Preparation of HEMA scaffold using concave glass Jin-Oh You

trextemperMechanics

Feb 22, 2014 (3 years and 3 months ago)

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Preparation of HEMA scaffold using concave glass

Jin
-
Oh You



1.
PMMA microspheres in diH
2
O

(20 wt

%)
are

mixed homogeneously, and 200 µL of
the microsphere suspension
is

placed on a
25 mm

concave glass cover
.



2.
After the PMMA microspheres
are
settled
, the water
i
s evaporated at 65
o
C

in the
oven
.



3. The polymer solution is prepared with
HEMA, TEGDMA,
d
iH
2
O,
ethylene glycol
,
and DMPAP

(the ratio can be changed)
.


4.
70 µL of HEMA solution
i
s added in the dried PMMA lattice and then pla
ced

under
UV
light (21.7 mW/cm
2
, 365 nm) for 90 seconds.



5.
After photopolymerization, the gel
i
s immersed in 10 mL of dichloromethane while
shaking vigorously on an orbital shake
r

for 48 hours to remove the PMMA
microspheres.



6.
The scaffold
i
s placed in diH
2
O

f
or 24 hours and washed.


7.
The scaffold
i
s cut to the desired shape using a cork borer.




S
ed
imentation
-
evaporation method for s
caffold fabrication


1.

PMMA
microspheres dispersed in water were allowed to settle to the

bottom of a 50mL centrifuge tube


2.

B
y gently

shaking the sample, the PMMA

microspheres could be arranged into

a closely packed structure during water evaporation.


3.

Place lattice in
an oven heated at
145°
C (low
er than the melting point of PMMA
,

160°C) for 50 min. (Please check PMMA particle
melting point on the spec
sheet)


4.

After dropping a mixture of water and ethanol (5:5, v/v) to wet the

surface of the PMMA

microspheres,
add polymer solution and UV for 90 s.


5.

The lattice pellet with polymer
solution was then frozen at

-
20
°
C for 3 h and
fin
ally freeze
-
dried.


6.

Proceed with our previous scaffold protocol. (water o/n, punch, drying o/n, DCM
3 nights, etc).


7.

Freeze
-
dry again and prepare sample for SEM.