3-Case Study

Case Studies in Chemical and Environmental Engineering 8 (2023) 100372
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Case Report
Nanomaterials as transmitters of non-viral gene vectors: A review

Ahmad Khalf Alkhawaldeh a, Ahmed Mahdi Rheima b, Mustafa M. Kadhim c,
Zainab sabri Abbas d, Alaa dhari jawad al-bayati e, Zainab Talib Abed f,
Firas mohamed dashoor Al-Jaafari g, Asala Salam Jaber h, Safa K. Hachim i, j, Farah K. Ali k,
Zaid H. Mahmoud l, Ghobad Behzadi pour m, Ehsan kianfar n, o, p, *
a
Department of Allied Medical Sciences, Zarqa University College/ Al-Balqa Applied University, Zarqa, Jordan
b
Department of Chemistry, College of Science, Mustansiriyah University, Baghdad, Iraq
c
Medical Laboratory Techniques Department, Al-Farahidi University, Baghdad, 10022, Iraq
d
Wasit University, College of Science, Department of Chemistry, Iraq
e
Department of Chemical Engineering and Petroleum Industries, Al-Mustaqbal University College, Hilla, 51001, Iraq
f
College of Pharmacy, The University of Mashreq, 10021, Baghdad, Iraq
g
Wasit University, College of Science, Department of Physics, Iraq
h
Department of Pharmacy, Mazaya University College, Thi-Qar, Iraq
i
College of Technical Engineering, The Islamic University, Najaf, Iraq
j
Department of Medical Laboratory, College of Health and Medical Technology, National University of Science and Technology, Thi-Qar, 64001, Iraq
k
Medical Laboratory Techniques Department, Al-Turath University College, Baghdad, Iraq
l
University of Diyala, College of Sciences, Department of Chemistry, Diyala, Iraq
m
Department of Physics, East Tehran Branch, Islamic Azad University, Tehran, Iran
n
Department of Chemical Engineering, Arak Branch, Islamic Azad University, Arak, Iran
o
Young Researchers and Elite Club, Gachsaran Branch, Islamic Azad University, Gachsaran, Iran
p
Mechanical Engineering Department, Faculty of Engineering and Pure Sciences Istanbul Medeniyet University, Istanbul, Turkey
A R T I C L E I N F O A B S T R A C T
Keywords: With the rapid development of nanotechnology in the recent decade, novel DNA and RNA delivery systems for
Block copolymer gene therapy have become available that can be used instead of viral vectors. These non-viral vectors can be
Polymer nano particles made of a variety of materials, including inorganic nanoparticles, carbon nanotubes, liposomes, protein and
Solid lipid particles
peptide-based nanoparticles, as well as nanoscale polymeric materials. They have as advantages over viral
Non-virus gene
PLGA
vectors a decreased immune response, and additionally offer flexibility in design, allowing them to be func
Silica tionalized and targeted to specific sites in a biological system with low cytotoxicity.gene therapy keeps hopes a
Gold nano particles life for the treatment of a wide range of diseases such as cancer, nano particles are now known as promising
carriers for the effective and safe vectors of genes to specific cells or tissues. This could provide alternative
therapies for conventional approaches that use viruses as gene carriers. The expression of genetic material such
as DNA, RNA into cells and tissues has raised considerable hopes for therapeutic and diagnostic purposes. But
getting nucleic acids into the cell also faces challenges. These challenges are less for non-virus carriers as a gene
and drug vectors method than for viral or free vectors and are therefore considered less risky and more
appropriate. of expanding nonverbal nano carriers, we will look at a few of these nano carriers, penicillin, PEI,
PLGA, silica, block copolymer, Quantum dot, gold nano particles, and common carbon nano tubes. Problems
include the use of nano particles such as polymer nano particles, liposomes, solid lipid particles, in targeted gene
vectors will be investigated. Gene-based therapy is the intentional modulation of gene expression in specific cells
to treat pathological conditions. This modulation is accomplished by introducing exogenous nucleic acids such as
DNA, mRNA, small interfering RNA (siRNA), microRNA (miRNA) or antisense oligonucleotides. Given the large
size and the negative charge of these macromolecules, their delivery is typically mediated by carriers or vectors.
In this Review, we introduce the biological barriers to gene delivery in vivo and discuss recent advances in
* Corresponding author. Department of Chemical Engineering, Arak Branch, Islamic Azad University, Arak, Iran.
E-mail address: ehsan_kianfar2010@yahoo.com (E. kianfar).
https://doi.org/10.1016/j.cscee.2023.100372
Received 10 April 2023; Received in revised form 5 May 2023; Accepted 7 May 2023
Available online 8 May 2023
2666-0164/© 2023 The Authors. Published by Elsevier Ltd. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-
nc-nd/4.0/).
A.K. Alkhawaldeh et al. Case Studies in Chemical and Environmental Engineering 8 (2023) 100372
material sciences, nanotechnology and nucleic acid chemistry that have yielded promising non-viral delivery
systems, some of which are currently undergoing testing in clinical trials. The diversity of these systems high
lights the recent progress of gene-based therapy using non-viral approaches.
1. Introduction reticuloendothelial system (RES) agents and is specific to the target cell
or tissue [54–57]. The nano particle process is expected to enter the
Gene therapy has the potential to cure human diseases caused using endocytosis cell, so the carrier system must be able to escape the en
defective genes. Some of these diseases include sac fibrosis, macular dosome and deliver DNA to the nucleus [58–62]. As shown in Fig. 1, a
degeneration, Parkinson’s disease, and virus cancers [1–5]. Gene ther fine machine should include a difference of building blocks for definite
apy has also been used to express different genes or to turn them off in behaviors. Clinical applications of genotypes haven’t grown signifi
animal tissues such as the central nervous system, lungs, liver, etc cantly because of the insecurity within body liquids their down tissue
[6–10]. The effective development of gene therapy involves the efficient absorption due unto their above relative molecular mass, macromole
vectors of therapeutic genes to the cell to displace or silence defective cule pregnancy [63–67]. Therefore, it is need to style an appropriate
disease-causing organisms in humans [11–16]. Virus vectors similar delivery machine which will deliver DNA or oligonucleotide strand to
adenovirus and retrovirus are commonly occupied in gene therapy due the target. Two general approaches to accomplish this goal [68–73]: 1.
in their high efficiency in gene delivery [17–22]. However, some Use of virus vectors, 2. Non-virus vectors. Despite the high efficiency of
problems have led to a review of the use of virus vectors in human virus vectors for transmitting DNA to a wide variety of cells, there are
clinical trials. Recently, non-virus particles have received widespread major problems, Including the danger of safety answer to virus vectors,
attention in gene therapy [23–27]. This is because they can overcome a as well as the Possibility of accidental mixing of the virus vectors with
variety of toxicities caused by virus delivery [28–32]. Some common the wild-type virus [74–80]. So, thither is a necessity for a more reliable
non-virus vectors that allow genetic material to pass-throng cellular carrier to deliver DNA [48–50]. This carrier must have the surface
barriers are discussed in this article. Gene therapy as A potential method charge density and hydrophobicity required to Interact with the lipid
for the treatment of genetic disorders and others malignancies has components of the cell as well as the DNA [81–85]. Due to the negative
attracted very regard in recent years [33–38]. In DNA plasmid gene Intrinsic charge of DNA (due to The Presence of phosphate structural
therapy (plasmids are usually in the form a circular double-stranded groups), Positively charged materials are usually used as carriers to
DNA molecule (although there are linear types of plasmids that are deliver DNA so that the carrier can be attached to DNA using electro
replicated independently of the nucleus) to express specific therapeutic static interaction (Fig. 2). Cationic Polymers, Cationic Phospholipids Are
Proteins or oligonucleotides to silence the gene that causes disease the two main types of non-virus gene delivery carriers currently under
(antisense therapy) are used [39–43]. The main importance of pure DNA Investigation [86–91]. Due to the positive surface load, both kinds react
plasmid is for the development and production of vaccines that a small electrostatic via the negative DNA, combine [92–94]. Despite the easy
amount of Protein can lead to the desired immune response [44–48]. construction of lipid compounds, Low gene vectors and toxicity of most
Because of the degradation of DNA by serum nucleases, Pure DNA is of them Limit their use [95–98]. Cationic Polymeric Compounds are
rarely suitable for the vectors of genetic material in virus. Therefore, a often more stable than cationic Lipid compounds, but generally have
carrier system is usually recommended That is biocompatible and lower gene vectors than virus carriers [99–102]. These carriers make the
immunogenicity [49–53]. (non-immunogenic), protects DNA against Polyelectrolyte with the combined DNA and Protect it from nuclease
degradation [103–106]. These Polymeric vectors have good adaptability
and structural variability that allow the attachment of specific compo
nents to target the vectors in gene expression Through specific receptors
[107–110]. Studies have been Performed on other nanostructures for
DNA vectors, A: targeting the delivery system for specific cells, B: vec
tors from cell membrane, C: cell absorption, setback in endosomes, D:
Fig. 2. Schematic of the process of interaction of a positively charged polymer

Fig. 1. The imager of the biological barriers than the gene vectors carrier must carrier with a negative DNA. Figures reproduced with permission
overcome. Figures reproduced with permission from Ref.[117]. from Ref. [118].
2
Intracellular vectors of Plasma DNA to nucleus [111–115,116]. 3. Non-virus nanoparticles being studied and expanded
2. Nanoparticles for gene and drug vectors 3.1. Chitosan
Speiser et al. [116,119] for the first time created colloidal particles Chitosan’s are the very natural amino Polysaccharide found in the
less than 1 μm in diameter that contained macromolecular compounds. cell wall of fungi, the hard skeletons of crustaceans such as crabs,
Since then, much work has been done in using nano Particles the arrival shrimp, as well as in Insects [156–160]. It is estimated that its annual
of medicine and gene delivery. Initially, most nano Particles were Production is as much as cellulose. Chitin is of great Interest, not because
designed to carry vaccines and anti-cancer drugs [120–124]. However, it is an available source, but because of Its very high potential in virus
the use of these carriers for the Possibility of using drugs and vaccines applications [161–165]. Chitosan is an amino Polysaccharide obtained
orally and jocularly is currently under investigation [125–129]. Drugs or from the distillation of chitin under alkaline conditions and is very
biologically active molecules are trapped, encapsulated in NPTS, similar to cellulose (Fig. 3). Chitosan is a non-toxic biocompatible
chemically attached to NPTS or absorbed to the Particle surface Polymer that has many applications in drug delivery, more recently, In
[130–134]. The choice of the appropriate method for The Preparation of gene delivery [116,119].
the drug carried by the nano Particle depends on the Properties of the Used chitosan and chitosan depolymerized oligomers to deliver the
drug and the nano particle [135–138]. Two different kinds of systems Plasmid in the virus. The resulting solution is stirred vigorously for
with different Internal structures are Possible [139–144]. 180–300 seconds, then left at room temperature for 1800 seconds to
Combine the Particle with the Plasmid well [167–170]. Gene Vectors
1. Matrix-type systems consisting of the entanglement of oligomers, was Performed in virus with Cos-1 Cells and in, vivo studies were Per
Polymer units known as nano spheres or nano particles [145–148]. formed on intestinal tissue [171–173]. High levels of gene expression
2. Storage systems that contain a core coated with a Polymer wall and were observed in the chitosan-Plasmid Formulation relative to Pure
are defined as nano capsules [149–154,155]. DNA in the upper small Intestine. In this Study, the Parameters affecting
the Particle size and stability of the carrier System, chitosan molecular
weight, Plasmid Concentration and Charge Ratio were determined
Fig. 3. Structural Similarities Between Cellulose, Chitin, and Chitosan. Figures reproduced with permission from Ref. [166].
3
[113–115,120,121,174–178]. The Combination of High molecular by the effect of Polyethylene mine (PEI) %wt. on gene delivery in This
weight Plasmids and Chitosan Was shown to be more Stable against Salt method. Their results showed that Low molecular weight Polyethylene
and the challenges Posed by Interaction with Serum [179–182]. mine (PEI) was more effective than extra molecular weight Polyethylene
They examined Parameters affecting Particle Size and the Physico mine (PEI) in virus gene delivery. other researchers have shown that the
chemical Properties of nano Particles and demonstrated DNA Protection method is effective in vivo gene delivery [229–234].
by chitosan [183–187]. They also Investigated PEGylating Chitosan
nano Particles (nano Particles Coated with Polyethylene Glycol Poly 3.2. Solid Lipid Nanoparticles (SLN)
mer) (Fig. 4) to Improve storage stability as well as Formulation effi
ciency [116,119,188–191]. The tissue distribution of chitosan nano They Found that Solid Lipid Nanoparticles (SLN), in addition to Drug
Particles containing DNA and chitosan nano Particles containing Pel Delivery, could also be a good Carrier for plasmid DNA Vectors
leted DNA was Investigated and it was observed That the Clearance of [237–244]. This group Produced highly Cationic Solid fat nano Particles
Pelleted nano Particles was much slower in the First 15 minutes. It with a zeta Potential of more than +40 and a Size of 100 nm Through the
should be noted that the efficiency of gene vectors Also Depends on the hot homogenization Technique [245–249,250–254]. They Found that
Type of Cell [192–196]. Sato and his Team Found that the Gene Vectors solid fat nano Particles fused together after binding to Plasmid DNA,
efficiency of Chitosan depends on the molecular weight, pH of the cul producing larger Particles measuring between 300 and 800 nm (Fig. 5).
ture medium, Serum, and Stoichiometry of the Designed System [117, The gene vectors efficiency of this Carrier has been demonstrated in
155,197–200]. Since Then, various types of lipid compounds have been virus in the Cos-1 cell line [249,255–262].
used as carriers of gene vectors. Due to Their amphiphilic nature, these
lipids easily Form micelles in water and Interact with DNA rapidly and
3.3. Polyethylene mine and penicillin
efficiently due to Their Positive charge [201–205]. Cationic liposomes
are Prepared by evaporating an organic solvent from a mixture of
Structure Penicillin (Fig. 6) shows. It has high biodegradability and,
Cationic lipids and then coating the resulting lipid film in aqueous me
of course, high toxicity, which limits its in-body use. “Preliminary
dium under high stirring Conditions, resulting in the formation of
studies show that if the right molecular weight” “Polyethylene mine
multi-layer vesicles (MLVs) [206–212]. Single-wall Vesicles are also
(PEI) (Fig. 6) has been the first carrier " “PLA and PLGA particle syn
obtained by sonication or extraction of multi-wall vesicles. Adding DNA
thesis methods are considered and, in this method,”, 100 nm can be
to Cationic Liposomes Causes Dramatic Changes in liposomes and DNA
made that can be removed by the cell in small quantities [160–162,
[213–217]. This changes the Initial Structure of the Liposome and cre
201-2254-260]. The reason for this is the lack of amine groups at the
ates an adapted structure. The reported results show that changing the
particle level (low positive charge). However, by using cellular targeting
surface Characteristics of liposomes Improves the biological distribution
groups as well as endosomal lysis agents such as Chloroquine, this defect
as well as the targeted delivery of DNA to liposomes [218–222]. The
of low gene Vectors perception can be eliminated. In an effort to improve
structure of Some of the cationic lipids used in DNA delivery is shown in
gene transfer efficiency and reduce toxicity [263–267]. In this method,
Fig. 5. They made PCL by adding Polyethylene imine Polycation (Pol
PEG first binds to Penicillin and then interacts with DNA, and then binds
ycation liposome system (PCL)) To the liposome [223–228]. The
to the Phosgene Peptide [118,268–272]. The resulting Particle has a
resulting PCL Does not require Phosphatidylethanolamine or cholesterol
positive charge and as a result the resulting particles have good stability
as components of a normal liposome. The researchers also Investigated
and of course the use of PEG and Peptide reduces toxicity and Increases
Fig. 4. Some Cationic Lipids Used in gene delivery. Figures reproduced with permission from Ref. [235].
4
Fig. 5. Solid Lipid nanoparticles (SLN)with plasmid DNA (upper half) made from a combination of several smaller particles. A small particle can be seen at the
bottom of the image. Figures reproduced with permission from Ref. [236].
3.4. Poly (lactide-Co-glycolide)
Various polymers Have Been Used for Drug Delivery and gene De
livery applications, but PLA = poly (lactic acid and PLGA) are the only
FDA-approved Polymers that are of great interest. The emulsion-solvent
evaporation method is one of the most common [250,279–283]. PLA
and PLGA particle synthesis methods are considered and, in this method,
Polyvinyl alcohol (PVA) is used as a common stabilizer, with this tech
nique, both large and small particles are obtained, and if 100 nm dialysis
membranes are obtained [251–254,284]. The tiny particles with an
average size of 702 ± 2 nm is separated and the larger impermeable
particles have an average Size of 202 ± 9 nm. Smaller particles are 27
Equal larger in gene Vectors to Cos-7 cells and 4 Equal larger in HEK-293
Cells. The surface load, cell harvest, and DNA release of both groups are
approximately Equal [172,173,285–291].
Others have studied the residual role of PVA on cellular uptake as
well as on PLGA nano Particles. Removal of PVA during the Preparation
Process is very difficult, so some PVA will remain with the Particles,
which depends on the amount of initial PVA, the Type of organic Solvent
Used in The Emulsion, several other factors. PVA affects Particle Size,
Zeta Potential, and Poly disparity index (PDI), nano Particle surface
hydrophobicity [119,241–245]. Particles with high residual PVA,
although smaller in size, also have low Cellular uptake, which may be
due to Further reduced hydrophobicity of the nano Particle Surface
[174–176,292–294].
3.5. Silica
Fig. 6. Structure of (a) PEI [274] (b) Penicillin. Figures reproduced with The use of silica nanoparticles for gene delivery applications has not
permission from Ref. [273]. received much attention. A series of studies have shown that superfi
cially charged silica particles have the ability to interact with plasma
the Improvement of gene Vectors [163–165,166,235,236,273].
DNA and be used in virus gene Vectors. These particles are produced by
Polyethylene mine (PEI) (Fig. 6) has been the first carrier of gene
changing the level of silica on the market. Compared to PEI particles, no
Vectors and has been the subject of many studies. of course, the toxicity
specific toxicity is observed for these particles at the required concen
of Polyethyleneimine is the biggest drawback of this carrier [167,168,
trations [177,178,246–249,295–301]. Fig. 7 shows the structure of
223–225,274–278].
silica.
Sagara and Kim [116] reported the use of a Combination of Galac
tose, Polyethylene Glycol, and polyethylene mine (Glc-PEG-PEI) for
Gene Delivery to Hepatocyte cells, and showed better Transmission of 3.6. Block copolymers
this system than similar PEI carriers [116,119]. They introduce this
system as a suitable system for liver gene delivery. Co-polymers are combinations of the cationic and hydrophobic
Rudolph et al. [169] report the use of PEG-coated PEI to Deliver DNA moieties that react spontaneously with DNA polyanion to form micelles.
to the lungs through the use of nebulizers or intratracheal injections. The Kataoka et al. are pioneers in this field of research [179,180]. They
author reports that the use of high molecular weight PEG reduces the have studied various aspects of the construction of these cationic bun
efficiency of gene vectors. This may be due to the PEG Space barrier to dles, physicochemical properties, and biological aspects such as cell
the carrier interacting with the particle [169–171,226–229]. harvesting and endosomal escape. Fig. 8 shows the synthesis and
structure of block copolymers.
5
Fig. 7. Structure of (A) silica (B) silica nanoparticles(C) Silica particles in various morphologies were observed by SEM and TEM images (D) Different forms of silica
nanoparticles. Figures reproduced with permission from Refs. [119,292,293].
6
Fig. 7. (continued).
7
Fig. 7. (continued).
Fig. 8. The Synthesis and structure of Block Copolymers. Figures reproduced with permission from Ref. [195].
3.7. Quantum dots attached the Quantum Dots to the plasmid DNA using a melamine linker
containing a disulfide bond. This system showed very low toxicity and
Burgess et al. showed that plasmid DNA could be covalently attached high DNA delivery to the cell nucleus. Bhatia et al. also linked the siRNA
to Quantum Dots. They encapsulated the quantum points with TOPO = to Quantum Dots using an endosome escape agent (lipofectamine) and
triocytylphosphine oxide and TOP (triocytylphosphine), and then were able to turn off the EGFP (Enhanced green fluorescent protein)
8
gene by up to 29% [177–182,302–308,309–315]. Various applications Particles was Higher than PEI Particles.
and advantages of bio functionalized quantum dots have been tabulated Rosi et al. [116,119] attached antisense oligonucleotides with thiol
in Table 1 [183–186,194]. groups to 13 nm gold nano Particles and obtained good Results for EGFP
gene silencing. In order to improve the Vectors of oligonucleotides with
nano Particles, Efforts have been made to make combinations of nano
3.8. Gold nanoparticles particles and DNA with unstable bonds, which are broken in the cell
cytoplasm and the oligonucleotide strand is separated from the nano
DNA Coated on metal nano particles (for gene transfer) was first used particles. For this purpose, in one of the methods, by increasing the
by Particle acceleration Devices (gene guns). In the Initial Studies, DNA concentration of intracellular glutathione, it is possible to release the
was deposited on tungsten nano Particles and these nano Particles were oligonucleotide filament that is attached to the particle by binding thiol
deposited on Large Polypropylene Particles [255–259,316–322]. The to the particle [116,119,192,275].
device’s explosive Pressure is used to Push the Particles, and large
Particles Are trapped on the Polycarbonate disk at the End of the ma
chine Tube, while tungsten nano Particles pass through the disk and 3.9. Carbon nanotubes
enter the cells [323–328,329–334]. Sometime later, devices with helium
gas Pressure that generated a shock wave were used to Vectors gold nano The use of carbon nano tubes for gene therapy dates back to recent
Particles into tissues. These devices Vectors 4 times more luciferase gene years. These carbon Structures are highly Insoluble and Covalent and
to the skin of mice than Previous series of Devices, but the shock caused non-Covalent Functionalization Techniques have been Developed to
by the Pressure of helium gas could cause cell Damage [187,188, Increase the Solubility of These Structures [260–262,355,356–359].
335–341,342,343]. Therefore, later models used helium gas Pressure to Civilianization Involves two methods. Oxidation of Carbon nano tubes
launch a bullet. The Pressure from the Bullet stopped the bullet on the Under Acidic Conditions Produces Acidic Ends [360–365,366–370].
disk at the end of the Device Tube, but like Previous models that did not Non-Covalent Functionalization Typically Involves hydrophobic or
use helium gas pressure, the DNA Particles went to the other side Step-by-Step Interactions between Carbon nano tubes and Surfactants,
[344–349]. Discs were thrown, causing minimal damage to the cells. In Amino Acids, or nucleotide Strands.
addition, Arc-Discharge guns have been Used as Particle accelerators. Bianco et al. used carbon nanotubes covalently attached to cationic
These systems contain gold Particles coated on a Mylar film (polyester functional groups to bind DNA. Cell harvesting of these compounds
layer). The electric arc created by the two electrodes accelerates the film contained DNA in the Hella cell line through endocytosis [263–267,
on a screen. The Film stops on the screen, but the gold Particles Pass 371–373]. Microscopic results showed that DNA binding to multicol
through the screen and reach the cells. In this way, the volume of Par ored carbon nanotubes was much stronger than binding DNA to
ticles Vectors by the Device Can Be changed by changing the Voltage, single-walled nanotubes. Special has been done using carbon nanotubes.
The Density of Au-DNA Particles [189,190,350–354]. Particle Dai and colleagues used phospholipid-PEG-functionalized nanotubes to
bombardment is one of the Primary methods of DNA Vectors that can be react with thiol-terminated siRNA [268–272]. The terminal amine
used for Immunization applications. Recently, researchers have been attached to the nanoparticle reacts with the sulfur group attached to the
thinking about changing the level of gold nano Particles to Vectors their DNA. These structures have been shown to have excellent siRNA de
endocytosis to cells. livery and gene silencing capabilities. In-body tests performed with
Rotello et al. [116,119] synthesized monolayer-Protected gold carbon nanotubes also provide a clear perspective for this method
clusters (MMPCs = mixed monolayer protected gold Clusters) by [176–184,374–376].
Reacting alkanotiols with 2 nm Gold Nano Particles. Positively Charged First described in 1991 [44], carbon nanotubes (CNTs), formed from
Clusters had the highest gene Vectors capacity (68%) and this Group one or more sheets of graphene shaped in a cylindrical structure, have
Showed that Gene Vectors capacity was Directly related to the alkyl unique chemical and physical properties. Iijima [44,45] initially
chain length. Klibanov and colleagues synthesized PEI-conjugated gold discovered multiwalled carbon nanotubes (MWNTs), which have a
nano Particles. The Synthesized Particles were able to Vectors 15 to 6 diameter of 4–30 nm, with two or more graphene cylinders centrically
times more Genes than PEI particles, although the Toxicity of These arranged. Two years later Iijima and Ichihashi [45,46] reported on
Table 1
Different approach of bio functionalized QD.
Cognition rate Conjugated to purpose approach Benefit References
QD
Streptavidin analysis of Biotinylated all Employed in the pathology Ability to resist light and Top sensitivity [160-162]
human DNA in metaphase of medical genetic diagnosis
chromosomes
Angiotensin 2 The Chinese Hamster Ovary – QD Yield And Photostability Levels are Higher Than The [163]
(CHO) Cells Is Used For Angiotensin Dye Conjugate
Incubation
CdS QD With Carboxyl Group DNA Molecules Used in marking DNA DNA is quantitatively examined in a more advanced manner, [164]
Molecules which is associated with stable QD fluorophores
Adenine and AMP bacteria is used for incubation bacteria is marked through – [145]
mechanisms dependent on
Purine
QDs-labeled – The syphilis is checked Lateral flow test, which is has lower cost, is sensitive, and can be [170]
through a new point of care easily used
based on QD
Cy3-SSDNA – – to detect solution-based fluorescence, minor DNA changes are [165]
necessary
One End of DNA Hybrid Bound – – This method in highly efficient in terms of detection potentials, in [166]
to Streptavidin Conjugated more appropriate to deal with temperature, and has shorter
605QD analysis time as a result of enhanced hybridization efficiency and
higher diffusion coefficient
3-mercaptoropionic acid – – – [173]
9
single walled carbon nanotubes (SWNTs), which have a diameter of choosing the right molecular weight, they can be optimized for specific
0.4–3 nm, and which are composed of a single graphene sheet. While applications. By attaching targeting agents to specific tissues or cells or
their small size and chemical inertness are attractive properties for de other changes, their biological and physicochemical properties can be
livery of DNA, their hydrophobic nature makes them poorly soluble in improved. Once you are fully aware of the proper carrier structure, it is
aqueous solutions, which places limitations on their application in bio usually easy to industrialize carrier manufacturing, but it is clear that all
logical systems. To increase their solubility and dispersion, CNTs can be of this is only possible for non-virus carriers [116,119].
functionalized through either covalent or non-covalent functionaliza
tion [47,48]. The two most common types of covalent functionalization 6. Conclusion
reactions are oxidations and cyclo-additions. While covalent function
alization can improve the biocompatibility and solubility of pristine As is evident from this review, many new and innovative approaches
(chemically unmodified) carbon nanotubes, many of their intrinsic to therapeutic gene delivery have emerged in recent years. Nano
physical properties are either destroyed or weakened [47]. Alterna materials appear especially well suited as delivery vectors for small RNA
tively, non-covalent functionalization can be carried out by coating the molecules. Given that the vast majority of studies with nanomaterials
carbon nanotube with amphiphilic molecules, such as sodium dodecyl have been conducted in cell culture systems, it is also clear that addi
sulfate [SDS] [49] or proteins [50], which preserves the chemical tional tests with whole organisms will be necessary before we can expect
structure, allowing the CNTs to maintain their unique physical proper to see a trend away from the use of viral vectors. Viral vectors are still
ties [51–54]. MWNTs are generally considered less attractive than considered the primary choice for gene delivery, evident by their use in
SWNTs because of their larger diameter. However, the large diameter is 67% of current clinical trials versus less than 1% of non-viral vectors. For
beneficial for the delivery of larger payloads of DNA. Additionally, the novel nanomaterials, techniques and gene delivery approaches continue
toxicity is not based solely on the size of the nanotube, but is also to be developed, the key challenge will be to balance transfection effi
determined by the method of functionalization [47]. ciency, targeting specificity, particle size, biodegradability, and cyto
Liu et al. [361] covalently functionalized MWNTs with chitosan-folic toxicity, as well as their short- and long-term fates in the environment.
acid nanoparticles (CS-FA NPs) to deliver plasmid DNA. The group This will typically require a mechanism to prevent the removal of vec
demonstrated that shorter MWNTs have a higher transfection efficiency, tors by the reticulo-endothelial system (RES). Many nanomaterials can
but as a consequence, also have a higher cytotoxicity. Yet, when func avoid the RES system by shielding their charged groups with PEG,
tionalized with CS-FA NPs, the transfection efficiency was increased, proteins or polysaccharides, as highlighted with a number of examples.
while the cytotoxicity decreased [361]. Additionally, the smaller the nanoparticles are, the lower the probability
of surface neutralization by serum proteins and subsequent RES clear
4. Strategies to improve cellular uptake of non-virus carrier ance. Quantum dots are of special interest in this respect. The co-
systems administration of two different nanomaterials to target and deliver
gene payloads to create a synergistic effect and the use of theranostic
Non-virus gene delivery is used to express different genes or to turn nanoparticles are especially interesting developments that show great
them off in animal tissues such as the central nervous system, lungs, promise. Furthermore, the use of standardized assays that can be used to
liver, etc. Gene delivery with these carriers is often inadequate determine the efficacy and fate of a variety of gene delivery vectors
compared to virus carriers, and gene survival is often poor in virus. would enable head-to-head comparisons. Following such additional
Further advances in non-virus Gene Delivery Technology Could Expand research, and detailed characterization of their interaction with the
Its applications and provide a reliable replacement for viral carriers patient host, it is likely that some nanomaterials will be approved for use
[117,155,194–196] For in vivo therapies, it is important to target the in humans in the near future, thus expanding the therapeutic repertoire
carrier for a specific cell to prevent effects on other non-target tissues for gene therapy. In the past decades, substantial advances have been
and cells. Active tagging is often done for a series of cells. However, in achieved in different areas related to gene-based therapy, including the
some cases, depending on the physiological condition, passive targeting development of new delivery materials, as well as improved potency and
of the carrier is also done passively [197–202]. For example, the pres stability of nucleic acids. Moreover, advances in genomics have greatly
ence of irregular pores in the endothelial tissue of tumor vessels enhanced our understanding of the genetic basis of disease and provided
(Irregular endothelial fenestration) allows the entry of nanoparticles of a a range of new targets for genetic medicine. Many recently identified
certain size or composition. They bring (EPR Effect). To achieve active disease targets are considered ‘undruggable’ using small-molecule in
targeting in-body conditions, the carrier must minimize nonspecific in hibitors. The potential of exogenous nucleic acids may provide an
teractions by being Coated with a Polymer Coating Such as PEG and avenue to translate our knowledge obtained from biomedical research
Have specific cell-specific targeting agents. Different ligands are used for into therapeutics. The broad application of non-viral gene-based therapy
this purpose, which depending on the structure and type of target cell will require continued efforts not only to understand the structur
can include folate, transferrin, EGF, FGF, aphrodisiacs, Nano bodies, e–function relationship and the biology behind delivery vectors but also
different antibodies [116,119]. to expand the list of tissues and organs that can be successfully targeted.
In addition, non-viral delivery of genome editing systems could facilitate
5. Target prospects and future both precise and permanent correction of disease genes. The develop
ment of advanced delivery vectors should increase the potential of gene-
Today, gene delivery research is looking for better carriers that based therapy to treat a range of different tissues and to silence, correct
match the patient’s characteristics and type of disease. Therefore, the or introduce specific genes with minimal adverse effects. More than a
possibility of designing specific carriers for various diseases in the future decade after the initial discovery of RNAi, the promise of RNAi-based
is high. Most genetic diseases require carriers who can transfer genes of drugs seems imminent. Several therapeutic approaches based on
medium and large size. Gene injection is not the only problem with gene siRNA have shown promising results in clinical trials and have illumi
delivery. In some cases, the carrier must provide the conditions for the nated the challenges associated with the application of RNAi in humans.
gene to be activated in the target tissue. Therefore, the carrier must have With the recent development of nanoparticle and conjugate formula
a regulatory device that allows the doctor to turn the gene on and off and tions, in vivo delivery of siRNAs and/or miRNAs can be achieved with
change the level of protein therapy. Among these tools, we can mention high efficiency and low toxicity, and efficacy has been shown in humans.
the promoters with large size and complex shape, but the placement of Although non-viral gene-based therapies have yet to be approved by the
these promoters in the vector is a major problem. It is very clear That in FDA, recent progress in the clinical development of RNAi has generated
The Future, polymers will play a key role in advancing gene carriers. By considerable excitement. As discussed, several clinical programmes
10
have reported targeted knockdown in humans following non-viral de University, Arak, IranYoung Researchers and Elite Club, Gachsaran
livery of siRNAs. One compound, ALN-TTR02, has now transitioned into Branch, Islamic Azad University, Gachsaran, Iran.
Phase III clinical trials. More recently, modified siRNAs conjugated to
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104981. Conference on Engineering Technology and its Applications (IICETA), 2022, May,
[326] G. Fadhil Smaisim, A.M. Abed, S.K. Hadrawi, A. Shamel, Parametric investigation pp. 77–84.
of thermal behaviour of salt-gradient solar pool for climatic conditions, Clean [349] G.F. Smaisim, N.M. Prabu, A.P. Senthilkumar, A.M. Abed, Synthesis of biodiesel
Energy 6 (5) (2022) 693–704. from fish processing waste by nano magnetic catalyst and its thermodynamic
[327] G.F. Smaisim, M. Gholami, D. Toghraie, M. Hashemian, A.M. Abed, Numerical analysis, Case Stud. Therm. Eng. (2022), 102115.
investigation of the flow and heat transfer of Al2O3/water nanofluid in a tube [350] W.A. AbdulHussein, A.M. Abed, D.B. Mohammed, G.F. Smaisim, S. Baghaei,
equipped with stationary and self-rotating twisted tapes, Prog. Nucl. Energy 151 Investigation of boiling process of different fluids in microchannels and
(2022), 104335. nanochannels in the presence of external electric field and external magnetic field
[328] Y. Jiang, G.F. Smaisim, M.Z. Mahmoud, Z. Li, H.Ş. Aybar, A.M. Abed, using molecular dynamics simulation, Case Stud. Therm. Eng. (2022), 102105.
Simultaneous numerical investigation of the passive use of phase-change [351] S. Ahamad, M. Mohseni, V. Shekher, G.F. Smaisim, A. Tripathi, J. Alanya-Beltran,
materials and the active use of a nanofluid inside a rectangular duct in the A detailed analysis of the critical role of artificial intelligence in enabling high-
thermal management of lithium-ion batteries, J. Power Sources 541 (2022), performance cloud computing systems, in: 2022 2nd International Conference on
231610. Advance Computing and Innovative Technologies in Engineering (ICACITE),
[329] S.A. Jasim, M.H. Ali, Z.H. Mahmood, M. Rudiansyah, F.H. Alsultany, Y. IEEE, 2022, April, pp. 156–159.
F. Mustafa, A. Surendar, Role of alloying composition on mechanical properties of [352] A.N. Doss, D. Shah, G.F. Smaisim, M. Olha, S. Jaiswal, A comprehensive analysis
CuZr metallic glasses during the nanoindentation process, Met. Mater. Int. 28 (9) of internet of things (IOT) in enhancing data security for better system integrity-A
(2022) 2075–2082. critical analysis on the security attacks and relevant countermeasures, in: 2022
[330] D.O. Bokov, Y.F. Mustafa, Z.H. Mahmoud, W. Suksatan, M.A. Jawad, T. Xu, Cr- 2nd International Conference on Advance Computing and Innovative
SiNT, Mn-SiNT, Ti-C70 and Sc-cnt as effective catalysts for CO2 reduction to Technologies in Engineering (ICACITE), IEEE, 2022, April, pp. 165–167.
CH3OH, Silicon 14 (14) (2022) 8493–8503. [353] A. Lefteh, M. Houshmand, M. Khorrampanah, G.F. Smaisim, Optimization of
[331] S.A. Jasim, W.K. Abdelbasset, K. Hachem, M.M. Kadhim, G. Yasin, M.A. Obaid, Z. modified adaptive neuro-fuzzy inference system (MANFIS) with artificial bee
H. Mahmoud, Novel Gd2O3/SrFe12O19@ Schiff base chitosan (Gd/SrFe@ SBCs) colony (ABC) algorithm for classification of bone cancer, in: 2022 Second
nanocomposite as a novel magnetic sorbent for the removal of Pb (II) and Cd (II) International Conference on Distributed Computing and High Performance
ions from aqueous solution, J. Chin. Chem. Soc. 69 (7) (2022) 1079–1087. Computing (DCHPC), IEEE, 2022, March, pp. 78–81.
[332] A.A. Mansoor Al Sarraf, H. Alsultany, F.H. Mahmoud, Z.S. Shafik, S, A. [354] A.S. Sallal, G.F. Smaisim, S.M. Thahab, The heat transfer from fined perforated
I. Mashhadani, Z. I, A. Sajjadi, Magnetic nanoparticles supported zinc (II) complex pipe improved due to nano-fluid, J. Phys. Conf. 1973 (2021, August), 1, p.
(Fe3O4@ SiO2-Imine/Thio-Zn (OAc) 2): a green and efficient magnetically 012075). IOP Publishing.
reusable zinc nanocatalyst for synthesis of nitriles via cyanation of aryl iodides, [355] Ning Zhao, Libo Deng, Dawei Luo, Peixin Zhang, One-step fabrication of biomass-
Synth. Commun. 52 (9–10) (2022) 1245–1253. derived hierarchically porous carbon/MnO nanosheets composites for symmetric
[333] Z. Hameed Mahmood, Y. Riadi, H.A. Hammoodi, A.F. Alkaim, Y. Fakri Mustafa, hybrid supercapacitor, Appl. Surf. Sci. 526 (2020), 146696.
Magnetic Nanoparticles Supported Copper Nanocomposite: A Highly Active [356] H. Al-Madhhachi, G.F. Smaisim, Experimental and numerical investigations with
Nanocatalyst for Synthesis of Benzothiazoles and Polyhydroquinolines, Polycyclic environmental impacts of affordable square pyramid solar still, Sol. Energy 216
Aromatic Compounds, 2022, pp. 1–19. (2021) 303–314.
[334] Z.H. Mahmoud, R.A. Al-Bayati, A.A. Khadom, The efficacy of samarium loaded [357] G.F. Smaisim, Investigation on heat transfer augmentation using continuous and
titanium dioxide (Sm: TiO2) for enhanced photocatalytic removal of rhodamine B broken ribs on a plate of heat exchanger, Int. J. Energy Environ. 9 (3) (2018).
dye in natural sunlight exposure, J. Mol. Struct. 1253 (2022), 132267. [358] G.F. Smaisim, Augmentation of heat transfer in corrugated tube using four-start
[335] M.W. Tian, A.M. Abed, S.R. Yan, S.M. Sajadi, M.Z. Mahmoud, H.Ş. Aybar, G. spiral wall, Al-Qadisiya. J. Eng. Sci. ournal for Engineering Sciences 10 (4) (2017)
F. Smaisim, Economic cost and numerical evaluation of cooling of a cylindrical 451–467.
lithium-ion battery pack using air and phase change materials, J. Energy Storage [359] G.F. Smaisim, Enhancement heat transfer of Cu-water nanofluids with
52 (2022), 104925. thermophysical properties modeling by artificial neural network, J. Univ. Bombay
[336] K.A.M. Alharbi, G.F. Smaisim, S.M. Sajadi, M.A. Fagiy, H.Ş. Aybar, S.E. Elkhatib, 25 (5) (2017) 1721–1735.
Numerical study of lozenge, triangular and rectangular arrangements of lithium- [360] Ehsan Kianfar, Synthesis of Characterization Nanoparticles Isophthalic Acid/
ion batteries in their thermal management in a cooled-air cooling system, Aluminum Nitrate (CAU-10-H) Using Method Hydrothermal. Advances in
J. Energy Storage 52 (2022), 104786. Chemistry Research, Nova Science Publishers, Inc., NY, USA, 2020.
[337] W. Wu, G.F. Smaisim, S.M. Sajadi, M.A. Fagiry, Z. Li, M.A. Shamseldin, [361] Ehsan Kianfar, CO2 capture with ionic liquids: a review, Adv. Chem. Res. 67
H.Ş. Aybar, Impact of phase change material-based heatsinks on lithium-ion (2020). Nova Science Publishers, Inc., NY, USA.
battery thermal management: a comprehensive review, J. Energy Storage 52 [362] Ehsan Kianfar, Enhanced light olefins production via methanol dehydration over
(2022), 104874. promoted SAPO-34, Adv. Chem. Res. 63 (2020). Chapter: 4, Nova Science
[338] M.W. Tian, G.F. Smaisim, S.R. Yan, S.M. Sajadi, M.Z. Mahmoud, H.Ş. Aybar, A. Publishers, Inc., NY, USA.
M. Abed, Economic cost and efficiency analysis of a lithium-ion battery pack with [363] Ehsan Kianfar, Gas hydrate: applications, structure, formation, separation
the circular and elliptical cavities filled with phase change materials, J. Energy processes, Thermodynamics, in: James C. Taylor (Ed.), Advances in Chemistry
Storage 52 (2022), 104794. Research, Edition vol. 62, Nova Science Publishers, Inc., NY, USA, 2020. Chapter:
[339] W. Brontowiyono, W.A. AbdulHussein, G.F. Smaisim, M.Z. Mahmoud, S. Singh, H. 8.Publisher.
A. Lafta, S. Aravindhan, Annealing temperature effect on structural, magnetic [364] Mehran Kianfar, Farshid Kianfar, Ehsan Kianfar, The effect of nano-composites on
properties and methyl green degradation of Fe2O3 nanostructures, Arabian J. Sci. the mechanic and morphological characteristics of NBR/PA6 blends, Am. J. Oil
Eng. (2022) 1–8. Chem. Technol. 4 (1) (2016) 29–44.
[340] Y. Tian, I. Patra, H.S. Majdi, N. Ahmad, R. Sivaraman, G.F. Smaisim, [365] Ehsan Kianfar, The effect of nano-composites on the mechanic and morphological
M. Hekmatifar, Investigation of atomic behavior and pool boiling heat transfer of characteristics of NBR/PA6 blends, Am. J. Oil Chem. Technol. 4 (1) (2016) 27–42.
water/Fe nanofluid under different external heat fluxes and forces: a molecular [366] G. Smaisim, O. Fatta, A. Valera-Medina, A. Rageb, N. Syred, Investigation of heat
dynamics approach, Case Stud. Therm. Eng. 38 (2022), 102308. transfer and fluid mechanics across a heated rotating circular cylinder in
[341] G.F. Smaisim, H. Al-Madhhachi, A.M. Abed, Study the thermal management of Li- crossflow, in: 54th AIAA Aerospace Sciences Meeting, 2016, 0494.
ion batteries using looped heat pipes with different nanofluids, Case Stud. Therm. [367] G. Smaisim, O. Fatla, A. Valera Medina, A.M. Rageb, N. Syred, Experimental and
Eng. 37 (2022), 102227. theoretical investigation of the effect of rotating circular cylinder speed on the lift
[342] I. Raya, A.A. Mansoor Al Sarraf, G. Widjaja, S. Ghazi Al-Shawi, M. F Ramadan, Z. and drag forces, Int. J. Energy Environ. 7 (1) (2016) 23–36.
H. Mahmood, H. Ghaleb Maabreh, ZnMoO4 nanoparticles: novel and facile [368] N. Nasajpour-Esfahani, H. Garmestani, M. Rozati, G.F. Smaisim, The role of phase
change materials in lithium-ion batteries: a brief review on current materials,
17
thermal management systems, numerical methods, and experimental models, [374] A.M. Abed, A.K. Yakoob, G.F. Smaisim, H.T. Gatea, Design and sizing of stand-
J. Energy Storage 63 (2023), 107061. alone photovoltaic (PV) system for powered mobile cleaning and disinfection
[369] M. Niknejadi, A.A. Alizadeh, H. Zekri, B. Ruhani, N. Nasajpour-Esfahani, G. chamber system, AIP Conf. Proc. 2776 (2023, April). No. 1, p. 050001). AIP
F. Smaisim, Numerical simulation of the thermal-hydraulic performance of solar Publishing LLC.
collector equipped with vector generators filled with two-phase hybrid nanofluid [375] H. Wang, A.A. Alizadeh, A.M. Abed, A. Piranfar, G.F. Smaisim, S.K. Hadrawi,
Cu-TiO2/H2O, Eng. Anal. Bound. Elem. 151 (2023) 670–685. M. Hekmatifar, Investigation of the effects of porosity and volume fraction on the
[370] X. Dai, H.T. Andani, A.A. Alizadeh, A.M. Abed, G.F. Smaisim, S.K. Hadrawi, atomic behavior of cancer cells and microvascular cells of 3DN5 and 5OTF
D. Toghraie, Using Gaussian Process Regression (GPR) models with the Matérn macromolecular structures during hematogenous metastasis using the molecular
covariance function to predict the dynamic viscosity and torque of SiO2/Ethylene dynamics method, Comput. Biol. Med. (2023), 106832.
glycol nanofluid: a machine learning approach, Eng. Appl. Artif. Intell. 122 [376] X. Liu, Y. Zhang, D. Ma, H. Tang, L. Tan, Q. Xie, S. Yao, Biocompatible multi-
(2023), 106107. walled carbon nanotube-chitosan–folic acid nanoparticle hybrids as GFP gene
[371] Y.X. Zhang, A.A. Alizadeh, A.M. Abed, N. Nasajpour-Esfahani, G.F. Smaisim, S. delivery materials, Colloids Surf., B 111 (2013) 224–231.
K. Hadrawi, M.X. Wang, Investigating the effect of size and number of layers of [377] Z.H. Mahmoud, R.A. Al-Bayati, A.A. Khadom, Electron transport in dye-sanitized
iron nanochannel on the thermal behavior and phase change process of calcium solar cell with tin-doped titanium dioxide as photoanode materials, J. Mater. Sci.
chloride/sodium sulfate hexa-hydrate with molecular dynamics simulation, Mater. Electron. 33 (8) (2022) 5009–5023.
J. Energy Storage 62 (2023), 106762. [378] A. Bahadoran, M.K. Jabarabadi, Z.H. Mahmood, D. Bokov, B.J. Janani, A. Fakhri,
[372] J. Tang, A. Ahmadi, A.A. Alizadeh, R. Abedinzadeh, A.M. Abed, G.F. Smaisim, Quick and sensitive colorimetric detection of amino acid with functionalized-
D. Toghraie, Investigation of the mechanical properties of different amorphous silver/copper nanoparticles in the presence of cross linker, and bacteria detection
composites using the molecular dynamics simulation, J. Mater. Res. Technol. 24 by using DNA-template nanoparticles as peroxidase activity, Spectrochim. Acta
(2023) 1390–1400. Mol. Biomol. Spectrosc. 268 (2022), 120636.
[373] A.A. Alizadeh, A.M. Abed, H. Zekri, G.F. Smaisim, B. Jalili, P. Pasha, D.D. Ganji,
Numerical investigation of the effect of the turbulator geometry (disturber) on
heat transfer in a channel with a square section, Alex. Eng. J. 69 (2023) 383–402.
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Case Studies in Chemical and Environmental Engineering 8 (2023) 100372

Contents lists available at ScienceDirect

Case Studies in Chemical and Environmental Engineering

Nanomaterials as transmitters of non-viral gene vectors: A review

Fig. 2. Schematic of the process of interaction of a positively charged polymer

2. Nanoparticles for gene and drug vectors 3.1. Chitosan

3.4. Poly (lactide-Co-glycolide)

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