INTERSTITIAL DISTRIBUTION OF COLLAGEN &HYALURONIC ACID

Project: Research project

Description

The objectives of this study are to define the distribution of collagen and
hyaluronic acid in the interstitium of the mesentery and, with the same
data, evaluate sensitivity of UV-IR absorption for estimating protein and
fluid. Because two molecules cannot occupy the identical space, collagen
and hyaluronic acid exclude protein and fluid in the interstitium. This
exclusion modulates the volume of free fluid in the interstitium and thus
edema formation. The experimental procedure estimates protein mass and fluid volume by using
differential absorption at 280 and 320nm (corrected for collagen
scattering) and 1000 and 1500nm, respectively, in accordance with the
Lambert-Beer relationship. Sampling a TV image provides an array of 512 x
512 bytes of data. Using a 20X objective in our TV-microscope system yields
a microregion sample of about 2um2. TV images are grabbed, digitized,
integrated and stored within 5 seconds for each wavelength. The
experimental protocol involves obtaining 3 sets of control data, then
superfusing with either collagenase or hyaluronidase or a combination. TV
sampling will be done every 15 minutes for I hour, followed by 30 minute
intervals until enzyme action is completed. Enzyme application causes degeneration of the interstitial constituents
thereby reducing the extent of exclusion and permitting a more uniform
distribution of protein and fluid to occur. Comparing the distribution of
protein and fluid before and after the enzyme should reveal the
distribution of the interstitial constituents. Microregions which exhibit
increases in fluid and protein following collagenase treatment will
represent regions of high collagen content. Similarly, microregions which
exhibit disproportionate increases in protein following hyaluronidase
treatment will reflect regions which had high hyaluronic acid content. The
ability of the UV-IR absorption procedure to detect these changes of
distribution will reveal the sensitivity and specificity of the method. This study may serve to identify the true nature of the perivascular
protein concentration critical to the evaluation of the mechanisms of
transvascular protein and fluid transport and possibly identify the extent
to which alterations of the distribution and integrity of the interstitial
matrix may contribute to edema formation.
StatusFinished
Effective start/end date7/1/906/30/95

Funding

  • National Institutes of Health
  • National Institutes of Health: $88,160.00

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Hyaluronic Acid
Collagen
Proteins
Collagenases
Enzymes
Hyaluronoglucosaminidase
Mesentery
Protein Transport
Edema
Sensitivity and Specificity
Acids

ASJC

  • Medicine(all)