Multiorgan microdevices

 Guided By:-
Prof. P. V. Devrajan
 By:-
Ashish Singh Rajput
14PHP2003
M.Pharm(Pharmaceutics)
Institute of Chemical Technology, Mumbai.
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Contents
•Introduction
•Design and Fabrication
•Microfabricated Organ Models
Lungs on Chip
Liver on chip
Gut Epithelium on chip
Cardiac System on chip
•Contibution in Drug development process.
•Challenges
•Bibliography.
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 Introduction:-
 Multiorgan micro-devices are in-vitro set up of animal cells to simulate the same physiological
environment and study the effect of drug on different cells and organs.
 These systems are capable of simulating human metabolism.
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 The devices have the potential to predict potential toxic side effects with higher accuracy before a drug
enters the expensive and time consuming phase of clinical trials.
 Since single organ devices are testing platforms for tissues that can later be combined with other
tissues within multi-organ devices
 Multi-organ micro-devices can be seen as physical representations of Physiologically based
pharmacokinetic models in which the organs are represented by an actual compartment.
 Devices could be a way for the development of individualized medicine.
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Human PBPK model showing different Organs.

Why use microfluidics?
The science of manupulating small amounts of fluids in
Microfabricated hollow channels.
 Sample savings – nL of enzyme, not mL
 Faster analyses – can heat, cool small volumes quickly
 Integration – combine lots of steps onto a single device
 Novel physics – diffusion, surface tension, and surface effects dominate
This can actually lead to faster reactions!
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 Device Development and Fabrication:-
 Photolithography is a core microfabrication technique used to transfer microscale patterns to
photosensitive materials by selective exposure to optical radiation.
 A silicon wafer is spin coated with a thin uniform film of a photosensitive material
( Photoresist )
 Photomask with a pattern defined covers the photosensitive material.
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 Exposure of the photoresist to high-intensity UV light through the photomask which protects
some regions and exposes others based on the design of the pattern.
 Soft lithography involves fabrication of elastomeric stamps using liquid prepolymer of PDMS is
cast against the pattern of photoresist.
 the PDMS stamp is inked with protein solution, dried and brought in conformal contact with a
surface for a period ranging from 30 s to several minutes

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 Upon removal of the stamp, a pattern is generated on
the surface that is defined by the raised structure of the stamp.
 In 3d Cell culture different ECM gel , Hydrogels and
Agarose are used as base mould which enable them to
grow equivallently in all direction.
PDMS:Poly dimethylsiloxane
ECM : Fibronectin, collagen

 Microfabricated Organ Models:-
 Lungs on Chip
 Liver on chip
 Gut Epithelium on chip
 Cardiac System on chip
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 Lungs on chip:-
 The device is made using human lung and blood vessel cells and it can predict absorption of
airborne nanoparticles and mimic the inflammatory response triggered by microbial pathogens.
 Human lung alveolar pithelial cells are cultured at an air–liquid interface on one side while
human lung capillary endothelial cells are grown on the opposite side .
 Human lung-on-a-chip device consists of two PDMS microfluidic channel layers separated by a
thin (10 mm), flexible, ECM-coated PDMS membrane with micro engineered pores (10 mm in
diameter) that mimics the alveolar–capillary interface of the living lung

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• Three PDMS layers are aligned and irreversibly
bonded to form two sets of three parallel
microchannels separated by a 10-mm-thick
PDMS membrane containing an array of
through-holes with an effective diameter of 10
mm.
• After bonding, PDMS etchant is flowed through
the side channels. Etching of the membrane
layers produces two large side chambers to
which vacuum is applied to cause mechanical
stretching.
• Actual lung- on-a-chip microfluidic device can
be seen on Fig. E

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Casting
PDMS
Membrane
Pre
polymered
the layers
Photolithography
of microchannels
Coat with the binding
layer and incubate at
65 “c overnight
Bound
irreversibly
with the two
layers
Etching the
membrane
with TBAF
& NMP
Apply
Hydrostatic
Pressure
&Vaccume
Run the
etchant
solution
Upper
chamber is
Alveolar
chamber
Lower
chamber
Blood flow Workflow

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A Microfluidic device was used to model the airway
architecture to simulate abnormal obstruction of
airways and to study the effect of liquid propagation
and rupture on the alveolar epithelial cells lining the
alveoli.
 The two layer device was designed to allow
controlled mechanical stretching of the
endothelial–epithelial bilayer, mimicking the
mechanical cues present in the lung during
breathing.

Liver on Chip
 Liver and kidney are the major organ responsible for detoxification of toxins and
metabolism of drug.
 Organ to organ interaction often seen with liver which change the metabolite of drug.
 The Co-culture pattern of Rat primary hepatocytes and stromal cells improved various
liver-specific functions which were very close to actual liver physiology.
 The functional unit of the liver, the acinus, produces different set of proteins depending
on the locations within the unit as per the O2 gradient in tissue.
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Liver and Kidney
Interaction
(Liver cell
models HepaRG
was used)
Ca+2 release in
Kidney
Active metabolite
shows Anticancer
activity
Liver and Kidney
Interaction
(Liver cell line
HepG2/C3a was
used)
Less Bioactivation of
Drug and
Perturbation in cell
differentiation
Ifosfamide activated by CYP450)
(Ifosfamide activated by CYP450)
Less Ca+2
release.
Some special types of liver
cells are responsible for
bioactivation of drug

 GIT on Chip
 The in vivo environment of the GI tract is extremely complex consisting of circular
tissues and metered length.
 The lumen is separated by several layers of tissues containing mucosa, muscle, and
blood vessels.
 The inside lining of the epithelial layer is covered with villi, which increase the
absorptive surface area.
 The two major in vitro methods for predicting drug absorption are the Caco-2 model
and the parallel artificial membrane permeability assay (PAMPA).They mainly test the
permeability of drugs based on passive diffusion.
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 The Caco-2 cell monolayer model was able to predict the
absorption coefficient of rapidly and completely absorbed
drugs, while the prediction for slowly and incompletely
absorbed drugs were inaccurate.
 Sung et al. developed a novel hydrogel microfabrication
method to create collagen scaffold mimicking the shape of
intestinal villi, and cultured Caco-2 cells into a 3-
dimensional villi shape.
 Using this 3D villi scaffold, permeability coefficients were
measured and were shown to be closer to in vivo values than
the conventional 2D model.

 Heart On chip
 The Muscular Thin Film (MTF) assay measures contractile stresses generated by
anisotropic muscular tissue engineered on top of a deformable elastic thin film.
 The contractility of the engineered tissue is derived from the observation of the three-dimensional
(3D) deformations of MTFs
 The assay has been employed for evaluating contractility and tissue structure from
multiple tissues .
 The microdevice has a heated metallic base for maintaining physiological temperatures,
transparent top for optically recording MTF deformation and embedded electrodes for
electrical field stimulation of the tissue

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An MTF chip was brought from the incubator and placed in the
aluminum chamber.
The polycarbonate top was tightly fastened creating a fluidic seal
between the top and PDMS coated MTF chip.
The Infusions of 10 ml at a flow rate of 1 ml /min were employed
for complete flush out of the system.
Microtissues were electrically stimulated at 2 Hz with 10– 15 V of a
bipolar square pulse of 10 ms duration to stimulate membrane
depolarization.
After completion of contractility experiments, MTF chip is
immunostained for nuclei, actin, and sarcomeric Actin to directly
compare the stress generated by each tissue on the chip relative to
its sarcomere organization.

 Cost effective drug discovery
 Predicting Drug efficacy and Toxic side Effects.
 Testing drug Interaction & combinations.
 Predicting the bioavailability of Drug.
 Drug specific treatment and Individualized medicine.
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1) Development of suitable Culture Media.
 Typical cell culture media contain a mixture of defined nutrients dissolved in a buffered
physiological saline solution.
 Cell-cultures are designed to mimic the relevant in vivo environment. A temperature of
37 °C relevant to body temperature, and a controlled humidified gas mixture of 5% CO2
and 95% O2 are the standard physical conditions.
 Media depends upon the organ and cells because each organ is specific in terms of
nutrients intake .
E.g.- The Promo Cell Skeletal Muscle Cell Growth Medium is a low-serum (5% V/V)
medium optimized for the expansion of human skeletal muscle cells.
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 One media could not be suitable for two different type of cells so it is very hard to select
the commom media in multiorgan devices.
E.g.- The tissues were stimulated with TGF-ß1, . TGF-ß1( transforming growth factor
beta ) supported the growth of A549 lung cells, but inhibited the growth of HepG2/C3A
liver cells.
 This response highlights the difficulty of finding a common medium with growth
factors that support the viability of all cell types.
 Some common culture medias are
Low serum (5% v/v )or Serum free medium
Fetal bovine Serum
ES cult Basal medium (Marketed Product)
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Cell Sources:-
 Primary human cell , such as skin ,
skeletal muscle , and blood ,are
relatively easy to obtain. Acquisition of
others, such as neurons, is more
problematic .
 In such cases investigators are often
limited to cadaver tissue as a cell source.
 Alternative methods like iPPC culture,
stem cells propagation and 3D cell
culture best suited for culture of cell and
highlights the importance of novel in
vitro platforms for developing new
therapies
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Induced
Pluirepotent
cell culture
technique
Methods
to cell
culture
Genetic
Engineering
3D cell
culture
Techniques
Stem cell
propagation

 Bibliography:-
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(1995) 111–119
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 D. Huh, B.Matthews, A.Mammoto,M.Montoya-Zavala, Reconstituting organ-level lung functions on a chip, Science
(2010) 1662–1668.
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