From Microfluidics to 3D Bioprinting for Human/Organs-

on-Chips DevelopmentAmsterdam, 11-12th, December, 2018

Dinh Ngoc Duy

Department of Biomedical Engineering

National University of Singapore

Outline

Reconstruction of Human/Organs Function

Microfluidic Technology

3D Bioprinting

Part 1: Brain/neuron-on-Chip (Miniaturisation for Life Sciences Group, Institute of

Analytical Science, Dortmund, Germany.

Collaboration with Dr. Jean-Michel. Peyrin

(neurobiologist, Institute of Biology Paris-Seine,

France) who founded the MicroBrain

Biotech company and Neurotoxicology and

Chemosensation Group, Leibniz, Germany)

Part 2: NIR Laser

Bioprinting- Light

assembly (NUS, Singapore)

Why microfluidic devices are

great tool for cell neurobiology?

The neuron system to investigate

disease propagation:

infectious agents, pronsm herpes

and rabies virus

Degeneration , alpha synuclein in

Parkinson and amyloid beta in

Alzheimers

Toxicity nanoparticles as manganese

propagation

Part 1:

PNAS,1977, CAMPENOT, Department of Neurobiology, Harvard Medical

Taylor et al Nature Methods, 2,599–605 (2005)

Microfluidic chips: transparent,

to precisely control, monitor

and manipulate cellular

microenvironments, fluidic

perfusion

Limitations of current Microfluidic Chips

for Neuron

Tens of thousands of neurons to

attain significant numbers of inter-

compartment connections.

Neurons (<5%) extend outgrowths

between compartments.

High throughput studies would

require the sacrifice of multiple

animals with data analysis

complicated by inter-individual

variance.

Brain/Neuron-on-Chip–ISAS, Germany

Microfluidic Circuit Meniscus Pinning – Water MaskChip Image

Precise control the number of cells in each compartment by differential flow

microfluidics

Integrated a novel biomaterial co-patterning method for cell patterning inside

the microfluidic chip, promote neuron interconnection

Fluidic isolation using hydrostatic pressure control and aspiration methods.

Define a neuronal network high level intercompartment connectivity

Cell number is defined and minimal (dopamine cells and substantia nigra cells

for Parkinson’s disease or peripheral neurons) (10–100-fold less than existing

systems)

Dinh Ngoc Duy et al Lab Chip, 2013,13, 1402-1412; JoVE, 2014, Springer Book, 2015

Co-Patterning with PLL-g-PEG-TRITC

1. PLL-FITC

2. Water Mask /

Plasma

3. PLL-g-PEG-

TRITC

1. PLL

2. Water Mask /

Plasma

3. PLL-g-PEG-

TRITC

4. Avidin FITC

Illustration of the water masking concept and results

Biomaterial Co-Patterning

Dinh Ngoc Duy et al Lab Chip, 2013,13, 1402-1412; JoVE, 2014, Springer Book, 2015

Dinh Ngoc Duy et al Lab Chip, 2013,13, 1402-1412; JoVE, 2014, Springer Book, 2015

Neuron Cells Loading

Dinh Ngoc Duy et al Lab Chip, 2013,13, 1402-1412; JoVE, 2014, Springer Book, 2015

Cells Loading

Dinh Ngoc Duy et al Lab Chip, 2013,13, 1402-1412; JoVE, 2014, Springer Book, 2015

Neurite in

Patterning chip

go through

microgroove

channel more

than

unpatterning

Dinh Ngoc Duy et al Lab Chip, 2013,13, 1402-1412; JoVE, 2014, Springer Book, 2015

Neuron culture

on chip cells

day 14

2,3-compartment

chip

Dinh Ngoc Duy et al Lab Chip, 2013,13, 1402-1412; JoVE, 2014, Springer Book, 2015

Applications: disease propagation: infectious agents,

Parkinson, Alzheimer, manganese propagation

Dinh Ngoc Duy et al JoVE, 2014

Impacts & Potential Application

-The dissemination of aggregated molecules from one neuron to the

others and protein transport between neurons and glial cells. (Prof

Tsuneya Ikezu, MD, PhD, Alzheimer's Disease Center, Boston University

School of Medicine)

- To understand the mechanisms involved in the propagation of

hyperphosphorylated tau, β-amyloid and long-range BNDF-mediated

signalling.

Dinh Ngoc Duy et al Lab Chip, 2013,13, 1402-1412; JoVE, 2014, Springer Book, 2015

Integrate to Neurovascular unit

I. Investigate the dissemination of aggregated molecules

from one neuron to the others and protein transport

between neurons and glial cells

II. Integrate Cerebral Organoid (Glioma stem cell) in

microfluidic neurovascular model to study how thepatient’s tumor develop and react to treatment

III. To study how immune cell enter the brain and interact with neuron

IV. To evaluate immunotherapy for brain cancer and evaluating

therapeutic antibodies, which are delivered through BBB

Part 2: 3D bioprinting for Reconstruction of

Human/Organs Function

Computer designed 3D

structures

Automated process,

repeatability.

Programmable

approach

Challenge of 3D Bio-Printing:

Resolution

Liver Lobule

Adv. Healthcare Mater. 2015,

Schematic view of bottom-up assembled organs from

modular tissue composed of diverse cell types

Rebuilding the Body, Nature

Outlook Technology: The

promise of printing

Herb Brody et al Nature, 2016

Trends in Biotechnology May 2015, Vol. 33, No. 5

Corresponding technologies for assembling

building blocks at different scales

A NIR Laser Bioprinting - Light

Directed Assembly Building Blocks

in Microscale

Ultra precise, High-

throughput control in

Micro-Objects (~ 100

µm)

Low laser power,

less damage

biological sample

Dinh Ngoc-Duyet al Small, 2017

Part 2:

Photothermal Convection: solve differentiation

equations: electromagnetic (EM), heat transfer (HT) and

fluid mechanics (FM)

Dinh Ngoc-Duyet al Small, 2017

High spatial control- Single Micro-

Particles manipulation

Scale bar:100 µm

Dinh Ngoc-Duyet al Small, 2017

High throughput Assembly- Microparticle Hydrogel

Writing Patterns

Scale bars:

(b, e, f, h, i, k) 6 mm,

(d) 200 µm, (l) 400

µm

Dinh Ngoc-Duyet al Small, 2017

NIR Laser Assembling building blocks forming

Desired Patterns

Dinh Ngoc-Duyet al Small, 2017

Bottom up Tissue Engineering Approach

Mesenchymal stem cells seeded hydrogel particle

assembly

Scale bars:

(a) 30 µm,

(d) 120

µm.

Dinh Ngoc-Duyet al Small, 2017

Conclusions and Outlooks : potential to

revolutionize the 3D bioprinting technologies

We are negotiating with Fluxbio, a 3D Bioprinting start-up company based on

Electrospray technology for commercialization

Featured in Small, Wiley AdvancedScienceNews , 3D printer and 3D printing

news(3ders.org); 3Dprint.com; NanoHybrids, 3dprintingindustry.com

NIR Laser 3D-Bioprinting

(developing)

Commercial Companies

Lab Chip, 2017,17,2395-2420

Summary

Precise control the number of

cells in each compartment

Define a neuronal network high

level intercompartment connectivity

Cell number is defined and

minimal

Neuron Microfluidic Chip

NIR Light Bioprinting

High resolution and high

throughput control

Potential to revolutionize the 3D

bioprinting technologies

Thank you for your attention