See discussions, stats, and author profiles for this publication at: https://www.researchgate.

net/publication/324020830

Applications of zeolites in biotechnology and medicine – a review

Article  in  Biomaterials Science · March 2018

DOI: 10.1039/C8BM00028J

CITATIONS READS

58 3,561

4 authors, including:

Marta Vandrovcova Ivana Nemcakova

Ministry of Education, Youth and Science, the Czech Republic The Czech Academy of Sciences
53 PUBLICATIONS   1,053 CITATIONS    15 PUBLICATIONS   245 CITATIONS   

SEE PROFILE SEE PROFILE

Ivan Jirka
The Czech Academy of Sciences
60 PUBLICATIONS   769 CITATIONS   

SEE PROFILE

Some of the authors of this publication are also working on these related projects:

Biocompatibility of doped DLC layers prepared by hybrid laser technology View project

Strain engineering of electronic structure in graphene View project

All content following this page was uploaded by Ivan Jirka on 12 December 2018.

The user has requested enhancement of the downloaded file.

Biomaterials
Science
REVIEW

Applications of zeolites in biotechnology and

Cite this: Biomater. Sci., 2018, 6, 974
medicine – a review
Lucie Bacakova, *a Marta Vandrovcova, a
Ivana Kopova a
and Ivan Jirka *b

Received 8th January 2018,
Accepted 25th March 2018
DOI: 10.1039/c8bm00028j
rsc.li/biomaterials-science
Zeolites are microporous tectosilicates of natural or synthetic origin, which have been extensively used in
various technological applications, e.g. as catalysts and as molecular sieves, for separating and sorting
various molecules, for water and air purification, including removal of radioactive contaminants, for har-
vesting waste heat and solar heat energy, for adsorption refrigeration, as detergents, etc. These appli-
cations of zeolites were typically related with their porous character, their high adsorption capacity, and
their ion exchange properties. This review is focused on potential or already practically implemented
applications of zeolites in biotechnology and medicine. Zeolites are promising for environment protec-
tion, detoxication of animal and human organisms, improvement of the nutrition status and immunity of
farm animals, separation of various biomolecules and cells, construction of biosensors and detection of
biomarkers of various diseases, controlled drug and gene delivery, radical scavenging, and particularly
tissue engineering and biomaterial coating. As components of scaffolds for bone tissue engineering, zeo-
lites can deliver oxygen to cells, can stimulate osteogenic cell differentiation, and can inhibit bone resorp-
tion. Zeolites can also act as oxygen reservoirs, and can improve cell performance in vascular and skin
tissue engineering and wound healing. When deposited on metallic materials for bone implantation,
zeolite films showed anticorrosion effects, and improved the osseointegration of these implants. In our
studies, silicalite-1 films deposited on silicon or stainless steel substrates improved the adhesion, growth,
viability and osteogenic differentiation of human osteoblast-like Saos-2 cells. Zeolites have been clinically
used as components of haemostatics, e.g. in the Advanced Clotting Sponge, as gastroprotective drugs,
e.g. Absorbatox® 2.4D, or as antioxidative agents (Klinobind®). Some zeolites are highly cytotoxic and
carcinogenic, e.g. erionite. However, in other zeolites, the antiproliferative and pro-apoptotic effects can
be used for tumor therapy.

1. Introduction blocks) into a corner-sharing network (secondary building

Zeolites are hydrated tectosilicates appearing as minerals in
nature, and also as artificial materials synthesized in the
laboratory.1–4 (Fig. 1). The term “zeolites” was introduced in
1756 by Swedish mineralogist Axel Fredrik Crønsted, who dis-
covered stilbite. Crønsted observed that upon rapid heating,
this material produced steam from water that had been
adsorbed by the material. Based on this, he called the material
zeolite, from the Greek ζέω (zéō), meaning “to boil” and λίθoς
(líthos), meaning “stone”.5
Zeolites are microporous crystalline aluminosilicates com-
posed of TO4/2 tetrahedra, where T stands for Si or Al.
Crystallization of zeolites proceeds under hydrothermal con-
ditions by linking these Al (Si) tetrahedra ( primary building
a
Institute of Physiology of the Czech Academy of Sciences, Videnska 1083, 14220
Prague 4, Czech Republic. E-mail: Lucie.Bacakova@fgu.cas.cz
b
J. Heyrovsky Institute of Physical Chemistry of the Czech Academy of Sciences,
Dolejskova 3, 18223 Prague 8, Czech Republic. E-mail: ivan.jirka@jh-inst.cas.cz
blocks) through bridging oxygen (O) atoms constituted by
rings and prisms of various sizes. These units combine to
generate frameworks with a regular distribution of molecular-
sized pores and cavities with pore sizes ranging between
4–12 Å.1,8,9 Over 40 naturally occurring zeolite frameworks are
known, and synthesized zeolites are known in the form of 229
unique zeolite frameworks.4
In the zeolite framework, Si is tetravalent and Al is trivalent.
SiO4/2 tetrahedra are therefore electro-neutral, and AlO4/2 tetra-
hedra are negatively charged. This charge is compensated by
some extra-framework cation (typically by alkali and alkali
earth metals). The chemical composition of hydrated zeolite
can be expressed by the following formula:
jMx=n ; ðH2 OÞz j½ðAlO2 Þx ðSiO2 Þy 
where M is an extra-framework cation with valence n, and x, y
are pertinent values of molar concentrations of Al and Si in
the zeolite framework, and z is molar concentration of H2O.

974 | Biomater. Sci., 2018, 6, 974–989 This journal is © The Royal Society of Chemistry 2018
Biomaterials Science
Fig. 1 Natural zeolites clinoptilolite6 (A) and stilbite with bluish
cavansite7 (B), and synthetic zeolite silicalite-1 (C, D) in the form of
films on a Si(100) substrate (C) or on an AISI-316L stainless steel sub-
strate (D). C: A continuous layer of b-oriented crystals covered by iso-
lated a-oriented crystals; D: a highly defective film composed of
a-oriented crystals.
Ivana Kopova, Lucie Bacakova, Ivan Jirka and Marta
Vandrovcova (from left to right)
Lucie Bacakova (LB) graduated from the Faculty of General
Medicine, Charles University, Prague, Czechoslovakia in 1984.
She completed her CSc studies (“candidatus scientiarum”, a PhD
equivalent), from the Czechoslovak Academy of Sciences, and
became an Associate Professor at the 2nd Medical Faculty, Charles
University in Prague. Since 2005, she has been Head of the
Department of Biomaterials and Tissue Engineering, Institute of
Physiology of the Czech Academy of Sciences. She is a specialist in
studies on the interaction of differentiated and stem cells with
various biomaterials, and in vascular, bone and skin tissue engin-
eering. She has published ∼170 papers in reputed journals
(4348 citations in total, 3643 citations without self-citations,
h-index 33).
This journal is © The Royal Society of Chemistry 2018
Review
The distribution of Al in the zeolite framework obeys the
Loewenstein rule8 (i.e. no framework Al–O–Al linkages are
present). Some examples of zeolite framework structures are
depicted in Fig. 2. Al in the zeolite framework can be isomor-
phously substituted by some other element, e.g. Ti, Zr, B.
Some zeolites can also be purely siliceous (Si/Al = ∞).
Zeolites have great potential for a wide range of technical,
industrial, agricultural, commercial and also biomedical appli-
cations. For this reason, zeolites have also been referred to as
“magic stones”.10 The applications of zeolites are typically
related to their porous character (i.e. their inner surface, which
is as large as ∼102 m2 g−1) and their ion exchange properties.
The adsorption properties of zeolites are governed by the
ability of adsorbate molecules to penetrate into zeolite voids,
which is limited by their dimensions and by zeolite pore sizes
(ranging from 0.4 nm up to 1.3 nm). Adsorption may proceed
on both the inner and the outer zeolite surface. An interesting
intermediate case – adsorption of molecules with dimensions
comparable with zeolite pore sizes – is also observed. Pure
silica zeolites do not obey ion exchange properties. These
materials can adsorb neutral molecules.
Zeolites are utilized as catalysts, e.g. in the petrochemical
industry,11 as molecular sieves for separating and sorting
molecules according their dimensions12,13 as adsorbents, e.g.
Marta Vandrovcova (MV) graduated from the Faculty of Sciences
of Charles University in Prague in 2004. In 2007 she joined the
group of Lucie Bacakova at the Institute of Physiology of the Czech
Academy of Sciences, Prague. She was awarded her PhD in human
physiology and pathophysiology in 2011. Her main research inter-
ests include interactions of bone-derived cells in cultures with
hard biocompatible artificial materials developed for replacements
of damaged bone tissue. She is a (co-) author of 36 papers in jour-
nals with an impact factor (h-index 13).
Ivana Kopova (IK) graduated from the Faculty of Sciences,
Charles University in Prague, where she was also awarded her
PhD in molecular biology in 2016. Her main research interest is in
the biocompatibility and cytotoxicity of materials used in bone
tissue engineering, with special emphasis on potential cellular
stress and DNA damage response. So far, she has (co-) authored 6
papers in journals with an impact factor (h-index 4).
Ivan Jirka (IJ) was awarded his CSc diploma by the J. Heyrovský
Institute of Physical Chemistry of the Czech Academy of Sciences
(former Czechoslovak Academy of Sciences). His doctoral studies
dealt with the characterization of copper clusters on carbon by
X-ray photoelectron spectroscopy. After the political liberation in
Czechoslovakia, he worked as a post-doctoral research fellow at
ETH, Zürich, Switzerland (Professor R. Prins) and at Lehigh
University in the USA (Professor K. Klier). Recently he became a
senior scientist in the J. Heyrovský Institute of Physical Chemistry
of the Czech Academy of Sciences. His field of interest is materials
science (catalytic systems, biomaterials, zeolites). He is a (co-)
author of 46 papers in journals with an impact factor
(h-index 14).
Biomater. Sci., 2018, 6, 974–989 | 975
Review
Fig. 2 Scheme of the structure of sodalite, zeolite A and zeolites X, Y.9
for water, soil and air purification,14–23 for removing radio-
active contaminants,24 for harvesting waste heat and solar heat
energy,25 for adsorption refrigeration,26 and as detergents, e.g.
for laundry,27 just to mention the most frequent applications
(Fig. 3).
In the last decade, much attention has focused on the syn-
thesis and characterization of supported zeolite films.29,30
They can be prepared using suitable reaction conditions on
the surface of various supports in the form of continuous
layers composed of inter-grown variously-oriented zeolite crys-
tals with dimensions and thicknesses varying typically from
∼102 to ∼103 nm.30 Zeolite coatings are under extensive investi-
gation for their potential applications as selective membranes,
chemical sensors, components of microelectronic devices,31,32
and particularly for surface modification of bone
implants.33–35
Fig. 3 Overview of the formation of zeolite structures and the broad
field of zeolite applications.28
976 | Biomater. Sci., 2018, 6, 974–989
Biomaterials Science
This review is focused on applications of zeolites in various
biotechnologies and in medicine, namely protection of the
living environment ( purification of water, soil and air, includ-
ing the removal of radioactive contaminants), detoxification of
living organisms, applications in agriculture, veterinary medi-
cine and zootechnology, separation of various biomolecules
and cell types, drug and gene delivery, construction of bio-
sensors and detection of biomarkers, creation of novel antioxi-
dants, haemostatics, scaffolds for tissue engineering and
implant coatings, and wound dressings. Most of these appli-
cations are under experimental development, but some of
them are already in clinical use, e.g. in haemostatics or in
gastroprotective drugs. Examples of the significance and the
potential bio-applications of various zeolites are given in
Table 1.
It should be taken into account that some zeolites are cyto-
toxic. In some cases, this cytotoxicity is highly undesirable,
because it can cause cancer, e.g. in the case of erionite.
However, in other cases, zeolite cytotoxicity can be utilized for
the treatment of tumors, if the use is based on the antiproli-
ferative and pro-apoptotic action of zeolites.
2. Application of zeolites in
environment protection
Zeolites can be applied effectively in environment protection,
e.g. in water, soil and air purification and in decontamination
of the environment from radioactivity. These applications are
based on the high sorption and ion-exchange activity of zeo-
lites. The most widely-used zeolite in these applications is
naturally occurring clinoptilolite. For example, clinoptilolite
modified by a surfactant (hexadecyltrimethylammonium) was
used for simultaneous removal of ammonium and nitrates
from wastewater.14 Another system for removing ammonium
and nitrogen from groundwater was based on a passive per-
meable reactive barrier with an oxygen-releasing compound
and clinoptilolite.15 Metallic ions with potential adverse effects
on biological systems can also be removed from water with the
use of zeolites. Clinoptilolite coated with magnetite nano-
particles is promising for the removal of molybdenum from
industrial wastewater streams.16 Clinoptilolite-rich zeolite
powders modified with a bio-inspired adhesive agent, i.e., poly-
dopamine, have been systematically studied as adsorbents for
Cu(II) cations from an aqueous solution.17 Natural clinoptilo-
lite-rich zeolitic tuff was investigated for the ion-exchange
kinetics of other divalent heavy metal ions, namely Mn2+, Zn2+
and Pb2+.18 Clinoptilolite tuff was also mechanically converted
to micro- and nanoparticles, which were then modified with
hexadecyltrimethyl ammonium bromide and dithizone, and
were used for removing Pb2+ from aqueous solutions.19 The
adsorption properties of chitosan/zeolite conjugate films were
evaluated for removing Cr6+ ions, which are potentially carci-
nogenic, from industrial wastewater.20 The cleaning properties
of zeolites can be further enhanced by immobilizing
Pseudomonas arsenicoxydans in order to remediate arsenite-
This journal is © The Royal Society of Chemistry 2018
Biomaterials Science Review

Table 1 Significance and applicability of various zeolites in biotechnology and medicine

Code Zeolite Significance, examples of applications

ANA Analcime Antioxidant effects in humans36

PHI Phillipsite Antioxidant effects in humans36
LTL Zeolite L Cell separation,37 detection of cancer cells,38 DNA delivery39
FAU Faujasite Drug delivery (5-fluorouracil),40 wound healing41–43
Studies on endocytosis44
FER Ferrierite Antioxidant45
ERI Erionite Highly carcinogenic46–48
HEU Clinoptilolite The most widely used zeolite:
• Purification of water,14–21 soil22 and air23
• Removal of radioactive contaminants24
• Detoxification of organisms49–51
• Positive effects on nutrition and the digestive tract,52–54 gastroprotective effects55
• Drug delivery (diclofenac,56 doxorubicin57)
• Construction of biosensors58,59
• Antioxidant, antiapoptotic, anti-inflammatory activity60,61
• Antitumor activity62
MFI Zeolite Socony Mobil-5 (ZSM-5), Drug delivery (gentamycin), antibacterial properties, bone implants63,64
Silicalite-1
ABW Zeolite A Antimicrobial wound-healing dressing,65 antimicrobial coating of bone implants,66 inhibition of
osteoclasts67

contaminated waters.21 Zeolites are also promising agents for
water softening. Disk membranes generated from high-purity
natural clinoptilolite mineral rock have shown promising
water desalination and de-oiling performance.68
Zeolites are also potent agents for soil purification. For
example, together with other natural minerals, i.e. bentonite
and goethite, clinoptilolite was found to be a promising adsor-
bent that can be utilized for removing toxic Cr3+ and Cr6+ from
contaminated soil.22
In air purification, clinoptilolite as an adsorbent bed, com-
bined with Fe(0) and Cu2O nanoparticles as catalysts, was used
for removing benzene, toluene and xylene from polluted air.23
In the decontamination of environments from radioactivity,
clinoptilolite was evaluated as sorbents for remediating land
contaminated with radiocaesium and radiostrontium in order
to return it to farming use.24 In decontaminating water from
radionuclides, e.g. 134Cs, 137Cs and 90Sr, the cation-exchange
capacity of a clinoptilolite was substantially enhanced by
exposing it to high-energy (8 MeV) electrons.69 Clinoptilolite
was also used for alimentary decontamination, e.g. for remov-
ing radioactive caesium (137Cs) from the meat of broilers and
pheasants.70
3. Organism detoxification using
zeolites
In addition to environmental decontamination, animal and
human organisms can also be decontaminated from toxic
agents. For example, mice exposed to Pb and supplemented
with clinoptilolite exhibited 2.3-fold lower frequency of
chromosome aberrations, a 2.5-fold higher mitotic index, a
1.5-fold higher percentage of normal erythrocytes, and a 1.3-
fold higher body weight in comparison with Pb-exposed and
unsupplemented animals.49,50 Clinoptilolite in the form of
porous nanoparticles is also promising for the treatment of
children with attention deficit hyperactivity disorder (ADHD)
by trapping Pb in their organism, and also by delivering DOPA
(i.e., L-3,4-dihydroxyphenylalanine, a precursor of dopamine)
into their central nervous system after injection of the zeolite
clinoptilolite in suspension into the blood. However, this treat-
ment has not yet been clinically approved.51,71 Consumption
of ethanol together with clinoptilolite decreased the
absorption of ethanol and its blood level in healthy alcohol
drinkers. This effect was more pronounced in males.72
Microencapsulated urease-zeolite W oral sorbent was proposed
for the removal of urea from the organism during uremia.73
It is problematic to eliminate toxins from the blood in
patients suffering from the final stages of uremia. Small
uremic toxin molecules can be removed using zeolites.
Elimination of urea, uremic acid, creatinine, p-cresol and
indoxyl sulphate was investigated in a study by Wernert et al.74
75% of creatinine can be eliminated by adsorption onto acidic
mordenite, and 60% of p-cresol can be eliminated by adsorp-
tion onto a silicalite-1. The efficiency of this elimination pro-
cedure is similar to or even higher than the conventional dialy-
sis system.74 These results were further confirmed by another
study by the same group,75 performed on pure silica MFI zeo-
lites and alumino-silicate MFI zeolites (Si/Al = 30), with H+,
Na+, K+ and Mg2+ as charge compensating cations. When
exposed to aqueous solutions of p-cresol at 37 °C, all tested
types of MFI zeolites showed high adsorption capacity for
p-cresol, but this capacity was highest for pure silica MFI.75
4. Use of zeolites in veterinary
medicine and zootechnology
Zeolites can improve animal health via several mechanisms,
including direct positive effects on the morphology, function

This journal is © The Royal Society of Chemistry 2018 Biomater. Sci., 2018, 6, 974–989 | 977
Review
and microbial flora of the digestive tract, delivery of metaboli-
cally important ions, improving the nutrition status, strength-
ening immunity, decontamination of food and drinking water
from toxins. For these applications, mainly natural zeolite clin-
optilolite is used, and it is the most widespread compound in
the medical market.
Adding clinoptilolite into the diet of poultry improved the
morphology of the gut (e.g., induced a greater height of villi)
and the composition of physiological microbial flora in the
gut,52 reduced intestinal mucosa damage and the inflamma-
tory response to bacterial lipopolysacharide,53 increased the
antioxidant capacity of the organism,54 and increased the
content of omega-3-polyunsaturated fatty acid in the body and
in the eggs.76,77 These positive effects of dietary clinoptilolite
were further enhanced by ion exchange with Zn. In poultry, Zn
deficiency can cause reduced weight gain, skeletal malfor-
mations, poor bone mineralization and immunological dys-
functions. Zn delivered through clinoptilolite reached higher
bioavailability than ZnSO4, as evidenced by enhanced tissue
Zn accumulation and the expression of Zn transporter genes
in laying hens78 and in broilers.79 Zinc-bearing clinoptilolite
also showed protective effects against Salmonella pullorum
infection in chicken.80 The health of farm animals ( poultry,
cattle) can also be improved significantly by removing nitrates
and mycotoxins (aflatoxin, ochratoxin) from their food and
drinking water with the use of clinoptilolite;81–83 for a review,
see ref. 3.
The positive effects of zeolites on the digestive tract,
observed in domestic animals, can also be projected to human
medicine. A clinoptilolite-based drug, Absorbatox® 2.4D,
reduced the severity of clinical symptoms and signs associated
with endoscopically negative gastroesophageal reflux disease
and non-steroidal anti-inflammatory drug medication. This
gastroprotective effect of clinoptilolite is probably due to its
binding to hydrogen ions and biologically active amines and
nitrates.55
5. Use of zeolites for separating
biomolecules and cells
Zeolites are promising for the development of new chromato-
graphic carriers for purification of proteins and fractionation
of cellular components such as proteins and nucleic acids.84,85
Various proteins can also be immobilized on the surface of
zeolite L crystals using mild radical nitroxide exchange reac-
tions. Exposure of these crystals to aqueous protein solutions
leads to site-selective immobilization of proteins onto the crys-
tals.86 In addition, zeolites can be used for refolding various
proteins from their inactive form to their active form, which is
important e.g. in preparing recombinant proteins.87–89
Another interesting application is for separating various
types of microbial and mammalian cells. Specific microbial
cells were shown to absorb selectively to certain zeolites. This
selectivity depends on the physical and chemical properties of
the outer surface of the zeolite, e.g. its wettability and particu-
978 | Biomater. Sci., 2018, 6, 974–989
Biomaterials Science
larly the surface charge governed by pH. With the use of Na-
BEA and H–Y zeolites, three representative microbes,
Escherichia coli, Bacillus subtilis and Staphylococcus aureus,
were separated from their mixture.90 The separation of mam-
malian cells, e.g., cell lines and cells in primary cultures, was
based on the different adhesion behavior of these cells on
zeolite L substrates functionalized with two enantiomers of
penicillamine (PEN) – L-PEN and D-PEN.37 Disk-shaped zeolite
L particles, coated with the cyclic integrin ligand c[RGDfK] via
an isocyanate linker, were used for rapid detection of cancer
cells, based on the fast adhesion ability of these cells.38
6. Zeolites as promising carriers for
drug and gene delivery
Zeolites are promising for therapeutical delivery of various
drugs, e.g. anti-inflammatory, anticancer and antimicrobial
drugs, and also for delivery of nucleic acids. Clinoptilolite was
proposed as a microporous carrier for pH-controlled oral deliv-
ery of aspirin (acetylsalicylic acid), i.e. a non-steroidal anti-
inflammatory drug with an anticoagulant property, in order to
prevent gastric irritation due to inhibition of cyclooxygenase I
located in the gastric mucosa.91 Similar problems are associ-
ated with the oral delivery of other non-steriodal anti-inflam-
matory drugs, namely diclofenac and piroxicam, and these
problems too can be solved with the use of zeolitic carriers,
e.g. clinoptilolite56 or zeolites X and Y.92 Microporous zeolites
of distinct framework types, textural properties and crystal
morphologies (BEA, ZSM and NaX) are promising for oral
delivery of indomethacin, another poorly water-soluble non-
steroidal anti-inflammatory drug, as demonstrated in cultures
of the Caco-2 human colorectal adenocarcinoma cell line.93
Zeolites and their composites were also proved to be suit-
able carriers for anticancer drugs, such as 5-fluorouracil, doxo-
rubicin and mitoxantrone. Faujasite (NaY) or zeolite L nano-
particles, loaded with 5-flourouracil, were internalized by cells
of two human colorectal carcinoma cell lines, i.e. HCT-15 and
RKO cells, where the drug was released and markedly
decreased the cell viability. At the same time, control unloaded
zeolite particles presented no toxicity to both cancer cell
lines.40 Similar effects on human colon adenocarcinoma Caco-
2 cells were observed when 5-fluorouracil was encapsulated
into porous NaX or BEA zeolitic particles. This effect was more
pronounced in NaX particles.94 In another in vitro study, mag-
netite-zeolite 4A composite nanoparticles loaded with 5-flouro-
uracil efficiently inhibited the proliferation of human gastric
carcinoma AGS cells through apoptotic mechanisms.95 A
smart system for the delivery of doxorubicin was recently devel-
oped, consisting of zeolitic imidazole framework nanospheres.
This system enabled easy encapsulation and controlled release
of the drug.96 The Zn2+-loaded clinoptilolite/graphene oxide
nanocomposite is also a promising carrier for doxorubicin.57
Another cytostatic, i.e. mitoxantrone, was loaded into the pores
of zeolite beta polymorph disordered framework (BEA) nano-
This journal is © The Royal Society of Chemistry 2018
Biomaterials Science
particles, and showed considerable cytotoxicity when applied
to cancer cells.97
Zeolites have been tested for delivering various anti-
microbial drugs, such as antibiotics, chemotherapeutics,
metallic ions and nitric oxide (NO). ZSM-5 nanoparticles
showed high loading efficiency and sustained release of genta-
micin, which significantly reduced the adhesion of
Staphylococcus epidermidis and prevented biofilm formation.
These particles are therefore promising for preventing infec-
tion of bone implants.63 Porous zeolite/chitosan composites
proved to be promising for the delivery of cephalexin.98 Zeolite
Y was used for sustained release of sulfadiazine for the prepa-
ration of topical antibacterial formulations, and the antibacter-
ial effect was further enhanced by encapsulation of Ag nano-
particles.99 Other metallic ions, such as Zn2+ and Cu2+, have
also been loaded into zeolites (types X and A) in order to incor-
porate their antibacterial, anticandidal and antifungal pro-
perties.100 NO is important for the regulation of various bio-
logical processes, such as vascular tone, neurotransmission,
inflammatory cell responsiveness, and also for defence against
invading pathogens and wound healing. Composites of
polytetrafluoroethylene and NO-storing Zn2+-exchanged zeolite
A significantly reduced the growth of clinically relevant strains
of bacteria, namely Gram-negative Pseudomonas aeruginosa
and Gram-positive methicillin-sensitive and methicillin-resist-
ant Staphylococcus aureus and Clostridium difficile.101
For potential gene delivery into cells, zeolite L crystals have
been developed as multifunctional nanocontainers to deliver
simultaneously DNA oligonucleotides and other organic mole-
cules into living cells. Multifunctional zeolite L was prepared
by filling the pore system with guest molecules, whilst DNA
was adsorbed electrostatically on their surface.39,102 The
efficiency of cell transfection can also be enhanced by zeolites.
Zeolite-based silicalite nanoparticles markedly enhanced (by
over 150%) the transfection efficiency of poly(ethylene imine)-
plasmid DNA complexes applied to human embryonic kidney
HEK-293 cells.103
Zeolite nanoparticles carrying drugs, DNA and other bio-
active molecules are usually internalized into cells by endo-
cytosis. Zeolite nanoparticles can therefore be used as tools for
studying the endocytosing mechanisms and pathways in cells.
For example, de-aluminated zeolite Y nanoparticles were used
for studying endocytosis in human peripheral dendritic cells,
i.e. important cells of the immune system, in correlation with
the maturation status of these cells.44
7. Construction of biosensors and
detection of biomarkers using zeolites
Zeolites are promising materials for creating biosensors, and
also systems for harvesting and detecting biomarkers of
serious diseases, particularly tumors.
In biosensing, clinoptilolite and zeolite beta are promising
for developing potentiometric biosensors based on co-immo-
bilization of enzymes with various types of zeolite on pH-ion-
This journal is © The Royal Society of Chemistry 2018
Review
sensitive field-effect transistors.58 A citrate-selective sensor was
prepared by modifying a polyvinyl chloride membrane with
modified nano-clinoptilolite particles by a hexadecyltrimethyl
ammonium surfactant.59 A Cu-A zeolite/graphene-modified
glassy carbon electrode was constructed for simultaneous
electrochemical determination of dopamine and ascorbic
acid.104 Ascorbic acid and Acetaminophen A were also deter-
mined voltammetrically by a novel carbon paste electrode
modified with Fe3+-exchanged clinoptilolite nano-particles.105
ZSM-5 zeolite was applied for in situ real-time determi-
nation of superoxide anion, based on direct electron transfer
of Mn2+ phosphate (Mn3(PO4)2), which acted as a superoxide
dismutase. Mn2+ was ion-exchanged into
ZSM-5 microstructures, and was further coated with poly
(diallyldimethylammonium chloride).106 Recently, zeolite NaX
synthesized at various temperatures (from 60 to 100 °C)
immobilized with urease was tested for the potential construc-
tion of a urea biosensor.32
In addition, a strategy for harvesting and detecting tumor
secretory proteins was developed using zeolites. Proteins
secreted from the human hepatocellular carcinoma cell line
were enriched by zeolite L nanocrystals, followed by 1-D
sodium dodecyl sulfate-polyacrylamide gel electrophoresis
(SDS-PAGE) for protein fractionation, and then by liquid
chromatography-electrospray ionization mass spectrometry
coupled with tandem mass spectrometry (LC-ESI-MS/MS) for
protein identification and further proteomic and secretomic
analysis.107
8. Antioxidative, antiapoptotic and
anti-inflammatory effects of zeolites
Zeolites have significant antioxidant activity, which can be uti-
lized in human and veterinary medicine. Clinoptilolite showed
positive effects on hepatocytes after partial hepatectomy in rats
on the basis of the antioxidative effects of this zeolite. After
oral administration, clinoptilolite decreased the level of malon-
dialdehyde, a marker of oxidative stress, in the liver tissue. At
the same time, the activity of superoxide dismutase and the
level of glutathione, i.e. antioxidant molecules, significantly
increased in this tissue.60
Zeolites also have a protective effect on hepatocytes during
anticancer therapy using Adriamycin (doxorubicin). In primary
cultures of liver cells derived from rats treated with
Adriamycin, subsequent treatment with clinoptilolite signifi-
cantly reduced the production of imflammatory cytokines, i.e.
tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β) by
hepatocytes. In addition, this treatment led to reduced apoptosis
of hepatocytes, which was detected using the TdT-dUTP
Terminal Nick-End Labeling (TUNEL) method. These results
were attributed to the antioxidant effects of clinoptilolite.61 In
tumor-bearing animals (mice and dogs) treated with doxorubi-
cine, too, clinoptilolite decreased the lipid peroxidation and
enhanced the effect of doxorubicine. This was manifested by a
strong reduction in the pulmonary metastasis count.108
Biomater. Sci., 2018, 6, 974–989 | 979
Review
Other natural zeolites, such as chabazite, phillipsite and
analcime, administered perorally, also increased the blood
levels of antioxidant enzymes, namely gluthatione peroxidase,
superoxide dismutase and gluthatione reductase, and reduced
the lipid peroxidation in clinically healthy men, both non-
smokers and smokers.36 Faujasite and ferrierite zeolites
limited the effects of reactive oxygen species (ROS) on human
albumin under in vitro conditions. This was attributed to the
conversion of ROS to water molecules within the zeolite void
system, which generates H+ ion transport.45
The antioxidative properties of zeolites could also be utilized
in the treatment of Alzheimer’s disease. Studies on neuronal-
like SH-SY5Y cells in vitro, and on APPswePS1dE9 transgenic
mice in vivo (i.e. an animal model of ageing-related neurodegen-
eration), revealed that dietary zeolites (i.e. tribomechanically
micronized clinoptilolite tuff) reduced the production of mito-
chondrial ROS, increased the activity of superoxide dismutase
in the hippocampus, and reduced the amyloid levels and the
plaque load in mouse brain.109 In an ex vivo study performed
on homogenized brains, taken from mice after feeding with a
diet containing clinoptilolite, this zeolite showed a beneficial
effect on the ligand binding to serotonergic receptors.110
The antioxidative effect of zeolites can also protect the skin
from damage caused by UV irradiation. For example, NaY zeolites
decreased the photocatalytic activity of TiO2, a component of
some sunscreens, and thus prevented damage to cultured human
skin fibroblasts by ROS generated by UV-irradiated TiO2.111
Zeolites could therefore be used for creating novel sunscreens.
As was mentioned above, the antioxidant properties of clin-
optilolite can also be used in veterinary medicine, e.g. for
improving the antioxidative capacity of broiler chicken, mani-
fested by an increase in glutathione peroxidase, catalase and
superoxide dismutase, and by a decrease in malondialdehyde.54
Antioxidant properties of zeolites have also been utilized in
human medicine. In a randomized double-blind, controlled
clinical trial, a natural zeolite, Klinobind®, especially when
treated with mechanical micronization and with tribochemical
and thermal activation, suspended in water and administered
perorally, increased antioxidant enzymes (catalase) and
decreased ROS (H2O2) in blood plasma taken from smokers.112
Tribomechanically-activated clinoptilolite also proved to be ben-
eficial for the treatment of dyslipidemia. In a phase I/IIa pro-
spective, open-label, uncontrolled, dose/granule size-ranging
study, this mineral significantly improved the lipid profiles in
the blood of dyslipidemic patients, evaluated by the concen-
trations of total cholesterol, low-density lipoprotein cholesterol,
high-density lipoprotein cholesterol and triglycerides.113
9. Zeolites in haemostasis
Due to their adsorption capacity, zeolites can be used as
potent haemostatics. Zeolites can entrap exothermically large
volumes of water in the open porous internal space of zeolites.
At the same time, zeolites concentrate coagulation factors and
platelets in haemorrhaging blood (for a review, see ref. 114).
980 | Biomater. Sci., 2018, 6, 974–989
Biomaterials Science
Accordingly, zeolite-based haemostat QuickClot® (i.e., a com-
mercial material composed of zeolite beads) accelerated blood
coagulation and shortened the clot formation time, when added
to blood taken from human volunteers and from sheep in
vitro.115 Another zeolite-based haemostat is Advanced Clotting
Sponge (ACS), which is similar to QuickClot®, but consists of
larger beads of the zeolite, which are packaged in a gauze.114,116
However, the haemostatic effect of zeolites is associated
with an exothermic reaction, which can increase the temperature
of zeolites to as much as 90 °C, and can cause severe damage
(i.e., burns) to the adjacent tissues. This problem was addressed
by pre-hydrating the zeolites, so that they could absorb water less
exothermically. This modified zeolite formulation was commer-
cialized as Advanced Clotting Sponge plus (ACS+®).114,116
Another possible way to reduce the exothermic reaction of zeo-
lites and the temperature at the wound site is by adding another
mineral, bentonite, to the zeolite (in a bentonite : zeolite weight
ratio of 2 : 1). This formulation, commercialized as CoolClot®,
significantly reduced the bleeding time in dogs and the clotting
time in human blood samples in vitro,117 and supported the
healing of experimental wounds in rats in vivo, without a signifi-
cant increase in the wound surface temperature.118
Other minerals applied as haemostatic agents were phyllo-
silicates like smectite (WoundStat119,120), kaolin (e.g. QuikClot
Combat Gauze® 114) and halloysite.121 The effect of phyllosili-
cate smectite on haemostasis was reported to be even better
than the effect of zeolites.119,120 However, practical application
of zeolites in haemostasis is desirable, as phyllosilicates are
nano-materials which could get into the blood circuit.
10. Zeolites in tissue engineering,
implant coating and wound healing
Zeolite-based materials can be considered as a kind of bio-
active ceramics, and thus they can be used effectively in the
construction of scaffolds for bone tissue engineering, in which
zeolites can mimic the mineral component of the natural bone
matrix. For example, zeolites (specifically zeolite NaY) were com-
bined with poly(vinylidene fluoride), an electroactive material
showing a piezoelectric effect and considered to be suitable for
bone tissue engineering. The resulting composites promoted
the proliferation of human foreskin fibroblasts and mesenchy-
mal stem cells (MSCs), and also the differentiation of MSCs
towards osteoblasts. In addition, these composites did not
evoke any inflammatory reaction after subcutaneous implan-
tation in C57Bl/6 mice in vivo, as indicated by N-acetyl-β-D-gluco-
saminidase and NO assays performed in blood plasma.122
Other important zeolite-containing biomaterials are zeolite-
A/chitosan hybrid composites with zeolite contents of
20–55 wt%, prepared by in situ transformation of silica/chito-
san mixtures in a sodium aluminate alkaline solution through
impregnation–gelation–hydrothermal synthesis. These compo-
sites were prepared in various shapes, including cylinders,
plates and thin films. They contained macropores ranging in
size from 100 μm to 300 μm, and showed suitable mechanical
This journal is © The Royal Society of Chemistry 2018
Biomaterials Science
properties for bone tissue engineering. In addition, when
immersed in simulated body fluid (SBF), they acted as bio-
active, i.e. they promoted the formation of hydroxyapatite
(HAp) by their ion exchanging properties, and, when enriched
with Ag+, they showed antimicrobial activity.123
Similar antimicrobial and HAp-forming activity in SBF was
also observed in Ag ion-exchanged nanocrystalline zeolite
ZSM-5 (Ag-Nano-ZSM-5) and in Ag ion-exchanged conventional
zeolite (Ag-ZSM-5). Biocompatibility assays suggested that Ag-
Nano-ZSM-5 and HAp grown in Ag-Nano-ZSM-5 did not
impose any toxicity on mouse RAW 264.7 macrophage-like
cells and human colon adenocarcinoma Caco-2 cells. This
suggested that these materials have considerable potential for
biomedical applications, e.g. for constructing bone implants.64
In another study, ZSM-5 zeolite monocrystals with a disk-like
shape (∼350 nm in diameter and ∼165 nm in thickness), pro-
vided good support for the spread and proliferation of human
bone marrow stromal cells. In addition, in comparison with
conventional HAp particles, ZSM-5 zeolites possessed greater
drug-loading efficiency and a greater drug-sustained release
property because of the ordered micropores, larger Brunauer–
Emmett–Teller (BET) surface areas, and a higher content of
chemical functional groups. For gentamicin-loaded ZSM-5 zeo-
lites, the sustained release of gentamicin significantly reduced
the bacterial adhesion and prevented biofilm formation.63
Zeolites also enhanced the growth and osteogenic differen-
tiation of osteoblasts, when added directly to cell culture
media. Specifically, synthetic Na-A zeolite added into a cell
culture medium enhanced the autocrine synthesis of trans-
forming growth factor-β (TGF-β), DNA synthesis, the activity of
alkaline phosphatase and the production of osteocalcin in
normal human osteoblast-like hOB cells.124 Zeolite A also
inhibited osteoclast-mediated bone resorption in vitro.67
In addition, zeolites A and ZSM-5 were applied as coatings
on stainless steel and zeolite ZSM-5 was applied as a coating
on an aluminum alloy, where they decreased the colonization
of the material surface with Halomonas pacifica g, a marine
bacterium, in comparison with bare uncoated surfaces. This
effect was explained by the hydrophobicity of the zeolite coat-
ings and their electrostatic interactions with the bacteria.66
Silicalite-1, i.e. a siliceous analog of ZSM-5 zeolite, can form
a compact film (SF) which is highly corrosion-resistant.33–35
For example, the SF synthesized on Ti-6Al-4 V alloy prevented
the release of cytotoxic Al and V ions, even after exposure to
highly corrosive solutions (in 0.856 M NaCl at pHs of 7.0 and
1.0). Thus, the anti-corrosive properties of dense SF eliminate
a substantial drawback of metal supports in their application
as a prosthetic material, i.e. the evolution of toxic ions when
they interact with tissue.34,35 Another beneficial property of SF
is in this extent its mechanical properties, i.e. the elastic
modulus value (30–40 GPa), which is close to that of bone
(∼30 GPa).35 This reduces the resorption of implant materials
covered by SF. At the same time, SF significantly increased the
adhesion and proliferation of pluripotent mouse embryonic
cells in vitro, in comparison with a glass surface. This was
attributed to the three-dimensional structure of the zeolite
This journal is © The Royal Society of Chemistry 2018
Review
coating.34,35,125,126 Moreover, the SF has been shown to be
osteoconductive, i.e. to promote the proliferation of human
fetal osteoblasts (hFOBs), in comparison with the bare titanium
alloy, Ti6Al4V. In addition, SF promoted the differentiation of
hFOBs into mature osteoblasts, which was manifested by the
expression of the BMP-2 gene, and by the production of a
mineralized matrix at earlier times in culture, indicating higher
osteoinductive properties of the SF than of Ti6Al4V alone.34,35
Similarly, a SF on Ti-6Al-4V alloy stimulated osteogenic differen-
tiation of mouse MC3T3-E1 preosteoblasts, as demonstrated by
the increased expression and activity of alkaline phosphatase,
i.e. an enzyme participating in bone matrix mineralization.
Moreover, this coating significantly enhanced osteointegration
and bone regeneration after the implantation of Ti-6Al-4V
samples into rabbit femoral condylar defects.127
In our own studies, the a- and b-oriented SFs prepared on
Si(100) substrates or on stainless steel substrates supported
the adhesion, subsequent growth, viability and osteogenic
differentiation of human osteoblast-like Saos-2 cells.125,126
These cells adhered in higher numbers and proliferated to
higher population densities than the cells on uncoated sub-
strates, on microscopic glass coverslips and on standard cell
culture polystyrene dishes. This was more apparent for SF with
a more developed three-dimensional structure of the outer
surface, i.e. for SF deposited on AISI-316L stainless steel sub-
strates (Fig. 4). However, on all tested materials, the Saos-2
cells formed well-developed vinculin-containing focal
adhesion plaques and an F-actin cytoskeleton (Fig. 5). In
addition, SFs increased the activity of alkaline phosphatase, a
marker of osteogenic cell differentiation.
Despite the hardness of zeolite crystals, zeolites also have
potential for soft tissue engineering and regeneration, namely
vascular and skin tissue engineering and wound healing.
Fluorinated zeolite Y crystals were embedded in 3D scaffolds
made of polyurethane, i.e. a polymer suitable for the construc-
tion of vascular replacements. These scaffolds significantly
increased the proliferation activity of human coronary artery
smooth muscle cells and the depth of their penetration into
the scaffolds, in comparison with the control zeolite-free
scaffolds. The reason was that the zeolite particles acted as a
reservoir of oxygen, and enhanced the oxygen delivery to the
cells.128 Zeolite-Y powder functionalized with ferric iron-di-
ethyldithiocarbamate complexes is able to trap NO radicals in
liquids and in biological systems.129 It can therefore be uti-
lized as a radical scavenger, as a component of biosensors,
and also as a reservoir of NO, i.e. an important vasodilator, in
artificial vascular prostheses. In addition, NO released from
Co-ion exchanged zeolite-A inhibited platelet aggregation and
adhesion of human platelets in vitro.130 Platelet aggregation
and platelet adhesion are serious complications when vascular
replacements are used, and can lead to stenosis and oblitera-
tion of the lumen of the prosthesis. However, long-term
contact with blood undoubtedly led to leaching of highly toxic
Co ions from zeolite A. A further investigation is required.
In skin tissue engineering and wound healing, hybrid
scaffolds containing pectin, gelatin or gelatin/hyaluronan
Biomater. Sci., 2018, 6, 974–989 | 981
Review
Fig. 4 Number of human osteoblast-like Saos-2 cells on days 1 and 3
after seeding on SFs deposited on Si(100) and stainless steel (SS) sub-
strates, on bare Si(100) and stainless steel (SS), and on control micro-
scopic glass coverslips (Glass) and polystyrene dishes (PS). Mean ± S.E.
M. from 48–64 measurements for each experimental group and time
interval. ANOVA, Student–Newman–Keuls Method. Statistical signifi-
cance ( p ≤ 0.05) of the differences in comparison with other groups is
indicated by the labels of these groups above the columns. All: signifi-
cantly different in comparison with all other groups.
Fig. 5 Immunofluorescence staining of vinculin, a protein of focal adhesion plaques (green) in human osteoblast-like Saos-2 cells on day 3 after
seeding on SFs deposited on Si(100) (A), on SFs deposited on stainless steel (B), on bare Si(100) (C), on bare stainless steel (D) and on control micro-
scopic glass coverslips (E). The actin cytoskeleton is stained in red with phalloidin conjugated with tetramethylrhodamine (TRITC), and the cell nuclei
are stained in blue with 4’,6-diamidine-2’-phenylindole dihydrochloride (DAPI). Leica TCS SPE DH 2500 confocal microscope. Scale bar = 50 μm.
982 | Biomater. Sci., 2018, 6, 974–989
Biomaterials Science
matrix with copper-activated particles of faujasite proved to be
promising materials. These scaffolds had a highly porous
architecture, which supported the ingrowth and the viability of
NIH3T3 fibroblasts in vitro, and improved skin regeneration in
rats in vivo. These beneficial effects were also mediated by
increased oxygen supply to the cells, because faujasite par-
ticles acted as oxygen reservoirs. At the same time, the
scaffolds acted as antimicrobial substances, due to copper that
was released from the faujasite particles.41–43 For antimicrobial
effects in wound healing applications, zeolites can also be
loaded with NO. An ointment containing NO-loaded, zinc-
exchanged zeolite A showed considerable bactericidal activity
in relatively low concentrations, which supported the viability
of 3T3 fibroblasts.65 Another suitable component of dressings
for a full thickness experimental cutaneous wound was clinop-
tilolite. The wound healing rate and the healing quality after
the use of these dressings were even better than in commer-
cially available and clinically-used Biobran® biosynthetic
wound dressings.131 Zeolites can also be used for skin whiten-
ing. For example, zeolite 4A inhibited melanin synthesis
induced by alpha-melanocyte-stimulating hormone in mela-
noma B16F10 cells.132
11. Cytoxicity of zeolites
Despite all the favorable effects of zeolites described above,
some zeolites can act as highly cytotoxic and carcinogenic. The
most important of these is erionite, a naturally occurring
fibrous mineral (e.g. in the Cappadocian region of Turkey, in
central Mexico, in the USA and in Northern Italy). Erionite
This journal is © The Royal Society of Chemistry 2018
Biomaterials Science
forms brittle, wool-like fibrous masses in the hollows of rock
formations, and thus has similar properties to asbestos (Fig. 6),
causing lung cancer and malignant mesothelioma.46–48 The
mechanism of the cytotoxicity of erionite was further investi-
gated in experiments in vitro, where human monocyte U937
cells internalized erionite fibers, suspended in the cell culture
medium, through their cytoplasmic membrane. These fibers
were then found in the cytosol and at the nucleus. In a period
of 24 hours, first erionite fibers rich in sodium and potassium,
and then calcium-rich erionite fibers, induced cell necrosis.133
Other fibrous zeolites, such as offretite and scolecite, also acted
as cytotoxic. They perturbed the cell structure and induced for-
mation of swollen mitochondria and squared cells.133
Other adverse effects of zeolites were described after dietary
supplementation of calves with sodium zeolite A (Na-A). This
zeolite disturbed the mineral metabolism and the tissue
mineral composition of the animals. Retention of Al was
observed in all tissues, and there were increased concen-
trations of Si in the aorta, spleen, lung, muscle, and kidney of
the calves exposed to the Na-A, and Al was increased in all Na-
A-exposed tissues. The concentrations of elements physiologi-
cally present in the organism were also altered by Na-A. There
were increased calcium concentrations in aorta, liver, muscle,
and tendon; there were increased phosphorus concentrations
in aorta, but decreased concentrations in plasma; there were
increased magnesium concentrations in aorta, heart, kidney,
liver, and pancreas, but decreased concentrations in plasma;
and there were decreased iron concentrations in kidney and
liver.136
Even clinoptilolite, a natural zeolite considered as non-toxic
and with great potential for various biomedical applications,
reduced cell viability and DNA synthesis and increased cell
apoptosis when added in the form of nanoparticles into a cell
culture medium. These effects were more apparent in tumor
cell lines than in diploid fibroblasts. In tumor cell lines, clin-
optilolite reduced the activity of epidermal growth factor (EGF)
receptors, and also the activity of PKB/Akt and NFκB, i.e. key
proteins involved in cell survival and apoptosis. These effects
were attributed to the adsorptive and ion-exchange properties
of clinoptilolite, which can clear some components of the
serum supplement of the cell culture medium, e.g. EGF, and
Fig. 6 Morphology and chemical structure of erionite. A. Erionite-K,
formula (K2,Ca,Na2)2[Al4Si14O36]·15H2O. North railway cut, Malpais Hill,
Dudleyville, Pinal Co., Arizona, USA.134 B. The silicate framework of erio-
nite consists of alternating single rings (blue) and double rings ( pink) of
tetrahedra. Running down the holes formed by the rings are potassium
(yellow) and calcium (green) atoms.135
This journal is © The Royal Society of Chemistry 2018
Review
can affect Ca levels and Ca-dependent signalling pathways.137 In
addition, finely-ground clinoptilolite inhibited protein kinase B
(c-Akt), induced the expression of p21WAF1/CIP1 and p27KIP1
tumor suppressor proteins, and blocked the growth in several
cancer cell lines. It can therefore be utilized as an adjuvant in
anticancer therapy.138 In addition, excessive supplementation of
food for poultry with clinoptilolite can lead to an inflammatory
response in the intestine and damage to the intestinal barrier.
This inflammatory reaction is mediated by the increase in CD4-
positive and CD25-positive T and B lymphocytes and by higher
serum concentrations of IL-2 and IL-10.139
As was mentioned above, the as-synthesized SFs, deposited
on stainless steel, supported the adhesion, proliferation and
osteogenic differentiation of human osteoblast-like Saos-2
cells. However, DNA damage in human osteoblast-like U-2 OS
cells cultured on their surface calcined to 300 °C and 500 °C
was observed.140,141 U-2 OS cells are considered as a suitable
model for studies of markers of DNA damage response (DDR),
because, unlike Saos-2 and MG-63 human osteoblast-like cell
lines, U-2 OS cells retain p53 (for a review, see ref. 140 and
141). An important marker of the DDR is phosphorylated
histone H2AX (gamma H2AX), an indicator of breaks in DNA
molecules, which was present in cells on heat-treated SFs
(Fig. 7). Moreover, as revealed by fluorescence staining, the
cells on SFs treated at 500 °C were vacuolized, with a disturbed
F-actin cytoskeleton and with less-developed focal adhesion
plaques than on the control microscopic glass coverslips, non-
heat-treated SFs and SFs treated at 300 °C (Fig. 7). At the same
time, the initial adhesion and proliferation of Saos-2 cells was
significantly lower on SFs treated at 500 °C than on non-
treated SFs and on SFs treated at 300 °C, as manifested by
lower cell numbers on days 1 and 3 after cell seeding.125
Fig. 7 Left: DNA damage in human osteoblast-like U-2 OS cells, evalu-
ated by the percentage of cells positive for phosphorylated histone
gamma H2AX, measured by flow cytometry after 3-day cultivation on
control microscopic glass coverslips – Glass, on SFs deposited on stain-
less steel and stored at room temperature – SF(RT), and on SFs de-
posited on stainless steel and then treated at 300 °C – SF(300) or
500 °C – SF(500) ANOVA, Student–Newman–Keuls method, ***p ≤ 001
in comparison with Glass and SF(RT). Right: Morphology of human
osteoblast-like Saos-2 cells after 3-day cultivation on the materials.
Vinculin is stained in green using immunofluorescence, F-actin is
stained in red with phalloidin conjugated with TRITC, and the cell nuclei
are stained in blue with DAPI. Leica TCS SPE DH 2500 confocal
microscope.
Biomater. Sci., 2018, 6, 974–989 | 983
Review
The SFs in our studies were synthesized using tetrapropyl
ammonium hydroxide (TPAOH) as a template. According to
our recent results, the heat-treated SF at 500 °C is covered by
products of TPAOH degradation, which are partially volatile
polyaromatic condensed hydrocarbons (PAHs), including
naphthalene, phenanthrene, anthracene, pyrene and fluor-
anthene in micromole concentrations. Toxicity of PAHs is well-
documented.142 Some aromatic species appear on the outer
surface of SFs already at 300 °C.143 Complete elimination of
carbonaceous residues is difficult, even at high temperatures
and with prolonged time of treatment.143
12. Conclusion
Zeolites are natural or synthetic materials which have proved
to be effective in a broad spectrum of applications, including
applications in various biotechnologies and medicine. These
applications are related to the adsorption and ion-exchanging
properties of zeolites, and include decontamination of the
living environment from harmful elements and molecules,
including radioactive contaminants, detoxication of organ-
isms, improvements to the nutrition status and the immunity
of animals, separation of biomolecules and various cell types,
construction of biosensors and detection of biomarkers, deliv-
ery of drugs, nucleic acids and other bioactive molecules, and
scavenging of oxygen and other radicals. Zeolites have been
clinically used as components of potent haemostatics and
gastroprotective drugs. They are also promising as components
of scaffolds for bone, vascular and skin tissue engineering, and
for anticorrosive and bioactive coatings (e.g., silicalite-1 compact
films, SFs) for bone implants. Our studies have shown that SFs
promoted the adhesion, growth and osteogenic differentiation
of human bone-derived cells. However, after thermal treatment,
these films can cause DNA damage. Preparation of calcined SFs
with an open microporous structure and a non-toxic outer
surface is a challenging problem. Some other zeolites, particu-
larly erionite, can also act as cytotoxic and carcinogenic. Even
clinoptilolite, one of the most promising zeolites for biomedical
applications, can act as a pro-inflammatory agent and, in the
form of free nanoparticles, it can reduce cell viability and DNA
synthesis. Certain “two-faced” features of zeolites need to be
taken into account when they are used in biotechnology and in
medicine, although the antiproliferative and pro-apoptotic
effects of zeolites can be utilized in tumor therapy.
Conflicts of interest
The authors report no conflicts of interest in this work.
Acknowledgements
This study was supported by the Grant Agency of the Czech
Republic (Grant No. 16-02681S) and by the Ministry of
Education, Youth and Sports of the Czech Republic within the
984 | Biomater. Sci., 2018, 6, 974–989
Biomaterials Science
project No. LM2015062 (Czech-BioImaging). Mr Robin Healey
(Czech Technical University in Prague) is gratefully acknowl-
edged for his language revision of the manuscript.
References
1 R. M. Barrer, Zeolites and clay minerals as sorbents and
molecular sieve, Academic Press, London, New York, 1978.
2 R. M. Barrer, Zeolite Synthesis: An Overview, in Surface
Organometallic Chemistry: Molecular Approaches to Surface
Catalysis, eds. J. M. Basset et al. NATO ASI Series (Series C:
Mathematical and Physical Sciences), Springer,
Dordrecht, 1988, vol. 231, pp. 221–244.
3 N. Eroglu, M. Emekci and C. G. Athanassiou, Applications
of natural zeolites on agriculture and food production,
J. Sci. Food Agric., 2017, 97, 3487–3499.
4 International Zeolite Association, Database of Zeolite
Structures, News from the Structure Commission, http://
www.iza-structure.org/, (accessed January 2018); Zeolite
Framework, http://asia.iza-structure.org/IZA-SC/ftc_table.
php, (accessed January 2018).
5 A. F. Crønsted, Introduction to Zeolite Science and Practice, ed.
H. V. Bekkum, E. M. Flanigen, M. Jacobs and J. C. Jansen,
Akad. Handl. Stockholm, Elsevier Science, Amsterdam, 2001.
6 Clinoptilolite, https://commons.wikimedia.org/wiki/File:
D09AD0BBD0B8D0BDD0BED0BFD182D0B8D0BBD0BED0-
BBD0B8D182.jpg, (accessed March 2018).
7 Geotimes, Mineral resource of the month: natural and
synthetic zeolites, http://www.geotimes.org/june08/article.
html?id=nn_zeolites.html, (accessed January 2018).
8 W. Loewenstein, The distribution of aluminium in the tetra-
hedral of silicates and aluminates, Am. Mineral., 1954, 39, 92.
9 Porous materials, Research – Zeolites, http://www.uni-
muenster.de/Chemie.pc/Koller/zeolites.html, (accessed
January 2018).
10 C. Laurino and B. Palmieri, Zeolite: “The magic stone”;
Main nutritional, environmental, experimental and clini-
cal fields of application, Nutr. Hosp., 2015, 32, 573.
11 C. Perego and A. Carati, Zeolites: From model materials to
industrial catalysts, ed. J. J. Cejka, J. Paréz-Pariente and
W. J. Roth, 2008, 357–389.
12 F. J. Chen, Y. Xu and H. B. Du, An extra-large-pore zeolite
with intersecting 18-, 12-, and 10-membered ring chan-
nels, Angew. Chem., Int. Ed., 2014, 53, 9592.
13 Z. Song, Y. Huang, W. L. Xu, L. Wang, Y. Bao, S. Li and
M. Yu, Continuously adjustable, molecular-sieving “gate”
on 5A zeolite for distinguishing small organic molecules
by size, Sci. Rep., 2015, 5, 13981.
14 Q. Tao, M. Hu, X. Ma, M. Xiang, T. C. Zhang, C. Li, J. Yao
and Y. Liang, Simultaneous removal of ammonium and
nitrate by HDTMA-modified zeolite, Water Sci. Technol.,
2015, 72, 1931.
15 G. Huang, F. Liu, Y. Yang, W. Deng, S. Li, Y. Huang and
X. Kong, Removal of ammonium-nitrogen from ground-
water using a fully passive permeable reactive barrier with
This journal is © The Royal Society of Chemistry 2018
Biomaterials Science
oxygen-releasing compound and clinoptilolite, J. Environ.
Manage., 2015, 154, 1.
16 B. Verbinnen, C. Block, D. Hannes, P. Lievens,
M. Vaclavikova, K. Stefusova, G. Gallios and
C. Vandecasteele, Removal of molybdate anions from
water by adsorption on zeolite-supported magnetite,
Water Environ. Res., 2012, 84, 753.
17 Y. Yu, J. G. Shapter, R. Popelka-Filcoff, J. W. Bennett and
A. V. Ellis, Copper removal using bio-inspired polydopa-
mine coated natural zeolites, J. Hazard. Mater., 2014, 273,
174.
18 M. Jovanovic, N. Rajic and B. Obradovic, Novel kinetic
model of the removal of divalent heavy metal ions from
aqueous solutions by natural clinoptilolite, J. Hazard.
Mater., 2012, 233–234, 57.
19 M. Anari-Anaraki and A. Nezamzadeh-Ejhieh,
Modification of an Iranian clinoptilolite nano-particles by
hexadecyltrimethyl ammonium cationic surfactant and
dithizone for removal of Pb(II) from aqueous solution,
J. Colloid Interface Sci., 2015, 440, 272.
20 A. C. Batista, E. R. Villanueva, R. V. Amorim, M. T. Tavares
and G. M. Campos-Takaki, Chromium(VI) ion adsorption
features of chitosan film and its chitosan/zeolite conju-
gate 13X film, Molecules, 2011, 16, 3569.
21 C. Valenzuela, R. Moraga, C. Leon, C. T. Smith,
M. A. Mondaca and V. L. Campos, Arsenite oxidation by
pseudomonas arsenicoxydans immobilized on zeolite and
its potential biotechnological application, Bull. Environ.
Contam. Toxicol., 2015, 94, 667.
22 A. Molla, Z. Ioannou, S. Mollas, E. Skoufogianni and
A. Dimirkou, Removal of chromium from soils cultivated
with maize (Zea mays) after the addition of natural min-
erals as soil amendments, Bull. Environ. Contam. Toxicol.,
2017, 98, 347.
23 R. Rostami and A. Jonidi Jafari, Application of an
adsorptive-thermocatalytic process for BTX removal
from polluted air flow, J. Environ. Health Sci. Eng., 2014,
12, 89.
24 A. V. Voronina, M. O. Blinova, V. S. Semenishchev and
D. K. Gupta, Returning land contaminated as a result of
radiation accidents to farming use, J. Environ. Radioact.,
2015, 144, 103.
25 D. I. Tchernev, Natural zeolites in solar energy heating,
cooling, and energy storage, Rev. Mineral. Geochem., 2001,
45, 589.
26 R. Z. Wang and R. G. Oliveira, Adsorption refrigeration –
an efficient way to make good use of waste heat and solar
energy, International Sorption Heat Pump Conference, June
22-24, Denver, CO, USA, 2005, pp. 1–22.
27 C. Fruijtier-Pölloth, The safety of synthetic zeolites used
in detergents, Arch. Toxicol., 2009, 83, 23.
28 S. J. Reitmeier(09.07.2009), Investigation of hydrocarbon
transport on surface modified zeolites, https://www.elite-
netzwerk.bayern.de/elitenetzwerk-home/forschungsarbeiten/
naturwissenschaften/2009/reitmeier-zeolites/, (accessed
January 2018).
This journal is © The Royal Society of Chemistry 2018
Review
29 E. V. Rebrov, Sol-gel synthesis of zeolite coatings and their
application in catalytic microstructured reactors, Catal.
Ind., 2009, 1, 322.
30 J. Caro and M. Noack, Zeolite membranes – Recent devel-
opments and progress, Microporous Mesoporous Mater.,
2008, 115, 215.
31 C. M. Lew, R. Cai and Y. Yan, Zeolite thin films: from com-
puter chips to space stations. Zeolite thin films: from
computer chips to space stations, Acc. Chem. Res., 2010,
43, 210.
32 D. E. Anderson, S. Balapangu, H. N. A. Fleischer,
R. A. Viade, F. D. Krampa, P. Kanyong, G. A. Awandare and
E. K. Tiburu, Investigating the influence of temperature
on the kaolinite-base synthesis of zeolite and urease
immobilization for the potential fabrication of electro-
chemical urea biosensors, Sensors, 2017, 17, 1831.
33 G. Chaw, R. S. Bedi, Y. Yan and J. Wang, Zeolites as wear
resistant coatings, Microporous Mesoporous Mater., 2012,
151, 346.
34 R. S. Bedi, L. P. Zanello and Y. Yan, Osteoconductive and
osteoinductive properties of zeolite MFI coatings on tita-
nium alloys, Adv. Funct. Mater., 2009, 19, 3856.
35 R. S. Bedi, D. E. Beving, L. P. Zanello and Y. Yan,
Biocompatibility of corrosion-resistant zeolite coatings for
titanium alloy biomedical implants, Acta Biomater., 2009,
5, 3265.
36 G. Dogliotti, A. E. Malavazos, S. Giacometti, U. Solimene,
M. Fanelli, M. M. Corsi and E. Dozio, Natural zeolites cha-
bazite/phillipsite/analcime increase blood levels of anti-
oxidant enzymes, J. Clin. Biochem. Nutr., 2012, 50, 195.
37 J. El-Gindi, K. Benson, L. De Cola, H. J. Galla and
N. S. Kehr, Cell adhesion behavior on enantiomerically
functionalized zeolite L monolayers, Angew. Chem., Int.
Ed., 2012, 51, 3716.
38 A. Greco, L. Maggini, L. De Cola, R. De Marco and
L. Gentilucci, Diagnostic implementation of fast and
selective integrin-mediated adhesion of cancer cells on
functionalized zeolite L monolayers, Bioconjugate Chem.,
2015, 26, 1873.
39 A. Bertucci, H. Lülf, D. Septiadi, D. Manicardi,
R. Corradini and L. De Cola, Intracellular delivery of
peptide nucleic acid and organic molecules using zeolite-
L nanocrystals, Adv. Healthcare Mater., 2014, 3, 1812.
40 N. Vilaça, R. Amorim, A. F. Machado, P. Parpot,
M. F. Pereira, M. Sardo, J. Rocha, A. M. Fonseca, I. C. Neves
and F. Baltazar, Potentiation of 5-fluorouracil encapsulated
in zeolites as drug delivery systems for in vitro models of
colorectal carcinoma, Colloids Surf., B, 2013, 112, 237.
41 N. Ninan, M. Muthiah, I. K. Park, A. Elain, T. W. Wong,
S. Thomas and Y. Grohens, Faujasites incorporated tissue
engineering scaffolds for wound healing: in vitro and in
vivo analysis, ACS Appl. Mater. Interfaces, 2013, 5, 11194.
42 N. Ninan, M. Muthiah, N. A. Bt Yahaya, I. K. Park,
E. Elain, T. W. Wong, S. Thomas and Y. Grohens,
Antibacterial and wound healing analysis of gelatin/
zeolite scaffolds, Colloids Surf., B, 2014, 115, 244.
Biomater. Sci., 2018, 6, 974–989 | 985
Review
43 N. Ninan, M. Muthiah, I. K. Park, A. Elain, T. W. Wong,
S. Thomas and Y. Grohens, In vitro and in vivo evaluation
of pectin/copper exchanged faujasite composite mem-
branes, J. Biomed. Nanotechnol., 2015, 11, 1550.
44 L. I. Andersson and H. Eriksson, De-aluminated zeolite Y
as a tool to study endocytosis, a delivery system revealing
differences between human peripheral dendritic cells,
Scand. J. Immunol., 2007, 66, 52.
45 P. Pellegrino, B. Mallet, S. Delliaux, Y. Jammes, R. Guieu
and O. Schäf, Zeolites are effective ROS-scavengers in
vitro, Biochem. Biophys. Res. Commun., 2011, 410, 478.
46 M. A. Ortega-Guerrero and G. Carrasco-Núñez,
Environmental occurrence, origin, physical and geochem-
ical properties, and carcinogenic potential of erionite near
San Miguel de Allende, Mexico, Environ. Geochem. Health,
2014, 36, 517.
47 M. A. Ortega-Guerrero, G. Carrasco-Núñez, H. Barragán-
Campos and M. R. Ortega, High incidence of lung cancer
and malignant mesothelioma linked to erionite fibre
exposure in a rural community in Central Mexico, Occup.
Environ. Med., 2015, 72, 216.
48 M. Giordani, M. Mattioli, P. Ballirano, A. Pacella,
M. Cenni, M. Boscardin and L. Valentini, Geological
occurrence, mineralogical characterization, and risk
assessment of potentially carcinogenic erionite in Italy,
J. Toxicol. Environ. Health, Part B, 2017, 20, 81.
49 M. Beltcheva, R. Metcheva, N. Popov, S. E. Teodorova,
J. A. Heredia-Rojas, A. O. Rodríguez-de la Fuente,
L. E. Rodríguez-Flores and M. Topashka-Ancheva, Modified
natural clinoptilolite detoxifies small mammal’s organism
loaded with lead I. Lead disposition and kinetic model for
lead bioaccumulation, Biol. Trace Elem. Res., 2012, 147, 180.
50 M. Topashka-Ancheva, M. Beltcheva, R. Metcheva,
J. A. Rojas, A. O. Rodriguez-De la Fuente, T. Gerasimova,
L. E. Rodríguez-Flores and S. E. Teodorova, Modified
natural clinoptilolite detoxifies small mammal’s organism
loaded with lead II: genetic, cell, and physiological effects,
Biol. Trace Elem. Res., 2012, 147, 206.
51 M. Delavarian, A. Hassanvand and S. Gharibzadeh,
Increasing performance in children with ADHD by trap-
ping lead with a nano-zeolite, J. Neuropsychiatry Clin.
Neurosci., 2013, 25, E23.
52 Q. J. Wu, L. C. Wang, Y. M. Zhou, J. F. Zhang and
T. Wang, Effects of clinoptilolite and modified clinoptilo-
lite on the growth performance, intestinal microflora, and
gut parameters of broilers, Poult. Sci., 2013, 92, 684.
53 Q. J. Wu, Y. M. Zhou, Y. N. Wu, L. L. Zhang and T. Wang,
The effects of natural and modified clinoptilolite on intes-
tinal barrier function and immune response to LPS in
broiler chickens, Vet. Immunol. Immunopathol., 2013, 153,
70.
54 Y. Wu, Q. Wu, Y. Zhou, H. Ahmad and T. Wang, Effects of
clinoptilolite on growth performance and antioxidant
status in broilers, Biol. Trace Elem. Res., 2013, 155, 228.
55 W. Potgieter, C. S. Samuels and J. R. Snyman, Potentiated
clinoptilolite: artificially enhanced aluminosilicate
986 | Biomater. Sci., 2018, 6, 974–989
Biomaterials Science
reduces symptoms associated with endoscopically nega-
tive gastroesophageal reflux disease and nonsteroidal
anti-inflammatory drug induced gastritis, Clin. Exp.
Gastroenterol., 2014, 7, 215.
56 B. de Gennaro, L. Catalanotti, P. Cappelletti, A. Langella,
M. Mercurio, C. Serri, M. Biondi and L. Mayol, Surface
modified natural zeolite as a carrier for sustained diclofe-
nac release: A preliminary feasibility study, Colloids Surf.,
B, 2015, 130, 101.
57 M. Khatamian, B. Divband and F. Farahmand-Zahed,
Synthesis and characterization of zinc(II)-loaded zeolite/
graphene oxide nanocomposite as a new drug carrier,
Mater. Sci. Eng., C, 2016, 66, 251.
58 O. O. Soldatkin, M. K. Shelyakina, V. N. Arkhypova, E. Soy,
S. K. Kirdeciler, B. Ozansoy Kasap, F. Lagarde, N. Jaffrezic-
Renault, B. Akata Kurç, A. P. Soldatkin and
S. V. Dzyadevych, Nano- and microsized zeolites as a per-
spective material for potentiometric biosensors creation,
Nanoscale Res. Lett., 2015, 10, 59.
59 M. Hasheminejad and A. Nezamzadeh-Ejhieh, A novel
citrate selective electrode based on surfactant modified
nano-clinoptilolite, Food Chem., 2015, 172, 794.
60 K. Saribeyoglu, E. Aytac, S. Pekmezci, S. Saygili, H. Uzun,
G. Ozbay, S. Aydin and H. O. Seymen, Effects of clinoptilo-
lite treatment on oxidative stress after partial hepatectomy
in rats, Asian J. Surg., 2011, 34, 153.
61 H. Yapislar, E. Taskin, S. Ozdas, D. Akin and E. Sonmez,
Counteraction of apoptotic and inflammatory effects of
adriamycin in the liver cell culture by clinopitolite, Biol.
Trace Elem. Res., 2016, 170, 373.
62 K. Pavelić, M. Hadzija, L. Bedrica, J. Pavelić, I. Dikić,
M. Katić, M. Kralj, M. H. Bosnar, S. Kapitanović,
M. Poljak-Blazi, S. Krizanac, R. Stojković, M. Jurin,
B. Subotić and M. Colić, Natural zeolite clinoptilolite: new
adjuvant in anticancer therapy, J. Mol. Med., 2001, 78,
708.
63 Y. P. Guo, T. Long, Z. F. Song and Z. A. Zhu, Hydrothermal
fabrication of ZSM-5 zeolites: biocompatibility, drug deliv-
ery property, and bactericidal property, J. Biomed. Mater.
Res., Part B, 2014, 102, 583.
64 B. Kaur, R. Srivastava, B. Satpati, K. K. Kondepudi and
M. Bishnoi, Biomineralization of hydroxyapatite in silver
ion-exchanged nanocrystalline ZSM-5 zeolite using simu-
lated body fluid, Colloids Surf., B, 2015, 135, 201.
65 M. Neidrauer, U. K. Ercan, A. Bhattacharyya, J. Samuels,
J. Sedlak, R. Trikha, K. A. Barbee, M. S. Weingarten and
Z. M. Joshi, Antimicrobial efficacy and wound-healing
property of a topical ointment containing nitric-oxide-
loaded zeolites, J. Med. Microbiol., 2014, 63, 203.
66 G. Chen, D. E. Beving, R. S. Bedi, Y. S. Yan and
S. L. Walker, Initial bacterial deposition on bare and
zeolite-coated aluminum alloy and stainless steel,
Langmuir, 2009, 25, 1620.
67 N. Schütze, M. J. Oursler, J. Nolan, B. L. Riggs and
T. C. Spelsberg, Zeolite A inhibits osteoclast-mediated
bone resorption in vitro, J. Cell. Biochem., 1995, 58, 39.
This journal is © The Royal Society of Chemistry 2018
Biomaterials Science
68 S. Adamaref, W. An, M. O. Jarligo, T. Kuznicki and
S. M. Kuznicki, Natural clinoptilolite composite mem-
branes on tubular stainless steel supports for water soft-
ening, Water Sci. Technol., 2014, 70, 1412.
69 H. Yeritsyan, A. Sahakyan, V. Harutyunyan,
S. Nikoghosyan, E. Hakhverdyan, N. Grigoryan,
A. Hovhannisyan, V. Atoyan, Y. Keheyan and C. Rhodes,
Radiation-modified natural zeolites for cleaning liquid
nuclear waste (irradiation against radioactivity), Sci. Rep.,
2013, 3, 2900.
70 B. M. Mitrović, G. Vitorović, M. Vićentijević, D. Vitorović,
G. Pantelić and M. Lazarević-Macanović, Comparative
study of (137)Cs distribution in broilers and pheasants
and possibilities for protection, Radiat. Environ. Biophys.,
2012, 51, 79.
71 M. Delavarian, A. Hassanvand and S. Gharibzadeh, Using
natural zeolite as a transporter of dopamine,
J. Neuropsychiatry Clin. Neurosci., 2013, 25, E21.
72 A. Federico, M. Dallio, A. G. Gravina, C. Iannotta,
M. Romano, G. Rossetti, F. Somalvico, C. Tuccillo and
C. Loguercio, A pilot study on the ability of clinoptilolite
to absorb ethanol in vivo in healthy drinkers: effect of
gender, J. Physiol. Pharmacol., 2015, 66, 441.
73 M. V. Cattaneo and T. M. Chang, The potential of a micro-
encapsulated urease-zeolite oral sorbent for the removal
of urea in uremia, ASAIO Trans., 1991, 37, 80.
74 V. Wernert, O. Schaef, H. Ghobakhar and R. Denoyel,
Adsorption properties of zeolites for artificial kidney
applications, Microporous Mesoporous Mater., 2005, 83,
101.
75 D. Bergé-Lefranc, H. Pizzala, J. L. Paillaud, O. Schäf,
C. Vagner, P. Boulet, B. Kuchta and R. Denoyel,
Adsorption of small uremic toxin molecules on MFI type
zeolites from aqueous solution, Adsorption, 2008, 14, 377.
76 I. Fendri, L. Khannous, Z. Mallek, A. I. Traore,
N. Gharsallah and R. Gdoura, Influence of Zeolite on fatty
acid composition and egg quality in Tunisian Laying
Hens, Lipids Health Dis., 2012, 11, 71.
77 Z. Mallek, I. Fendri, L. Khannous, A. Ben Hassena,
A. I. Traore, M. A. Ayadi and R. Gdoura, Effect of zeolite
(clinoptilolite) as feed additive in Tunisian broilers on the
total flora, meat texture and the production of omega 3
polyunsaturated fatty acid, Lipids Health Dis., 2012, 11, 35.
78 L. Li, P. Li, Y. Chen, C. Wen, S. Zhuang and Y. Zhou, Zinc-
bearing zeolite clinoptilolite improves tissue zinc accumu-
lation in laying hens by enhancing zinc transporter gene
mRNA abundance, Anim. Sci. J., 2015, 86, 782.
79 Z. G. Tang, G. Y. Chen, L. F. Li, C. Wen, T. Wang and
Y. M. Zhou, Effect of zinc-bearing zeolite clinoptilolite on
growth performance, zinc accumulation, and gene
expression of zinc transporters in broilers, J. Anim. Sci.,
2015, 93, 620.
80 L. C. Wang, T. T. Zhang, C. Wen, Z. Y. Jiang, T. Wang and
Y. M. Zhou, Protective effects of zinc-bearing clinoptilolite
on broilers challenged with Salmonella pullorum, Poult.
Sci., 2012, 91, 1838.
This journal is © The Royal Society of Chemistry 2018
Review
81 P. D. Katsoulos, M. A. Karatzia, Z. Polizopoulou, P. Florou-
Paneri and H. Karatzias, Effects of prolonged consump-
tion of water with elevated nitrate levels on certain meta-
bolic parameters of dairy cattle and use of clinoptilolite
for their amelioration, Environ. Sci. Pollut. Res. Int., 2015,
22, 9119.
82 B. Kabak, A. D. Dobson and I. Var, Strategies to prevent
mycotoxin contamination of food and animal feed: a
review, Crit. Rev. Food Sci. Nutr., 2006, 46, 593.
83 J. Nedeljković-Trailović, S. Trailović, R. Resanović,
D. Milićević, M. Jovanovic and M. Vasiljevic, Comparative
investigation of the efficacy of three different adsorbents
against OTA-induced toxicity in broiler chickens, Toxins,
2015, 7, 1174.
84 H. Chiku, M. Matsui, S. Murakami, Y. Kiyozumi,
F. Mizukami and K. Sakaguchi, Zeolites as new chromato-
graphic carriers for proteins–easy recovery of proteins
adsorbed on zeolites by polyethylene glycol, Anal.
Biochem., 2003, 318, 80.
85 K. Sakaguchi, M. Matsui and F. Mizukami, Applications of
zeolite inorganic composites in biotechnology: current
state and perspectives, Appl. Microbiol. Biotechnol., 2005,
67, 306.
86 M. Becker, L. De Cola and A. Studer, Site-specific immo-
bilization of proteins at zeolite L crystals by nitroxide
exchange reactions, Chem. Commun., 2011, 47, 3392.
87 H. Chiku, A. Kawai, T. Ishibashi, M. Takehara, T. Yanai,
F. Mizukami and K. Sakaguchi, A novel protein refolding
method using a zeolite, Anal. Biochem., 2006, 348, 307.
88 T. Y. Nara, H. Togashi, C. Sekikawa, M. Kawakami,
N. Yaginuma, K. Sakaguchi, F. Mizukami and T. Tsunoda,
Use of zeolite to refold a disulfide-bonded protein,
Colloids Surf., B, 2009, 68, 68.
89 T. Y. Nara, H. Togashi, C. Sekikawa, K. Sakaguchi,
F. Mizukami and T. Tsunoda, High-throughput protein
refolding screening method using zeolite, Biotechnol.
Prog., 2009, 25, 1071.
90 M. Kubota, T. Nakabayashi, Y. Matsumoto, T. Shiomi,
Y. Yamada, K. Ino, H. Yamanokuchi, M. Matsui,
T. Tsunoda, F. Mizukami and K. Sakaguchi, Selective
adsorption of bacterial cells onto zeolites, Colloids Surf.,
B, 2008, 64, 88.
91 M. Tondar, M. J. Parsa, Y. Yousefpour, A. M. Sharifi and
S. V. Shetab-Boushehri, Feasibility of clinoptilolite appli-
cation as a microporous carrier for pH-controlled oral
delivery of aspirin, Acta Chim. Slov., 2014, 61, 688.
92 E. Khodaverdi, H. A. Soleimani, F. Mohammadpour and
F. Hadizadeh, Synthetic zeolites as controlled-release
delivery systems for anti-inflammatory drugs, Chem. Biol.
Drug Des., 2016, 87, 849.
93 C. Karavasili, E. P. Amanatiadou, E. Kontogiannidou,
G. K. Eleftheriadis, N. Bouropoulos, E. Pavlidou,
I. Kontopoulou, I. S. Vizirianakis and D. G. Fatouros,
Comparison of different zeolite framework types as car-
riers for the oral delivery of the poorly soluble drug indo-
methacin, Int. J. Pharm., 2017, 528, 76.
Biomater. Sci., 2018, 6, 974–989 | 987
Review
94 M. Spanakis, N. Bouropoulos, D. Theodoropoulos,
L. Sygellou, S. Ewart, A. M. Moschovi, A. Siokou, I. Niopas,
K. Kachrimanis, V. Nikolakis, P. A. Cox, I. S. Vizirianakis
and D. G. Fatouros, Controlled release of 5-fluorouracil
from microporous zeolites, Nanomedicine, 2014, 10, 197.
95 T. Sağir, M. Huysal, Z. Durmus, B. Z. Kurt, M. Senel and
S. Isık, Preparation and in vitro evaluation of 5-flouro-
uracil loaded magnetite-zeolite nanocomposite (5-FU-MZNC)
for cancer drug delivery applications, Biomed. Pharmacother.,
2016, 77, 182.
96 C. Adhikari, A. Das and A. Chakraborty, Zeolitic imidazole
framework (ZIF) nanospheres for easy encapsulation and
controlled release of an anticancer drug doxorubicin
under different external stimuli: A way toward smart drug
delivery system, Mol. Pharm., 2015, 12, 3158.
97 S. Grund, T. Doussineau, D. Fischer and G. J. Mohr,
Mitoxantrone-loaded zeolite beta nanoparticles: prepa-
ration, physico-chemical characterization and biological
evaluation, J. Colloid Interface Sci., 2012, 365, 33.
98 Y. Zhang, W. Yan, Z. Sun, C. Pan, X. Mi, G. Zhao and
J. Gao, Fabrication of porous zeolite/chitosan monoliths
and their applications for drug release and metal ions
adsorption, Carbohydr. Polym., 2015, 117, 657.
99 V. Mavrodinova, M. Popova, K. Yoncheva, J. Mihály and
A. Szegedi, Solid-state encapsulation of Ag and sulfadia-
zine on zeolite Y carrier, J. Colloid Interface Sci., 2015, 458,
32.
100 S. Demirci, Z. Ustaoğlu, G. A. Yılmazer, F. Sahin and
N. Baç, Antimicrobial properties of zeolite-X and zeolite-A
ion-exchanged with silver, copper, and zinc against a
broad range of microorganisms, Appl. Biochem.
Biotechnol., 2014, 172, 1652.
101 S. Fox, T. S. Wilkinson, P. S. Wheatley, B. Xiao,
R. E. Morris, A. Sutherland, A. J. Simpson, P. G. Barlow,
A. R. Butler, I. L. Megson and A. G. Rossi, NO-loaded Zn
(2+)-exchanged zeolite materials: a potential bifunctional
anti-bacterial strategy, Acta Biomater., 2010, 6, 1515.
102 H. Lülf, A. Bertucci, D. Septiadi, R. Corradini and L. De
Cola, Multifunctional inorganic nanocontainers for DNA
and drug delivery into living cells, Chemistry, 2014, 20,
10900.
103 M. E. Pearce, H. Q. Mai, N. Lee, S. C. Larsen and
A. K. Salem, Silicalite nanoparticles that promote trans-
gene expression, Nanotechnology, 2008, 19, 175103.
104 P. He, W. Wang, L. Du, F. Dong, Y. Deng and T. Zhang,
Zeolite A functionalized with copper nanoparticles and
graphene oxide for simultaneous electrochemical deter-
mination of dopamine and ascorbic acid, Anal. Chim.
Acta, 2012, 739, 25.
105 S. Sharifian and A. Nezamzadeh-Ejhieh, Modification of
carbon paste electrode with Fe(III)-clinoptilolite nano-par-
ticles for simultaneous voltammetric determination of
acetaminophen and ascorbic acid, Mater. Sci. Eng., C,
2016, 58, 510.
106 J. Zhou, Y. Luo, A. Zhu, Y. Liu, Z. Zhu and Y. Tian, A
reliable and durable approach for real-time determination
988 | Biomater. Sci., 2018, 6, 974–989
Biomaterials Science
of cellular superoxide anion based on biomimetic super-
oxide dismutase stabilized by a zeolite, Analyst, 2011, 136,
1594.
107 J. Cao, Y. Hu, C. Shen, J. Yao, L. Wei, F. Yang, A. Nie,
H. Wang, H. Shen, Y. Liu, Y. Zhang, Y. Tang and P. Yang,
Nanozeolite-driven approach for enrichment of secretory
proteins in human hepatocellular carcinoma cells,
Proteomics, 2009, 9, 4881.
108 N. Zarkovic, K. Zarkovic, M. Kralj, S. Borovic, S. Sabolovic,
M. P. Blazi, A. Cipak and K. Pavelic, Anticancer and anti-
oxidative effects of micronized zeolite clinoptilolite,
Anticancer Res., 2003, 23, 1589.
109 M. Montinaro, D. Uberti, G. Maccarinelli, S. A. Bonini,
G. Ferrari-Toninelli and M. Memo, Dietary zeolite sup-
plementation reduces oxidative damage and plaque gene-
ration in the brain of an Alzheimer’s disease mouse
model, Life Sci., 2013, 92, 903.
110 D. Mück-Šeler and N. Pivac, The effect of natural clinopti-
lolite on the serotonergic receptors in the brain of mice
with mammary carcinoma, Life Sci., 2003, 73, 2059.
111 B. Shen, J. C. Scaiano and A. M. English, Zeolite encapsu-
lation decreases TiO2-photosensitized ROS generation in
cultured human skin fibroblasts, Photochem. Photobiol.,
2006, 82, 5.
112 A. Atitlán-Gil, M. M. Bretón-de la Loza, J. C. Jiménez-
Ortega, H. Belefant-Miller and G. Betanzos-Cabrera,
Activated and micronized zeolite in the modulation of cel-
lular oxidative stress in Mexican smokers: a randomized
clinical trial, Rev. Invest. Clin., 2017, 69, 146.
113 M. Cutovic, M. Lazovic, V. Vukovic-Dejanovic, D. Nikolic,
I. Petronic-Markovic and D. Cirovic, Clinoptilolite for
treatment of dyslipidemia: preliminary efficacy study,
J. Altern. Complement. Med., 2017, 23, 738.
114 S. Pourshahrestani, E. Zeimaran, I. Djordjevic, N. A. Kadri
and M. R. Towler, Inorganic hemostats: The state-of-the-
art and recent advances, Mater. Sci. Eng., C, 2016, 58,
1255.
115 J. Li, W. Cao, X. X. Lv, L. Jiang, Y. J. Li, W. Z. Li, S. Z. Chen
and X. Y. Li, Zeolite-based hemostat QuikClot releases
calcium into blood and promotes blood coagulation in
vitro, Acta Pharmacol. Sin., 2013, 34, 367.
116 F. Arnaud, T. Tomori, W. Carr, A. McKeague, K. Teranishi,
K. Prusaczyk and R. McCarron, Exothermic reaction in
zeolite hemostatic dressings: QuikClot ACS and ACS+,
Ann. Biomed. Eng., 2008, 36, 1708.
117 S. Mortazavi, A. Tavasoli, M. Atefi, N. Tanide, N. Radpey,
P. Roshan-Shomal, H. Moradi and S. Taeb, CoolClot, a
novel hemostatic agent for controlling life-threatening
arterial bleeding, World J. Emerg. Med., 2013, 4, 123.
118 H. Khoshmohabat, B. Dalfardi, A. Dehghanian,
H. R. Rasouli, S. M. Mortazavi and S. Paydar, The effect of
CoolClot hemostatic agent on skin wound healing in rats,
J. Surg. Res., 2016, 200, 732.
119 K. R. Ward, M. H. Tiba, W. H. Holbert, C. R. Blocher,
G. T. Draucker, G. A. Proffitt, G. L. Bowlin, R. R. Ivatury
and R. F. Diegelmann, Comparison of a new hemostatic
This journal is © The Royal Society of Chemistry 2018
Biomaterials Science
agent to current combat hemostatic agents in a swine
model of lethal extremity arterial hemorrhage, J. Trauma,
2007, 63, 276.
120 J. W. Carraway, D. Kent, K. Young, A. Cole, R. Friedman
and K. R. Ward, Comparison of a new mineral based
hemostatic agent to a commercially available granular
zeolite agent for hemostasis in a swine model of lethal
extremity arterial hemorrhage, Resuscitation, 2008, 78, 230.
121 M. Alavi, A. Totonchi, M. A. Okhovat, M. Motazedian,
P. Rezaei and M. Atefi, The effect of a new impregnated
gauze containing bentonite and halloysite minerals on
blood coagulation and wound healing, Blood Coagulation
Fibrinolysis, 2014, 25, 856.
122 R. Costa, C. Ribeiro, A. C. Lopes, P. Martins, V. Sencadas,
R. Soares and S. Lanceros-Mendez, Osteoblast, fibroblast
and in vivo biological response to poly(vinylidene fluor-
ide) based composite materials, J. Mater. Sci. Mater. Med.,
2013, 24, 395.
123 L. Yu, J. Gong, C. Zeng and L. Zhang, Preparation of
zeolite-A/chitosan hybrid composites and their bioactiv-
ities and antimicrobial activities, Mater. Sci. Eng., C, 2013,
33, 3652.
124 P. E. Keeting, M. J. Oursler, K. E. Wiegand, S. K. Bonde,
T. C. Spelsberg and B. L. Riggs, Zeolite A increases pro-
liferation, differentiation, and transforming growth factor
beta production in normal adult human osteoblast-like
cells in vitro, J. Bone Miner. Res., 1992, 7, 1281.
125 I. Jirka, M. Vandrovcova, J. Plsek, M. Bousa, L. Brabec,
H. Dragounova and L. Bacakova, Interaction of human
osteoblast-like Saos-2 cells with stainless steel coated by
silicalite-1 films, Mater. Sci. Eng., C, 2017, 76, 775.
126 I. Jirka, M. Vandrovcova, J. Plsek, M. Bousa and
L. Bacakova, Interaction of silicalite-1 film with human
osteoblast-like Saos-2 cells: The role of micro-morphology,
Mater. Lett., 2017, 190, 229.
127 Y. Li, Y. Jiao, X. Li and Z. Guo, Improving the osteointegra-
tion of Ti6Al4 V by zeolite MFI coating, Biochem. Biophys.
Res. Commun., 2015, 460, 151.
128 D. G. Seifu, T. T. Isimjan and K. Mequanint, Tissue engin-
eering scaffolds containing embedded fluorinated-zeolite
oxygen vectors, Acta Biomater., 2011, 7, 3670.
129 E. van Faassen and A. Vanin, NO trapping in biological
systems with a functionalized zeolite network, Nitric
Oxide, 2006, 15, 233.
130 P. S. Wheatley, A. R. Butler, M. S. Crane, S. Fox, B. Xiao,
A. G. Rossi, I. L. Megson and R. E. Morris, NO-releasing
zeolites and their antithrombotic properties, J. Am. Chem.
Soc., 2006, 128, 502.
131 M. Uraloğlu, M. Livaoğlu, O. Agdoğan, S. Mungan,
A. Alhan and N. Karaçal, An evaluation of five different
dressing materials on split-thickness skin graft donor site
This journal is © The Royal Society of Chemistry 2018
View publication stats
Review
and full-thickness cutaneous wounds: an experimental
study, Int. Wound J., 2014, 11, 85.
132 Y. J. Shin, C. S. Han, C. S. Lee, H. S. Kim, S. H. Ko,
S. K. Hwang, S. G. Ko, J. W. Shin, S. K. Ye and
M. H. Chung, Zeolite 4A, a synthetic silicate, suppresses
melanogenesis through the degradation of microphthal-
mia-associated transcription factor by extracellular signal-
regulated kinase activation in B16F10 melanoma cells,
Biol. Pharm. Bull., 2010, 33, 72.
133 M. Cangiotti, S. Salucci, M. Battistelli, E. Falcieri,
M. Mattioli, M. Giordani and M. F. Ottaviani, EPR, TEM
and cell viability study of asbestiform zeolite fibers in cell
media, Colloids Surf., B, 2018, 161, 147.
134 Erionite, http://www.mindat.org/min-1399.html, (accessed
January 2018).
135 Erionite Structure, http://www.uwgb.edu/dutchs/petrology/
Erionite%20Structure.HTM, (accessed January 2018).
136 K. K. Turner, B. D. Nielsen, C. I. O’Connor-Robison,
F. H. Nielsen and M. V. Orth, Tissue response to a sup-
plement high in aluminum and silicon, Biol. Trace Elem.
Res., 2008, 121, 134.
137 M. Katic, B. Bosnjak, K. Gall-Troselj, I. Dikic and
K. Pavelic, A clinoptilolite effect on cell media and the
consequent effects on tumor cells in vitro, Front. Biosci.,
2006, 11, 1722.
138 K. Pavelić, M. Hadzija, L. Bedrica, J. Pavelić, I. Dikić,
M. Katić, M. Kralj, M. H. Bosnar, S. Kapitanović,
M. Poljak-Blazi, S. Krizanac, R. Stojković, M. Jurin,
B. Subotić and M. Colić, Natural zeolite clinoptilolite: new
adjuvant in anticancer therapy, J. Mol. Med., 2001, 78,
708.
139 L. Jarosz, D. Stepien-Pysniak, Z. Gradzki, M. Kapica and
A. Gacek, The effect of feed supplementation with
Zakarpacki zeolite (clinoptilolite) on percentages of T and
B lymphocytes and cytokine concentrations in poultry,
Poult. Sci., 2017, 96, 2091.
140 I. Kopova, L. Bacakova, V. Lavrentiev and J. Vacik, Growth
and potential damage of human bone-derived cells on
fresh and aged fullerene C60 films, Int. J. Mol. Sci., 2013,
14, 9182.
141 I. Kopova, V. Lavrentiev, J. Vacik and L. Bacakova, Growth
and potential damage of human bone-derived cells cul-
tured on fresh and aged C60/Ti films, PLoS One, 2015, 10,
e0123680.
142 Y. Guo, K. Wu, X. Huo and X. Xu, Sources, distribution,
and toxicity of polycyclic aromatic hydrocarbons,
J. Environ. Health, 2011, 73, 22.
143 I. Jirka, P. Sazama, A. Zikanova, P. Hrabanek and
M. Kocirik, Low-temperature thermal removal of template
from high silica ZSM-5. Catalytic effect of zeolitic frame-
work, Microporous Mesoporous Mater., 2011, 137, 8.
Biomater. Sci., 2018, 6, 974–989 | 989