Jopi J.W. Mikkonen

The defence architecture of the superficial cells of the oral mucosa

The oral epithelium together with the saliva and its components forms a complex structure which is the first line of defence in the oral cavity. The surface of superficial cells of the oral epithelium contains ridge-like folds, microplicae (MPL), which are typical of the surfaces of areas covered with protective mucus. The role of MPL seen on the upper surface of the oral epithelial cells is still unknown. The salivary mucus gel performs a protective diffusion membrane against harmful substances and this membrane is built up by epithelial cells covered by a highly hydrated and viscous gel, where mucins constitute the scaffold. The interaction between the MPL-structure and the mucins is shown in cornea, so that mucins are expressed on the tips of the MPL of the epithelial cells. We hypothesized that the MPL architecture of oral superficial epithelial cells provides the underlying basis for mucinss protective function as well as in ocular surface. The salivary mucous barrier is required to protect the superficial cells and the MPL-structure together with membrane anchored mucin binding protein (MBP) forms the ground to this mucous barrier. So, oral mucosal barrier complex (OMBC) contains both the MBP-mucin - complex and the MPL-structure of the superficial cells. In the future, studies of the alterations of the salivary mucins and that of the MPL-structure may yield therapeutic opportunities for burning mouth syndrome and perhaps for mucositis causing by irradiation. Focus on cell surface microplication and mucins in oral mucosal biology and oral mucosal diseases is a promising avenue for future research in several ways.

Microstructure of the Superficial Epithelial Cells of the Human Oral Mucosa

Abstract Background: The apical cell membrane of the oral mucosa adjacent to the saliva interface is thrown into membrane folds, termed microplicae (MPL) with variation in morphology. The present study classifies morphological changes undergone by MPL into qualitative and quantitative categories. Material and Methods: Oral mucosal specimens were obtained from 32 healthy patients. Half of each specimen was prepared routinely for light microscopy, and the other part for scanning and transmission electron microscopy. Different measurements of cell structure were presented: the density of MPL, the width and height of MPL, the width of furrows between two adjacent MPL and the distance of the centre of MPL. Morphometric measurements were obtained using a semiautomatic ImageJ analysis software (W Rasband, National Institutes of Health, Bethesda, MD). Results: Parallel and branching MPL was common observation in the area of lining mucosa and in the tongue between the filiform papillae. The density of MPL was higher in the cells of the buccal mucosa than in the cells of the tongue, 43.69 + 11.43% and 31.68 + 10.32%, respectively. The difference was significant (p < 0.001). The width of MPL was 0.16 µm in cells of the buccal mucosa and 0.12 µm in cells of the tongue. Conclusions: Our findings support the idea that MPL structure is a determining factor for the functionality of the oral epithelium since the values of the MPL were kept relatively stable. The role of MPL structure of the oral mucosal cells is discussed.

Microstructure of Oral Epithelial Cells as an Underlying Basis for Salivary Mucosal Pellicle

Abstract Background: Salivary mucosal pellicle forms the structural basis of the local innate immune defense mechanism of the oral mucosa. At the surface of the oral mucosa, the apical cell membrane adjacent to the saliva interface contains short membrane folds, termed microplicae (MPL). This MPL structure of oral epithelial cells and its function as a basis to the salivary mucosal pellicle is unclear. In this preliminary study, we describe the ultrastructural morphology of cell membrane of superficial cells of the oral mucosa and study the membrane-associated mucins (MAMs), MUC1 and MUC4, with immunohistological methods. Materials and methods: Oral mucosal specimens were obtained from six healthy patients. Half of each specimen was prepared routinely for light microscopy, and the other part for scanning and transmission electron microscopy. The presence of MUC1 and MUC4 were studied by immunohistochemical methods in oral mucosal specimens. Results: Morphologically, the cell membrane of MPL is partly discontinuous and membrane-associated molecules extrude from the cell membrane. MUC1 expression was detected in the superficial part of the buccal epithelium, while MUC4 had no expression in the oral squamous epithelium. Conclusions: The novel of this study is that the membrane-tethered molecules seem to occur onto the cell membrane of the superficial epithelial cells of the oral mucosa. Furthermore, the stratified squamous epithelium of the buccal mucosa produces MUC1 for the surface-saliva pellicle interface. The interaction between MPL structure, MUC1 mucin, and salivary mucosal pellicle is discussed.

Metabolic Profiling of Saliva in Patients with Primary SjögrenâÂÂs syndrome

Objective: To investigate the feasibility of 1 H-NMR spectroscopy for metabolic profiling of human saliva samples and to determine whether the concentration of certain salivary metabolites, mainly representing small organic acids and amino acids, differ between patients with primary Sjogren´s syndrome (pSS) and healthy controls. Methods: Stimulated whole-mouth saliva (SWMS) was collected from female pSS patients (n =15, all fulfilling the revised European Community proposed criteria). Salivary flow rate was immediately determined, samples were then frozen and subsequently analyzed by 1 H-NMR spectroscopy in comparison with samples collected from healthy individuals (n=15). Results: From each sample, up to 24 metabolites could be identified and quantified. Choline and taurine concentrations were very significantly higher in the pSS patients compared to healthy controls (p<0.001), but their concentrations correlated negatively with salivary flow rate. Alanine and glycine concentrations were significantly higher (p=0.004, p=0.007, respectively), whereas butyrate (p= 0.034), phenylalanine (p=0.026) and proline (p=0.032) were only slightly higher in pSS saliva samples than in controls. Conclusions: NMR spectroscopy has a potential for quantitative metabolic profiling of saliva samples. NMR spectroscopy is suitable for the analysis of NAAs in saliva and it can bypass the other methods, which are normally suitable for analysis of just one metabolite.

Surface Morphology of Superficial Cells in Irradiated Oral Mucosa: An Experimental Study in Beagle Dog

Abstract Objectives: The aim of the present study is to investigate if radiation induces changes in the superficial cells of the oral mucosa and secondly to describe morphological characteristics of the cell surface structure by scanning electron microscopy (SEM). Materials and methods: Ten beagle dogs aged 1–2 years were used in this study. One side of each mandible was irradiated in two sessions, each lasting 1 week. The total dosage was 40 Gy (Group A; 5 dogs) and 50 Gy (Group B; 5 dogs), in five fractions of 4 Gy. The other side of mandible (non-irradiated) served as a control. The specimen was harvested with a scalpel from the alveolar mucosa of the irradiated area 1 year after irradiation and studied with SEM. Results: In the control side, the surface structure of the cell contains straight parallel or branched microplicae (MPL), which were equally spaced over the cell surfaces. Discontinuous and short MPL were typical cell structure of irradiated mucosa. In 50 Gy group, the surface structure of epithelial cell was pitted and the cell boundaries were thick. Conclusions: The novelty of the present study is that radiation disrupts superficial cells of the oral mucosa. The role of the MPL structure of the superficial cells in mucositis development is discussed.

Oral mucosal epithelial cells express the membrane anchored mucin MUC1

OBJECTIVE The presence of a stable salivary pellicle (SP) is essential to provide a wet surface for the oral mucosal epithelia. The oral mucosa is covered by the SP which is suggested to be a mixed film of both salivary and epithelial components. Our aim was to analyse the presence of membrane-anchored mucin MUC1 in the oral mucosal epithelia. DESING The presence of MUC1 was studied by immunohistochemical and immunoelectron microscopical methods in 19 buccal mucosal specimens. The localization and intensity of the epithelial expression were analyzed. RESULTS Strong staining of MUC1 was found in the epithelial cells of intermediate and superficial layers. Some basal cells were shown faint expression. In the intermediate and superficial layers, the MUC1 expression was seen mainly on the upper cell surface. Furthermore, the expression of MUC1 was noted in the cytoplasm near the nucleus and in the rough granules. By electron microscopy, extracellular domain of membrane-anchored molecules extruded about 15-30nm above the cell surface in the apical cells of the oral epithelium. Immunoelectron microscopic examination shows that MUC1 is mainly localized in the plasma membrane of epithelial cells and also in small vesicles (75-100nm) just below the plasma membrane. CONCLUSION The membrane-anchored MUC1 is expressed in the superficial layer of the oral mucosal epithelium, especially on the upper surface of epithelial cells. MUCI may be the anchoring protein of the salivary pellicle stabilization.

Fissured tongue: a sign of tongue edema?

Fissured tongue (FT) is a condition frequently seen in the general population. Clinically, FT is characterized by grooves that vary in depth and are noted along the dorsal and/or dorsolateral aspects of the tongue. Furthermore, FT presents many enlarged, smooth filiform papillae and subepithelial inflammatory infiltration. Despite of many studies, the etiology of FT remains obscure. FT is believed to be a congenital anomaly associated with several disorders and with geographic tongue (GT). We hypothesize that FT is not a congenital anomaly, and FT with swollen filiform papillae may represent edema in the subepithelial tissue of the tongue. According to the literature, the difference in prevalence among different age groups indicates that FT is not a congenital disorder. FT appears to occur more commonly in adults, and it is very rare or not at all in children younger than 10 years old. An association between FT and GT is well established in the literature, supporting the results of previous authors suggesting that FT might be a consequence of GT. The most remarkable finding in the region of swollen papillae of FT samples has been the subepithelial infiltrates of polymorphonuclear leucocytes and lymphocytes causing the subepithelial edema. The clinically visible grooves and large edematic papillae clustered on the region of the fissures might be caused by the inflammation and edema underneath the epithelium. In the future, FT and GT must be researched together as two different entities of the same disease so that GT is a prestage of FT. The diagnosis of FT must be taken to consideration whether the tongue surface have smooth and swollen papillae or normal-appearing filiform papillae.

Microplicae - Specialized Surface Structure of Epithelial Cells of Wet-Surfaced Oral Mucosa

ABSTRACT The surface structure of the superficial cells of the oral mucosa is decorated with numerous membrane ridges, termed microplicae (MPLs). The MPL structure is typical of the epithelial surfaces that are covered with protective mucus. Cell membrane MPLs are no longer seen as passive consequences of cellular activity. The interaction between MPLs and the mucins has been demonstrated, however the role of MPL structure seen on the upper surface of the oral epithelial cells is speculative. The cell surface is of potentially great significance, as it harbors many markers for refined prognosis and targets for oral mucosal diseases and cancer therapy. With these aspects in mind, we conducted the present review of the MPL structure and function in order to form the basis for further studies of MPLs of the oral epithelial cells.

Acid-etching technique of non-decalcified bone samples for visualizing osteocyte-lacuno-canalicular network using scanning electron microscope

ABSTRACT The aim of this study was to define the acid-etching technique for bone samples embedded in polymethyl metacrylate (PMMA) in order to visualize the osteocyte lacuno-canalicular network (LCN) for scanning electron microscopy (SEM). Human jaw bone tissue samples (N = 18) were collected from the study population consisting of patients having received dental implant surgery. After collection, the bone samples were fixed in 70% ethanol and non-decalcified samples embedded routinely into polymethyl metacrylate (PMMA). The PMMA embedded specimens were acid-etched in either 9 or 37% phosphoric acid (PA) and prepared for SEM for further analysis. PMMA embedded bone specimens acid-etched by 9% PA concentration accomplishes the most informative and favorable visualization of the LCN to be observed by SEM. Etching of PMMA embedded specimens is recommendable to start with 30 s or 40 s etching duration in order to find the proper etching duration for the samples examined. Visualizing osteocytes and LCN provides a tool to study bone structure that reflects changes in bone metabolism and diseases related to bone tissue. By proper etching protocol of non-decalcified and using scanning electron microscope it is possible to visualize the morphology of osteocytes and the network supporting vitality of bone tissue.

Alveolar bone remodeling after tooth extraction in irradiated mandible: An experimental study with canine model

ABSTRACT Objectives: The aim of the present study is to investigate the morphological and cellular changes in dental extraction socket that has been irradiated after the tooth extraction and to describe morphological characteristics of the osteocytes and osteocyte-lacunar-canalicular network (LCN) by scanning electron microscopy (SEM). Material and methods: Five beagle dogs aged 1–2 years were used in this study. One side of each mandible was irradiated in two sessions and the other side of mandible (non-irradiated) served as a control. The mandible bone blocks were processed by bulk staining en bloc in basic fuchsin and the specimens were embedded routinely in polymethyl methacrylate resin without preliminary decalcification. All blocks were subjected to micro-CT imaging, after that the specimens were prepared for light microscopy and SEM. Results: Alterations in bone macrostructure are minimal in irradiated bone, but the changes in LCN are clear. In the area of the tooth extraction socket, the connections of osteocytes to the vessels and to neighboring osteocytes were not observed both in irradiated and nonirradiated bone. However, osteoclasts were located in the bone surface entering inside to the bone between osteons. In the lamellar bone of lateral sides, a decrease in canalicular connections between osteocytes and periosteum was found in irradiated bone as compared to the non-irradiated side. Conclusions: The novelty of the present study is that radiation disrupts osteocytes and their dendrites.