Title page for ETD etd-04102008-202907


Type of Document Master's Thesis
Author Vasanjee, Sunil C.
Author's Email Address vasanjee@vetmed.lsu.edu
URN etd-04102008-202907
Title Phenotypic Characterization of Canine Cranial Cruciate Ligament Associated Synoviocytes
Degree Master of Science (M.S.)
Department Veterinary Clinical Sciences
Advisory Committee
Advisor Name Title
Giselle Hosgood Committee Chair
Mandi J Lopez Committee Co-Chair
Jeffrey Gimble Committee Member
Keywords
  • immunohistochemistry
  • synovium
  • CrCL
  • Canine
Date of Defense 2008-02-07
Availability unrestricted
Abstract
Identification of synoviocytes surrounding the canine cranial cruciate ligament (CrCL) has not been investigated.

Objectives

1) Develop and validate a technique to identify and quantify normal canine CrCL associated synoviocytes.

2) Compare synoviocyte phenotype proportions surrounding normal and abnormal canine CrCLs

Design

In vitro experimental

Animals

Cranial cruciate ligaments from 4 intact female and 6 intact adult male mixed-breed dogs (objective 1) and from 8 adult female hound dogs (objective 2)

Methods

Objective 1) Normal CrCLs - CD18 and HSP25 epitopes were colocalized using immunohistochemistry. Sagittal sections were quantified in the proximal, middle, and distal aspects of each section. Western blot, RT-PCR and immunoelectron microscopy was used to confirm the presence of CD18 and HSP25 in the canine CrCL.

Objective 2) Normal, artificially stretched and naturally partially disrupted canine CrCLs -

CD18 and HSP25 epitopes were colocalized using fluorescent immunohistochemistry. Sagittal sections were prepared from the central aspect of each CrCL and phenotypes were quantified in the proximal, middle, and distal aspects of each section.

Results

Objective 1) Synoviocyte populations stained positive for CD18 (CD18+) or HSP25 (HSP25+), and a small population of cells stained for both epitopes (DS+). The proportion (mean SEM) of HSP25 + synoviocytes (57 7.5 %) was significantly greater than the proportion of CD18 + synoviocytes (27 8.2 %), which was significantly greater than the proportion of DS+ synoviocytes (16 3.5%). Western blot, RT-PCR and immunoelectron microscopy confirmed the presence of CD18 and HSP25 epitopes in the canine CrCL.

Objective 2) The pixel count for HSP25 + synoviocytes (57 7.5 %) was significantly greater than the proportion of CD18 + synoviocytes (27 8.2 %), which was significantly greater than the proportion of DS+ synoviocytes (16 3.5%) in all groups. There was no significant difference in the proportions of each of the phenotypes between CrCLs.

Conclusion

Three synoviocyte phenotypes were identified using immunohistochemical staining. Synoviocyte phenotype proportions did not differ between normal and abnormal CrCLs, however the HSP25+ synoviocytes were the predominant phenotype in all CrCLs.

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