[The immunophenotyping and distribution of myofibroblasts in liver with fibrosis in fallow deer naturally infected with Fascioloides magna. [Doctoral dissertation]] | Imunofenotipizacija i distribucija miofibroblasta u jetri sa fibrozom kod jelena izazvanom Fascioloides magna. Doktorska disertacija
2011
Kukolj, V.
The subject of this dissertation was to examine liver samples from 20 adult fallow deer (Dama dama) of both sexes, naturally infected with the giant liver fluke (Facioloides magna). Pathohistological and immunohistological analyses were perfumed on liver samples that were fixed in 10% formalin, processed in an automatic tissue processor, and embedded in paraffin. Paraffin sections around 3-5 micro m thick were stained using heamatoxin eosin, Masson trichrome and Weigert van Gieson. The immunophenotyping of the cells that synthetize extracellular matrix was carried out using the streptavidin biotin (LSAB2) method and the double immunohistochemical staining technique, using antibodies for alpha-smooth muscle actin (alpha-SMA), desmin, transforming growth farcto beta1 (TGF-beta 1), receptor for transforming growth factor beta 1 (TGF-beta 1 RI), plateplet-derived growth factor A (PDGF-A), and cytokeratin. Semiquantitative analysis was used for the evaluation of necroinflammatory activity, ductular reaction, vascular lesions, and intensity of alpha-SMA expression. The number of perisinusoidal cells in livers with different degrees of fibrosis was determined using morphometric analysis. In addition to descriptive statistical parameters, ANOVA and Tukey test were also applied. Changes corresponding to different degrees of liver fibrosis, with pigmented areas and parasitic cysts in the parenchyma, were established by macroscopic examination in all 20 examined fallow deer. The obtained results suggest that all populations of liver myofibroblasts, in association with vascular and extravascular smooth muscle cells, participate in the stepped-up forming of extracellular matrix in the course of the development of parasitic liver fibrosis in fallow deer naturally infected with Fascioloides magna.
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