Immunohistochemical Demonstration of Cathepsins B and L in the Periodontal Ligament (PDL) of the Rat Molar.

Takeyama, Masaki; Irie, Kazuharu; Nakamura, Hiroaki; Kominami, Eiki; Hanada, Kooji; Ozawa, Hiroki; Ejiri, Sadakazu

doi:10.2330/joralbiosci1965.43.194

Cited by 4 publications

(3 citation statements)

References 24 publications

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“…Periodontal ligament cells may be involved in both production and degradation of collagenous matrices by expressing COLs I, III, and MMP-8 during the formation of periodontal tissues. Cathepsins B and L have been localized in mature rat periodontal ligaments (Takeyama et al, 2001). Cultured human periodontal ligament cells express cathepsins B and L (Goseki et al, 1996).…”

Section: Discussionmentioning

confidence: 99%

Expression of MMP-8 and MMP-13 mRNAs in Rat Periodontium during Tooth Eruption

et al. 2002

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The present study was designed to investigate mRNA expression of matrix metalloproteinase-8 (MMP-8) and MMP-13 in forming periodontium during tooth eruption in the rat. RT-PCR for the decalcified paraffin sections indicated expression of MMP-8 and MMP-13 in the periodontal tissues. In situ hydridization demonstrated expression of MMP-8 in osteoblasts, osteocytes, periodontal ligament cells, cementoblasts, and cementocytes along with collagen types I and III. In contrast, transcripts of MMP-13 were confined to a small population of osteoblasts and osteocytes in alveolar bone. The results suggested that MMP-8 may be involved in remodeling the periodontium during tooth eruption, and its expression may be coordinated with that of collagen types I and III, whereas the participation of MMP-13 may be rather limited.

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Section: Discussionmentioning

confidence: 99%

Expression of MMP-8 and MMP-13 mRNAs in Rat Periodontium during Tooth Eruption

et al. 2002

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“…Cathepsins B and L are lysosomal cysteine proteinases found in osteoclasts, osteoblasts and periodontal ligament (Goto et al, 1993;Aisa et al, 2003;Takeyama et al, 2001). In this study, TNF-a did not affect the mRNA levels of cathepsins B and L in Saos2 cells (Fig.…”

Section: Discussionmentioning

confidence: 50%

“…One-step RT-PCR was performed using a SuperScript one-step RT-PCR kit. Primers were synthesized on the basis of the reported human cDNA sequences for BSP (Kim et al, 1994), Cbfa1 (Komori et al, 1997), osterix (Nakashima et al, 2002), a1 (I) collagen (Bernard et al, 1983), cyclooxygenase-2 (COX-2) (Hla and Neilson, 1992), interleukin-6 (IL-6) (Ishimi et al, 1990), cathepsin B (Aisa et al, 2003), cathepsin L (Goto et al, 1993;Takeyama et al, 2001), TIMP-1 (Rifas et al, 1994) and glyceraldehyde-3-phosphate dehydrogenase (GAPDH). Sequences of the primers used for RT-PCR and the expected size of the PCR products were as follows:…”

Section: Rt-pcrmentioning

confidence: 99%

Effect of TNF‐α on human osteosarcoma cell line Saos2—TNF‐α regulation of bone sialoprotein gene expression in Saos2 osteoblast‐like cells

Nakayama

Kato

Nakajima

et al. 2004

Cell Biology International

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Tumor necrosis factor-alpha (TNF-alpha) is a major mediator of inflammatory response in many diseases. It inhibits bone formation and stimulates bone resorption. To determine the molecular mechanisms involved in the regulation of gene expression of osteoblast-like cells, we analyzed the effects of TNF-alpha on the human osteosarcoma cell line Saos2. We used RT-PCR to examine the effects of TNF-alpha on bone sialoprotein (BSP), core binding factor a1 (Cbfa1), osterix, alpha 1 (I) collagen, cyclooxygenase-2 (COX-2), interleukin-6 (IL-6), cathepsin B, cathepsin L and tissue inhibitors of metalloproteinase-1 (TIMP-1). TNF-alpha (10ng/ml) increased BSP, IL-6 and COX-2 mRNA levels after 3h, reaching maximal levels at 12 h. Cbfa1 mRNA levels increased after 3 h, but decreased by 24 h. Osterix, cathepsin B, cathepsin L and TIMP-1 mRNA levels did not change after stimulation with TNF-alpha. On the other hand, alpha 1 (I) collagen mRNA expression was suppressed by TNF-alpha at 24 h. Transient transfection analyses were performed using chimeric constructs of the rat BSP gene promoter linked to a luciferase reporter gene. TNF-alpha (10 ng/ml) had no effect on the promoter activities of BSP transfected into Saos2 cells. The results of gel mobility shift assays using radiolabeled double-stranded cAMP response element (CRE) and FGF2 response element (FRE) oligonucleotides in the proximal promoter of the rat BSP gene showed increased binding of nuclear proteins at 6 h. Gel mobility shift assays with radiolabelled COX-2-CRE and COX-2-NF kappa B oligonucleotides revealed an increase in the binding of nuclear proteins from TNF-alpha-stimulated Saos2 cells. These studies, therefore, showed that TNF-alpha indirectly increased BSP expression, and that it could be mediated through COX-2 and Cbfa1 expression in Saos2 osteoblast-like cells.

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Cathepsins B and L Increased during Response of Periodontal Ligament Cells to Mechanical Stress In Vitro

Yamaguchi

Ozawa

Nogimura

et al. 2004

Connective Tissue Research

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Cathepsin is a typical and well-characterized lysosomal cysteine protease that, under pathological conditions, is involved in tissue destruction. A recent immunocytochemical study demonstrated that cathepsins B (CAB) and L (CAL) were localized in the periodontal ligament (PDL) of the rat molar, and they were expressed in compressed sites during experimental tooth movement. Further, we demonstrated previously that the levels of CAB and CAL in gingival crevicular fluid increased during orthodontic tooth movement. Therefore, CAB and CAL may play important roles in the process of collagen degradation during orthodontic tooth movement, and our in vitro study examined the secretion of CAB and CAL in PDL cells following mechanical stress. PDL cells were subjected to 0.5, 1.0, 2.0, or 3.0 g/cm2 of compression force or an increase in surface area by tension force of 0.28%, 0.95%, 1.72%, or 2.50% for 24 hr. For detection of CAB and CAL in conditioned medium, commercially available ELISA kits were used. We found compression and tension significantly increased the secretions of both CAB and CAL in PDL cells, which were exhibited in a time- and force magnitude-dependent manner. The compression-stimulated secretion of CAB was increased approximately 3-fold and that of CAL 4-fold, as compared with the control. Further, tension-stimulated secretion of CAB was increased by 1.5-fold and that of CAL 2-fold compared with the control. When analyzed using a semiquantitative polymerase chain reaction assay, CAB and CAL mRNA were increased in response to both compression and tension forces. These findings demonstrated that mechanical stress (compression and tension forces) causes an increase in secretion of CAB and CAL in PDL cells in vitro.

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Immunohistochemical Demonstration of Cathepsins B and L in the Periodontal Ligament (PDL) of the Rat Molar.

Cited by 4 publications

References 24 publications

Expression of MMP-8 and MMP-13 mRNAs in Rat Periodontium during Tooth Eruption

Expression of MMP-8 and MMP-13 mRNAs in Rat Periodontium during Tooth Eruption

Effect of TNF‐α on human osteosarcoma cell line Saos2—TNF‐α regulation of bone sialoprotein gene expression in Saos2 osteoblast‐like cells

Cathepsins B and L Increased during Response of Periodontal Ligament Cells to Mechanical Stress In Vitro

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