Jarkko Hietanen

Oral and salivary parameters in patients with rheumatic diseases.

We studied the presence of secondary Sjögrens syndrome (SS) and the composition of saliva, prevalence of oral pathogens, periodontitis, mouth mucosa, and teeth in patients with various rheumatic diseases and in healthy controls. The hypothesis was that different rheumatic diseases might cause differences in oral health characteristics because of the liability of secondary SS in the patients. The study involved 77 patients and 77 age-matched and sex-matched controls. Twenty patients were suffering from spondylarthropathy (SPA), 18 from ankylosing spondylitis (AS), 24 from rheumatoid arthritis (RA), and 15 from mixed connective tissue disease (MCTD). Clinical and radiographic oral health status was recorded and salivary flow rates were measured. Selected salivary proteins and immunoglobulins were analysed by routine methods. Minor salivary gland biopsy samples were taken from the patients for assessment of inflammatory focus scores. Differences between patients and controls and in between the different rheumatic diseases were analysed statistically. Secondary SS was diagnosed in 39% (30/77) of the patients. A severe periodontal condition (community periodontal index of treatment needs score 3 or 4) occurred in 58% (45/77) of the rheumatic patients compared with only 26% (20/77) of the controls (p<0.0001). The severity of focal sialadenitis (focus score) correlated significant with salivary IgA, IgG, and IgM concentrations. Salivary albumin, total protein, IgG, and IgM concentrations were higher in all patient groups than in the controls. The number of patients with low salivary flow rates was higher in all patient groups compared to controls. Oral yeast counts were significantly higher in the patients than in the controls (p<0.001). In a subgroup analysis, patients with SS had higher values for salivary IgA and IgM than patients without SS. Dental caries and oral lactobacilli were more frequent in patients with SS, but SS was not associated with periodontitis. No major differences were noted in other salivary biochemical parameters between these two groups. Patients with rheumatic diseases, irrespective of specific diagnosis, thus had various alterations in salivary flow and composition and oral health. The findings may reflect the autoimmune inflammation of the salivary glands frequently observed in these patients.

Gelatinase A (MMP-2), Collagenase-2 (MMP-8), and Laminin-5 γ2-Chain Expression in Murine Inflammatory Bowel Disease (Ulcerative Colitis)

Dextran sulfate sodium-induced inflammatory bowel disease in mice resembles human ulcerative colitis. In inflammatory bowel diseases matrix metalloproteinases contribute to tissue degradation. Laminin-5 is an anchoring filament protein in the basement membrane area that can be cleaved by matrix metalloproteinases. We investigated the expression of matrix metalloproteinases-2 and -8 and laminin-5 γ2-chain in dextran sulfate sodium-induced mice by immunohistochemistry and in situ hybridization. Matrix metalloproteinase-8 expression was evidenced in the colon surface epithelial cells and the protein was more abundant in dextran sulfate sodium-induced mice colon. Matrix metallproteinase-2 and laminin-5 γ2-chain colocalized in the colon surface epithelial cells and in the basement membrane zone as demonstrated by double immunostaining. In dextran sulfate sodium-induced colon, matrix metalloproteinase-2 immunoreactivity was detected in epithelial cells in the lower parts of the crypt and surrounding the degraded crypts. Matrix metalloproteinase-2 and -8 could participate in the local epithelial inflammatory processes and tissue destruction. The presence of laminin-5 γ2-chain indicates alternative anchoring mechanisms in the colon, a compartment devoid of hemidesmosomes.

Soluble membrane‐type 1 matrix metalloproteinase (MT1‐MMP) and gelatinase A (MMP‐2) in induced sputum and bronchoalveolar lavage fluid of human bronchial asthma and bronchiectasis

The aim of this study was to investigate the involvement of the MT1‐MMP/MMP‐2 cascade in induced sputum (IS) and bronchoalveolar lavage fluid (BALF) from bronchial asthma (BA) and bronchiectasis (BE) patients and healthy controls. The molecular forms and cellular origins of MT1‐MMP and MMP‐2 were determined by Western immunoblotting, immunohistochemistry and in situ hybridization. Elevated levels of soluble activated and autocatalyzed MT1‐MMP species as well as activated forms of MMP‐2 in IS and BALF samples from BA and BE patients were evidenced. The activation degrees of soluble MT1‐MMP and MMP‐2 were significantly correlated in BA and BE IS and BALF. Only low levels of both these MMPs were observed in healthy control IS and BALF. The co‐expression of MMP‐2 with MT1‐MMP was evidenced by double immunostaining in bronchial epithelial cells, submucosal glandular cells, smooth muscle cells and monocyte/macrophages. The MT1‐MMP/MMP‐2 cascade is present and active in human inflammatory lung disease fluid and tissue samples. This cascade seemingly reflects the active destructive phases of these chronic lung diseases.

HLA-DRB1* alleles and temporomandibular joint erosion in patients with various rheumatic diseases

Objective: To investigate the association between HLA antigens and temporomandibular joint (TMJ) erosion, salivary composition, and focal sialadenitis in patients with rheumatic diseases. Methods: Eighty‐four patients, 24 with rheumatoid arthritis (RA), 19 with mixed connective tissue disease (MCTD), 19 with ankylosing spondylitis (AS), and 22 with spondyloarthropathy (SPA) were studied. Each patient underwent clinical examination of the masticatory system, unstimulated and stimulated saliva collection, and minor salivary gland biopsy. Radiographs (OPTG) of the TMJ were obtained, and HLA allele (A, B, C and DRB1*) analysis was performed. Erosion in OPTG was scored from 0 (no erosion) to 4 (condyles totally eroded). In the analysis, scores 0–2 were grouped as normal or mild changes, and scores 3–4 as distinct erosions. One hundred healthy blood donors served as controls for HLA typing. Results: Distinct erosion of the TMJ in OPTG was observed in 22 (27%) patients. It affected four (17%) of the 24 patients with RA, three (17%) of the 18 with MCTD, seven (37%) of the 19 patients with AS and eight (38%) of the 21 with SPA non‐significant (NS). The mean erosion scores were 1.7 for RA, 1.3 for MCTD, 2.5 for SPA, and 1.6 for AS patients [probability (p)=0.04]. The frequency of HLA‐B27 antigen was higher in the AS and SPA patients, and that of HLA‐DRB1*04 allele higher in RA patients than in control subjects. In the whole patient population, HLA‐DRB1*01 allele was significantly associated with erosions 16/36 (44%) versus 6/46 (13%) (p=0.0014). In the SPA group, patients with HLA‐DRB1*01 allele had a significantly higher occurrence of distinct erosions than patients without this allele [8/10 (80%) versus 0/11 (0%) (p=0.0002)], whereas DRB1*06 was protective [0/8 (0%) versus 8/13 (62%) (p=0.018)]. HLA‐DRB1*04 was associated with increased salivary IgG in the RA patients. Conclusion: HLA antigens are significantly associated with the development of destructive lesions in the TMJ, as well as composition of saliva in patients with various rheumatic diseases.

Acantholytic cells in pemphigus: A scanning and transmission electron microscopic study

Scanning electron microscopy (SEM) was performed on oral scrapings from seven patients with pemphigus vulgaris and one with pemphigus vegetans as well as on skin samples from one of those with pemphigus vulgaris. The unstained, fixed, critical-point dried cells were identified with the aid of a light microscope. On the basis of the surface appearance, three main types of acantholytic cells were identified: smooth-surfaced, wrinkled, and microvillous. The acantholytic cells were rounded to ovoid and some showed either a central bulging or a concavity. Small, rounded to ovoid holes or indentations (diameter 0.1-0.3 mumol) were observed on most cell surfaces. The surfaces of acantholytic cells were occasionally occupied by a few micro-organisms. Neutrophilic polymorphonuclear leucocyte/acantholytic cell rosettes were studied by SEM. No gap between acantholytic cell and surrounding leucocytes could be demonstrated. It is postulated that neutrophils may mediate the cytolysis of antibody-coated acantholytic cells. Oral scrapings from one patient with pemphigus vulgaris were collected for transmission electron microscopy (TEM). The acantholytic cells were rounded or ovoid and usually had centrally placed nuclei. Tonofilaments were either randomly distributed or concentrically arranged around the nucleus. In other acantholytic cells there was a halo containing vesicles of varying size around the nucleus. TEM findings suggest that acantholytic cells represent dead or dying cells. Both TEM and SEM findings suggest that when acantholytic cells first separate they may show variable numbers of microvilli, which are probably lost in older acantholytic cells.

Oral lichen planus and chronic junctional stomatitis: differences in lymphocyte subpopulations

Objective. Oral lichen planus (OLP) is an oral counterpart or oral manifestation of the common skin disease lichen planus. Chronic junctional stomatitis (CJS) is a relatively unknown condition characterized by a stromal lymphocyte infiltrate, which is also a diagnostic feature of OLP. The differential diagnosis of OLP and CJS is unclear and they have been suggested to represent variants of the same disease. Material and methods. To investigate possible differences in lymphocyte (sub)populations between these two conditions, we immunostained 10 OLP and 10 CJS specimens for CD1-a, and the lymphocyte markers, CD3, CD4, CD5, CD8, and CD20. We scored the staining results by a four-step grading system and used the Fisher exact test to analyze them statistically. Results. The proportional amount of (CD20 positive) B lymphocytes was higher in CJS than in OLP and the predominance of CD4 positive T lymphocytes over CD8 positive T lymphocytes was stronger in OLP than in CJS. The differences were statistically significant. Conclusion. The results reflect differences in the lymphatic infiltrate between OLP and CJS. Their significance needs further investigation.

Clinical and cytological features of oral pemphigus

Nine patients with pemphigus vulgaris and one with pemphigus vegetans were studied. All ten patients had oral lesions, five of them skin involvement as well. The patients were followed up for variable periods at different intervals. At each visit, their clinical condition was assessed as mild, moderate, severe or remissive. At examinations, oral light microscopical smears and blood samples for indirect immunofluorescence were taken. From one patient with pemphigus vulgaris, smears from skin blister fluid were also studied. The highest intercellular antibody titers were found in severely affected patients, and those at the mild stage of the disease usually gave low or negative titers. A correlation between the number of acantholytic cells in smears and the clinical condition of the patient was usually found: improvement of the clinical state was followed by decreased numbers of acantholytic cells. Acantholytic cells were rounded or ovoid, having an enlarged, hyperchromatic, centrally or eccentrally situated nucleus. The cytoplasm was usually eosinophilic. Neutrophilic polymorphonuclear leucocyte/acantholytic cell rosettes were sometimes seen in the smears, as well as eosinophil/acantholytic cell rosettes. Variable numbers of leucocytes (from 3 to 11) were seen surrounding the acantholytic cell. Both binucleated epithelial and acantholytic cells and multinucleated giant cells were found in the smears. Multinucleated giant cells were sometimes seen engulfing cells resembling acantholytic cells.

Freeze-fracturing of oral epithelial cells in pemphigus

Biopsies from the oral mucosa of three patients (two with pemphigus vulgaris and one with pemphigus erythematosus) were taken for both freeze-fracturing and thin-sectioning studies. Oral mucosa from a clinically healthy volunteer was used as control. The samples collected from both the involved and uninvolved oral mucosa of pemphigus patients showed widened intercellular spaces and fewer than normal desmosomes. Thin sections showed a random distribution of tonofilaments. However, in freeze-fracture replicas tonofilaments were arranged in thick bundles with a slightly coiled orientation. In freeze-fracture replicas of samples from a pemphigus patient in remission no difference was found in the number of particles on both fracture faces.