Slides from the Pistoia Alliance Debates webinar on 18th June 2015, on 3D-printing and nanotechnology in pharma R&D.

1. 18 June, 2015
Printing therapeutic nanobots:
Nanoscale 3D-printing in
pharmaceutical R&D
a Pistoia Alliance Debates webinar
chaired by Jim McGurk

2. This webinar is being recorded

3. ©PistoiaAlliance
Panellists
Nanoscale 3D-printing in pharmaceutical R&D 318 June, 2015
Jim McGurk is Senior Director and Global Head of Data Architecture for Daiichi-
Sankyo R&D Informatics. His current position involves development and
implementation of master data management and governance for the Global R&D
organization. He is also responsible for creating and implementing a 'Big Data' strategy;
providing internal capabilities for accessing, managing and analyzing external/public
data sources. Jim has more than 20 years’ experience in software and system
development in pharmaceutical discovery and development, as well as developing
strategic directions driving architectural, software and hardware decisions.
Ahmed A. Busnaina, Ph.D. is the William Lincoln Smith Chair Professor, University
Distinguished Professor, and founding Director of National Science Foundation’s
Nanoscale Science and Engineering Center (NSEC) for High-rate Nanomanufacturing
and the NSF Center for Nano and Microcontamination Control at Northeastern
University, Boston, MA. Prior to joining Northeastern University in 2000, he was a
professor and a director of the Microcontamination Control Lab at Clarkson University
from 1983-2000. Dr. Busnaina specializes in nanoscale printing of 2D and 3D
structures for devices and sensors. He is an associate editor of the Journal of
Nanoparticle Research, and is a fellow of the American Society of Mechanical
Engineers and the Adhesion Society.
Dr. Utkan Demirci is currently an associate professor at Stanford University School of
Medicine, Canary Center Early Cancer Detection. He leads a group of 20+ researchers
focusing on micro- and nano-scale technologies. The Demirci Bio-Acoustic MEMS in
Medicine Labs (BAMM) laboratory specializes in applying micro- and nanoscale
technologies to problems in medicine at the interface between micro/nanoscale
engineering and medicine. Our major research theme focuses on creating new
microfluidic technology platforms targeting broad applications in medicine. In this
interdisciplinary space at the convergence of engineering, biology and material science,
our goal is to create novel technologies for bioprinting, disposable point-of-care (POC)
diagnostics and monitoring of infectious diseases, cancer and controlling cellular
microenvironment and microscale tissue engineering applications.

4. Scalable Nanoscale Printing for Sensors,
Electronics, Medical and Materials Applications
Ahmed Busnaina,
William Lincoln Smith Professor and Director, Northeastern University
NSF Nanoscale Science and Engineering center
for High-rate Nanomanufacturing
www.nano.neu.edu
www.nanomanufacturing.us

5.  Printing offers an excellent approach to making structures and devices using
nanomaterials.
 Current electronics and 3D printing using inkjet technology, used for printing
low-end electronics, flexible displays, RFIDs, etc. are very slow (not scalable)
and provide only micro-scale resolution (20 microns).
 20 microns was the silicon electronics line width in 1975.
 Even with these scale limitations, the cost of a currently printed sensor is
1/10th to 1/100th the cost of current silicon-based sensors.
 Printed Electronics is a $50B industry projected to reach $290B in 2025.
Current Technologies Using Nanomaterials
to Make Electronics and Sensors?

6. Directed Assembly of Nanoparticles,
Carbon Nanotubes and Polymers
50 nm50 nm 50 nm
copper
fluorescent PSL fluorescent silica
30 µm 5 µm
Metal
II
Metal I
SWNT
Bundles
Multiple polymer systems,
Rapid Assembly, multi-scales CNTs Rapid, multi-scale Assembly
Nanoparticle Rapid, multi-scale Assembly (ACS Nano 2014)

7. WSiO2
Damascene Templates for Nanoscale Offset Printing
Silicon-based
Hard
Templates
PEN PIPolymer-based
Templates
Assembled SWNT Assembled Particles
Advanced Materials, 2015

8. •3-D nanostructure manufacturing process
•using nano-colloids (Northeastern U.)
•Manufacturing of 3-D nanostructures using directed nanoparticle assembly process. (A and B) NPs suspended in
aqueous solution are (A) assembled and (B) fused in the patterned via geometries under an applied AC electric field. (C)
Removal of the patterned insulator film after the assembly process produces arrays of 3-D nanostructures on the
surface. (D) Scanning electron microscopy (SEM) image of gold nanopillar arrays. Economic approach.
•MC Roco, Nov 2 2014
ACS Nano, April 2014

9.  Leveraging the directed assembly and transfer processes developed at
the CHN, Nanoscale Offset Printing has been developed.
The ink is made of nanoparticles, nanotubes, polymers or other nano-
elements that are attracted to the printing template using directed
assembly.
Introducing Nanoscale Offset Printing
Nanoscale Offset
Printing Template Nanoscale Offset Printing System
NanoOPS Includes Six
Modules:
1. Hexagon Frame Module
2. Template Load Port Module
3. Directed Assembly
Module
4. Mask Aligner Module
5. Transfer Module
6. Template Load Port Module
1
2
3
4
5
6
This approach offers faster printing with a 1000 times smaller that inkjet or 3D printing.
NanoOPS Videos on Youtube:
From Lab to Fab: Pioneers in Nano-Manufacturing: https://www.youtube.com/watch?v=tZeO9I1KEec
NanoOPS at Northeastern University: https://www.youtube.com/watch?v=2iEjIcog774

10. Nanoscale Science
Directed
Assembly and
Transfer
Energy Electronics
Flexible
Electronics
CNTs for
Energy
Harvesting
Assembly of
CNTs and NPs
for Batteries
What Could We manufacture with Mul scale Offset Prin ng?
SWNT & NP
Interconnects
SWNT NEMS &
MoS2 devices
Multi-
biomarker
Biosensors
1
0
m
m
Drug
Delivery
Antennas, EMI
Shielding,
Radar,
Metamaterials
Materials Bio/Med
2-D Assembly of
Structural Apps.

11. Bio and Chem Sensors
Functionalized
SWNTs
4
μ
m
25
0
μm
Band-Aid Sweat Glucose & lactate sensor
Chemical H2S
Sensor
Cancer and
cardiac diseases.
Detection is 200
times lower
than
Current
technology
Sensors for E. coli bacteria, viruses, and other
pathogens
Supporting
printed
electronics for
sensor systems

12. Flexible CNT Bio Sensors for Glucose,
Urea and Lactate in Sweat or Tears
2 μm200 nm
Functionalized
SWNTs Gold
PEN
4 μm
250 μm
1 μm
200 nm
CNT Sensors have been also used to detect viruses, bacteria and
antibiotics (in Blood, water or air)

13. in-vivo biosensor
(0.1 mm x 0.1 mm)
Tested for detected with biomarkers for prostate (PSA), colorectal (CEA),
ovarian (CA125) and cardiac diseases. Detection limit: 15 pg/ml
Current technology detection limit is 3000 pg/ml
Publications: Langmuir Journal, 27, 2011 and Lab on a Chip Journal, 2012
US Patents: Multiple biomarker biosensor: (US 2011/0117582 A1), 2 more filed patents
 Multiple-biomarker detection
 High sensitivity
 Low cost
 Low sample volume
 In-vitro and In-vivo testing
Cancer and Cardiac Disease Optical Biosensors

14. Novel Drug Manufacturing for Oral Delivery of Poorly Soluble Drugs
1) Directed assembly of dissolved drugs into
nanorods
2) Delivery of drug in the blood with
subsequent tumor targeting
14The eudragit polymer can be dissolved at a specific pH
value to release the drugs in the desired region of intestine.
Problem:
 More than 40% of new chemical entities (NCEs) and anti-cancer agents
are insoluble in water and cannot be taken orally.
Solution:
 Fabrication of drug-loaded micelles into 3-D nanorods.

15. What is Unique about our Approach?
15
Chauhan V.P. et al., Angew Chem Int Ed Engl . 50, 11417–11420 (2011).
Ryan M. Pearson et al. MRS Bulletin, 39, 2014.
K. Huang et al., ACS Nano 6 , 4483 ( 2012 ).
H. Cabral et al. , Nat. Nanotechnol. 6 , 815 ( 2011 ).
A. Pluen et al., Biophys. J. 77 , 542 ( 1999 ).
 Effective penetration of NPs through the tumor is determined by the particle size
and shape.
 Most studies focus on the oral drug delivery of drug in the form of nanoparticles,
which have limited penetration into tumors.
 It has been shown that nanorods would penetrate tumors faster and more efficient
than nanoparticles.

16. Novel Drug Manufacturing for Oral Delivery of Poorly Soluble Drugs
16
100nm
Paclitaxel drug loaded micellar
nanorod
Acetaminophen drug assembled into nanoscale vias
Advantages:
 Poorly water-soluble drugs  orally acceptable
 Small size nanorods (~100nm)  high permeability, effective transport through
intestines
 Controlled size and shape  sufficiently high and fixed drug dosage
 Controlled aspect ratio  enhanced cell uptake  better bioavailability
 Composite drugs  overcome multi drug resistance
Before Assembly After Assembly

17. Utkan Demirci, PhD
Department of Radiology
Stanford University School of Medicine
Canary Center for Cancer Early Detection
Electrical Engineering (by Courtesy)
Scaffold-free 3-D cellular assembly
utkan@stanford.edu

18. Background-Tissue Engineering Strategies
• Top-down strategy
– Sculpting a large piece of
biomaterial into a well-shaped
and size-controllable scaffold
for tissue constructs
– Challenge: Hard to control cell
distribution and
microenvironment.
• Bottom-up strategy
– Assemble microscale cell
encapsulating building blocks into
large tissue constructs
– Challenge: Require biocompatible
intrinsic/extrinsic mechanism to
assemble building blocks
Soft Matter, 2009,5, 1312-1319

19. Trends in Biotechnology, 2015

20. 3D Micro-Masonary of Tissue Constructs
UV light
Photomask
Prepolymer
Solution
Glass
Slides
TMS/PMA
coated glass
Hydrogel Unit

21. 3-D Microgel Assembly Technologies
Nat. Comm. 2014 Adv. Mater. 2013 Adv. Mater. 2013 Adv. Mater. 2012 Biomaterials 2012 Biomicrofluidics 2012
Can we build complex 3-D constructs that would mimic native tissues,
e.g. for applications in drug testing?

22. Glass slide
500um GelMA without neuron
Neural Experiments Over Gel Size Control
100um GelMA with neuron
axon
Control

23. Spatial Size Control

24. Robots to Manipulate Single Gels
In collaboration with Dr. Metin Sitti-CMU.

25. Inspiration: Faraday waves (FWs)
Faraday waves: Standing waves arise through a parametric instability on the surface of a
vertically oscillated fluid layer.
 Wavelengths: micrometer scale to millimeter scale
 Diverse forms: stripes, squares, hexagons, quasi-periodic forms etc.
 Easy control of topography by frequency and amplitude
Source: Miles et al., Annu.Rev.Fluid Mech,22:143,1990
M Faraday

26. Spheroid Assembly

27. Typical Patterns

28. High throughput assembly system
System Throughput Max freq Feature
Old 1/ batch 200 Hz
New 6/batch >300 Hz Compatible with
multiwell plate
Old system New system

29. Retrieval of Constructs for Long-term culture

30. Biopreserving Microfluidic Bioprinted 3-D constructs

31. We can grow, maintain and cryopreserve embryoid bodies a microfluidic chip
Guven et al, Stem Cell Trans. Med. 2015
Microfluidic Systems: A Platform for Precision Medicine
Viability
Before cryopreservation After cryopreservation BrdU prolieration assay

32. Osteocalcin GFP HUVEC GFP HUVEC Osteocalcin
Bioprinting pre-vascularized osteogenic constructs
100µm
200µm
100µm
Mimicking the native histology

33. 3-D bioprinted and patterned EGFR2 mutated (red) and naïve
(green) lung cancer cells.
Migration Studies in Cancer Microenvironment

34. Conclusions & Future Directions
(i) Precision bioengineering method to create and analyze development and
function of specific neural circuits
(ii) Our laboratory’s mission is applying nano and micro-scale technologies to
real problems in medicine with impact at the clinic.
(iii) Non-invasive methods including Magnetic and Acoustic Fields are
promising technologies to build more complex 3D architecture to mimic
the native tissue microenvironment.
(iv) We can preserve function of stem cells on chip differentiated to ovarian
(v) We need the ability to preserve FUNCTIONAL vascularized cell, tissue,
organs preserving morphology, mechanics, and biology.

35. Panellist debate

36. Audience Q&A
Please use the chat / question / hand-raise functions in
GoToWebinar

37. ©PistoiaAlliance
We will set up all attendees on IP3
http://ip3.pistoiaalliance.org/
Nanoscale 3D-printing in pharmaceutical R&D18 June, 2015

38. Sustaining translational data for
secondary use
Join us for the next Pistoia Alliance Debates webinar,
Wednesday 15th July @ 11am-midday Eastern
https://attendee.gotowebinar.com/register/7562549094912217345

39. info@pistoiaalliance.org @pistoiaalliance www.pistoiaalliance.org
Thank you for joining us!