeta potential), aggregation propensity of particle, stability, biodegrad-
ability, state of dispersion, rate of dissolution, hydration and functionality of
Fig. 1.7 Depiction of nano–

bio interface. Reproduced
with permission from [94].
Copyright © 2009, Rights
Managed by Nature
Publishing Group
1.5 Nanoparticle Interfacial Interaction ... 19
interface depend on the characteristics of the suspending medium [94].

Additionally, ionic strength, pH, temperature, and the presence of organic mole-
cules also affect the particle properties [95]. The acquired properties of nanopar-
ticles contribute actively to the interaction of nanoparticles with the biological
medium. However, forces like long-range forces, which arise from van der Waals
(attractive in nature) and electrostatic double layer interactions, and short-range
forces, which arise from charge, solvent interaction, steric hindrance, depletion,
etc., are found at the nanoparticle media interface responsible for interaction with
biomolecules [94].
1.5.1 Nanoparticle–Biological Membrane Interaction
Increasing bacterial resistance toward traditional/conventional antibiotics is a major

global health concern in the current era [96, 97]. Some bacterial strains have the
potential to produce slime, which facilitates the adhesion and formation of biofilms
on any artificial surfaces or implantable devices. The formation of biofilms
enhances the possibility of bacterial survival by preventing antibiotic action [96,
97]. Additionally, the interaction of nanoparticle to bacteria is a research focus of
nanoecotoxicity; bacteria play a major role for maintaining normal ecosystem [98].
Moreover, bacteria are generally used as representative model for single-cell
organisms. However, the nanoparticle interaction with bacteria helps, in broad
view, us to understand the adhesion of nanoparticle on a cell membrane, and its
uptake. Although the toxicity of nanoparticles toward microbes is a well-established
research area [99, 100]. However, to explore whether the toxicity caused upon
interaction with membrane or internalization or both is essential to understand the
mechanism of nanotoxicity. Different non-specific interactions like electrostatic,
dipole–dipole, H-bond, hydrophobic, and van der Waals interactions are respon-
sible for adhesion of bacteria on any material surfaces, triggering bacterial biofilm
formation [96]. Hence, before screening any nanoparticle-mediated approach as
possible antibiotics, the material must have antimicrobial property to reduce the
microbial adhesion. The NPs with photocatalytic properties have the potential to
reduce biofilm formation [96]. Inside microbial medium, the interactions between
accessible functional groups of nanoparticle and biomolecules like lipopolysac-
charide (LPS), phospholipid, protein, and lipoteichoic acid (LTA) present over the
bacterial envelop contribute to interaction pattern at the interface. The functional
groups of biomolecules enhance adhesion of bacteria to different surfaces and help
their proliferation [98]. Hence, the accessible functional groups present at bacterial
envelop and nanoparticle surfaces, along with the physicochemical property of
nanoparticle, determine the fate of bacteria as well as the nanoparticle (whether
nanoparticle will be compatible or toxic to bacteria) in biological milieu.
20 1 Nanoparticle
Fig. 1.8 Nanoparticle internalization pathways. Reproduced with permission from [101].
Copyright © 2015, American Chemical Society
1.5.2 Cellular Internalization of Nanoparticles
Nanoscale materials are being formulated for different biological applications. For
successful biomedical applications, sometimes nanoparticles are often functional-
ized with different ligands, coated with different biopolymers to target a specific cell
[101]. Upon conjugation/encapsulation with drug molecules, cell internalizes the
nanoparticle through different pathways (Fig. 1.8) and releases the drug molecule
successfully. However, the efficacy of nanoparticle-based approaches depends on
physicochemical parameters of the nanoparticle like shape and size, surface
potential, surface functionalities. In addition, different physicochemical properties
of the membrane such as (i) stretching and elasticity of membrane, (ii) thermal
fluctuation of cell membrane, and (iii) hydrophobic exclusion of polar surfaces by
membrane resist the nanoparticle against the permeabilization [94, 102].
Interestingly, the nanoparticles upon interaction produce sufficient energy at the
interface to overcome these forces [94].
1.5.3 Nanoparticle–Nucleic Acid Interaction
Exploring the interaction between the nanoparticle and biomolecule is an active

research area, in the current era. Nanoparticles, having high surface free energy,
show high affinity to interact with biomolecules, especially biopolymers resulting in
various applications like development of novel biosensor for clinical diagnosis,
Another random document with
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marriage of long ago, and what had come of it. Had they had
children, did their blood still run on the far worlds? But even if it did,
what was that now to those two of long ago?
There had been a poem about flowers at the end of the old book on
flowers Haller had lent him, and he remembered some of it.
"All are at one now, roses and lovers,

Not known of the winds and the fields and the sea,
Not a breath of the time that has been hovers
In the air now soft with a summer to be."
Well, yes, Kellon thought, they were all at one now, the Rosses and
the Jennies and the things they had done and the things they had
thought, all at one now in the dust of this old planet whose fiery final
summer would be soon, very soon. Physically, everything that had
been done, everyone who had lived on Earth, was still here in its
atoms, excepting the tiny fraction of its matter that had sped to other
worlds.
He thought of the names that were so famous still through all the
galactic worlds, names of men and women and places.
Shakespeare, Plato, Beethoven, Blake, the old splendor of Babylon
and the bones of Angkor and the humble houses of his own
ancestors, all here, all still here.
Kellon mentally shook himself. He didn't have enough to do, that was
his trouble, to be brooding here on such shadowy things. He had
seen all there was to this queer little old place, and there was no use
in coming back to it.
But he came back. It was not, he told himself, as though he had any
sentimental antiquarian interests in this old place. He had heard
enough of that kind of gush from all the glittering phonies in the ship.
He was a Survey man and all he wanted was to get back to his job,
but while he was stuck here it was better to be roaming the green
land or poking about this old relic than to have to listen to the
endless babbling and quarrelling of those others.
They were quarrelling more and more, because they were tired of it
here. It had seemed to them a fine thing to posture upon a galactic
stage by helping to cover the end of Earth, but time dragged by and
their flush of synthetic enthusiasm wore thin. They could not leave,
the expedition must broadcast the final climax of the planet's end,
but that was still weeks away. Darnow and his scholars and
scientists, busy coming and going to many old sites, could have
stayed here forever but the others were frankly bored.
But Kellon found in the old house enough interest to keep the waiting
from being too oppressive. He had read a good bit now about the
way things had been here in the old days, and he sat long hours on
the little terrace in the afternoon sunshine, trying to imagine what it
had been like when the man and woman named Ross and Jennie
had lived here.
So strange, so circumscribed, that old life seemed now! Most people
had had ground-cars in those days, he had read, and had gone back
and forth in them to the cities where they worked. Did both the man
and woman go, or just the man? Did the woman stay in the house,
perhaps with their children if they had any, and in the afternoons did
she do things in the little flower-garden where a few bright, ragged
survivors still bloomed? Did they ever dream that some future day
when they were long gone, their house would lie empty and silent
with no visitor except a stranger from far-off stars? He remembered
a line in one of the old plays the Arcturus Players had read. Come
like shadows, so depart.
No, Kellon thought. Ross and Jennie were shadows now but they
had not been then. To them, and to all the other people he could
visualize going and coming busily about the Earth in those days, it
was he, the future, the man yet to come, who was the shadow. Alone
here, sitting and trying to imagine the long ago, Kellon had an eery
feeling sometimes that his vivid imaginings of people and crowded
cities and movement and laughter were the reality and that he
himself was only a watching wraith.
Summer days came swiftly, hot and hotter. Now the white sun was
larger in the heavens and pouring down such light and heat as Earth
had not received for millennia. And all the green life across it
seemed to respond with an exultant surge of final growth, an act of
joyous affirmation that Kellon found infinitely touching. Now even the
nights were warm, and the winds blew thrilling soft, and on the
distant beaches the ocean leaped up in a laughter of spray and
thunder, running in great solar tides.
With a shock as though awakened from dreaming, Kellon suddenly
realized that only a few days were left. The spiral was closing in fast
now and very quickly the heat would mount beyond all tolerance.
He would, he told himself, be very glad to leave. There would be the
wait in space until it was all over, and then he could go back to his
own work, his own life, and stop fussing over shadows because
there was nothing else to do.
Yes. He would be glad.
Then when only a few days were left, Kellon walked out again to the
old house and was musing over it when a voice spoke behind him.
"Perfect," said Borrodale's voice. "A perfect relic."
Kellon turned, feeling somehow startled and dismayed. Borrodale's
eyes were alight with interest as he surveyed the house, and then he
turned to Kellon.
"I was walking when I saw you, Captain, and thought I'd catch up to
you. Is this where you've been going so often?"
Kellon, a little guiltily, evaded. "I've been here a few times."
"But why in the world didn't you tell us about this?" exclaimed
Borrodale. "Why, we can do a terrific final broadcast from here. A
typical ancient home of Earth. Roy can put some of the Players in
the old costumes, and we'll show them living here the way people
did—"
Unexpectedly to himself, a violent reaction came up in Kellon. He
said roughly,
"No."
Borrodale arched his eyebrows. "No? But why not?"
Why not, indeed? What difference could it possibly make to him if
they swarmed all over the old house, laughing at its ancientness and
its inadequacies, posing grinning for the cameras in front of it,
prancing about in old-fashioned costumes and making a show of it.
What could that mean to him, who cared nothing about this forgotten
planet or anything on it?
And yet something in him revolted at what they would do here, and
he said,
"We might have to take off very suddenly, now. Having you all out
here away from the ship could involve a dangerous delay."
"You said yourself we wouldn't take off for a few days yet!" exclaimed
Borrodale. And he added firmly, "I don't know why you should want
to obstruct us, Captain. But I can go over your head to higher
authority."
He went away, and Kellon thought unhappily, He'll message back to

Survey headquarters and I'll get my ears burned off, and why the
devil did I do it anyway? I must be getting real planet-happy.
He went and sat down on the terrace, and watched until the sunset
deepened into dusk. The moon came up white and brilliant, but the
air was not quiet tonight. A hot, dry wind had begun to blow, and the
stir of the tall grass made the slopes and plains seem vaguely alive.
It was as though a queer pulse had come into the air and the ground,
as the sun called its child homeward and Earth strained to answer.
The house dreamed in the silver light, and the flowers in the garden
rustled.
Borrodale came back, a dark pudgy figure in the moonlight. He said
triumphantly,
"I got through to your headquarters. They've ordered your full
cooperation. We'll want to make our first broadcast here tomorrow."
Kellon stood up. "No."
"You can't ignore an order—"
"We won't be here tomorrow," said Kellon. "It is my responsibility to
get the ship off Earth in ample time for safety. We take off in the
morning."
Borrodale was silent for a moment, and when he spoke his voice had
a puzzled quality.
"You're advancing things just to block our broadcast, of course. I just
can't understand your attitude."
Well, Kellon thought, he couldn't quite understand it himself, so how
could he explain it? He remained silent, and Borrodale looked at him
and then at the old house.
"Yet maybe I do understand," Borrodale said thoughtfully, after a
moment. "You've come here often, by yourself. A man can get too
friendly with ghosts—"
Kellon said roughly, "Don't talk nonsense. We'd better get back to the
ship, there's plenty to do before take off."
Borrodale did not speak as they went back out of the moonlit valley.
He looked back once, but Kellon did not look back.
They took the ship off twelve hours later, in a morning made dull and
ominous by racing clouds. Kellon felt a sharp relief when they
cleared atmosphere and were out in the depthless, starry blackness.
He knew where he was, in space. It was the place where a
spaceman belonged. He'd get a stiff reprimand for this later, but he
was not sorry.
They put the ship into a calculated orbit, and waited. Days, many of
them, must pass before the end came to Earth. It seemed quite near
the white sun now, and its Moon had slid away from it on a new
distorted orbit, but even so it would be a while before they could
broadcast to a watching galaxy the end of its ancestral world.
Kellon stayed much of that time in his cabin. The gush that was
going out over the broadcasts now, as the grand finale approached,
made him sick. He wished the whole thing was over. It was, he told
himself, getting to be a bore—
An hour and twenty minutes to E-time, and he supposed he must go
up to the bridge and watch it. The mobile camera had been set up
there and Borrodale and as many others of them as could crowd in
were there. Borrodale had been given the last hour's broadcast, and
it seemed that the others resented this.
"Why must you have the whole last hour?" Lorri Lee was saying
bitterly to Borrodale. "It's not fair."
Quayle nodded angrily. "There'll be the biggest audience in history,
and we should all have a chance to speak."
Borrodale answered them, and the voices rose and bickered, and
Kellon saw the broadcast technicians looking worried. Beyond them
through the filter-window he could see the dark dot of the planet
closing on the white star. The sun called, and it seemed that with
quickened eagerness Earth moved on the last steps of its long road.
And the clamoring, bickering voices in his ears suddenly brought
rage to Kellon.
"Listen," he said to the broadcast men. "Shut off all sound
transmission. You can keep the picture on, but no sound."
That shocked them all into silence. The Lee woman finally protested,
"Captain Kellon, you can't!"
"I'm in full command when in space, and I can, and do," he said.
"But the broadcast, the commentary—"
Kellon said wearily, "Oh, for Christ's sake all of you shut up, and let
the planet die in peace."
He turned his back on them. He did not hear their resentful voices,
did not even hear when they fell silent and watched through the dark
filter-windows as he was watching, as the camera and the galaxy
was watching.
And what was there to see but a dark dot almost engulfed in the
shining veils of the sun? He thought that already the stones of the
old house must be beginning to vaporize. And now the veils of light
and fire almost concealed the little planet, as the star gathered in its
own.
All the atoms of old Earth, Kellon thought, in this moment bursting
free to mingle with the solar being, all that had been Ross and
Jennie, all that had been Shakespeare and Schubert, gay flowers
and running streams, oceans and rocks and the wind of the air,
received into the brightness that had given them life.
They watched in silence, but there was nothing more to see, nothing
at all. Silently the camera was turned off.
Kellon gave an order, and presently the ship was pulling out of orbit,
starting on the long voyage back. By that time the others had gone,
all but Borrodale. He said to Borrodale, without turning,
"Now go ahead and send your complaint to headquarters."
Borrodale shook his head. "Silence can be the best requiem of all.
There'll be no complaint. I'm glad now, Captain."
"Glad?"
"Yes," said Borrodale. "I'm glad that Earth had one true mourner, at
the last."
THE END
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Textbook Interfacial Phenomena On Biological Membranes 1St Edition Manoranjan Arakha Ebook All Chapter PDF

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Textbook Interfacial Phenomena On Biological Membranes 1St Edition Manoranjan Arakha Ebook All Chapter PDF

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Interfacial Phenomena on Biological

Membranes 1st Edition Manoranjan

Physics of Biological Membranes Patricia Bassereau

Permeability of Biological Membranes 1st Edition Gaspar

Biomembrane Simulations Computational Studies of

Transport Across Natural and Modified Biological

Interfacial Physical Chemistry of High-Temperature

Minorities and Small Numbers from Molecules to

Handbook on Biological Warfare Preparedness S.J.S.

My Thoughts on Biological Evolution Motoo Kimura

More information about this series at http://www.springer.com/series/10358

ISSN 2196-8861 ISSN 2196-887X (electronic)

© Springer International Publishing AG 2018

Printed on acid-free paper

This Springer imprint is published by Springer Nature

1.6 Applications of Nanoparticle–Biomolecular Interactions

3.3.3 Enhanced ROS Production in Presence

6 Effect of Interfacial Assembly of Antimicrobial Peptide

© Springer International Publishing AG 2018 1

science and technology. Hence, to do so, the synthesis of different nanoparticles

1.1 Synthesis of Nanoparticles

Different protocols have been optimized to synthesize different types of nanopar-

Fig. 1.1 Protein nanoparticles prepared by a coacervation or phase separation, b emulsion/solvent

homogenizer/ultrasonic shear is being used to emulsify the aqueous solution of

1.2 Different Kinds of Nanoparticles

According to different applications of nanotechnology, different methods have been

1.2.1 Zinc Oxide Nanoparticle (ZnONP)

1.2.2 Iron Oxide Nanoparticle (IONP)

1.2.3 Silver Nanoparticle (AgNP)

sulfadiazine is being replaced by AgNP to treat wounds [27–29]. Due to enhanced

Liposome is considered as one of the ﬁrst nanoparticles formulation, which was

1.2.5 Albumin-Functionalized NPs

Albumin-bound nanoparticles are important class of nanoparticles which carry the

1.2.6 Polymeric NPs

1.2.7 Quantum Dot

1.3 Physicochemical Properties of Nanoparticles

1.3.1 Shape, Size, and Curvature

Shape, size, and curvature of nanomaterials greatly influence the internalization of

1.3.2 Surface Concentration

1.3.3 Surface Functionality

1.3.4 Surface Potential

1.4 Physicochemical Properties of Biological Membranes

1.4.1 Cell Membrane

mechanical stress to cell, cell membrane helps in separating the intracellular

1.4.2 Bacterial Cell Wall

Fig. 1.4 Structure of

membrane of these bacteria is linked to a thinner peptidoglycan layer by lipoproteins.

1.4.3 Eukaryotic Cell Membrane

1.4.4 Nucleic Acid Biomacromolecules

1.4.5 Protein Biomacromolecules

1.4.5.1 Protein Folding

Protein folding is one of the most fundamental and spontaneous phenomena

interactions. The reduced number of possible conformations at each stage simpliﬁes

shifts the equilibrium toward protein aggregation, where intermolecular interactions

1.4.5.2 Protein Misfolding and Diseases

1.5 Nanoparticle Interfacial Interaction with Biological

In the current era, nanoparticle is considered as promising tool with different

Fig. 1.7 Depiction of nano–

interface depend on the characteristics of the suspending medium [94].

1.5.1 Nanoparticle–Biological Membrane Interaction

Increasing bacterial resistance toward traditional/conventional antibiotics is a major

1.5.2 Cellular Internalization of Nanoparticles

1.5.3 Nanoparticle–Nucleic Acid Interaction