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9 Bioprinting in Pharmaceuticals 301
Fig. 9.2 (a) Fabrication technology via sacrificial 3D printing with a gelatin-fibrin extracellular
matrix hydrogel, (b) proximal tubule completely epithelialized after post-seeding and cultured
proximal tubule epithelial cells reprinted from (Fransen et al.
2021)
aspect that needs to be considered while performing bioprinting is the selection of
either SB or SF technique. SBB offers really commercial availability, applicability,
and cost efficiency (Yu et al.
2016). On the contrary, after prolonged culture,
parenchymal cells in scaffold-based 3D bioprinted constructs decreased stability and
functional markers. However, in SFB, cells can be incorporated at high densities,
and they self-assemble into native-like tissues. Compared to immobilization and
interaction inside a scaffold, greater level of cell-cell interaction can occur in an
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SFB. These attributes enable the generation of tissues models that resemble native
tissue and prolong the time for which the phenotype and functionality of the cells
can be preserved (Gopinathan and Noh
2018). Based on the objectives of a particular
pharmaceutical study, for example drug discovery, development, and absorption, the
optimal bioprinting approach should be chosen. For instance, SBB is preferred in
drug delivery research because hydrogel degradation can be modulated to control
drug release. The biomechanical signals essential for the tests specify therapeutic
screening (Zhang et al.
2019b). SFB has gained more popularity in disease research
and in the screening of new anticancer drugs because the hypoxic centers in
the micro-tissues produced by these technologies more closely mimic the cancer
environment (Knowlton et al.
2015). For the bioprinting of undifferentiated cells,
SBB is preferred. Hydrogels should be picked based on the needs of the individual
cells in scaffold-based bioprinting. Since high viscosity of hydrogel scan easily
clogs nozzles, it is advised to fabricate cell-based microarrays with SFB (typically
by inkjet bioprinting) (Khoshnood and Zamanian
2020). Another possible solution
for this problem could be the use of less viscous hydrogels or a tactile controlledvalve extrusion technique to extrude more viscous spheroids (Parfenov et al.
2020).
The bio-printability, biocompatibility, cross-linking mechanism (physical and
chemical) mechanical properties (firmness, strength, versatility), and pore size
serve a crucial role during t he selection of hydrogel (Do et al.
gelatin, agar, collagen, hyaluronic acid, fibrinogen, and Matrigel
2017). Alginate,
™
are examples
of natural hydrogels that are preferable than synthetic hydrogels like polyethylene
glycol (PEG) and polyethylene oxide-polypropylene oxide-polyethylene oxide
(Pluronic) (Unagolla and Jayasuriya
ples of protein hydrogels that are biocompatible (Pajoum Shariati et al.
2020). Collagen, gelatin, and fibrin are exam-
2015).
The biocompatibility of synthetic hydrogels can be improvised further by adding
an arginine-aspartate-glutamate (RGD) sequence. Additionally, the target tissue
characteristics should be considered while choosing hydrogels (Cui et al.
2020). For
example, bone cells can be bioprinted via collagen type I and type II cells, whereas
endothelial cells are best printed with fibrin. Matrigel
™
, made up of the ECM of
Engelbreth-Holm-Swarm (EHS) murine cancer cells, is preferred for growing many
mammalian cells, including cancer and cardiac cells (Bakunts et al.
2008).
All bioprinting mechanisms are perhaps used for controlled drug delivery
studies, even though SBB is effortlessly executed using EBB, DBB, and LBB. For
experiments involving the administration of drugs at gradient concentrations, DBB
is helpful (Ker et al.
2011).
Similarly, thermal based inkjet bioprinting is preferential for delivering genetic
materials like DNA or oligonucleotides because it guarantees highly efficient gene
transfection by forming provisional membrane pores without significantly harming
cells (Xu et al.
2009; Cui et al. 2010). EBB approaches can be used for low-
throughput drug testing to model tissue from bioprinting containing single cells
or tissue spheroids, irrespective of efficiency or toxicity assays. Most HTS was
carried out without a scaffold, and with DBB, HTSs with hydrogel binding of
cells were perhaps accomplished with EBB (Naghieh and Chen
methods (Yang et al.
2021). DBB has historically been the method of choice for
2021) or else LBB
9 Bioprinting in Pharmaceuticals 303
pharmaceutical applications. Still, LBB may be an available alternative because
it does not present any significant issues with nozzle clogged up and stressinduced cellular damages. Nevertheless, LBB is a high-priced as well as complex
technique that needs more scientific advancement to become widely accessible
for the pharmaceutical industry. Although bioprinted tissue engineering construct
requires fabrication with many cells, related paradigms are not obligatory, which
are not necessary in pharmaceutical-related studies (Cui et al.
al.
2017).
Wherever possible, the smallest construct dimensions should be taken into
account. HTS droplets can be bioprinted on gene chip or micro-wells up to a
resolution of one cell per dot. On cast molds, SF microspheres with a surface
area of about 100 m can be bioprinted, which mature into organoids suitable for
drug high-throughput testing (Hwang et al.
wells for bioprinting should be scaled appropriately. The use of manual pipetting
for media replacement or the addition of new drugs or doses is often involved in
transient assays to study acute toxicological studies as well as the assessment of
new anticancer agents (Mazzocchi et al.
bioreactors with a computer-controlled perfusion system are recommended for
research studies comprising long-term use of the bioprinted construct.
2021). Cell positioning within micro-
2019). However, distinct or duplicated
2010; Leberfinger et
9.9.2 Models of Heterocellular Co-culture
Cell-cell interactions significantly impact physiological function and drug response.
Bioprinting has an extra advantage over other 3D fabrication techniques due
to its ability to perform controlled co-culturing of various cell types. In 3D
printed chitosan nanofibrous scaffold-based culture, co-culturing rodent ventricle
cardiomyocytes with fibroblasts led to polarized preservation of shape and function.
In contrast, cardiomyocyte polarity and isolated contractions were found to be
destroyed (Hussain et al.
(such as hepatocytes, stellate, Kupffer cells) elicits the production of inflammatory
receptors or reactive oxygen species (ROS). The correct proportion of supporting
cells should be selected for bioprinting based on the desired pharmaceutical
outcomes. Bioprinted tumor cells are frequently co-cultured for studying the
mechanism of tumor progression, emigration, and metastatic phases of endothelial
cells (Zhang et al.
constructs stable (Roberts et al.
2013). Cross-interaction amongst various kinds of cells
2016). Fibroblasts are used as stromal cells to keep the 3D
2016).
9.10 Bioprinting in Pharmaceutical Research
9.10.1 Drug Delivery
As stated earlier, gene transfection has been done using thermal inkjet printing. Due
to the temporary disruption of the cell membranes caused by the thermal inkjet
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printing process, DNA plasmids can enter the cells through these pores. Shortterm micro disruption of the cell membrane brought on by shear stress and heat
enables the passage of DNA plasmids and cells through the printhead ink conduits
throughout the bioprinting procedure. This method is safe because the apertures
close just in time to preserve cell survival. Plasmids were subsequently delivered
into the cells, and droplets having genetically modified cells were equally distributed
to specific spots of a 2D or 3D matrix (Cui et al.
2010; Owczarczak et al. 2012). The
overexpression of plasmid DNA into cells is also accomplished using extrusionbased bioprinting constructs. Alginate mixed with calcium phosphate particles and
multipotent stromal cells (MSCs) were used to create bioprinted structures that
could be solid or porous. Cells were found to be successfully transfected with
DNA from a nonviral vector expressing bone morphogenic protein-2 (BMP-2).
Osteocalcin (OCN) and ALP activity and levels were higher in the BMP-2 plasmidcontaining bioprinted constructs than in the non-transfected control after 14 days of
in vitro culture (Loozen et al.
2013).
Heparin-binding growth factors such as BMP-2 and insulin-like growth factorII (IGF-II) have been spatially deposited on fibrin-coated coverslips in various
concentration ratios using a piezoelectric DOD printhead, generating various growth
factor gradient patterns. A combinatorial square pattern with different surface concentration levels of BMP-2 and fibroblast growth factor-2 (FGF-2) was also made
using the bioprinting technique, which leads to cell differentiation. This method
has also been used for fundamental cell biology and prosecution into patterning
and growth factor delivery (Miller et al.
2009). To create physiologically relevant
tissue models, controlled growth factor delivery is thought to be crucial. There
have been some compelling investigations that address this challenge. For instance,
when gelatin microparticles (GMPs) encapsulating vascular endothelial growth
factor (VEGF) were incorporated into 3D bioprinted human endothelial progenitor
cells (hEPCs)-Matrigel
™
/alginate constructs (Poldervaart et al. 2014), the ability
to prolong VEGF activity at the specific position was employed. It was revealed
that VEGF was continuously released from GMPs within 3 weeks. Bioprinted
designs were implanted subcutaneously in rat models for studying vascularization.
Histological studies exposed that slow VEGF release from GMPs resulted in much
greater vascular formation than rapid VEGF release by incorporating VEGF to the
media.
It should be observed that the polymer microspheres or water-oil (w/o) emulsions
did not allow effective control to transfer growth factors with specific differentiation
potential (Du et al.
2015). TKKTLRT, a short collagen-binding domain (CBD)
derivative of mammalian collagenase, was used to form growth factors that bind
to collagen. Bone mesenchymal stem cell (BMSC)-filled methacrylamide gelatin
frameworks coupled with CBD-BMP-2 collagen microfibers were printed by Dai’s
team using a specially designed bioprinter. Contrary to BMP-2 rapid-release
behavior, collagen microfibers were used to achieve a controlled release of CBDBMP-2. By increasing the expression of osteogenic indicators like ALP/BSP/OCN,
COLLA1, and alizarin, CBD-BMP-2 collagen microfibers were also discovered
9 Bioprinting in Pharmaceuticals 305
to endorse osteogenic differentiation of BMSC-loaded methacrylamide gelatin
scaffolds (Li et al.
2021).
9.10.2 Screening of Drugs for Efficiency and Toxicological Test
Bioprinted tissues with various ECM components and cells mimic the in-vitro
animal models for high- or low-throughput screening of therapeutic effectiveness
or toxicity of drugs. For such applications, biomimetic architecture and quantifiable
endpoints should be included in a seamless in vitro pharmacologic model for drug
screening. An integrated biological microelectromechanical system (bio-MEMS)
that has been printed has been shown as a promising functional biosensor for rapid
drug analysis (Cui et al.
printer was improved to accurately design and straighten C2C12 cells adjacent to
cantilevers at a resolution of 300 dpi. On the fourth day after bioprinting, cells
aligned into confluent myotubes over cantilevers, whereas non-bioprinted cells
were dispersed on a cantilever after 7 days without producing myofibrils. Myotube
contraction was demonstrated after being excited by an electrical pulse. Veratridine
(VTD) is an alkaloidal neurolysin that shows effective nerves and muscle sheaths
by controlling the voltage-gated sodium channel. The cells which are incapable
to contract can be undergone myotube contraction, where it can be excited by
an electrical pulse. Further, veratridine (VTD), an alkaloidal neurolysin that slow
conduction of nerves and muscle sheaths by controlling the voltage-gated sodium
channel, was administered to myotubes to stop their contraction. After the VTD
was removed, the myofibers could again contract synchronously in response to an
electric stimulus. The synchronized, spontaneous chemical stimulation from t he
bioprinted bio-MEMS devices proposed that this method may combine valuable
biosensors, motors, and actuators (Brodie and Sampson
Nguyen and his team have performed bioprinting to study drug-induced toxicity
at the organ level by developing primary 3D liver tissue, comprising human
umbilical vein endothelial cells (HUVECs), hepatocytes, and hepatic stellate cells
(Nguyen et al.
2016). A commercially accessible extrusion-based bioprinter was
employed to directly print 3D models onto 24-transwell culture. The model was
split into the core and the periphery, in which parenchymal and non-parenchymal
cells were incorporated, respectively. Metabolic and toxicity studies were performed
for the cells for 28 days. According to the study result, it was concluded that 3D
bioprinted scaffolds s how tissue- and cell type-specific responses that allow more
precise histological evaluation during a period compared to the results of the 2D
culture cytotoxicity study. The research has demonstrated the advantages of a 3D
culture microenvironment and the inclusion of several tissue-specific cell types
instead of altering primary cells (Chaji
Additionally, compared to cells in 2D, hMSCs in 3D matrices displayed sophisticated expression of angiogenic markers like CD31 or CD105. The same group has
shown that inhibiting ROCK promotes the release of VEGF or epidermal growth
factor (EGF) in bioprinted 3D constructs of matrix metalloproteinase (MMP)sensitive peptide, which boosts angiogenic sprouting and vascularization in rat
2013). In that research, Design Jet 500 thermal inkjet
1990).
2019).
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tissue in vitro (Hong et al. 2016). Human articular chondrocytes suspended in
a hydrogel consisting of poly (ethylene glycol) dimethacrylate (PEGDMA) were
bioprinted using a modified Design Jet 500 thermal inkjet printer to evaluate
bioprinted cartilage tissue models and differentiation. FGF-2/transforming growth
factor (TGF) co-treatment resulted in better chondrogenic characteristics in comparison to each factor acting singly (Cui et al.
2012).
Bioprinted 3D malignant tissues have been examined for sensitivity and resistance to chemotherapy (anticancer) agents. Many 3D bioprinted carcinoma cell
scaffolds demonstrated the ability to mimic in vivo anticancer drug responses. For
example, fibrinogen/gelatin/alginate hydrogels containing cells were extruded with
90% viability. The bioprinted cells generated spheroids inside 3D scaffolds. By
analyzing morphological characteristics and metabolic and MMP activity, it was
founded that 3D printed constructs were more chemoresistant to paclitaxel than 2D
planar samples. Organovo’s NovoGen Bioprinter
™
platform was used to bioprint SF
humanoid breast cancer cells to test the therapeutic outcome of tamoxifen (King et
al. 2014). A bio-inspired ECM of MSC-derived mammary fibroblasts, endothelium,
and fat cells was employed in the co-culture with the malignant cells (Li et al.
2020).
Histopathological examination of the bioprinted tissues revealed that they formed
distinct compartments for adipose, stromal, and epithelial constituents while developing micro-capillaries. The tissues remained viable in vitro for 2 weeks. Evaluating
the chemotherapeutic effects using the ATP luciferase assay revealed that isolated
2D cancer cells were more susceptible to tamoxifen-induced toxicity than cells
grown in 3D bioprinted constructs. For testing drug toxicity in the liver, Organovo
also developed a 3D bioprinted liver tissue model called “exVive3D” (Robbins
et al.
2013; Vaidya 2015). SF liver tissue was fabricated using a bioprinted
human liver cell pellet of stellate cells, hepatocytes, and endothelial cells on a
provisional structural mold framework. It was found that within the tissue, microcapillaries formed after 60 h of incubation. The bioprinted tissues expressed hepatic
enzyme markers, developed liver-secreted proteins like albumin and fibrinogen, and
maintained cell viability over 7 weeks. Levofloxacin (LVT) and trovafloxacin were
employed as two agents to authenticate the performance of bioprinted liver for
toxicological assays (Nguyen et al.
2016).
Amongst them, LVT which is used clinically for a long time was found to be
safe in bioprinted construct as shown in Fig.
9.3a, b, whereas trovafloxacin which
had failed in phase III trials was identified to have hepatic toxicity. Organovo’s
bioprinted 3D technology successfully evaluated the toxicity of the failed medicine
with the effectiveness of the commercial drug.
9.10.3 High-Throughput Screening (HTS) and Microarrays
In the 1980s, the pharmaceutical industry adopted HTS technologies to boost the
extent of productivity (Gribbon and Andreas
tion method that can be used for high-throughput studies and has the benefits of a
high yield, a short production time, and the ease of culture medium replenishment.
To enable the corresponding examinations of the drug toxicity and efficacy, several
2005). Bioprinting provides a fabrica-
9 Bioprinting in Pharmaceuticals 307
Fig. 9.3 Effect of drug in a bioprinted model illustrated by (a) untreated liver tissue model
fabricated by extrusion-based technique and (b) the same model treated with 100 μmof
trovafloxacin, reprinted from (Nguyen et al.
2016)
arrays have already been bioprinted for HTS. DBB is the earliest and most widely
used method for creating microarrays for high throughput (Gudapati et al.
Peng et al.
2016). In one of the earliest studies, high-throughput printing of bovine
2016;
serum albumin (BSA)-conjugated oligonucleotide arrays was achieved with good
bioprinting accuracy (Hsieh et al.
2004). Recently, an HTS miniature drug screening
platform was created using a modified HP model 5360 compact disc printer to create
an inkjet-based bioprinter with a picolitre resolution in a single drop (RodríguezDévora et al.
2012). The same area was successively printed on a glass slide with
three layers. The initial deposit contained an agar and bacterial blend solution, the
second layer contained 0.25% alginate, and the tertiary coating had CaCl
.The
2
findings showed that the inkjet bioprinted construct had cell viability, functionality,
and antibiotic effects comparable to those of the micropipetted models. Bioink
is a small drop or microwave-based technique to solve problems with inkjet
bioprinting, such as cell damage and ink clogging. For HTS applications, Demirci’s
team created acoustic-based bioprinting using a variety of cells, such as mouse
embryonic stem cells, AML-12 hepatocytes, fibroblasts, human Raji cells, and HL1 cardiomyocytes. Cell viability was maintained at a level close to 90% for various
cell types after nozzle expulsion of single cell in picolitre drops at a rate between 1
and 10,000 drops in a second (Demirci and Montesano
2007). This assembly also
created a power-driven cartridge ejector for high-throughput printing technology of
rat bladder smooth muscle cells immobilized in high-viscosity collagen (Moon et al.
2010;Xuetal. 2010). This stage enabled paradigms to be bioprinted with consistent
cell seeding, producing a layer-by-layer 3D cell design with constrained spatial
resolution and sustaining cell survival for extended cell culture periods. To generate
manageable, same-size embryonic bodies (EBs) from embryonic stem cells (ESCs),
the assembly combined micro-valve bioprinting with hanging drop technique. The
use of bioprinting led to the formation of EBs that were remarkably uniform in
size. The EBs were also more prominent as compared to manually created EBs.
The EBs created by this strategy were suitable for HTS of bioactive molecules
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Fig. 9.4 Effect of high-throughput screening of nano-shuttles. (a) Bioprinting of magnetized cells,
(b) 384- and 1536-well plates, reprinted from (Fernandez-Vega et al.
2022)
and for evaluating drug toxicity to embryos because the combined approach was
unsophisticated, reliable, and fast.
Hou’s team established primary pancreatic organoids for HTS by utilizing the
“Nano3D Biosciences magnetic 3D bioprinting system” in 384-well and 1536well plates as shown in Fig.
9.4a, b. Pancreatic and colon cancer cell lines were
used to form organoids with carcinoma-related fibroblasts. 3D experiments were
performed simultaneously with 2D, and the finding showed that 3D ECM-based
models have greater resistance towards cytotoxic agent as predicted. It was found
that the resistance differs between cell lines, and according to that finding, the
researchers developed a “resistance factor” to account for the variation in the IC50
between 3D models. Phenotypically, 3D models were demonstrated to mimic the
in vivo tumor surrounding accurately. Researchers recommended this application in
chemotherapeutic screening (Hou et al.
2018).
Tomi´c and group recently described a novel technique for creating 3D scaffolds
from ESC-filled hydrogels made of gelatin and alginate (Tomi´cetal.
2021). Instead
of aggregating after EBB, pluripotent, high-throughput EBs with stability and highdimensional center were obtained (Ouyang et al.
2015). Like Tomi´c and group,
Demirci’s team also created an anisotropic biomimetic fibrocartilage environment
in which a variety of methacrylate gelatin-encapsulated MSCs were printed with
some aspect of either BMP-2, TGF-β, or a combination of BMP-2 and TGF-β using
a valve-based droplet ejector (Gurkan et al.
2014).
The 3D bioprinted tissue with microscopic level anisotropy constructs demonstrated the capability to be utilized for high-throughput therapeutic validation
as a strong prediction for in vivo studies. In an attempt to bioprint a 3D co-
9 Bioprinting in Pharmaceuticals 309
culture prototype utilizing cancer cell normal fibroblasts (MRC-5) and (OVCAR-
5) micro-patterned on Matrigel
computerized cell bioprint (Xu et al.
™
, Demirci’s group developed a high-throughput
2011a). To produce droplets with a wide
nozzle (151 m diameter) and reduce local shear forces, a pulse generator was
used to control a nanoliter administration valve (solenoid valve ejector) having an
ejector for each cell. In a high-throughput and repeatable manner, the dual-expulsion
structures were coordinated, and two cell types were patterned in a spatially precise
microenvironment (e.g., cell compactness and cell-to-cell distance). Both cell types
survived printing and kept multiplying to produce 3D acini that could be cultured
for 3 weeks. This co-culture system with a bioprinting scaffold offered a biomimetic
means for high-throughput drug screening. 3D bioprinting has created a progressive
3D lung prototype for HTS for safety assessments and effective screening of drugs
(Horváth et al.
2015). The processing unit, which included a tool changer with
three workplaces and outfitted with design heads that could print up to three
diverse biomaterials or cells, was the central part of the bioprinter. “Human alveolar
epithelial type II cell line A54956 and EA. hy926 hybrid human cell line (derived
by fusing human umbilical vein endothelial cells (HUVECs) with A549 cells) were
bioprinted by droplet jetting process in contrast to Matrigel
™
,” which was printed
by contact dispensing.
A 3D air-blood interface model has also been bioprinted, and it was discovered
that the absorption coefficient or barrier functions of the bioprinted constructs were
comparable to those of the manual method. The bioprinting technique provided an
automated and repeatable construct that produced biomimetic cell deposits crucial
for an ideal air-blood tissue barricade. High-throughput drug screening can also
use a bioprinted microarray to examine cell-level reactions to ECM alteration. A
nanoliter cell-laden hydrogel array with a pressure-assisted DBB system and a
solenoid valve ejector was developed proposed by Xu’s group. To mimic human
periodontal ligament stem cell’s (PDLSC) reaction to ECM changes, cells were
loaded inside a gradient of PEG hydrogel/gelatin methacrylate (GelMA) matrices.
An increase in the PEG volume ratio was found to result in the decrease of cell
viability and spreadability (Ma et al.
2015). Studies on drug delivery, HTS, or dose-
effect relationships are expanding using this array model. A laser-based bioprinting
technique performed a high-throughput analysis of interactions between various cell
types and their environment. Endothelial colony-forming cells (ECFCs) and human
adipose-derived stem cells (ASCs) enclosed in hyaluronic acid/fibrinogen have been
bioprinted into 3D arrays. The 3D display array was optimized for the cell-to-cell
ratio, cell number (density), cell type grouping, layout, and elevation. Direct cell-cell
interactions were found to trigger the advancement of stable vascular-like networks
even in a VEGF-free medium, underscoring the potential of technique.
9.10.4 ADME Assay
Pharmacokinetics refers to administering drugs inside the body, including absorption, distribution, metabolism, and excretion. Drug ADME attributes are ideally
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examined during the preclinical discovery phase. Even though in vivo ADME
assays are performed on animals, biomimetic in vitro ADME assay models can
rapidly screen test for compounds with high drug-likeness, particularly while
examining the impact of drug metabolites over its effectiveness or cytotoxicity. For
in vitro pharmacokinetic analysis, Sun’s team has also created a 3D microfluidics,
microanalytical, and micro-organ device (Chang et al.
2010). A microfluidics tissue
compartment model made up of polydimethylsiloxane (PDMS) elastomer was
used to develop a bioprinted micro-liver using an automated syringe-based cellular
inscription procedure. The incorporated 3D tissue chamber unit was connected to a
syringe pump that supplied drugs and media in a sinusoidal flow pattern through
a convectional microchannel. The degree to which a non-fluorescence prodrug,
7-ethoxy-4-trifluoromethyl coumarin (EFC), transformed into a fluorescence 7hydroxy-4-trifluoromethyl coumarin, demonstrated the liver model to mimic the
biological function of drug metabolism (HFC).
In addition, the researchers sequentially merged HepG2 with human epithelial
™
cells covered in Matrigel
in a PDMS substrate. The microfluidic chips have been
used to support the bioprinted constructs with twofold micro-tissue microfluidic
chips and connected with a peristaltic pump (Snyder et al.
2011). The chips were
sealed under glass covers. Hepatocytes and epithelial cells have been utilized in
simulating drug absorption from GI lumen to liver. Additionally, the ability of
epithelial cells to metabolize the anti-radiation drug amifostine, a prodrug that is
transformed into an active drug by the epithelial cells, was tested. After marking
with the fluorescence 4
,6-diamidino-2-phenylindole (DAPI) as a marker, the
arrangement of binucleated cells with micronuclei as a sensor allowed researchers
to measure the damage to hepatocytes due to radiation. This micro-tissue is a
promising model for studying multicellular communication (Kramer et al.
2007).
Tang and his group established a glioblastoma model for tumor-ECM interaction
analysis by utilizing 3D bioprinted monoculture glioblastoma model (GBM).
Studies were conducted using gelatin-alginate-fibrinogen hydrogel by extrusionbased bioprinter. The finding showed that CD31, VEGFR2, CD133, and HIF-1a
were present in 3D bioprinted glioma model. Higher amounts of mitochondrion,
endoplasmic reticulum, and microvilli were observed as morphological change
(Tang et al.
as shown in Fig.
2021). Bioprinted 3D monoculture enables the tumor-ECM interaction
9.5a, b.
Madden’s team established an intestinal tissue model for ADME analysis
by utilizing human intestinal epithelial cells. 3D experiments were performed
simultaneously with Caco-2 monolayer (Madden et al.
2018). Findings showed that
3D intestinal model corresponds to endogenous levels, whereas Caco-2 monolayer
showed absence, overexpression, or reduced gene expression suggesting that bioprinted models mimic the native intestinal tissue as shown in Fig.
9.6.
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