An immunohistochemical study of feline myocardial fibrosis.

The aim of the present study was to investigate the pathology of feline myocardial fibrosis. The hearts from 40 cats with myocardial fibrosis were compared with the hearts from 25 normal cats. Clinical data were available in 11 cases. Hearts with myocardial fibrosis were hypomotile and there were hyperechoic areas in the ventricular wall on echocardiography. The presence of myocardial fibrosis was correlated significantly with hypertrophy of the ventricles, atrial dilation and angiosclerosis. Immunohistochemical studies demonstrated that normal feline cardiomyocytes expressed matrix metalloproteinase (MMP)-2, MMP-9, MMP-14, tissue inhibitor of matrix metalloproteinase (TIMP)-2 and transforming growth factor (TGF)-ß2. Fibroblasts in normal hearts expressed only TIMP-2. In the hearts with myocardial fibrosis, expression of MMP-2, TIMP-3 and TGF-ß2 by cardiomyocytes was significantly increased, but TIMP-2 expression was diminished. Fibroblasts in the affected hearts showed expression of MMP-14 in several cases. These findings suggest that a complex fibrotic remodelling of the feline myocardium occurs in this disease and that cardiomyocytes are involved in this process.

Distribution of extracellular matrix components in normal and degenerated canine tricuspid valve leaflets.

The aim of the present study was to investigate the composition and distribution of various extracellular matrix (ECM) components in normal canine tricuspid valves (TVs) and in TVs affected by chronic valvular disease (CVD). The parietal (pTV) and septal (sTV) leaflets of the TVs from 27 dogs were investigated immunohistochemically for expression of collagen types I, III, IV and VI, elastin, laminin, fibronectin and heparan sulphate. Normal pTV consisted mainly of elastin and collagen VI in the atrialis, fibronectin in the thin spongiosa and mixed collagens in the fibrosa. The layered structure was less distinct in sTV, with numerous adipocytes and proteoglycans in the spongiosa and collagen III predominating in the fibrosa. The earliest stages of CVD affecting the pTV were recognized in the spongiosa and progression to advanced disease was characterized by nodular accumulation of proteoglycans within the free edge of the leaflet. These nodular lesions of the pTV contained more fibronectin, elastin and collagens I and VI than those affecting the sTV. These findings contrast with those reported in CVD affecting the mitral valve (MV) in which the early lesions affect the atrialis and advanced disease involves the entire leaflet. The pathogenesis of CVD in TV may involve initial alterations of the tricuspid annulus that lead to early lesions within the spongiosa, resulting in further shear stress and proteoglycan accumulation at the free edge of the pTV.

Expression of genes encoding matrix metalloproteinases (MMPs) and their tissue inhibitors (TIMPs) in normal and diseased canine mitral valves.

The pathogenesis of canine chronic valvular disease (CVD) is not fully characterized. The present study investigates the expression of genes encoding matrix metalloproteinases (MMPs) and their tissue inhibitors (TIMPs) in normal and diseased mitral valves (MVs). Samples from normal (n=15) or diseased (n=10) canine MVs were subject to real-time polymerase chain reaction (PCR) for quantification of mRNA encoding MMP-1, -2, -9 and -14 and TIMP-2, -3 and -4. In normal valves there was low expression of mRNA encoding MMP-2, -9 and -14 and TIMP-3. In the valves from dogs with CVD there was significantly increased transcription of mRNA encoding MMP-1 and -14 and TIMP-2, -3 and -4, but no elevation in mRNA encoding MMP-2 and -9. MMPs and TIMPs are therefore likely to be involved in extracellular matrix metabolism in normal canine MVs and there are significant alterations in the expression of genes encoding these molecules during CVD.

Detection and characterization of chondroid metaplasia in canine atrioventricular valves.

The atrioventricular valves of 25 dogs of different breeds and age were examined grossly and microscopically following histochemical staining and immunohistochemical labelling for collagen types I, III and VI, and for fibronectin and laminin. Foci of cartilage were identified in the tricuspid septal leaflet within the fibrosa (n=21) or spongiosa (n=3). These were further characterized as either fibrocartilage, predominantly composed of collagens I and VI, or hyaline cartilage consisting of laminin and collagens III and VI. Eighteen of the dogs were of large breed and seven of small breed. Retrospective echocardiographic findings were available from five cases and in three of these a hyperechogenic structure was identified corresponding to the cartilage focus (0.1, 1.12 and 5.63 mm(2) in size). The clinical significance and mechanism of formation of these cartilaginous foci remain undetermined, although factors such as breed, size and concurrent chronic valvular disease may be significant.

Expression of transforming growth factor-beta1, -beta2 and -beta3 in normal and diseased canine mitral valves.

The pathogenesis of chronic valvular disease (CVD) in dogs remains unclear, but activation and proliferation of valvular stromal cells (VSC) and their transdifferentiation into myofibroblast-like cells has been described. These alterations may be influenced by transforming growth factor-beta (TGF-beta), a cytokine involved in extracellular matrix (ECM) regulation and mesenchymal cell differentiation. The present study investigates immunohistochemically the expression of TGF-beta1, -beta2, -beta3 and smooth muscle alpha actin (alpha-SMA) in normal canine mitral valves (MVs) (n=10) and in the valves of dogs with mild (n=7), moderate (n=14) and severe (n=9) CVD. In normal mitral valves there was no expression of alpha-SMA but VSC displayed variable expression of TGF-beta1 (10% of VSC labelled), TGF-beta2 (1-5% labelled) and TGF-beta3 (50% labelled). In mild CVD the affected atrialis contain activated and proliferating alpha-SMA-positive VSC, which strongly expressed TGF-beta1 and -beta3, but only 10% of these cells expressed TGF-beta2. In unaffected areas of the leaflet there was selective increase in expression of TGF-beta1 and -beta3. In advanced CVD the activated subendothelial VSC strongly expressed alpha-SMA, TGF-beta1 and -beta3. Inactive VSC within the centre of the nodules had much less labelling for TGF-beta1 and -beta3. TGF-beta1 labelling was strong within the ECM. These data suggest that TGF-beta plays a role in the pathogenesis of CVD by inducing myofibroblast-like differentiation of VSC and ECM secretion. Changed haemodynamic forces and expression of matrix metalloproteinases (MMPs) may in turn regulate TGF-beta expression.

Primary and secondary heart tumours in dogs and cats.

Primary and secondary neoplasms of the canine and feline heart are uncommon. During a 2-year period, 83 dogs suffering from primary cardiac (n=11), extracardiac benign (n=6) or malignant (n=66) tumours and 30 cats with primary cardiac (n=1) or extracardiac (n=29) malignant tumours were examined. Echocardiography revealed four cases of primary cardiac neoplasms in dogs, but secondary heart tumours were not detected. After necropsy, tissue samples from the heart and tumours were examined histologically and immunohistochemically. In dogs, primary neoplasms included seven haemangiosarcomas, two chemodectomas, one rhabdomyosarcoma, and one neurofibrosarcoma. In 24 of 66 dogs examined, metastases of extracardiac neoplasms were found in the heart (15 carcinomas, six malignant lymphomas, three haemangiosarcomas). In cats, one case of primary haemangiosarcoma of the pericardium and five cases of secondary cardiac tumours (two malignant lymphomas, three carcinomas) occurred. Cardiac neoplasms in cats were not identified clinically but were detected by detailed gross sectioning of the heart (n=2) or histopathological examinations (n=3). This study showed an unexpectedly high number (36%) of dogs with cardiac metastases.