EXTENDED REPORT

ß Irradiation decreases collagen type II synthesis and increases nitric oxide production and cell death in articular chondrocytes

J Ailland¹, W U Kampen², M Schünke¹, J Trentmann¹, B Kurz¹

¹ Institute of Anatomy, University of Kiel, Olshausenstr 40, D-24098 Kiel, Germany
² Clinic of Nuclear Medicine, Arnold-Heller-Str 9, University of Kiel, D-24105 Kiel, Germany

Correspondence to:
Correspondence to:
Dr B Kurz, Anatomisches Institut der CAU zu Kiel, Olshausenstr 40, D-24098 Kiel, Germany;
bkurz{at}anat.uni-kiel.de

Background: When synovitis is proved, intra-articularly injected ß emitting radionuclides like yttrium-90 (⁹⁰Y) are used to treat the inflamed synovium.

Objective: To study the viability, matrix production, and NO production during or after ⁹⁰Y treatment of chondrocytes.

Methods: Monolayer, alginate, and explant cultures of primary bovine articular chondrocytes as well as synoviocytes were incubated with 0–3 MBq ⁹⁰Y/ml medium for four days from culture day 3 onwards. Cell viability was demonstrated by light and electron microscopy or by trypan blue or ethidium bromide/fluorescein diacetate staining, membrane integrity by measurement of lactate dehydrogenase (LDH) activity in the culture supernatants. Biosynthetic activity was demonstrated by incorporation of [³H]proline and immunocytochemical staining of collagen type II. NO production was measured with the Griess reagent.

Results: In chondrocyte and synoviocyte monolayer cultures radiation caused a dose dependent increase in cell death and membrane destruction within four days. In alginate and explant cultures, where proliferation is low, no significantly increased LDH activity was seen, and cell viability was 100% for up to 14 days after irradiation. Collagen type II expression (alginate) and biosynthetic activity (alginate and explants) were decreased dose dependently while there was an increase in NO production. Light and electron microscopy data showed that five weeks after irradiation all cells in alginate and most cells in explants subjected to 3 MBq ⁹⁰Y/ml were dead, whereas after lower amounts of irradiation several morphologically intact cells were found.

Conclusions: ß Irradiation may influence the long term maintenance of cartilage tissue or the aetiology of degenerative joint diseases.

Keywords: radiosynoviorthesis; articular cartilage; matrix synthesis; cell viability

Abbreviations: ANOVA, analysis of variance; ETBr, ethidium bromide; FCS, fetal calf serum; FDA, fluorescein diacetate; HBSS, Hanks’s balanced salt solution; LDH, lactate dehydrogenase; NO, nitric oxide; PBS, phosphate buffered saline; RSYN, radiosynoviorthesis; TBS, Tris buffered saline

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