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7 4D Printing: The Next Dimension of Healthcare in Cancer Research 259
In Fig. 7.10a, the potential of dynamic scaffolds for enabling response to
the acidic environment was more obviously efficient than that of others. The
tumor volume induced by human triple-negative breast cancer cells (MDA-MB231 cells) in mice considerably decreased in the implantation treatment with IS,
compared with other scaffolds consisting of only DOX-HCL + FLU drug and DOXHCL + FLU + PLGA called PD5. Likewise, the scaffolds with core-shell, hollow,
and porous features have widely served as drug delivery and carriers. For example,
the sol–gel transition under the light stimulus has been utilized in controlling the ondemand drug release (Wei et al.
2020; Liu et al. 2021a).Weietal.(2020) utilized the
gel–sol transition of gel through the photothermal effect in controlling drug release.
The core-shell hydrogels were fabricated by a 3D printer with a co-axis nozzle and
comprised of PDA/Alg as the shell layer and DOX-HCL drug (1%w/w) loaded in gel
as the core layer. The hydrogels provided excellent and stable photothermal effects
after five cycles that were used to control on-demand drug release by triggered with
NIR laser irritation, which resulting to no drug leakage without irradiation. The
efficiency of cancer therapy was examined by fighting against 4 T1 cells with the
NIR light. No side effects of chemotherapy happened in vivo and the tumor volume
in mice was notably reduced. In the next year, core-shell scaffolds were further
developed as multilayer scaffolds for long-term drug incubation (Liu et al.
2021a).
The three-layered scaffolds based on PDA/PCL/Gel/Alg provided efficient control
of drug release, including DOX-HCL and dexamethasone (DEX), over 2 weeks and
also showed potential in vivo cancer damage, as seen in Fig.
7.10b, where the tumor
volumes were significantly reduced. In addition, the control of core structures in
drug delivery systems can be induced by an external magnetic field. For example,
Wang et al. (
2021) fabricated porous scaffolds consisting of alginate/Fe3O4 hollow
fibers using a coaxial 3D printer. DOX-HCL drug was encapsulated in an Algbased core part, and the alginate/Fe
hydrogel was prepared as the shell part. The
3O4
scaffolds provided stably on-demand drug release through the excellent deformation
and recovery process under magnetic stimulation with 0.42 T for 1 min as shown
in Fig.
7.10c. These scaffolds also enabled on-demand releases of proteins and
live cells like bovine serum albumin (BSA) and human mesenchymal stem cells,
respectively.
7.2.2.3 Millirobots
Not only are scaffolds widely used as a drug carrier platform for controllable drug
release, but mobile robots have also gained widespread attention in DDS with an
advanced feature of locomotion. To overcome challenges of limited tissue penetration, mobile robots have gained growing interest in drug delivery (Xu et al.
For instance, Hu et al. (
2022) exploited soft millirobots incorporating PNIPAM,
2018).
nanoclay and NdFeB. The millirobots were designed as double leptasterias-like
layers that have been expected as a good drug carrier which providing protection
of drug leakage and completed locomotion of carrier in a human stomach model
with various responses to thermal and magnetic fields as illustrated in Fig.
7.11.
The robot completely traveled on the rough surface of a stomach model and
provided controllable drug release on a targeted area with an 800-rpm frequency
260 A. Chinnakorn et al.
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Fig. 7.10 (a) A comprehensive image of intelligent scaffold used in drug delivery including
(i) schematic image showing a fabrication of the intelligent scaffold (IS) comprising of gelatin,
chitosan, 5-fluorouracil (named as FLU or 5-FU), and doxorubicin (DOX) by an electrohydrodynamic jet 3D printer, (ii) a graph of excellently controllable drug release of IS under pH
variables, (iii) visible light images and in vivo images of mice implanted with control, DOXFLU, DOX + FLU + PLGA (PD5) and IS, and (iv) potential therapeutic efficiency of recurrent
tumors after 30 day treatment (Shi et al.
including (i) schematic diagram of core-shell fiber scaffolds, which providing drug release after
NIR irradiation (0.6 Wcm
with different treatments: (control presented no treatment, + presented treatments with laser
irradiation, − presented treatments without laser irradiation), and (iii) the tumor volume with
different treatments after 15 day treatment (control presented no treatment, scaffolds presented
polydopamine/polycaprolactone/gelatin/alginate scaffolds, NIR presented treatments with NIR
irradiation) (Wei et al.
composite scaffold including (i) a schematic image illustrating a fabrication and application of the
alginate/Fe
after magnetic stimulation, (iv) released core gels from reopen of hollow scaffolds after magnetic
stimulation, and (v) efficiency of DOX drug release under the magnetic stimulation for 2 min
(Wang et al.
hollow scaffold in drug delivery, (ii) the alginate/Fe3O4 scaffolds before and (iii)
3O4
2021)
−2
and 808 nm), (ii) a graph of the tumor growth of tumors in mice
2020, Liu et al. 2021a), and (c) a comprehensive image of magnetic-
2020b), (b) applications of fiber scaffolds in drug delivery
of a rotating magnetic field. Levofloxacin (L830182) was loaded in the hydrogel at
a concentration of 20 mg/mL, and the drug-loaded hydrogel was heated in water
◦
C in order to achieve a contracted network, which prevented drug leakage
at 38
approximately 16% more than before heating with water. Moreover, the drug release
7 4D Printing: The Next Dimension of Healthcare in Cancer Research 261
Fig. 7.11 (a) Various response performances of leptasterias-like hydrogels, caused by the dif-
ferent configurations under thermal stimulation (scale bars = 5 mm), and (b) travel of the
leptasterias-like hydrogel with slipping, climping, rolling, and gripping abilities within a stomach
model (scale bars = 6 mm) (Hu et al.
2022)
at the targeted site was 35% increasingly activated with a mechanical centrifugal
force by the rotating magnetic field.
7.2.2.4 Microrobots
Indeed, the smaller robots, like microrobots can intensively manage smaller objects
and operate in hard-to-reach sites in body (Dabbagh et al.
2022). The size of
robots in DDS has been developed by downsizing to the level of microscale robots
(especially smaller than 100 μm), which offer manipulations of smaller objects like
single cells, bacteria, and narrow areas, along with supporting minimally invasive
medicine by focusing on targeted sites (Ceylan et al.
2019; Xin et al. 2021). For
example, microswimmers were applied in the cargo delivery with capabilities of
on-demand release and high precision. They were designed as a double-helical
geometry with a volume of 6 μm diameter and 20 μm length and fabricated
by a two-photon-based 3D printer (Bozuyuk et al.
Bozuyuk et al. (
2018) reported potential microswimmers that were based on ChMA,
2018; Ceylan et al. 2019).
superparamagnetic iron oxide nanoparticles (SPIONs) with 5 mg/mL concentration,
and DOX drug loading via an NHS-amine coupling reaction. These microswimmers
provided movable functions and controllable drug release by propelling with a
magnetic field and inducing with an external light stimulus, respectively. Their
controllable locomotion, including steering, optimally performed at 4.5 Hz under a
10 mT rotating magnetic field with an average forward velocity of 3.34 ± 0.71 μm/s.
These microswimmers succeeded in the control of on-demand drug release and
dose by on-off switchable irradiation with light at 365 nm wavelength and 3.4 10
−1
W/cm2 intensity as seen in Fig. 7.12a. There was only slight drug release without
light illustration, while around 15% of the drug was released when the light was
turned on. Moreover, these composites were biodegraded at 37
◦
C by the lysozyme
enzyme without producing a toxic degradation product in 204 h.
Ceylan et al. (2019) fabricated microswimmers based on GelMA and SPIONs.
These microswimmers were propelled by an external magnetic field and controlled
drug release with the MMP-2 enzyme. Their locomotion is optimally controlled
262 A. Chinnakorn et al.
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2018, Ceylan et al. 2019), (b) an overview image of fish-like microrobots
Fig. 7.12 (a) A comprehensive image of methacrylamide chitosan (ChMA)- and gelatin methacryloyl (GelMA)-based microswimmers incorporating with
superparamagnetic iron oxide nanoparticles (SPIONs) including (i) a fabrication of the ChMA/SPIONs microswimmer by using 3D two-photon direct laser
writing printer, (ii) drug release abilities of the microswimmer, (iii) illustrated images showing the swimming controls of the microswimmer under arotating
magnetic field (red line presented trajectory snapshot of pseudocoloredly green microswimmer), and (iv) a schematic diagram showing an idea concept of
therapeutic applications with GelMA/SPIONs microswimmer as following step1: microswimmer injection into body, step2: precise travel of microswimmer
to targeted site under magnetic stimulation, step3: activated drug release by MMP-2 enzyme, step4: the biodegradation process of microswimmer, and step5:
2021), and (c) an image of sperm-driven microrobot used to release drug upon cancer
in drug delivery systems including (i) a schematic illustration of a magnetic fish-like microrobot used to release drug at targeted cancer cell with shape-morphing
abilities, and (ii) HeLa-killing efficiency of the microrobots (Xin et al.
cell walls including (i) a schematically experimental image of drug-loaded delivery system that sperm and magnetic composite were used to drive the system
treatment assessment with antibody-modified magnetic contrast agent (Bozuyuk et al.
2018)
consisting of sperm cell and tetrapod microstructure, (ii) fluorescence image showing the DOX drug distribution in the area of HeLa cells after external magnetic
stimulation (Xu et al.
7 4D Printing: The Next Dimension of Healthcare in Cancer Research 263
under a rotating magnetic field with 20 mT magnetic field strength and 5 Hz
frequency, which moves with a velocity of 3.36 ± 0.71 μm/s. The microswimmers
rapidly responded to the MMP-2 enzyme by swelling and then affected an embedded drug’s release. Also, nontoxic products were found after completely degrading
the microswimmers in 118 h.
The fabrication of microrobots has been tremendously inspired by nature, such
as the opening/closing mouse or the movable fins of fish, the opening/closing claws
of crabs, the flyable ability of insects, and the sperm mechanism in a reproductive
system. In particular, Xin et al. (
2021) revealed fish-like microrobots with shape
morphing of the switchable open-close fish mouth under the pH variable and
controllable locomotion under the magnetic field, which facilitated drug release on
cancer cells as demonstrated in Fig.
7.12b. These fish-like microrobots employed
pH-responsive material in shape-morphing mechanism to achieve drug release, and
Fe
nanoparticles to propel microrobots. The microrobots efficiently maintained
3O4
drugs within structures and achieved controllable on-demand drug release under
environmental pH change, which resulted in HeLa cell death as manifested in Fig.
7.12b-(ii). Another group presented a sperm-driven microswimmer for a targeted
drug delivery system, consisting of DOX-HCL drug-loaded sperm cells and a
tetrapod microstructure (Xu et al. 2018). The tetrapod microstructure, fabricated
by 3D laser lithography, was used to guide and release the sperm. The tetrapod
structure was similar to a tubular body and had four flexible arched arms that acted
as liberating the sperm cell when they pushed against a tumor wall. The spermdriven microstructure achieved the sperm liberation to release drug which killed
HeLa cells under an effective magnetic field of around 5 mT as shown in Fig.
7.12c.
7.2.3 Pharmaceutical Models
As mentioned before, cancer is complex, and the patient’s response to treatment
can vary. However, a poor understanding of the cancer environment is possible and
could lead to inaccurate prognosis and diagnosis. 3D printing has a significant role
in cancer treatment, with aspects of diagnosis and treatment preparation, such as
anatomical model of superior mesenteric vessel used in colon cancer surgery (Chen
et al.
2020a), and an individual patient’s lung model (Chen et al. 2020b). These
3D-printed anatomical models were reported as enhanced methods that can support
surgeons and be potential assistance during surgery and treatment preparation,
compared with surgery without a 3D model (Tejo-Otero et al.
2020a). In the pharmaceutical aspect, the prediction of a patient’s response to a
drug is necessary in cancer treatments. The use of 3D bioprinting, which enables
products to combine constructs and cells, including single and multiple types,
enhance potential of models used in understanding tumor/cancer environment and
modeling the drug efficiencies (Wang et al.
2018; Kang et al. 2020).
4D printing is still a newborn technique for tumor and cancer models. Recently,
4D bioprinting has been utilized to facilitate the fabrication of dynamic architectures used in a drug-evaluated model. 4D bioprinting is the use of 4D printing
2022; Chen et al.
264 A. Chinnakorn et al.
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technology in a fabrication of biological materials like cells, or growth factor
cooperating with materials (Naniz et al.
2022). For example, Chadwick et al. (2020)
used self-transformable cell-culture arrays for the evaluation of drug responses
in glioblastoma patient-derived organoid (PDO) as illustrated in Fig.
7.13.The
arrays were fabricated from two main components that were poly(ethylene glycol)
diacrylate (PEGDA) and Bisphenol A ethoxylate dimethacrylate (BPADMA) by
a projection micro-stereolithography (PμSL). The 4D bioprinted arrays were
designed as interconnected wells of cell-culture that could contain biological
samples at the bottom and made them facilitated the histological process with onestep insert of entire array into a cassette through the shape deformation and recovery
effect, which inducing the shape recover at 50
◦
C.
7.3 Conclusion and Future Perspectives
Cancer is a priority health issue as it causes of millions of deaths annually. Although
the use of 3D printing in cancer treatment has been clearly manifested because
of advanced features like personalized fabrications with the precise positioning, it
cannot be denied that the limitations exist in the developments of 3D printing in
the cancer therapies (Kumari et al.
have appeared to help fight cancer. 4D printing is a potential innovation that
can produce dynamic constructs, which is explicitly beneficial for localized and
personalized cancer treatments with minimal invasions. The abilities of 4D printed
structures have been expected to encourage cancer treatments in many aspects, such
as reduced incision, precise treatment, and effective assistance to obtain insight into
and understanding of the cancer environment along with potential outcomes.
The reviewed reports prove the advantages of 4D printing in cancer treatments
to fabricate intelligent structures which provide controllable shape transformation
and locomotion upon specific conditions of stimuli. Indeed, dynamic structures
have been almost entirely operated by changing the value parameters of stimuli
such as lower/higher temperatures and pH changes. Therefore, those parameters
used to activate the dynamic structures in healthcare applications should be such
that they will not provide side effects to normal organs/tissues and toxic products
in the human body. For example, the temperature variable should not be lower or
higher until it is harmful to the human body. There are current efforts to prevent
the temperature issue such as reducing areas that relate to the temperature with
smaller structures like microrobots or modifying T
structures still lack studies of in vivo experiments because 4D printing is still a new
field of research. Though there are few experiments using models and small animals,
it is still required to understand the insight of 4D printed materials in biomedical
applications and explore the results of large-volume scales. The biocompatible study
of dynamic structures and in vivo experiments is certainly expected in the future in
order to enhance practical use or clinical applications.
2022). Some new treatments and innovations
. In other factors, the dynamic
trans
7 4D Printing: The Next Dimension of Healthcare in Cancer Research 265
Fig. 7.13 (a) A diagram of operating procedure of the programmable cell-culture array, (b) a shape deformation and recovery process of poly(ethylene glycol)
diacrylate (PEGDA)/Bisphenol A ethoxylate dimethacrylate (BPADMA)-based upon raised temperature, and (c) a diagram of shape-morphing function which
2020)
enabling the arrays to protect biological materials (Chadwick et al.
266 A. Chinnakorn et al.
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