Phase II Renewal Grant for the Nanoscale Science and Engineering Center (NSEC) for Affordable Nanoengineering of Polymeric Biomedical Devices (CANPBD) PIs: L.J. Lee, A.T. Conlisk, S.V. Olesik, D.L. Tomasko, R.J. Lee / NSF NSEC Grant: 0914790

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Phase II Renewal Grant for the Nanoscale Science
and Engineering Center (NSEC) for Affordable
Nanoengineering of Polymeric Biomedical Devices
(CANPBD) PIs L.J. Lee, A.T. Conlisk, S.V.
Olesik, D.L. Tomasko, R.J. Lee / NSF NSEC Grant
0914790
Intellectual Merits
For example, polymer and lipid-based molecular
and nanoparticle formulations to be developed for
ON delivery are shown in the second figure. The
lipid-ON conjugate through a disulfide bond is
designed for enhanced cell uptake and quick
release from endosomes. It can be used alone as a
molecular therapy agent or loaded into
nanoparticles. The hydrophobic molecule
conjugated cationic polymer (HMCCP) can form
stable micelles with ON. Like lipid-ON
conjugates, its small size is particularly
valuable for tumor penetration. Unlike lipid-ON
conjugates, micelles allow higher ON loading.
Again, they can be loaded in nanoparticles for
delivery. The two-stage liposomal nanoparticles
are designed for efficient release of cargos from
nanoparticles when they reach the targeted tissue
like a tumor. It is highly desirable to have a
synthesis method with which these formulations
can be precisely prepared and evaluated. As more
components such as multiple drugs, nuclear
localization signal (NLS) to enhance nuclei
permeation, integrase to integrate therapeutic
genes with host genes, and imaging agents for
additional functionalities are added, the only
viable approach to construct such multifunctional
complexes is to establish a nanofluidics-based
nanofactory assembly line.
2. Nanofiber-Based Nanofactory for Cell
Separation and Analysis The second nanofactory
integrates a foundation of electrospun nanofiber
with post-processing using femtosecond laser
ablation and supercritical fluid-based
impregnation. The figure below shows an example
of cell separations that can occur 2(A) stem
cell separation into different expansion chambers
based on relative adhesion into patterned
microwells and magnetic manipulation 2(B)
migratory cell separation on aligned nanofiber
arrays followed by expansion and subsequent
analysis to establish differences in
intracellular profiles. Electrospinning provides
synthetic analogs of biological structures found
in vivo. Femtosecond laser machining provides a
key benefit by shaping electrospun matrices at
the micron or submicron level to provide flow
channels, separation chambers and guided
assembly. Subcritical CO2 allows printed
additions of chemical functionality/bioactivity
in an efficient manner. A broad range of
envisioned medical uses includes
microenvironments for stem cell differentiation,
artificial organs and cancer treatment.

• The research vision of CANPBD is to revolutionize
  medical diagnosis and medicine by establishing an
  affordable multiscale synthesis and fabrication
  protocol leading to nanofluidic and polymer
  therapeutic devices for personalized
  nanomedicine.
• An important emphasis of Phase II is to
  commercialize the developed technologies in close
  collaboration with end users.
• The broader impacts of the activities planned for
  Phase II are to
• commercialize nanoengineered biomedical devices
  through affordable manufacturing methods and
  novel design
• extend research results from medical/biology
  applications to functional nanocomposites, water
  treatment, homeland security, environmental
  protection, and food industry toxicology
• establish new products and new industries to
  create high-paying jobs
• train the 21st century workforce in economically
  important and critical high-tech fields.

• Phase II System-Level Integration
• Nanofactory Assembly Lines
• In the following paragraphs, we describe two
  nanofactory assembly lines particularly useful
  for the aforementioned biomedical applications
  and how relevant CANPBD technologies and
  scientific studies will be applied to their
  design, fabrication, and bioevaluation.
• Nanofluidics-Based Assembly Line
• Based on nanofluidics, one nanofactory integrates
  nanomanufacturing and nanomanipulation arrays
  with a novel DNA combing and imprinting (DCI)
  process and nanowire electric circuit designs.
  Synthesis of multifunctional nanoparticles for
  gene delivery, drug/gene injection to living
  cells by nanochannel electroporation, nanowire
  biosensor arrays, and nanofluidics DNA separation
  are possible applications. In the following, we
  first describe the design of this nanofactory
  assembly line for the first two applications and
  then explain relevant nanotechnologies needed to
  realize the design.
• Example Synthesis of Multifunctional
  Nanoparticles for Gene Delivery. A
  multifunctional nanovector formulation that can
  increase blood circulation time, target specific
  tissues/cells and enhance intracellular release
  is shown in the schematic that follows. Although
  this is a relatively simple formulation, the
  actual structures of forming nanoparticles using
  the current synthesis methods are far from this
  ideal shape because of poor component
  distribution at the nanoscale. Consequently, the
  gene loading is very low (lt10 of the total
  nanoparticle weight), cytotoxicity is high and
  transfection efficiency is poor and inconsistent.
  Without the capability to prepare well-defined
  nanoparticles, our ability to design and evaluate
  new formulations is hindered.

Nanofactory 2(A) Adhesion Chromatography of Stem
Cells
Nanofactory 2(B) Migratory Chromatography of
Motile Cells
This material is based on work funded by the
National Science Foundation (Grant 0914790)