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PS-1 Main Body
PS-1 Main Body
PS-1 Main Body
Nano-porous materials containing pores between 2nm and 50nm in size are classified
as mesoporous particles according to IUPAC nomenclature. Their ordered pore
structure and high specific surface area render them desirable as adsorbents, for
catalysis, and as hosts for different kinds of molecules. Biological applications
include controlled drug delivery, enzyme encapsulation and phospholipid extraction
from biological matrices among others. Mesoporous Silica Nano-particles ( MSNs)
have been extensively studied as drug delivery agents, since 2001. “Their features
such as well-ordered internal mesopores (typically ca. 2–6 nm) with large pore
volume (0.6–1 cm3/g) and surface area (700–1000 m2/g), tunable size (50–200 nm) and
shape, robustness and easy surface modification, have revolutionised the field of
controlled drug delivery”. The downside however, is their aggregation in
physiological conditions and non-specific binding in protein containing solutions.
Collagen has been used with liposomes for drug delivery, to alter surface
properties of liposomes and as gel matrix to sequester liposomes. The present work
aims to include collagen in MPPs. The properties of collagen in non polar
environment and in the presence of lipids were studied, to better understand the
physical and chemical properties of collagen containing MPPs. Then MPPs containing
collagen were prepared and their properties were studied.
SDS-PAGE:
SDS-PAGE of collagen was performed to verify that the extract contained type-I
collagen. U.K. Laemmli’s protocol was followed.
6% Lechitin was added to each of the three solvents prepared above. True solution
was obtained by heating to 50 Degree Celsius.
Different volumes of water were added to each of the three prepared 6% Lechitin
solutions, and were incubated in a water bath at 50 degree Celsius. The maximum
volume of water that was soluble in the solutions were taken to be their water
holding capacities. The above determined concentrations were used to study the
behaviour of collagen in the presence of lipids.
The behaviour of collagen in Acetic acid, non polar solvents, and in the presence
of the lipid solvents, prepared as mentioned above were studied.
FIBRILLOGENESIS OF COLLAGEN:
A solution of 10% NaOH in Phosphate buffer of pH 7.2 was prepared. Equi-volume
collagen was added to study fibrillogenesis.
UV Spectroscopy:
The UV Spectrum of collagen in Acetic acid, in the three non polar solvents, and in
the presence of lipids were recorded, between 200 and 800nm. Water was used as the
blank for collagen in Acetic acid, while the non polar solvents were used as blank
for collagen in non polar solvents and in the presence of lipids. Fibrillogenesis
of collagen was recorded by measuring absorbance at 6 second time intervals at
313nm.
An alpha region consisting of two bands, a beta region consisting of two bands and
a gamma region consisting of a single band were obtained, conforming the presence
and type of collagen ( Collagen I) in the sample.
The spectra of collagen in the three non polar solvents and in the presence of
lipids were recorded and it was observed that there was a vertical shift in the
spectra and that there was no variation in the absorbance peak at around 210nm
( Fig 2). The more the upward shift, the less polar the solvent was. The polar
nature of collagen suggests that this maybe due to lesser solubility of collagen as
the polarity of solvent decreases.
Fig3: UV-Vis spectrum of collagen in the presence of lipids. (A) is the spectrum of
collagen in solvent 1 in the presence 1% and 6% Lechitin. (b) is the spectrum of
collagen in solvent 2 and in the presence of 6% Lechitin. ( C ) is the spectrum of
collagen in the presence of 6% and 1% Lipid.
The absorbance peak was found to be around 210nm in the presence of lipids also
( Fig 3). However, in the presence of lipids the absorbance values rose from 200 to
Absorbance maxima, unlike in the spectra recorded sans lipids. This suggests that