Takashi Kinumatsu

Involvement of laminin and integrins in adhesion and migration of junctional epithelium cells.

BACKGROUND AND OBJECTIVE The junctional epithelium attaches to the enamel surface with hemidesmosomes (of which laminin-5 and integrin-alpha(6)beta(4) are the main components) in the internal basal lamina. Laminin-5 is also involved in cell motility with integrin-alpha(3)beta(1), although their functions have not yet been clarified.The purpose of this study was to determine the functions of those adhesive components between the tooth and the junctional epithelium during cell migration.Because an idea has been proposed that directly attached to tooth cells (DAT cells) may not contribute to cell migration, 5-bromo-2-deoxyuridine staining was performed to confirm cell migration. MATERIAL AND METHODS We investigated laminin-gamma(2) (contained only in laminin-5), integrin-beta(4) (involved in cell-extracellular matrix contact) and integrin-alpha(3) (inducing cell migration) in the junctional epithelium, oral gingival epithelium and gingival sulcus epithelium of 6-wk-old ICR mice using laser microdissection, quantitative real-time reverse transcription-polymerase chain reaction, immunofluorescence and 5-bromo-2-deoxyuridine staining. RESULTS Laminin and integrins were clearly immuno-localized in the basal lamina of all epithelium. Quantitative analysis of laminin and integrin mRNAs by laser microdissection showed that they were more highly expressed in DAT cells than in basal cells in the oral gingival epithelium. In particular, a 12-fold higher expression of laminin-5 was observed in the junctional epithelium compared with the oral gingival epithelium. 5-Bromo-2-deoxyuridine staining showed rapid coronal migration of DAT cells. CONCLUSION These results suggest that the abundant expression of laminin-5 and integrin-alpha(6)beta(4) is involved in the attachment of DAT cells to teeth by hemidesmosomes. Abundant expression of laminin-5 and integrin-alpha(3)beta(1) might assist in DAT cell migration, confirmed by 5-bromo-2-deoxyuridine staining during the turnover of junctional epithelium.

Immunolocalization of laminin and integrin in regenerating junctional epithelium of mice after gingivectomy

BACKGROUND AND OBJECTIVE The expression patterns of adhesive proteins and extracellular matrix proteins in regenerating gingival epithelium after gingivectomy are unknown. The aim of this study was to examine the expression of laminin 1, laminin gamma(2) (a specific component of laminin 5), integrin beta(4) and integrin alpha(3) in the regenerating gingival epithelium in order to understand the mechanism of wound healing during reconstitution of the sulcular environment. MATERIAL AND METHODS The palatal gingivae of the maxillary molars of Institute of Cancer Research mice were excised, and the regenerating tissues were examined 1, 3, 5, 7 and 14 days later. Fresh, non-fixed and non-decalcified frozen sections were prepared and stained using immunofluorescence. RESULTS At 1 day post-surgery, intense expression of laminin gamma(2), integrin beta(4) and integrin alpha(3) was distinct in the frontal margin of the regenerating oral epithelium. Laminin gamma(2) was diffusely detected on the root surface and in connective tissues beneath the regenerating oral epithelium at 3 and 5 days. At 7 days, laminin gamma(2) was intermittently recognizable in the internal basal lamina (IBL) close to tooth-facing cells, while laminin gamma(2), integrin beta(4) and integrin alpha(3) were observed in the IBL and in the external basal lamina (EBL) of the regenerating junctional epithelium at 14 days. CONCLUSION These results suggest that secretion of laminin 5 in the connective tissue may induce epithelial cell migration, and that binding of laminin 5 to integrin alpha(6)beta(4) and integrin alpha(3)beta(1) in the IBL may provoke cell adhesion and migration of cells facing the tooth on the enamel surface of the regenerating junctional epithelium.

Effect of recombinant basic fibroblast growth factor on reimplanted teeth in beagle dogs

OBJECTIVES This study assessed the effect of recombinant fibroblast growth factor (FGF-2) on reimplanted teeth. STUDY DESIGN Premolars were reimplanted in dogs. Roots on the left side of the mandible were treated with FGF-2, whereas roots on right side served as controls. At 2, 4, or 8 weeks after treatment, animals were humanely killed and specimens collected and processed for histopathologic examination. RESULTS After 2 weeks, proliferating cell nuclear antigen-positive cell ratios in the experimental group were significantly greater than those in the control group (P < .01). After 4 and 8 weeks, the experimental group showed formation of new cementum. Moreover, incidence of replacement resorption in the experimental group was significantly lower than that in the control group (P < .01). CONCLUSION The results indicate that FGF-2 promotes formation of new periodontal ligament and prevents ankylosis and root resorption following reimplantation of teeth.

Treatment of Gingival Fenestration with Connective Tissue Graft: A Case Report

Here, we report a case of gingival fenestration requiring periodontal plastic surgery. The patient was a 32-year-old man presenting with the chief complaint of esthetic impairment and gingival twitching due to gingival fenestration. Baseline examination revealed localized periodontal breakdown, including gingival fenestration in the lower right central incisor (#41). Periodontal examination revealed 3% of sites with a probing depth of ≥4 mm and 8.9% with bleeding on probing. Radiographic examination revealed vertical bone loss in #15 and 36, together with buccal fenestration in #41. Based on a clinical diagnosis of chronic periodontitis with gingival fenestration, initial periodontal therapy comprised plaque control and scaling and root planing. Following suppression of inflammation, occlusal adjustment was performed in the anterior teeth. As plastic surgery, combined use of an elevated flap and a connective tissue graft was applied at #41. Following reevaluation, the patient was placed on maintenance care. The patients periodontal condition has remained stable over a 6-month period.