The purposes of this study is to evaluate the combination effects of TGF-beta1 and PDGF-BB on the periodontal ligament cells to use as a regeneration promoting agent of periodontal tissue. Human periodontal ligament cells were prepared from the first premolar tooth extracted for the orthodontic treatment and were cultured in DMEM/10% FBS at the 37degrees C, 5% CO2 incubator. Authors measured the DNA synthesis, total protein, collagen and noncollagenous protein synthesis according to the concentration of TGF-beta1, (1, 5ng/ml) and PDGF-BB (1, 10 ng/ml) in combination. To explore further this delayed effect of TGF-beta1, we preincubated human periodontal ligament cells with TGF-beta1 for 4 or 24 hours before PDGF-BB stimulation. The results were as follows: The DNA synthetic activity was increased dose dependently by TGF-beta1, PDGF-BB. The combination of TGF-beta1 and PDGF-BB consistently enhanced the DNA synthetic activity to PDGF-BB alone. The ability of TGF-beta1 to enhance DNA synthetic activity in PDGF-BB treated periodontal ligament cells was dose dependent. The maximum mitogenic effect was at the 5ng/ml of TGF-beta1 and 10ng/ml of PDGF-BB. Preincubation of cells with TGF-beta1 resulted in significantly greater response to PDGF-BB at all TGF-beta1 concentration studied, and may be useful for clinical application in periodontal regenerative procedures. The total protein, collagen and noncollagen synthesis was increased dose pendently by TGF-beta1, PDGF-BB. The % of collagen was slightly decresed according to the concentration of TGF-beta1, PDGF-BB. The effect of TGF-beta1, PDGF-BB were not specific for collagen synthesis since it also increased noncollagenous protein synthesis. This study demonstrates that PDGF-BB is major mitogens for human periodontal ligament cells in vitro, and supports a role for TGF-beta1 as a regulation of the mitogenic and total protein formation to PDGF-BB in these cells.
Bicuspid ; Collagen ; DNA ; Humans ; Incubators ; Mitogens ; Periodontal Ligament* ; Regeneration ; Tooth ; Transforming Growth Factor beta1*

Progressive dysphagia in a 53 year old man was caused by a giant polypoid tumor in the lower intrathoracic esophagus. Radical transthoracic esophagectomy and esophagogastrostomy were carried out. Microscopic examination of the tumor revealed a true carcinosarcoma, composed of a mixture of basaloid squamous cell carcinoma and chondrosarcoma with multiple cartilagenous productions. Carcinoma metastases were found in the subcarinal and perigastric lymph nodes. Immunohistochemically, squamous area displayed strong positive to cytokeratin, and basaloid area showed positive immunoreaction to high molecular weight cytokeratin (34beta E12). Spindle cell sarcoma reacted to vimentin and smooth muscle actin. Chondrosarcomatous area reacted to vimentin and S-100 protein. He received postoperative chemotherpy and radiotherapy. He has been free of disease for 11 months.
Actins ; Carcinoma, Squamous Cell ; Carcinosarcoma* ; Chondrosarcoma ; Deglutition Disorders ; Esophageal Neoplasms ; Esophagectomy ; Esophagus* ; Humans ; Keratins ; Lymph Nodes ; Middle Aged ; Molecular Weight ; Muscle, Smooth ; Neoplasm Metastasis ; Radiotherapy ; S100 Proteins ; Sarcoma ; Vimentin

Recently the transcriptional up-regulation of human beta-defensin 2 (HBD-2) by lipopolysaccharide (LPS) was found to be associated with NF-kappaB binding site. Although the general mechanisms of NF-kappaB activation by LPS stimulation are well understood, less is known about the signal transduction pathway leading to LPS-induced NF-kappaB activation in human corneal epithelial (HCE) cells. The aim of this study was to investigate the intracellular signals involved in LPS-induced HBD-2 mRNA expression in HCE cells. Pretreatments of inhibitors for NF-kappaB, protein tyrosine kinase, p38 mitogen activated protein kinase (MAPK), and c-Jun N-terminal kinase (JNK) attenuated the LPS-induced NF-kappaB DNA binding activity and HBD-2 mRNA expression. Furthermore, pretreatments with inhibitors for protein kinase C (PKC), phosphatidylcholine-phospholipase C, phosphatidylinositol-phospholipase C, or phosphatidate phosphohydrolase prevented LPS-induced HBD-2 mRNA expression and HBD-2 prmoter-driven luciferase activity. However, the increased expression of HBD-2 mRNA and the increased DNA binding activity of NF-kappaB induced by LPS were not changed by the blockage of extracellular signal-regulated kinase (ERK) and of addition of antioxidants. Forskolin, a protein kinase A (PKA) agonist did not induce HBD-2 mRNA expression. These data demonstrate that LPS-induced HBD-2 mRNA expression via NF-kappaB is, at least in part, dependent on PKC, p38 MAPK, JNK, and protein tyrosine kinase status, but appears to be independent on PKA, ERK and ROS in HCE cells. Taken together, there may be more than one signaling pathways that lead to LPS-induced up-regulation of HBD-2 mRNA expression in HCE cells.
Antioxidants ; Binding Sites ; Colforsin ; Cyclic AMP-Dependent Protein Kinases ; DNA ; Epithelial Cells* ; Humans* ; JNK Mitogen-Activated Protein Kinases ; Luciferases ; NF-kappa B ; p38 Mitogen-Activated Protein Kinases ; Phosphatidate Phosphatase ; Phosphotransferases ; Protein Kinase C ; Protein Kinases ; Protein-Tyrosine Kinases ; RNA, Messenger* ; Signal Transduction ; Up-Regulation