An electron microscopic study on the aging process of the lumbar intervertebral disc

It is well recognized that the intervertebral disc is composed of a nucleus pulposus, an annulus fibrosus, and a cartilage plate and may play a very important role in spinal stability and kinetics. Degeneration by aging of the lumbar intervertebral disc has been implicated as an important causal fac...

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Published inJournal of Nippon Medical School Vol. 52; no. 4; pp. 408 - 417
Main Author Nakase, Takeshi
Format Journal Article
LanguageJapanese
Published Japan The Medical Association of Nippon Medical School 01.08.1985
Subjects
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ISSN0048-0444
1884-0108
DOI10.1272/jnms1923.52.408

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Abstract It is well recognized that the intervertebral disc is composed of a nucleus pulposus, an annulus fibrosus, and a cartilage plate and may play a very important role in spinal stability and kinetics. Degeneration by aging of the lumbar intervertebral disc has been implicated as an important causal factor in the development of low back pain. This study has been undertaken to evaluate the ultrastructural changes of the nucleus pulposus and the annulus fibrosus of the lower lumbar intervertebral disc which has the most pressure from outside. Morphological alteration of the chondrocytes and the collagen fibrils have been carefully investigated. Twenty-eight intervertebral discs were obtained from autopsies in the department of pathology, Nippon Medical School. The subjects ranged from a 27-week old fetus to an 84-year old man. The intervertebral disc between the 4th and 5th lumbar vertebral body was excised and the nucleus pulposus and the annulus fibrosus were isolated. The annulus fibrosus was further sectionalized to anterior inner part, anterior outer part, lateral inner part, lateral outer part, posterior inner part and posterior outer part. Every part was fixed in 2.5% glutaraldehyde, post-osmificated, dehydrated in graded alcohols and embedded in Epok 812. Ultra-thin sections were cut, stained doubly with uranyl acetate and lead citrate and observed by the Hitachi 800 electron microscope. Moreover, the Periodic Acid Methenamine Silver (PAM) stain had been applied for further detailed analysis of collagen fibril. The results obtained were as follows: 1) In the nucleus pulposus, degeneration of chondrocytes was observed from the fetal stage. 2) In the inner part of the annulus fibrosus, degeneration of chondrocytes began from the second decade and progressed rapidly during the third decade. 3) In the outer part of the annulus fibrosus, degeneration of chondrocytes began from the third decade and progressed during the fourth decade. 4) Comparing the anterior, the lateral and the posterior part of an annulus fibrosus, there was no remarkable change among them. 5) In degenerated chondrocytes, the shape of the cell membrane and nucleus was irregular, and the lacunar space was wide, where cell debris and so-called banded structures were present. These banded structures had 800-1, 000Å periodicity and were negative for the PAM stain, which distinguished them from native collagen fibrils. 6) The collagen fibrils measured 200-800Å in diameter at the nucleus pulposus, 600-1, 200Å at the inner part, and 600-2, 000Å at the outer part of the annulus fibrosus.
AbstractList It is well recognized that the intervertebral disc is composed of a nucleus pulposus, an annulus fibrosus, and a cartilage plate and may play a very important role in spinal stability and kinetics. Degeneration by aging of the lumbar intervertebral disc has been implicated as an important causal factor in the development of low back pain. This study has been undertaken to evaluate the ultrastructural changes of the nucleus pulposus and the annulus fibrosus of the lower lumbar intervertebral disc which has the most pressure from outside. Morphological alteration of the chondrocytes and the collagen fibrils have been carefully investigated. Twenty-eight intervertebral discs were obtained from autopsies in the department of pathology, Nippon Medical School. The subjects ranged from a 27-week old fetus to an 84-year old man. The intervertebral disc between the 4th and 5th lumbar vertebral body was excised and the nucleus pulposus and the annulus fibrosus were isolated. The annulus fibrosus was further sectionalized to anterior inner part, anterior outer part, lateral inner part, lateral outer part, posterior inner part and posterior outer part. Every part was fixed in 2.5% glutaraldehyde, post-osmificated, dehydrated in graded alcohols and embedded in Epok 812. Ultra-thin sections were cut, stained doubly with uranyl acetate and lead citrate and observed by the Hitachi 800 electron microscope. Moreover, the Periodic Acid Methenamine Silver (PAM) stain had been applied for further detailed analysis of collagen fibril. The results obtained were as follows: 1) In the nucleus pulposus, degeneration of chondrocytes was observed from the fetal stage. 2) In the inner part of the annulus fibrosus, degeneration of chondrocytes began from the second decade and progressed rapidly during the third decade. 3) In the outer part of the annulus fibrosus, degeneration of chondrocytes began from the third decade and progressed during the fourth decade. 4) Comparing the anterior, the lateral and the posterior part of an annulus fibrosus, there was no remarkable change among them. 5) In degenerated chondrocytes, the shape of the cell membrane and nucleus was irregular, and the lacunar space was wide, where cell debris and so-called banded structures were present. These banded structures had 800-1, 000Å periodicity and were negative for the PAM stain, which distinguished them from native collagen fibrils. 6) The collagen fibrils measured 200-800Å in diameter at the nucleus pulposus, 600-1, 200Å at the inner part, and 600-2, 000Å at the outer part of the annulus fibrosus.
Author Nakase, Takeshi
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References 27) Antonopaulson, C.A.: Determination of glycosaminoglycan (mucopolysaccharides) from tissues on the microgram scale. Biochim. Biophys. Acta, 83, 1-19, 1964.
3) Böhmig, R.: Die BultgefäBversorgung der Wirbelbandscheiben, das Verhalten des intervertebralen Chordasegments und die Bedeutung beider fur die Bandscheibendegeneration; Zugleich ein Beitrag zur enchondralen Ossification der Wirbelkorper. Arch. f. Klin. Chir., 158, 374-424, 1929.
2) 浪越康夫: 崎形性脊椎炎の椎体椎間軟骨および棘状突起における組織学的研究. 日整会誌, 4, 42-83, 1929.
15) Sylven, B.: On the biology of the nucleus pulposus. Acta Orthop. Scand., 20, 275-279, 1951.
23) Hirsch, C. and Schajowitz, F.: Studies on structural changes in the lumbar annulus fibrosus. Acta Orthop. Scand., 22, 184-230, 1952.
29) 井上一: 整形外科領域におけるScanning Electron Microscopyの応用V. 脊椎椎間板. 臨整外, 8, 738-747, 1973.
41) Gardell, S. and Rastgeldi, S.: On the mucopolysaccharides of nucleus pulposus. Acta Chem. Scand. [B], 8, 362-374, 1954.
22) Malmgren, H. and Sylven, B.: Biophysical and physiological investigations on caltilage and other mesenchymal tissues. V. Identification of bovine nuclei pulposi. Biochim. Biophys. Acta, 9, 706-725, 1952.
16) 戸田昭士: 髄核の電子顕微鏡学的研究. 中部整災誌, 2, 556-572, 1960.
32) Armstrong, J.R.: Lumbar Disc Lesions. E. & S. Livingstone Ltd., 1958.
19) 樋口政法: 椎体軟骨板の組織発生, 光学ならびに電子顕微鏡的研究. 日整会誌, 55, 589-600, 1981.
6) Deucher, W.C. and Love, J.G.: Pathologic aspects of posterior protrusion of the intervertebral disc. Arch. Pathol., 27, 201-211, 1939.
25) Naylor, A.: The biophysical and biochemical aspects of intervertebral disc herniation and degeneration. Ann. R. Coll. Surg. Engl., 31, 91-114, 1962.
34) Luse, S.A., Zopf, P. and Cox, T.W.: An electron microscopic study of in vitro and in vivo long spacing collagen. Anat. Rec., 145, 254-255, 1963.
26) Buddecke, E. and Sziegoleit, M.: Isolierung, Chemische zusammensetzung und altersabbangige verteibung im mucopolysacchariden menschlicher zwischenwirbelscheiben. Hoppe-Seyler Z. Physiol. Chem., 337, 66-70, 1964.
44) Mitchell, P.E.G., Hendry, N.G.C. and Billewicz, W.Z.: The chemical background of intervertebral disc prolapse. J. Bone Joint Surg. [Br.], 43, 141-151, 1961.
13) 高田典彦: 腰椎椎間板ならびに椎間関節の加令変化に関する研究 (第1編). 日整会誌, 47, 333-349, 1973.
18) Buckwalter, J.A., Maynard, J.A. and Reginald, R.C.: Banded Structures in Human Nucleus Pulposus. Clin. Orthop., 139, 259-266, 1979.
12) 石川正士: 腰部椎間板ヘルニアの研究, 特にその病理学的研究. 千葉医会誌, 38, 3-27, 1962.
31) 矢島権八, 相原薫: Periodic Acid Methenamin Silver染色. 医のあゆみ, 76, 328-338, 1971.
38) 勝呂徹, 中村哲雄, 北原宏, 秋田徹, 土田豊実, 井上駿一: 脊柱側轡症における椎間板の組織学的電顕的検討. 整形外科基礎科学, 11, 193-194, 1984.
1) Schmorl, G.: Zur Kenntnis der Wirbelkörpelepiphyse und der an ihr vorkommenden Verletzungen. Arch. f. Klin. Chir., 153, 35-45, 1928.
40) Hashimoto, K., Yamanishi, Y., Maeyeus, E., Dabbous, M.K. and Kanzaki, T.: Collagenolytic activities of squamous cell carcinoma of the skin. Cancer Res., 33, 2790-2801, 1973.
17) Cornah, M.S., Meachim, G. and Parry, E.W.: Banded structures in the matrix of human and rabbit nucleus pulposus. J. Anat., 107, 351-362, 1970.
20) 渡辺庄司: 人椎間板の加令的変化, 組織化学及び電顕像の比較研究. 信州医誌, 33, 16-32, 1960.
39) 奥田隆司: EHDP短期大量投与ラットの脛骨近位骨端軟骨に認められた周期性横紋を有する特殊線維構造について. 整形外科基礎科学, 11, 193-194, 1984.
28) 吉田三夫: 腰部脊柱における椎間板, 前後縦靭帯および血管系の年令的変化の病瑠組織学的ならびに組織化学的研究. 日医大誌, 36, 61-80, 1969.
33) Jakus, M.A.: Further observations on the fine structure of the cornea. Invest. Ophthalmol. Vis. Sci., 1, 202-225, 1962.
9) Coventry, M.B., Ghormley, R.K. and Kernohan, J.W.: Intervertebral disc; Its microscopic anatomy and pathology; Anatomy, development, and physiology. J. Bone Joint Surg., 27, 105-112, 1945.
43) Bernardi, G., Happey, F. and Naylor, A.: Mucopolysaccharides from cartilarge and nucleus pulposus. Nature, 180, 1341-1342, 1957.
4) Püschel, J.: Der Wassergehalt normaler und degenerierter Zwischen wirbelscheiben. Beitr. z. Path. Anat. u. z. Allg. Path., 84, 123-130, 1930.
21) Keyes, P.S. and Compere, E.L.: The normal and pathological physiology of the intervertebral disc. J. Bone Joint Surg., 14, 897-938, 1932.
37) Ramsey, H.J.: Fibrous long-spacing collagen in tumors of the nervous system. J. Neuropathol. Exp. Neurol., 24, 40-48, 1965.
14) 中村哲雄: 特発性側弯症椎間板の病理組織学的研究. 日整会誌, 54, 523-538, 1980.
30) Takeda, T.: Three-dimensional observation of collagen framework of human lumbar discs. J. Jpn. Orthop. Ass. 49, 45-57, 1975.
42) Hall, D.A., Lloyd, P.F., Happey, F., Horton, W.G. and Naylor, A.: Mucopolysaccharides of human nuclei pulposi. Nature, 179, 1078-1079, 1957.
7) Saunders, J.B., de C.M. and Inman, V.T.: Pathology of the intervertebral disc. Arch. Surg., 40, 389-416, 1940.
10) Töndury, G.: Neuere Ergebnisse über die Entwicklangsphysiologie der Wirbelsaule. Arch. Orthop. Unfall Chir., 45, 313-322, 1952.
11) Harris, R.L.: Structural changes in the lumbar intervertebral discs. Their relationship to low back pain and sciatica. J. Bone Joint Surg. [Br.], 36, 304-322, 1954.
36) Silberberg, R., Silberberg, M. and Feir, D.: Occurrence of long-spacing (FLS) collagen in articular cartilage of the mouse. Path. Microbiol., 26, 779-783, 1963.
5) Übermuth, H.: Die Bedeutung der Altersveranderungen der menschlichen Bandscheiben fur die Pathologie der Wirbelsaule. Arch. f. Klin. Chir., 156 , 567-577, 1930.
8) 塚田一雄: 人類椎間体の年令による変化に就て. 日病会誌, 29, 338-339, 1939.
24) Davidson, E.A. and Woodhall, B.: Biochemical alterations in herniated intervertebral discs. J. Biol. Chem., 234, 2951-2954, 1959.
35) Raimondi, A.J., Mullan, S. and Evans, J.P.: Human brain tumors; An electron microscopic study. J. Neurosurg., 19, 731-753, 1962.
References_xml – reference: 9) Coventry, M.B., Ghormley, R.K. and Kernohan, J.W.: Intervertebral disc; Its microscopic anatomy and pathology; Anatomy, development, and physiology. J. Bone Joint Surg., 27, 105-112, 1945.
– reference: 15) Sylven, B.: On the biology of the nucleus pulposus. Acta Orthop. Scand., 20, 275-279, 1951.
– reference: 43) Bernardi, G., Happey, F. and Naylor, A.: Mucopolysaccharides from cartilarge and nucleus pulposus. Nature, 180, 1341-1342, 1957.
– reference: 41) Gardell, S. and Rastgeldi, S.: On the mucopolysaccharides of nucleus pulposus. Acta Chem. Scand. [B], 8, 362-374, 1954.
– reference: 40) Hashimoto, K., Yamanishi, Y., Maeyeus, E., Dabbous, M.K. and Kanzaki, T.: Collagenolytic activities of squamous cell carcinoma of the skin. Cancer Res., 33, 2790-2801, 1973.
– reference: 42) Hall, D.A., Lloyd, P.F., Happey, F., Horton, W.G. and Naylor, A.: Mucopolysaccharides of human nuclei pulposi. Nature, 179, 1078-1079, 1957.
– reference: 3) Böhmig, R.: Die BultgefäBversorgung der Wirbelbandscheiben, das Verhalten des intervertebralen Chordasegments und die Bedeutung beider fur die Bandscheibendegeneration; Zugleich ein Beitrag zur enchondralen Ossification der Wirbelkorper. Arch. f. Klin. Chir., 158, 374-424, 1929.
– reference: 5) Übermuth, H.: Die Bedeutung der Altersveranderungen der menschlichen Bandscheiben fur die Pathologie der Wirbelsaule. Arch. f. Klin. Chir., 156 , 567-577, 1930.
– reference: 36) Silberberg, R., Silberberg, M. and Feir, D.: Occurrence of long-spacing (FLS) collagen in articular cartilage of the mouse. Path. Microbiol., 26, 779-783, 1963.
– reference: 35) Raimondi, A.J., Mullan, S. and Evans, J.P.: Human brain tumors; An electron microscopic study. J. Neurosurg., 19, 731-753, 1962.
– reference: 44) Mitchell, P.E.G., Hendry, N.G.C. and Billewicz, W.Z.: The chemical background of intervertebral disc prolapse. J. Bone Joint Surg. [Br.], 43, 141-151, 1961.
– reference: 16) 戸田昭士: 髄核の電子顕微鏡学的研究. 中部整災誌, 2, 556-572, 1960.
– reference: 10) Töndury, G.: Neuere Ergebnisse über die Entwicklangsphysiologie der Wirbelsaule. Arch. Orthop. Unfall Chir., 45, 313-322, 1952.
– reference: 14) 中村哲雄: 特発性側弯症椎間板の病理組織学的研究. 日整会誌, 54, 523-538, 1980.
– reference: 38) 勝呂徹, 中村哲雄, 北原宏, 秋田徹, 土田豊実, 井上駿一: 脊柱側轡症における椎間板の組織学的電顕的検討. 整形外科基礎科学, 11, 193-194, 1984.
– reference: 34) Luse, S.A., Zopf, P. and Cox, T.W.: An electron microscopic study of in vitro and in vivo long spacing collagen. Anat. Rec., 145, 254-255, 1963.
– reference: 23) Hirsch, C. and Schajowitz, F.: Studies on structural changes in the lumbar annulus fibrosus. Acta Orthop. Scand., 22, 184-230, 1952.
– reference: 13) 高田典彦: 腰椎椎間板ならびに椎間関節の加令変化に関する研究 (第1編). 日整会誌, 47, 333-349, 1973.
– reference: 6) Deucher, W.C. and Love, J.G.: Pathologic aspects of posterior protrusion of the intervertebral disc. Arch. Pathol., 27, 201-211, 1939.
– reference: 32) Armstrong, J.R.: Lumbar Disc Lesions. E. & S. Livingstone Ltd., 1958.
– reference: 18) Buckwalter, J.A., Maynard, J.A. and Reginald, R.C.: Banded Structures in Human Nucleus Pulposus. Clin. Orthop., 139, 259-266, 1979.
– reference: 19) 樋口政法: 椎体軟骨板の組織発生, 光学ならびに電子顕微鏡的研究. 日整会誌, 55, 589-600, 1981.
– reference: 26) Buddecke, E. and Sziegoleit, M.: Isolierung, Chemische zusammensetzung und altersabbangige verteibung im mucopolysacchariden menschlicher zwischenwirbelscheiben. Hoppe-Seyler Z. Physiol. Chem., 337, 66-70, 1964.
– reference: 21) Keyes, P.S. and Compere, E.L.: The normal and pathological physiology of the intervertebral disc. J. Bone Joint Surg., 14, 897-938, 1932.
– reference: 31) 矢島権八, 相原薫: Periodic Acid Methenamin Silver染色. 医のあゆみ, 76, 328-338, 1971.
– reference: 30) Takeda, T.: Three-dimensional observation of collagen framework of human lumbar discs. J. Jpn. Orthop. Ass. 49, 45-57, 1975.
– reference: 7) Saunders, J.B., de C.M. and Inman, V.T.: Pathology of the intervertebral disc. Arch. Surg., 40, 389-416, 1940.
– reference: 33) Jakus, M.A.: Further observations on the fine structure of the cornea. Invest. Ophthalmol. Vis. Sci., 1, 202-225, 1962.
– reference: 4) Püschel, J.: Der Wassergehalt normaler und degenerierter Zwischen wirbelscheiben. Beitr. z. Path. Anat. u. z. Allg. Path., 84, 123-130, 1930.
– reference: 1) Schmorl, G.: Zur Kenntnis der Wirbelkörpelepiphyse und der an ihr vorkommenden Verletzungen. Arch. f. Klin. Chir., 153, 35-45, 1928.
– reference: 28) 吉田三夫: 腰部脊柱における椎間板, 前後縦靭帯および血管系の年令的変化の病瑠組織学的ならびに組織化学的研究. 日医大誌, 36, 61-80, 1969.
– reference: 8) 塚田一雄: 人類椎間体の年令による変化に就て. 日病会誌, 29, 338-339, 1939.
– reference: 24) Davidson, E.A. and Woodhall, B.: Biochemical alterations in herniated intervertebral discs. J. Biol. Chem., 234, 2951-2954, 1959.
– reference: 25) Naylor, A.: The biophysical and biochemical aspects of intervertebral disc herniation and degeneration. Ann. R. Coll. Surg. Engl., 31, 91-114, 1962.
– reference: 37) Ramsey, H.J.: Fibrous long-spacing collagen in tumors of the nervous system. J. Neuropathol. Exp. Neurol., 24, 40-48, 1965.
– reference: 20) 渡辺庄司: 人椎間板の加令的変化, 組織化学及び電顕像の比較研究. 信州医誌, 33, 16-32, 1960.
– reference: 39) 奥田隆司: EHDP短期大量投与ラットの脛骨近位骨端軟骨に認められた周期性横紋を有する特殊線維構造について. 整形外科基礎科学, 11, 193-194, 1984.
– reference: 11) Harris, R.L.: Structural changes in the lumbar intervertebral discs. Their relationship to low back pain and sciatica. J. Bone Joint Surg. [Br.], 36, 304-322, 1954.
– reference: 12) 石川正士: 腰部椎間板ヘルニアの研究, 特にその病理学的研究. 千葉医会誌, 38, 3-27, 1962.
– reference: 2) 浪越康夫: 崎形性脊椎炎の椎体椎間軟骨および棘状突起における組織学的研究. 日整会誌, 4, 42-83, 1929.
– reference: 29) 井上一: 整形外科領域におけるScanning Electron Microscopyの応用V. 脊椎椎間板. 臨整外, 8, 738-747, 1973.
– reference: 17) Cornah, M.S., Meachim, G. and Parry, E.W.: Banded structures in the matrix of human and rabbit nucleus pulposus. J. Anat., 107, 351-362, 1970.
– reference: 22) Malmgren, H. and Sylven, B.: Biophysical and physiological investigations on caltilage and other mesenchymal tissues. V. Identification of bovine nuclei pulposi. Biochim. Biophys. Acta, 9, 706-725, 1952.
– reference: 27) Antonopaulson, C.A.: Determination of glycosaminoglycan (mucopolysaccharides) from tissues on the microgram scale. Biochim. Biophys. Acta, 83, 1-19, 1964.
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Snippet It is well recognized that the intervertebral disc is composed of a nucleus pulposus, an annulus fibrosus, and a cartilage plate and may play a very important...
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StartPage 408
SubjectTerms Adolescent
Adult
Aged
Aging
aging process
banded structure
Child
Child, Preschool
chondrocyte
collagen fibril
Female
Humans
Infant
Infant, Newborn
intervertebral disc
Intervertebral Disc - ultrastructure
Lumbar Vertebrae - ultrastructure
Microscopy, Electron
Middle Aged
Pregnancy
Title An electron microscopic study on the aging process of the lumbar intervertebral disc
URI https://www.jstage.jst.go.jp/article/jnms1923/52/4/52_4_408/_article/-char/en
https://www.ncbi.nlm.nih.gov/pubmed/4055993
https://www.proquest.com/docview/76429775
Volume 52
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ispartofPNX Journal of Nippon Medical School, 1985/08/15, Vol.52(4), pp.408-417
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