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Increase of uPA receptor in gingival fibroblasts by LPS and IL-1b
254
P23-5 Miscellaneous
Results Histologically, clot and fibrin networks with inflammatory cell infiltration were formed in the deep area of the wound one day after operation.The wound was epithelialized at 3 days and angiogenesis was initiated at 3 days. Granulation tissue in the deep area was observed within 5 days. Immunohistochemically, endothelial nuclei of capillaries and some fibroblasts in the wound were often stained positive for PCNA in the early period and fibroblasts in the scar tissue were stained weakly for PCNA at 14 days. By RT-PCR, b F G F was found to be expressed fi'om 1 to 10 days, strongly from 3 to 5 days and decreasing gradually thereafter.
11. Increase of uPA Receptor in Gingival Fibroblasts by LPS and IL-lb
Ogura, N. 1, Tanaka, E l, Abiko, y..2, Nagura, H. 1 Departments of 1Oral Surgery and 2Biochemistry, Nihon University of Dentistry at Matsudo, Chiba, Japan Plasminogen activators, urokinase type plasminogen activator (uPA) and tissue type plasminogen activator (tPA), are serine proteases which convert plasminogen into an active serine protease, plasmin. This generation system has important roles in pathological processes including extracellular matrix degradation, cell migration, wound healing, inflammatory cell chemotaxis and tumor cell invasion and metastasis. Recently, it has been l~eported that an important step in the regulation of uPA activity involves binding to a high affinity cell surface receptor (uPAR) which is linked to the plasma membrane by a glycosylphosphatidylinositol (GPI) anchor. It is suggested that uPA binding to uPAR leads to an accelerated generation of plasmin activity. We have previously reported the activation of the PA/plasmin system in gingival fibroblasts (Gin cells) following treatment with lipopolysaccharide (LPS), which is associated with oral infection (J Periodont Res 1995: 30: 132). It was also suggested that the PA secreted by Gin cells is uPA rather than tPA. This study focuses on cell-surface-associated PA in Gin cells treated with the inflammatory factors, LPS and IL-lb. The PA activity in the cell lysate of Gin cells was increased by both LPS and IL-lb. The increased PA activity was inhibited by amiloride which is a specific inhibitor of uPA. Phosphatidylinositol-specific phospholipase C (PI-PLC)-releasable PA activity was also increased by LPS and IL-lbl To confirm this finding, expression of the uPAR gene was examined by RT-PCR. uPAR m R N A was increased in response to both LPS and IL-lb in Gin celts. We suggested that the increase in both uPA and uPAR in Gin cells in response to LPS or IL-lb is important for extracellular matrix degradation and inflammatory progression.
12. Effects of Biscoelaurin Alkaloid on Superoxide Anion Generation by Peripheral Blood and/or Salivary Neutrophils in Various Diseases of the Oral Mucosa
Harada, T., Kanaoka, 1t., Kingetsu, A., Yoshimura, E Department of OMFS, Shimane Medical Univ., Izumo, Japan The effects of biscoclaurin alkaloid on superoxide anion (02*) generation by peripheral blood and/or salivary neutrophils in various diseases of the oral mucosa were examined. 02* generation was assessed by the reduction of cytochrome C. Separation of peripheral blood neutrophils was performed by eentrifugation using Mono-poly Resolving Medium (Flow Laboratories, Irvine, Scotland). Salivary neutrophils were obtained from patients' saliva. The oral cavity of each donor was thoroughly washed 12 times with 15 ml of saline for 30 s. The pooled washings (180 ml) of each patient were centrifuged and passed individually through 300-mesh nylon sheets. The results were as follows. 0.2 laM, 1 ~tM, and 5 ~tM of biscoclaurin alkaloid (BA) suppressed 02* generation by peripheral blood and/or salivary neutrophils depending upon the concentrations of BA. 5 ~aM of BA completely suppressed 02* generation by both types of neutrophils. The level of 02* generation by peripheral blood neutrophils in various diseases of the oral mucosa (such as carcinoma in situ and .oral lichen planus) in the posttreatment period was lower than that in the pretreatment period. The administration of BA for various diseases of the oral mucosa was effective in most cases. The suppression of 02* generation by both peripheral blood and/or salivary neutrophils supports the efficacy of administration of BA for treatment of these diseases.
13. Postnatal Development of Motoneurons Supplying the Jaw Closing and Opening Muscle in the Rat
Shohara, E., Masuda, T., Sugimura, M. Department of Oral and Maxillofacial Surgery, Nara Medical University, Nara, Japan In the postnatal dietary life of mammals, sucking action precedes masticatory action in the food-taking behavior. This time gap may effect the postnatal development of the jaw closing and opening muscles and the supplying motoneurons. We have studied changes in growth of the motonenrons innervating the masseter and anterior digastric muscles after birth using 108 rats. Infant rats were given a horseradish peroxidase (30%) saline solution injection into the masseter or anterior digastric muscle to label the supplying motoneurons at 1, 3, 5, 7, 9, 10, 12, and 14 days, 3 weeks, and 1 month after birth. Conventional histological processing using the tetramethyl benzidine method was used to label cells. Labeled motoneurons were identified and their mean diameter was measured. In the developmental stage from postnatal day one to one
P23-5 Miscellaneous
Results Histologically, clot and fibrin networks with inflammatory cell infiltration were formed in the deep area of the wound one day after operation.The wound was epithelialized at 3 days and angiogenesis was initiated at 3 days. Granulation tissue in the deep area was observed within 5 days. Immunohistochemically, endothelial nuclei of capillaries and some fibroblasts in the wound were often stained positive for PCNA in the early period and fibroblasts in the scar tissue were stained weakly for PCNA at 14 days. By RT-PCR, b F G F was found to be expressed fi'om 1 to 10 days, strongly from 3 to 5 days and decreasing gradually thereafter.
11. Increase of uPA Receptor in Gingival Fibroblasts by LPS and IL-lb
Ogura, N. 1, Tanaka, E l, Abiko, y..2, Nagura, H. 1 Departments of 1Oral Surgery and 2Biochemistry, Nihon University of Dentistry at Matsudo, Chiba, Japan Plasminogen activators, urokinase type plasminogen activator (uPA) and tissue type plasminogen activator (tPA), are serine proteases which convert plasminogen into an active serine protease, plasmin. This generation system has important roles in pathological processes including extracellular matrix degradation, cell migration, wound healing, inflammatory cell chemotaxis and tumor cell invasion and metastasis. Recently, it has been l~eported that an important step in the regulation of uPA activity involves binding to a high affinity cell surface receptor (uPAR) which is linked to the plasma membrane by a glycosylphosphatidylinositol (GPI) anchor. It is suggested that uPA binding to uPAR leads to an accelerated generation of plasmin activity. We have previously reported the activation of the PA/plasmin system in gingival fibroblasts (Gin cells) following treatment with lipopolysaccharide (LPS), which is associated with oral infection (J Periodont Res 1995: 30: 132). It was also suggested that the PA secreted by Gin cells is uPA rather than tPA. This study focuses on cell-surface-associated PA in Gin cells treated with the inflammatory factors, LPS and IL-lb. The PA activity in the cell lysate of Gin cells was increased by both LPS and IL-lb. The increased PA activity was inhibited by amiloride which is a specific inhibitor of uPA. Phosphatidylinositol-specific phospholipase C (PI-PLC)-releasable PA activity was also increased by LPS and IL-lbl To confirm this finding, expression of the uPAR gene was examined by RT-PCR. uPAR m R N A was increased in response to both LPS and IL-lb in Gin celts. We suggested that the increase in both uPA and uPAR in Gin cells in response to LPS or IL-lb is important for extracellular matrix degradation and inflammatory progression.
12. Effects of Biscoelaurin Alkaloid on Superoxide Anion Generation by Peripheral Blood and/or Salivary Neutrophils in Various Diseases of the Oral Mucosa
Harada, T., Kanaoka, 1t., Kingetsu, A., Yoshimura, E Department of OMFS, Shimane Medical Univ., Izumo, Japan The effects of biscoclaurin alkaloid on superoxide anion (02*) generation by peripheral blood and/or salivary neutrophils in various diseases of the oral mucosa were examined. 02* generation was assessed by the reduction of cytochrome C. Separation of peripheral blood neutrophils was performed by eentrifugation using Mono-poly Resolving Medium (Flow Laboratories, Irvine, Scotland). Salivary neutrophils were obtained from patients' saliva. The oral cavity of each donor was thoroughly washed 12 times with 15 ml of saline for 30 s. The pooled washings (180 ml) of each patient were centrifuged and passed individually through 300-mesh nylon sheets. The results were as follows. 0.2 laM, 1 ~tM, and 5 ~tM of biscoclaurin alkaloid (BA) suppressed 02* generation by peripheral blood and/or salivary neutrophils depending upon the concentrations of BA. 5 ~aM of BA completely suppressed 02* generation by both types of neutrophils. The level of 02* generation by peripheral blood neutrophils in various diseases of the oral mucosa (such as carcinoma in situ and .oral lichen planus) in the posttreatment period was lower than that in the pretreatment period. The administration of BA for various diseases of the oral mucosa was effective in most cases. The suppression of 02* generation by both peripheral blood and/or salivary neutrophils supports the efficacy of administration of BA for treatment of these diseases.
13. Postnatal Development of Motoneurons Supplying the Jaw Closing and Opening Muscle in the Rat
Shohara, E., Masuda, T., Sugimura, M. Department of Oral and Maxillofacial Surgery, Nara Medical University, Nara, Japan In the postnatal dietary life of mammals, sucking action precedes masticatory action in the food-taking behavior. This time gap may effect the postnatal development of the jaw closing and opening muscles and the supplying motoneurons. We have studied changes in growth of the motonenrons innervating the masseter and anterior digastric muscles after birth using 108 rats. Infant rats were given a horseradish peroxidase (30%) saline solution injection into the masseter or anterior digastric muscle to label the supplying motoneurons at 1, 3, 5, 7, 9, 10, 12, and 14 days, 3 weeks, and 1 month after birth. Conventional histological processing using the tetramethyl benzidine method was used to label cells. Labeled motoneurons were identified and their mean diameter was measured. In the developmental stage from postnatal day one to one