Aims: Fourier transform infrared imaging (FTIRI) is a new quantitative imaging technique for direct visualization of chemical constituents. Our goal was to investigate the suitability of FTIRI to characterize material properties of articular cartilage (AC) and its ability to indirectly determine biomechanical characteristics of AC. Methods: Cylindrical AC samples (dia.=3.7 mm, n=6) with different stages of osteoarthrosis (OA) were prepared from bovine patellae and mechanical properties of AC were determined with a highresolution material testing device to determine Youngñs modulus (stiffness) at equilibrium (E). After biomechanical testing, one piece of the sample was processed for the histological grading of OA and the other piece was processed for FTIRI. Measurements were conducted from air-dried cryosections. Degree of cartilage degeneration was characterized by the integrated area of amide I and II absorbance. Water content of the specimen was determined from the remaining tissue by measuring the wet and dry weight of the sample. Results: Histological Mankinñs grades of the samples ranged from 0 to 7 indicating that cartilage samples showed only mild to moderate OA. FTIRI absorption showed high correlations with histological grading (r=−0.928) and water content (r=−0.980). Also, average infrared absorption of AC correlated highly linearly with E (r=0.826). Conclusions: Present results show that FTIRI offers a new tool for structural evaluation of AC quality and chemical composition. FTIR correlated well with the histological and biomechanical þndings. Technique offers a new approach to optically determine cartilage constituents. In addition to in vitro research FTIR can be coupled to arthroscopic þber optic probe in order to diagnose cartilage structure and composition in vivo.

Introduction: The evidence of genetic background as an important causative factor in disc degeneration and osteoporosis is increasing. Defects in the COL2A1 gene coding for type II collagen are known to lead to disturbed chondrogenesis and ossification. Retardation of growth, abnormal shape of vertebral bodies and intervertebral discs and occult spina bifida have been described in young mice with the defect. How the gene defect is manifested later in life has not been described.

Purpose of the study: The purpose of this study was to describe, at the microscopic level, the structure of intervertebral discs of transgenic Del1 mice carrying a deletion mutation in the Col2a1 gene, and the effect of the gene defect on the structural properties of bone. In addition, we wanted to see how the gene defect manifests in disc tissue and skeletal bone later in life and if there were differences between sexes.

Materials and methods: The study material consisted of transgenic male (n=27) and female (n=21) mice and their age-matched littermate controls (n=22 and 21, respectively). The transgenic mice were offspring of the transgenic founder mouse Del1 harbouring six copies of a mouse type II collagen transgene with a 150-bp deletion. The mice were divided into two age groups, the younger group being 3 to 13 months and the older 15 to 21 months of age. The two major macromolecules of the intervertebral discs, proteoglycans (PGs) and collagen, were studied. The PG concentration of the intervertebral discs’ nucleus pulposus, annulus fibrosus, and the vertebral bodies and end plates was measured from Safranin-O-stained sections using digital densitometry. Collagen orientation of these structures was evaluated using quantitative polarised light microscopy. Bone mineral density (BMD) was measured with dual energy x ray absorptiometry (DXA), and the breaking force of the femoral bone with three point bending test only for nine 14-month-old females (four control mice and five with gene defect) and fourteen 14-month-old male mice (six control mice and eight with gene defect).

Results: In the young mice, there were no changes in the measured parameters in the intervertebral discs due to the gene defect. However, Safranin-O density and thus PG concentration of the vertebral trabecular bone was 47 % lower in the young transgenic female mice than in the controls (p< 0.001). Ageing had a significant effect on the measured parameters. The Safranin-O density in the nucleus pulposus of the old transgenic male mice was 35 % higher than in the age-matched controls (p< 0.05). In the females, however, Safranin-O density in the nucleus pulposus was 53 % (p< 0.01) and in the vertebral bone 68 % (p< 0.01) lower in the transgenic mice than in the controls. The Safranin-O density in the annulus fibrosus of the transgenic female mice was not changed as compared to the controls. The collagen orientation in the nucleus pulposus of old transgenic male mice was 27 % higher than in the age-matched controls (p< 0.05). In the old females there was no difference in the collagen orientation of the nucleus pulposus between the transgenic mice and controls but in the annulus fibrosus the orientation was 41 % (p< 0.01) and in the vertebral bone 70 % (p< 0.05) lower in the transgenic mice than in the controls. There was no difference in the BMD and the breaking force of the femurs of 14-month-old male mice as compared with the age-matched controls. However, in the old transgenic female mice, the femoral BMD was 14 % (p=0.05) and the breaking force 27 % (p=0.09) lower than in the controls.

Conclusions: The transgene of the Col2a1 gene caused a decrease in the nucleus pulposus PG concentration and in the annulus fibrosus collagen orientation in the old female mice. These features can compromise the structural and load-bearing properties of the discs and thus predispose to disc degeneration. Interestingly enough, the male mice seemed to benefit from the genetic defect in this respect. In addition, in the old transgenic female mice, the PG concentration and the collagen orientation of the vertebral trabecular bone were decreased which contributed to the loss of BMD and breaking force of bone seen in these mice. The fact, that these differences in the bone were not seen in the male mice suggests that this animal model could possibly be used in studies of postmenopausal osteoporosis.