PMID- 14588300
OWN - NLM
STAT- MEDLINE
DCOM- 20031204
LR  - 20220316
IS  - 1529-9430 (Print)
IS  - 1529-9430 (Linking)
VI  - 1
IP  - 6
DP  - 2001 Nov-Dec
TI  - Electron microscopic observation of established chondrocytes derived from human 
      intervertebral disc hernia (KTN-1) and role of macrophages in spontaneous 
      regression of degenerated tissues.
PG  - 422-31
AB  - BACKGROUND CONTEXT: Biological and pathological cell processes during the 
      degeneration of intervertebral discs are as yet poorly understood. PURPOSE: An 
      electron microscope was used to observe disc hernia degeneration at the cellular 
      level as expressed in extruded tissue from a human intervertebral disc and in 
      cultured chondrocytes. The mechanism of spontaneous regression was analyzed in 
      order to investigate the effects of homologous macrophages, and the results of 
      this analysis may be developed into a clinical therapy. STUDY DESIGN/SETTING: 
      Extruded tissue specimens excised during surgery on human intervertebral disc 
      hernia and cultured chondrocytes isolated from the excised tissue were observed 
      by means of electron microscopy. Extracellular matrix metalloproteinase-3 (MMP-3) 
      and its antagonist, tissue inhibitor of metalloproteinases-1 (TIMP-1), were 
      observed by means of immune electron microscopy. Macrophages confirmed by CD68 
      immunostaining were added to the chondrocyte culture and observed by means of 
      electron microscopy. PATIENT SAMPLE: All control subjects and patients gave 
      written consent to the study. OUTCOME MEASURES: KTN-1 was directly observed 
      without culture, and nuclei degeneration, the development of chromatin granules, 
      changes in the osmotic pressure of the nuclear membrane and rough-surfaced 
      endoplasmic reticulum, and the development of fat droplets were observed. 
      METHODS: Tissues excised during surgery were divided, a part of the tissues were 
      fixed in various fixatives for electron microscopy and immune electron microscopy 
      analysis, and the other part was treated with collagenase. In addition, 
      chondrocytes were isolated and cultured. Human peripheral blood mononuclear cells 
      were separated using the Ficoll method. After culturing the cells, macrophages 
      were collected, added to the chondrocyte culture, and observed under an electron 
      microscope. CD68 positivity of the macrophages was confirmed by CD68 
      immunostaining. RESULTS: Freshly isolated chondrocytes in the hernia's extruded 
      region differed markedly from cultured chondrocytes. By means of immunoelectron 
      microscopy, MMP-3 and TIMP-1 were localized at the endoplasmic reticulum of the 
      cultured chondrocytes. Infiltration of macrophages among the chondrocytes was 
      observed in the mixed culture. CONCLUSIONS: The tissue extruded from the 
      intervertebral disc showed obvious signs of degeneration, such as changes in 
      osmotic pressure. Macrophages were observed to be the mechanism of spontaneous 
      regression.
FAU - Tsuru, M
AU  - Tsuru M
AD  - Department of Orthopedic Surgery, Kurume University School of Medicine, 67 
      Asahi-machi, 830-0011 Kurume City, Japan.
FAU - Nagata, K
AU  - Nagata K
FAU - Ueno, T
AU  - Ueno T
FAU - Jimi, A
AU  - Jimi A
FAU - Irie, K
AU  - Irie K
FAU - Yamada, A
AU  - Yamada A
FAU - Nishida, T
AU  - Nishida T
FAU - Sata, M
AU  - Sata M
LA  - eng
PT  - Journal Article
PL  - United States
TA  - Spine J
JT  - The spine journal : official journal of the North American Spine Society
JID - 101130732
RN  - EC 3.4.24.17 (Matrix Metalloproteinase 3)
SB  - IM
MH  - Cell Line
MH  - Chondrocytes/chemistry/*ultrastructure
MH  - Humans
MH  - Intervertebral Disc/*pathology
MH  - Macrophages/*pathology
MH  - Matrix Metalloproteinase 3/analysis
MH  - Microscopy, Immunoelectron
EDAT- 2003/11/01 05:00
MHDA- 2003/12/05 05:00
CRDT- 2003/11/01 05:00
PHST- 2003/11/01 05:00 [pubmed]
PHST- 2003/12/05 05:00 [medline]
PHST- 2003/11/01 05:00 [entrez]
AID - S1529943001000559 [pii]
AID - 10.1016/s1529-9430(01)00055-9 [doi]
PST - ppublish
SO  - Spine J. 2001 Nov-Dec;1(6):422-31. doi: 10.1016/s1529-9430(01)00055-9.