Beruflich Dokumente
Kultur Dokumente
of Biomaterials
MS 413100_Biomedical materials
Date: 2016.01.08
Tzu-Wei Wang, PhD.
MCD:
sintered apatite grit
H(OCH2CH2)nOH
Q:
Consider the following hypothetical protein and two
materials (A) and (B). Assume that in this case the water
contact angle correlates directly with the hydrophobicity
of the material surface. To which material would greater
adsorption of the given protein be expected? Why?
A:
Material B has a greater contact angle than A. Material B
is more hydrophobic than Material A. Consequently, one
would expect greater adsorption of the highly
hydrophobic protein X to the surface of Material B,
through hydrophobic interaction, assuming that the
surface charges of Materials A and B are comparable.
Plasma Treatment
Chemical Vapor Deposition
Physical Vapor Deposition
Radiation Grafting
Photografting
Self-Assembled Monolayers
Plasma Treatment
.
http://en.wikibooks.org/wiki/Microtechnology/Additive_Processes
(e-)
(N2)
(2N+)
(2e-)
()
(Plasma sputtering)
(Plasma coating)
(Ex: Acrylic Acid, N-Vinyl 2-Pyrrolidone, HEMA, NIPAAm, etc )
(
)
(Plasma grafting)
(Ex: -OH)
1. are conformal
2. are free of voids/pinhole defects
3. are easily prepared
4. are sterile when removed from the reactor
5. produce a low amount of leachable substances
6. demonstrate good adhesion to substrate
7. allow unique film chemistries to be produced
8. can be characterized relatively easily
disadvantages
REACTOR
R.F
GENERATOR
VACUUM
GAUGE
sample
MONOMER
SOURCE
GAS SOURSE
(nitrogen, argon, oxygen, water vapor)
DIFFUSION
PUMP
ROTARY
PUMP
Radiation Grafting
The substrate is exposed to a radiation source of high
energy, which forms reactive species at the surface to
create covalent bonding of the coating to the underlying
material.
These methods are often employed to bind hydrogels to
hydrophobic substrates.
This technique also provides a means to easily tailor the
properties of the coating since a mixture of monomers or
other precursors can be used.
For the removal of toxic heavy metals from industry waste water
Prof. Masao Tamada, Environmental and Industrial Materials Research Division, Japan Atomic Energy Agency
Photografting
Photografting is similar to radiation grafting, but, the
radiation is UV or visible light.
A number of photoresponsive chemical moieties have
been developed to facilitate this type of surface
modification (viz., phenyl azide or benzophenone
).
BP
Solution Coatings
In this technique, the substrate is dipped in
a solution containing the dissolved coating
material (usually a polymer or proteins
dissolved in an organic solvent or aqueous
solution). The substrate is then left to dry
and, as the solvent evaporates, the
coating is deposited on the surface.
1. Selective adhesion
Ridge
2. Guidance
valley
Microcontact printing
Fig. In microcontact printing, a mold of
the desired pattern is first created, often
via photolithographic techniques on a
silicon wafer (a-e). A silicone rubber
material (PDMS) is then polymerized in
the mold to make a positive "stamp (fh). The stamp is "inked with the
surface-modifying substance by
dipping it into a solution containing the
molecule of interest (i) and the inked
stamp is then pressed onto the
substrate (j). After carefully removing the
stamp, the process can be repeated to
modify the portion of the substrate that
was not stamped the first time, thus
creating multifunctional surfaces. In this
case, a biomaterial could be modified
to expose one or more proteins with a
well-controlled spatial orientation, thus
potentially modulating cell attachment and
improving interaction of the implant with
the native tissue.
PDMS: polydimethylsiloxane
(A)
UV
PDMS
70
(B)
12 1 1,2
1. ; 2.
ES cells
NIH-3T3 cells
NIH-3T3 cells
ES cells
Fig. 4. Patterned co-cultures of ES cells with fibroblasts.
(A) represents light (left) and fluorescent (right) images
after 1 day. (B) and (C) illustrate that co-cultures remained
stable for 3 and 5 days. (D) represents the reversal in the
order of cell seeding.
AML12 cells
(hepatocytes)
NIH-3T3 cells
Microfluidics
Fig. For the micro fluidics technique, (a) the mold is fabricated as described in
microcontact printing, but the mold is a positive, rather than a negative, image of the
desired design. (b) PDMS is then polymerized in the mold. (c) The formed PDMS is
then removed from the mold and pressed against a glass slide. This setup is
then plasma-treated to increase the hydrophilicity of inner areas of the
channels only, while the regions between channels remain hydrophobic to maintain
the integrity of each channel. Like for microcontact printing, (d) the PDMS form is
then pressed against the substrate and a small amount of a solution containing
the molecule of interest is injected or placed near the opening of a channel,
where capillary action pulls the liquid throughout the channels. The areas of the
substrate under the channels are appropriately modified. (e) After the molecule has
reacted with the surface, the PDMS form is removed and the surface is rinsed.
A distinct advantage of this process is that it takes very little fluid volume, so it can be
easily used with expensive or difficult-to-produce biologic reagents.
http://en.wikipedia.org/wiki/Microfluidics
Crosslinker
Direct coupling
poly-ethylenimine (PEI)
PEG
poly(divinylbenzene) (polyDVB)
poly tert-butyl acrylate (PtBA)
Journal of Polymer Science: Part A: Polymer Chemistry, Vol. 48, 99108 (2010)
Q:
A researcher has created a biodegradable polymeric
implant material that degrades through a surface
degradation mechanism. The material is intended to
serve as a tissue engineering scaffold, in which longterm cell adhesion to the material is important. Initial
studies have found that cells will not adhere to the
surface of the material. The researcher is considering
covalent attachment [with a poly(ethylene glycol)
spacer of 3400 Da] of a peptide sequence known to
improve cell attachment to other materials to the surface
of this material. The researcher asks if you support the
idea. Will you support the idea? Why or why not? Would
bulk modification with the peptide sequence be a more
or less appropriate method of modification for the
intended result?
H(OCH2CH2)nOH
A:
The idea is not feasible to yield the intended effect for the given
application and should not be supported. Recalling that degradation
by surface erosion involves the continual loss of the material at the
surface of the construct (much like a bar of soap disappears with
time of exposure to water as the surface continually erodes away), it
follows that any modification to the surface of this material would
only be viable for a short period of time until the modified surface
degrades away. Following the initial surface degradation, the cells
will be exposed to the bulk unmodified material, to which cell
attachment has been shown to be nil. Thus, although the cells may
attach initially, long-term cell attachment could not feasibly be
attained with this modification technique. Bulk modification, however,
would be a more appropriate technique because the cell adhesion
peptide would be present throughout the bulk of the material, unlike
surface modification. As a result, cells could potentially attach to the
material throughout the stages of degradation.
Immobilized Enzymes
enzymes are a subclass of proteins that act to promote specific
chemical reactions involving other biomolecules.
controlled release
Nanotechweb.org
Instrumentation
1. Holder for solid sample
2. Holder for liquid
3. Means to determine contact angle (may be automated)
Unlike many other characterization techniques, the output of
contact angle analysis is a single number ( or c ).
because the contact angle measurement is not always fully
automated, user bias can have a substantial effect on the results.
Fig. Different experimental setups
for determining the contact angle: (a)
sessile drop, (b) captive air bubble,
(c) capillary rise method, and (d)
Wilhelmy plate method. The circles
indicate where the contact angle is
measured.
Light Microscopy
Light microscopy is a relatively simple technique
that is used as a first approach to gain primarily
qualitative information about surface topography,
or to view thin sections of a sample.
For opaque samples, the light source can be
located above rather than below the sample.
The resolution of such instruments is limited by
the wavelength of white light, and in most cases
is around 0.2 m.
Instrumentation
Information Provided
Light microscopy is used exclusively for imaging, and provides in
itself only qualitative assessment. Images can be further analyzed
using specialized software to obtain semi-quantitative measurements.
While light microscopy has the advantage that vacuum is not required
to view samples, it remains difficult to see thick or hydrated samples.
XPS-ESCA
The binding energy gives an idea of how tightly bound the electron is
to the nucleus, and it changes in accordance with the type of atom as
well as interactions with the nuclei of neighboring atoms.
Ek: the kinetic energy of the electron
Eb: the binding energy of the electron
is the frequency
h is the Planck's constant (6.6 X 10-34 J-s)
(X-Ray)
(photoelectron)
Fig. Schematic of ESCA equipment. First, the sample is bombarded with Xrays. The resulting emitted electrons then enter the analyzer chamber.
Because of the difference in voltage between the two walls and the
geometry of the analyzer, only electrons with a certain kinetic energy
can be collected by the detector, with the remaining electrons striking
non-detectable areas. By altering the voltage difference between the walls
in a controlled manner, the electrostatic field is altered to permit the detector
to record the amount of electrons having various kinetic energies. At the
end of the voltage sweep, the entire spectrum is plotted for that sample.
Instrumentation
1. Source-produces X-rays with known wavelength.
2. Electron analyzer-uses an electrostatic field to separate
electrons based on kinetic energy.
3. Detector-converts impact by separated electrons into an
electrical signal.
4. Processor (computer)-translates the signal from the
detector into the appropriate spectrum.
Information Provided
ESCA methods are extremely sensitive and can detect all
elements except hydrogen and helium in the outermost
~100 of an organic or inorganic material at
concentrations down to 0.1 atomic percent.
ATR-FTIR
Attenuated Total Internal Reflectance Fourier Transform-Infrared Spectroscopy (ATR-FTIR)
Ewing G.E. (2004): Thin film water. J. Phys. Chem. B. 108, 15953-15961
SIMS
Secondary Ion Mass Spectrometry (SIMS)
Ionization begins when primary ions, such as O2+, Ar+, Xe+, or Cs+,
are ejected from an ion gun and strike the sample surface. This
causes the surface layer of atoms to be stripped off, or sputtered,
both as neutral species and as ions.
These emitted ions are called secondary ions and are drawn into
the analyzer for separation by mass in a similar manner to bulk
mass spectroscopy.
(Leibniz Institute for Solid State and Materials Research, Dresden, Germany)
Information Provided
SIMS provides information about the
structure and composition of the
outermost few of both inorganic and
organic materials, although the accuracy
of quantitative methods for this type of
spectroscopy is limited. With dynamic
SIMS, composition as a function of depth
can also be recorded.
Fig. A spectrum
produced by a type of
SIMS for fibronectin
adsorbed on a
poly(styrene) surface.
The various peaks
correspond to different
amino acids found in
the fibronectin protein.
By comparing the
relative intensities of
certain peaks as the
protein is adsorbed on
different surfaces, the
biomaterialist can
obtain information
about the orientation
of the protein on each
surface.
http://virtuallaboratory.net/OmniaCellula/Contents/Topic6-4_Signaling.htm
Keratinocytes
Because electrons readily interact with atoms in the air, all electron
microscopy must be completed under vacuum.
Tapping mode
Fig. Schematic of AFM instrumentation. Imaging begins when the sample is placed on a stage
and the tip is lowered until it contacts the surface. When this occurs, reflection of a laser off the
cantilever indicates that the cantilever is bent toward the sample. The tip/cantilever are then
moved across the sample surface and the cantilever deflection is monitored. In response to
changes in defection, the stage is moved up and down to maintain contact between the sample
and the tip. Alterations in stage position are recorded and processed by the appropriate software
to form a three-dimensional image.
http://niufood.niu.edu.tw/img.php?img=662_5112
1f09.png&dir=users_sharing/1
AFM Applications-DPN
Dip-Pen Nanolithography
Nanografting
Direct DPN
Indirect DPN
http://www.sciencedirect.com/science/article/pii/S0304416510001133#gr6