Effects of Culture Supernatants of Peripheral Blood Leukocytes on Bone-minerals Resorbing Activity by Human Monocytes

It has been reported that human peripheral blood monocytes in vitro release calcium from fetal rat calvaria and from synthetic hydroxyapatite (HA) (SATO et al., 1984). In this paper, we examined the effects of lymphokine on the release of calcium from HA by human monocytes. Peripheral blood was coll...

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Published inNihon Shishubyo Gakkai Kaishi (Journal of the Japanese Society of Periodontology) Vol. 27; no. 4; pp. 831 - 836
Main Authors SATO, Iwao, IWAKAWA, Yoshinobu, NAKAMURA, Yasutaka, MARUMOTO, Yoshiko, NISHIMOTO, Masazumi, FUJIHASHI, Hiroshi, IKEDA, Katsumi
Format Journal Article
LanguageJapanese
Published JAPANESE SOCIETY OF PERIODONTOLOGY 1985
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ISSN0385-0110
1880-408X
DOI10.2329/perio.27.831

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Abstract It has been reported that human peripheral blood monocytes in vitro release calcium from fetal rat calvaria and from synthetic hydroxyapatite (HA) (SATO et al., 1984). In this paper, we examined the effects of lymphokine on the release of calcium from HA by human monocytes. Peripheral blood was collected from patients with periodontal disease (PPD) and from clinically healthy individuals, and peripheral blood leukocytes (PBL) were isolated by differential centrifugation using Sodium metrizoate-Ficoll. Then the monocytes were separated by the method based on adherence to plastic. PBL were cultured with phytohemagglutinin or dental plaque and afterwards culture supernatants (PBL-sup) were separated and used as a source of lymphokines (Osteoclast-activating factor: OAF and others). Assay of calcium from HA was done as follows: PBL supernatants were to a mixed suspension of monocytes and HA mixed cultured in 5% CO2 at 37°C for 24 to 72 hours. The quantity of released calcium from HA was measured by atomic absorption analysis. Human monocytes induced the release of calcium from HA, and its level was increased more significantly by addition of PBL supernatants from PPD than from healthy subjects. And Nitro Blue Tetrazorium reduction activity and [3H] Thymidine incorporation activity of monocytes were increased too by addition of PBL-sup. These results suggest that bone resorption activity of human monocytes is activated by lymphokines, perhaps by what is called “OAF”.
AbstractList It has been reported that human peripheral blood monocytes in vitro release calcium from fetal rat calvaria and from synthetic hydroxyapatite (HA) (SATO et al., 1984). In this paper, we examined the effects of lymphokine on the release of calcium from HA by human monocytes. Peripheral blood was collected from patients with periodontal disease (PPD) and from clinically healthy individuals, and peripheral blood leukocytes (PBL) were isolated by differential centrifugation using Sodium metrizoate-Ficoll. Then the monocytes were separated by the method based on adherence to plastic. PBL were cultured with phytohemagglutinin or dental plaque and afterwards culture supernatants (PBL-sup) were separated and used as a source of lymphokines (Osteoclast-activating factor: OAF and others). Assay of calcium from HA was done as follows: PBL supernatants were to a mixed suspension of monocytes and HA mixed cultured in 5% CO2 at 37°C for 24 to 72 hours. The quantity of released calcium from HA was measured by atomic absorption analysis. Human monocytes induced the release of calcium from HA, and its level was increased more significantly by addition of PBL supernatants from PPD than from healthy subjects. And Nitro Blue Tetrazorium reduction activity and [3H] Thymidine incorporation activity of monocytes were increased too by addition of PBL-sup. These results suggest that bone resorption activity of human monocytes is activated by lymphokines, perhaps by what is called “OAF”.
Author NISHIMOTO, Masazumi
SATO, Iwao
MARUMOTO, Yoshiko
IWAKAWA, Yoshinobu
IKEDA, Katsumi
FUJIHASHI, Hiroshi
NAKAMURA, Yasutaka
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  fullname: NAKAMURA, Yasutaka
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  fullname: MARUMOTO, Yoshiko
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  fullname: IKEDA, Katsumi
  organization: Department of Periodontology, Josai Dental University
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References 1) Owen, M.: Histogenesis of bone cells. Calcif. Tissue Res., 25: 205-207, 1978.
6) 丸本淑子, 楠公仁, 藤橋弘, 岡田春夫, 中村安隆, 池田克已: 歯槽骨吸収におよぼす Osteoclast-activating factor (OAF) の効果, 1. OAFの産生とその精製の試み. 日歯周誌, 24: 121-127, 1982.
19) Jee, W. S. S. and Nolan, PD.: Origin of osteoclasts from the fusion of phagocytes. Nature, 200: 225-226, 1963.
13) 湊啓輔, 西田俊明, 斎藤博, 長谷哲夫, 下山正徳: リンパ球幼若化反応. 日本臨床, 37: 1574-1580, 1979.
17) 小野崎菊夫, 橋本達一郎: マクロファージを活性化するリンフォカイン, MIFの精製から細胞性免疫の発現まで. 蛋白質核酸酵素, 26: 1910-1924, 1981.
9) Löe, H. and Silness, J.: Periodontal disease in pregnancy. I. Prevalence and severity. Acta. Odont. Scand., 21: 533-551, 1963.
11) Kumagai, K., Itoh, K., Hinoma, S. and Tada, M.: Pretreatment of plastic petridishes with fetal calf serum. A simple method for macrophage isolation. J. Immunol. Methods, 29: 17-25, 1979.
7) 丸本淑子, 藤橋弘, 佐藤巌雄, 中村安隆, 西本正純, 池田克巳: 歯槽骨吸収におよぼす Osteoclast-activating factor (OAF) の効果, II. OAF産生細胞への単球の関与. 日歯周誌, 25: 823-829, 1983.
4) Mundy, G. R., Altman, A. J., Gondek, M. D. and Bandelin, J. G.: Direct resorption of bone by human monocytes. Science, 196: 1109-1111, 1977.
15) 佐藤巌雄, 藤橋弘, 岩川吉伸, 中村安隆, 丸本淑子, 池田克已: ヒト単球の骨塩への作用. 日歯周誌, 27: 146-152, 1985.
16) Dominguez, J. H. and Mundy, G. R.: Monocytes mediate osteoclastic bone resorption by prostaglandin production. Calcif. Tissue Int., 31: 29-34, 1980.
8) Russel, A. L.: A system of classification and scoring for prevalence surveys of periodontal disease. J. Dent. Res., 35: 350-359, 1956.
12) 山本きよみ, 米原ヤス子, 三輪史郎: エステラーゼ染色. 月刊 Medical Technology 編, 染色法のすべて, 医歯薬出版, 東京, 1980, 118-121.
5) 角田左武郎, 松本英彦, 古屋誠, 木村義孝, 南雲正男: マクロファージによる直接的骨吸収. 日臨免誌, 6: 160-166, 1983.
18) Fishman, D. A. and Hey, E. D.: Origin of osteoclasts from mononuclear leukocytes in regenerating newt limbs. Anat. Res., 143: 329-339, 1962.
14) Baehner, R. L. and Nathan, D. G.: Quantitative nitroblue tetrazolium test in chronic granulomatous disease. New Eng. J. Med., 278: 1971-1976, 1968.
3) Heersche, J. N. M.: Mechanism of osteoclastic bone resorption: A new hypothesis. Calcif. Tissue Res., 26: 81-84, 1978.
10) Thorsby, E, and Bartlie, A.: A rapid method for preparation of pure lymphocyte suspensions. In: Histocompatibility Testing 1970, ed., Terasaki, P. I., Munksgaard, Copenhagen, 1971, 655-656.
2) Mundy, G. R., Varani, J., Orr, W., Gondek, M. D. and Ward, P. A.: Resorbing bone in chemotactic for monocytes. Nature, 275: 132-134, 1978.
References_xml – reference: 18) Fishman, D. A. and Hey, E. D.: Origin of osteoclasts from mononuclear leukocytes in regenerating newt limbs. Anat. Res., 143: 329-339, 1962.
– reference: 12) 山本きよみ, 米原ヤス子, 三輪史郎: エステラーゼ染色. 月刊 Medical Technology 編, 染色法のすべて, 医歯薬出版, 東京, 1980, 118-121.
– reference: 16) Dominguez, J. H. and Mundy, G. R.: Monocytes mediate osteoclastic bone resorption by prostaglandin production. Calcif. Tissue Int., 31: 29-34, 1980.
– reference: 5) 角田左武郎, 松本英彦, 古屋誠, 木村義孝, 南雲正男: マクロファージによる直接的骨吸収. 日臨免誌, 6: 160-166, 1983.
– reference: 13) 湊啓輔, 西田俊明, 斎藤博, 長谷哲夫, 下山正徳: リンパ球幼若化反応. 日本臨床, 37: 1574-1580, 1979.
– reference: 19) Jee, W. S. S. and Nolan, PD.: Origin of osteoclasts from the fusion of phagocytes. Nature, 200: 225-226, 1963.
– reference: 1) Owen, M.: Histogenesis of bone cells. Calcif. Tissue Res., 25: 205-207, 1978.
– reference: 2) Mundy, G. R., Varani, J., Orr, W., Gondek, M. D. and Ward, P. A.: Resorbing bone in chemotactic for monocytes. Nature, 275: 132-134, 1978.
– reference: 17) 小野崎菊夫, 橋本達一郎: マクロファージを活性化するリンフォカイン, MIFの精製から細胞性免疫の発現まで. 蛋白質核酸酵素, 26: 1910-1924, 1981.
– reference: 10) Thorsby, E, and Bartlie, A.: A rapid method for preparation of pure lymphocyte suspensions. In: Histocompatibility Testing 1970, ed., Terasaki, P. I., Munksgaard, Copenhagen, 1971, 655-656.
– reference: 15) 佐藤巌雄, 藤橋弘, 岩川吉伸, 中村安隆, 丸本淑子, 池田克已: ヒト単球の骨塩への作用. 日歯周誌, 27: 146-152, 1985.
– reference: 11) Kumagai, K., Itoh, K., Hinoma, S. and Tada, M.: Pretreatment of plastic petridishes with fetal calf serum. A simple method for macrophage isolation. J. Immunol. Methods, 29: 17-25, 1979.
– reference: 3) Heersche, J. N. M.: Mechanism of osteoclastic bone resorption: A new hypothesis. Calcif. Tissue Res., 26: 81-84, 1978.
– reference: 6) 丸本淑子, 楠公仁, 藤橋弘, 岡田春夫, 中村安隆, 池田克已: 歯槽骨吸収におよぼす Osteoclast-activating factor (OAF) の効果, 1. OAFの産生とその精製の試み. 日歯周誌, 24: 121-127, 1982.
– reference: 7) 丸本淑子, 藤橋弘, 佐藤巌雄, 中村安隆, 西本正純, 池田克巳: 歯槽骨吸収におよぼす Osteoclast-activating factor (OAF) の効果, II. OAF産生細胞への単球の関与. 日歯周誌, 25: 823-829, 1983.
– reference: 8) Russel, A. L.: A system of classification and scoring for prevalence surveys of periodontal disease. J. Dent. Res., 35: 350-359, 1956.
– reference: 4) Mundy, G. R., Altman, A. J., Gondek, M. D. and Bandelin, J. G.: Direct resorption of bone by human monocytes. Science, 196: 1109-1111, 1977.
– reference: 9) Löe, H. and Silness, J.: Periodontal disease in pregnancy. I. Prevalence and severity. Acta. Odont. Scand., 21: 533-551, 1963.
– reference: 14) Baehner, R. L. and Nathan, D. G.: Quantitative nitroblue tetrazolium test in chronic granulomatous disease. New Eng. J. Med., 278: 1971-1976, 1968.
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SubjectTerms Human monocytes
Osteoclast-activating factor (OAF)
Periodontal disease
Synthetic hydroxyapatite (HA)
Title Effects of Culture Supernatants of Peripheral Blood Leukocytes on Bone-minerals Resorbing Activity by Human Monocytes
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