Biomedical Engineering Reference
In-Depth Information
IFN-γ production on allergen sensitization was also tested by giving the mice
an intranasal treatment of 10 µg of cIFN that constituted the prophylactic cIFN
regimen. Some mice were given cIFN treatments after OVA sensitization as
a therapeutic regimen. In both the prophylactic and therapeutic treatments, a
final challenge with OVA were given before the mice were euthanized, and the
lungs were perfused in situ with phosphate-buffered saline (PBS), removed,
and fixed in 4% buffered formalin. From each group, lungs were left unfixed
and homogenized for lymphocyte isolation.
73
The whole lungs that were for-
malin-fixed were paraffin-embedded and sectioned into 15-micron portions.
From each mouse two sections were dewaxed, rehydrated, and labeled with
either anti-IL-12Rb2-fluorescein isothiocyanate (FITC) (Th1) or anti-T1/
ST2L-rhodamine (Th2) that were examined with a Nikon TE300 fluorescence
microscope.
7.4.2 The Use of Ceramic Core NPs as Delivery Systems
for Immunotherapy
The group of Pandey
111
evaluated carbohydrate modified ultrafine ceramic core
based NPs (aquasomes) as delivery systems (adjuvants) for immunotherapy
with ovalbumin (OVA) as model allergen. Both humoral and cellular-induced
immune responses were studied in BALB/c mice using aquasomes and alumi-
num hydroxide as allergen-delivery systems following the protocol described
below.
7.4.2.1 Preparation of Hydroxyapatite NMs
The hydroxyapatite (HA) core was first prepared following published protocols
using the self-precipitation method.
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The process is as follows:
(1)
Add 0.5 M • (NH
4
)
2
HPO
4
to 0.5 M Ca(NO
3
)
2
• 4H
2
O ethanol solution with
vigorous stirring. The pH of both solutions has to be 10 by adding NH
4
OH
drop wise.
(2)
Stir the mixture vigorously for 1 h using mechanical stirrer (REMI C-24,
Mumbai, India).
(3)
Dry the mixture at 40 °C in oven overnight.
(4)
Collect and wash the precipitated hydroxyapatite (ceramic) particles with
ethanol and then with distilled DI water.
(5)
Dry the washed ceramic particles overnight at 40 °C.
(6)
Use Fourier transformed infrared spectroscopy (FT-IR) for structure analy-
sis. Prepare the KBr sample disk using 1% (w/w) of hydroxyapatite pow-
der and dry at 100 °C. Record the spectra at 4000 to 400 cm
−1
(resolution
4.0 cm
−1
).
(7)
Perform phase analysis using X-ray diffraction (XRD) with Cu Kα radia-
tion (45 kV × 40 mA) in a wide-angle X-ray diffractometer (XRD-6000,
Shimadzu, Japan).
