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Biomedical subjects

I Kiviranta

Publications and source records attributed to I Kiviranta.

At least 19 recordsLinked to original sources

Articular cartilage collagen birefringence is altered concurrent with changes in proteoglycan synthesis during dynamic in vitro loading.

INTRODUCTION: The articular cartilage collagen network and proteoglycans are subject to changes in deteriorating joint diseases. In this study, we exposed articular cartilage plugs to cyclic loading and investigated the properties of collagen network and proteoglycans in different zones of the articular cartilage. METHODS: Articular cartilage full-depth plugs were exposed in vitro to 4.1 MPa cyclic (0.5 Hz) loading for 1 to 20 hr and investigated using quantitative microscopic methods (i.e., polarized light microscopy, microspectrophotometry, and autoradiography). RESULTS: The loading caused packing or condensation of the tissue. In histological sections, the height of uncalcified articular cartilage decreased by an average of 12.8% (range, 4 to 19.7%). Loading increased the birefringence of collagen in the superficial cartilage (P < 0.05), with thickening of the zone up to 41.4% at 20 hr. The thickness of the intermediate zone increased also (22% at 1 hr and 434% at 20 hr). Concomitantly, the birefringence (P < 0.05) and the thickness of the deep zone decreased (18.5 to 27.8%). Loading for 4 hr increased the 35S-sulphate incorporation of the cartilage explants by an average of 67% (P < 0.05). The increase was most significant in the deep cartilage. A simultaneous increase was observed in the proteoglycan concentration of the cartilage; the staining intensity with safranin-O increased by 8.8% (P < 0.05). After 8 hr loading, this stimulation decreased; at 20 hr, loading caused a clear inhibitory effect on proteoglycan synthesis in the superficial zone. DISCUSSION: According to these results, the chosen loading regimen increased the thickness and collagen orientation in the superficial zone. In contrast, the thickness and birefringence in the deep cartilage were reduced. The proteoglycan metabolism of chondrocytes was first stimulated deep in the cartilage, but as the loading continued, the effect proved to be inhibitory (especially in the superficial part of uncalcified cartilage).

Animals

Markers of cartilage and synovial metabolism in joint fluid and serum of patients with chondromalacia of the patella.

OBJECTIVE: To further our understanding of the pathogenesis of chondromalacia of the patella (CM), we have studied the release into knee joint fluid and serum, obtained from patients with CM, of molecules associated with the metabolism of joint cartilage matrix and synovium. METHODS: Interleukin-1 alpha (IL-1 alpha), interleukin-1 beta (IL-1 beta), interleukin-6 (IL-6), stromelysin-1 (MMP-3), interstitial collagenase (MMP-1), tissue inhibitor for metalloproteinases-1 (TIMP-1), phospholipase activity A2 (PLA2), hyaluronan (HA), aggrecan fragments (AGN) and antigenic keratan sulfate (KS) were quantified in knee joint lavage fluid from 96 patients with CM; KS and HA also was measured in serum. Chondromalacia was graded on a scale of I to IV according to Outerbridge (1961). The histopathology of the synovial membrane close to the patellofemoral joint was evaluated. Control samples were obtained from nine patients with knee pain presenting with arthroscopically normal knee joints. RESULTS: The concentrations of MMP-3, MMP-1 and TIMP-1 proteins in joint lavage fluid were increased in advanced (grade IV) CM, compared with controls. Levels of MMP-1 in lavage fluid correlated with the severity of CM (r = 0.38, P < 0.01) and MMP-1 and MMP-3 concentrations correlated with each other (r = 0.45, P < 0.001). TIMP-1 was elevated in grade IV CM compared with grades II and III CM (P < 0.02, P < 0.01). Interleukins (IL-1 alpha, IL-1 beta and IL-6) showed no significant change in CM. The lavage fluid level of PLA2 increased with the severity of CM (r = 0.40, P < 0.001). Serum KS was higher in CM IV than in controls (P = 0.05), while lavage fluid KS concentration was elevated in CM I (P = 0.04). There were no differences in the lavage fluid levels of AGN and HA between the different study groups. Synovium showed slight or moderate histological signs of inflammation in 9% of CM patients. CONCLUSION: The changes in the release and activity of these marker molecules from serum and synovial fluid may reflect changes in the metabolism of articular cartilage and synovium in CM, that are consistent with those observed in early-stage tibiofemoral cartilage changes in OA.

Adult

Collagen crosslinks in chondromalacia of the patella.

The aim of the study was to determine collagen concentration and collagen crosslinks in cartilage samples from chondromalacia of the patella. To study the extracellular matrix alterations associated to chondromalacia, we determined the concentration of collagen (hydroxyproline) and its hydroxylysylpyridinoline and lysylpyridinoline crosslinks from chondromalacia foci of the patellae in 12 patients and 7 controls from apparently normal cadavers. The structure of the collagen network in 8 samples of grades II-IV chondromalacia was examined under polarized light microscopy. The full-thickness cartilage samples taken with a surgical knife from chondromalacia lesions did not show changes in collagen, hydroxylysylpyridinoline and lysylpyridinoline concentration as compared with the controls. Polarized light microscopy showed decreased birefringence in the superficial cartilage of chondromalacia lesions, indicating disorganization or disappearance of collagen fibers in this zone. It is concluded that the collagen network shows gradual disorganization with the severity of chondromalacia lesion of the patella without changes in the concentration or crosslinks of collagen.

Adult

Articular cartilage superficial zone collagen birefringence reduced and cartilage thickness increased before surface fibrillation in experimental osteoarthritis.

OBJECTIVES: To investigate articular cartilage collagen network, thickness of birefringent cartilage zones, and glycosaminoglycan concentration in macroscopically normal looking knee joint cartilage of young beagles subjected to experimental slowly progressive osteoarthritis (OA). METHODS: OA was induced by a tibial 30 degree valgus osteotomy in 15 female beagles at the age of 3 months. Fifteen sisters were controls. Cartilage specimens were collected seven (Group 1) and 18 months (Group 2) postoperatively. Collagen induced optical path difference and cartilage zone thickness measurements were determined from histological sections of articular cartilage with smooth and intact surface by computer assisted quantitative polarised light microscopy. Volume density of cartilage collagen fibrils was determined by image analysis from transmission electron micrographs and content of glycosaminoglycans by quantitative digital densitometry from histological sections. RESULTS: In the superficial zone of the lateral tibial and femoral cartilage, the collagen induced optical path difference (birefringence) decreased by 19 to 71% (p < 0.05) seven months postoperatively. This suggests that severe superficial collagen fibril network deterioration took place, as 18 months postoperatively, macroscopic and microscopic OA was present in many cartilage areas. Thickness of the uncalcified cartilage increased while the superficial zone became thinner in the same sites. In operated dogs, glycosaminoglycan content first increased (Group 1) in the lateral tibial condyle and then decreased (Group 2) (p < 0.05). CONCLUSION: In this OA model, derangement of the superficial zone collagen network was the probable reason for birefringence reduction. This change occurred well before macroscopic OA.

Animals

Elevated levels of synovial fluid PLA2, stromelysin (MMP-3) and TIMP in early osteoarthrosis after tibial valgus osteotomy in young beagle dogs.

We determined the concentration of markers in cartilage and synovium metabolism in the synovial fluid (SF) of the knee of young beagle dogs with slowly progressive osteoarthrosis. Osteoarthrosis (OA) was induced by a tibial 30 degrees valgus osteotomy to the right hindlimb of 16 dogs. The contralateral knee served as control. The animals were killed 7 (group I) and 18 months (group II) after operation. The levels in SF of chondroitin sulfate (CS), tissue inhibitor of metalloproteinases (TIMP-1), stromelysin (MMP-3), hyaluronan (HA), and the activity of phospholipase A2 enzyme (PLA2) were assayed. The first microscopic signs of cartilage degeneration were observed 7 months postoperatively and the lesions became more severe, including osteophyte formation during the following 11 months. The synovial fluid level of MMP-3 was higher (p = 0.04) at both time-points in the knee joint of the operated hindlimb than in the contralateral joint. On the operated side, 7 months postoperatively, synovial fluid PLA2 activity was higher (p = 0.02) than in the contralateral knee joint, but not 18 months postoperatively. The SF level of TIMP-1 was higher (p = 0.04) in the operated joint than in the contralateral joint 18 months after operation. The molar ratio of MMP-3 to TIMP-1 was higher (p = 0.001) in group II than in group I. The changes observed in the concentration of synovial fluid markers in this slowly progressive canine OA model suggest that activation of an inflammation-related process occurs at an early stage of the OA disease induced by unilateral tibial valgus osteotomy.

Animals

Specimen preparation and quantification of collagen birefringence in unstained sections of articular cartilage using image analysis and polarizing light microscopy.

To establish an optimal method for analysis of the collagen structures from unstained tissue sections, a computerized image analysis system using a charge coupled device camera coupled to a polarizing light microscope was used. Retardation values of birefringence, which are proportional to the content and fibril orientation of collagen in the extracellular matrix of articular cartilage, were determined from sections prepared in different ways. In the superficial zone of articular cartilage, the highest retardation values were recorded from sections cut parallel to the so-called split lines indicating the anisotropic arrangement of collagen. Complete digestion of glycosaminoglycans reduced the retardation value by approximately 6.0%, suggesting a minor, but not insignificant, contribution of glycosaminoglycans to the birefringence of the matrix. The use of a mounting medium with a refractive index close to that of the collagen (e.g. DPX) increased the specificity of the method, since the optical anisotropy of collagen derives predominantly from the intrinsic (structural) birefringence. In conclusion, analysis of unstained sections after careful removal of paraffin and glycosaminoglycans from the tissues provides a sensitive and rapid quantitative assessment of oriented collagen structures in articular cartilage.

Animals

Immobilisation causes longlasting matrix changes both in the immobilised and contralateral joint cartilage.

OBJECTIVE: The capacity of articular cartilage matrix to recover during 50 weeks of remobilisation after an atrophy caused by 11 weeks of immobilisation of the knee (stifle) joint in 90 degrees flexion starting at the age of 29 weeks, was studied in young beagle dogs. METHODS: Proteoglycan concentration (uronic acid) and synthesis ([35S]sulphate incorporation) were determined in six and three knee joint surface locations, respectively. Proteoglycans extracted from the cartilages were characterised by chemical determinations, gel filtration, and western blotting for chondroitin sulphate epitope 3B3. RESULTS: The proteoglycan concentrations that were reduced in all sample sites immediately after the immobilisation, remained 14-28% lower than controls after 50 weeks of remobilisation in the patella, the summit of medial femoral condyle, and the superior femoropatellar surface. In the contralateral joint, there was a 49% increase of proteoglycans in the inferior femoropatellar surface after remobilisation, while a 34% decrease was simultaneously noticed on the summit of the medial femoral condyle. Total proteoglycan synthesis was not significantly changed after immobilisation or 50 weeks' remobilisation in the treated or contralateral joint, compared with age matched controls. The chondroitin 6- to 4- sulphate ratio was reduced by immobilisation both in the radioactively labelled and the total tissue proteoglycans. In the remobilised joint, this ratio was restored in femur, while in tibia it remained at a level lower than controls. Neither immobilisation nor remobilisation induced epitopes recognised by the monoclonal antibody 3B3 on native (undigested) proteoglycans. CONCLUSION: These results show that the depletion of proteoglycans observed after 11 weeks of immobilisation was not completely restored in certain surface sites after 50 weeks of remobilisation. The significant changes that developed in the contralateral joint during the remobilisation period give further support to the idea that a permanent alteration of matrix metabolism results even from a temporary modification of loading pattern in immature joints.

Animals

Slowly progressive osteoarthritis after tibial valgus osteotomy in young beagle dogs.

A slowly progressive osteoarthritis model in the skeletally immature canine knee joint is described. Forty-four young female beagle dogs were chosen as experimental animals. In 15 dogs, a 30 degrees valgus angulation of the right tibia was created by operation. Fourteen dogs underwent sham operation. Fifteen dogs served as control subjects. Alterations in the knee joints were evaluated macroscopically and histologically 7 and 18 months after operation. Seven months after surgery, two of seven beagles that had valgus osteotomy had a lesion with discoloration of cartilage in the medial condyle of the femur. Eighteen months after operation, five of the eight dogs that had valgus osteotomy showed fibrillation of the femoral and tibial cartilages. Mankin's scoring of the knee joint cartilages indicated statistically significant changes as compared with control subjects 7 and 18 months after surgery. Biomechanical analysis revealed shift of the mechanical axis toward the lateral compartment of the knee by the valgus osteotomy, patellofemoral malalignment, and inclination of the tibiofemoral joint line. These biomechanical alterations brought about the most severe cartilage lesions to the medial condyle of the femur and the patellofemoral joint. Cartilage fibrillation took more than 7 months to develop. Thus, this model offers a slowly progressive, well standardized, and reproducible method for the study of early changes of osteoarthritis in young beagle dogs.

Animals

A controlled biochemical and immunohistochemical study of human synovial-type (group II) phospholipase A2 and inflammatory cells in macroscopically normal, degenerated, and herniated human lumbar disc tissues.

STUDY DESIGN: Group II phospholipase A2 enzyme activity was studied biochemically and immunohistochemically in tissue samples from disc prolapses, degenerated discs, and macroscopically normal discs. In parallel, phospholipase A2 and inflammatory cells were studied by indirect immunocytochemistry. OBJECTIVES: To compare phospholipase A2 activity in normal discs and abnormal discs by an identical assay for phospholipases A2, and to compare the occurrence of inflammatory cells with phospholipase A2 activity and immunoreactivity. SUMMARY OF BACKGROUND DATA: It has been suggested that a high phospholipase A2 enzyme activity in herniated disc tissue could be significant in abnormal states such as sciatica and discogenic pain. No comparison between healthy disc tissue and samples of abnormal discs (degenerated or herniated) has been carried out. In particular, an identical assay for phospholipase A2 for such tissue samples, supported by immunohistochemical staining data, has never been applied in parallel to normal and abnormal disc tissue, and neither have such results been compared with the demonstration of inflammatory cells. METHODS: Group II phospholipase A2 enzyme activity was determined, in parallel, using an identical assay for tissue samples from 11 macroscopically normal discs, 33 disc herniations, and six discs showing degeneration by discography. For determination of phospholipase A2 enzyme activity, a radioassay using 1-palmitoyl-2-(1-14C)linoleoyl-L-3-phosphatidylethanolamine as the phospholipid substrate was used. Total tissue DNA as an estimate of total tissue cell number was measured in parallel with phospholipase A2 activity. All tissue samples also were studied by indirect immunocytochemistry, locating phospholipase A2 and T and B lymphocytes. RESULTS: Neither degenerated nor herniated disc tissue samples demonstrated a higher phospholipase A2 activity than control disc tissue samples. Average phospholipase A2 activity was actually higher in the control samples than in herniated disc samples (Mann-Whitney test, P < 0.001), possibly a result of a higher total DNA (P < 0.005). The observed level of phospholipase A2 activity was lower than that of inflammatory human synovial fluid. Neither was there marked immunoreactivity for phospholipase A2, which was observed in chondrocytes in areas of cartilage and occasional disc cells, supporting the biochemical results. Lymphocytes were more numerous only in herniated disc samples (15%), and their presence showed little overlap with phospholipase A2 immunoreactivity. CONCLUSIONS: Synovial-type (Group II) phospholipase A2 enzyme activity is not particularly high in disc tissue and does not appear to be higher in herniated or degenerated discs than control disc tissue. Immunoreactivity to phospholipase A2 is seen only occasionally and is strong only when cartilage tissue is present. Neither are inflammatory lymphocytes commonly observed.

Adult

Application of selected cationic dyes for the semiquantitative estimation of glycosaminoglycans in histological sections of articular cartilage by microspectrophotometry.

Selected commonly used cationic dyes, viz. Thionin, Safranin O, Toluidine Blue O, Dimethylmethylene Blue, Cuprolinic Blue, Cupromeronic Blue, N,N'-Diethylpseudoisocyanine, and a modified PAS-method, and staining methods with a variety of alternative procedures, e.g., variation of pH, use of the critical electrolyte concentration method, and blocking reactions (methylation-saponification, carboxymethylation), were tested to select optimal staining procedures for the semiquantitative histochemical estimation of glycosaminoglycans by microspectrophotometry in sections of articular cartilage. The methods were carried out on 3 microns-thick paraffin and 1 microns-thick glycolmethacrylate sections of bovine articular cartilage. The staining intensity of the sections was measured from spots 25 microns apart using a Leitz MPV 3 microspectrophotometer, starting at the surface of the cartilage and ending up at the tidemark. The result was compared with the fixed-charge density graph determined from the adjacent articular cartilage. Of the dyes tested, Thionin and Safranin O proved to be excellent cationic dyes for the histochemical quantification of cartilage matrix proteoglycans, since the staining intensity curves showed a linear correlation (r = 0.900-0.995) with the fixed charge density curves from the adjacent cartilage. Also, the stain distribution was consistently uniform across the sections. In 1 microns-thick glycolmethacrylate sections, the Safranin O staining gradient showed almost perfect identity with the fixed-charge density curve. Cuprolinic Blue and Cupromeronic Blue combined with the critical electrolyte concentration technique were also useful for the microspectrophotometric assays of glycosaminoglycans, but the presence of metachromasia should be checked prior to the measurements. The reliability of blocking procedures for quantitative histochemical work was not convincing.

Animals

Safranin O reduces loss of glycosaminoglycans from bovine articular cartilage during histological specimen preparation.

The ability of Safranin O, added to fixation and decalcification solutions, to prevent the escape of glycosaminoglycans (GAGs) from small cartilage tissue blocks during histological processing of cartilage has been studied. GAGs in the fixatives and decalcifying solutions used and those remaining in the 1 mm3 cubes of cartilage were assayed biochemically. The quantity of GAGs remaining in the cartilage cubes were determined from Safranin O-stained sections using videomicroscopy or microspectrophotometry. A quantity (10.6%) of GAGs were lost during a conventional 4% buffered formaldehyde fixation (48 h) and a subsequent decalcification in 10% EDTA (12 days) at 4 degrees C. Roughly one-quarter of the total GAG loss occurred during the 48 h fixation, and three-quarters during the 12 days of decalcification. Inclusion of 4% formaldehyde in the decalcification fluid decreased the loss of GAGs to 6.2%. The presence of 0.5% Safranin O in the fixative reduced this loss to 3.4%. When 0.5% Safranin O was included in the fixative and 4% formaldehyde in the decalcification solution, Safranin O staining of the histological sections increased on average by 13.5%. After fixation in the presence of 0.5% Safranin O, there was no difference in the staining intensities when decalcification was carried out in the presence of either Safranin O or formaldehyde, or both. It took 24 h for Safranin O to penetrate into the deep zone of articular cartilage, warranting a fixation period of at least this long. In conclusion, the addition of Safranin O to the fixative and either Safranin O or formaldehyde in the following decalcification fluid, markedly reduces the loss of GAGs from small articular cartilage explants during histological processing. However, for immunohistochemical studies, Safranin O cannot be included in the processing solutions, because it may interfere.

Animals

Coordinated regulation of hyaluronan and aggrecan content in the articular cartilage of immobilized and exercised dogs.

OBJECTIVE: To study the influence of joint loading and immobilization on articular cartilage hyaluronan concentration and histological distribution in the knee joints of young dogs subjected to 11 weeks' immobilization by splinting, and 15 weeks' running exercise at a rate of 40 km/day. METHODS: The amount of hyaluronan in articular cartilage was determined by a competitive binding assay using a biotinylated hyaluronan binding complex (HABC) of aggrecan and link protein. Histologic sections were stained for the localization of hyaluronan with the HABC probe. Extracted proteoglycans were characterized by sodium dodecyl sulfate agarose gel electrophoresis. RESULTS: Immobilization significantly reduced the concentration of hyaluronan in all sites studied (tibial and femoral condyles, patellar surface of femur). The proportion of hyaluronan to total uronic acid (mainly from aggrecan) remained unchanged because of a concurrent decrease in aggrecan. The ratio of hyaluronan and aggrecan remained constant also in runners. The staining pattern of free hyaluronan in the tissue sections and the electrophoretic mobility of the extracted proteoglycans were not affected by the different loading regimes. CONCLUSION: Reduced joint loading due to splint immobilization significantly decreases both hyaluronan and aggrecan in the articular cartilage. The remarkably parallel changes in aggrecan and hyaluronan content suggest that joint loading exerts a coordinated influence on their metabolism.

Aggrecans

Differences in submicroscopic structure of the extracellular matrix of canine femoral and tibial condylar articular cartilages as revealed by polarization microscopical analysis.

The submicroscopic orientation patterns of sulfated glycosaminoglycan side chains of proteoglycan molecules and collagen fibrils were compared in different extracellular matrix areas of femoral and tibial articular cartilages of young adult beagle dogs using qualitative and quantitative polarization microscopic analytical methods. Paraffin sections were cut perpendicularly to the articular surfaces from the femoral and tibial condyles and stained. Picrosirius red F38 staining combined with an antecedent digestion with testicular hyaluronidase was used to enhance the optical anisotropy of collagen. Birefringence of sulfated glycosaminoglycan molecules was selectively amplified by a combination of carboxymethylation with CH3I and a subsequent staining with toluidine blue. The specimens were analysed in a polarization microscope equipped with compensator plates, and retardation values of birefringence were determined in territorial and interterritorial matrix areas of different zones using monochromatic plane polarized light. It was found that besides some similarities there were significant differences in the submicroscopic organization of extracellular matrix between femoral and tibial articular cartilages. Common structural features of the femoral and tibial cartilages were the sulfated glycosaminoglycans and collagen fibrils which were circularly oriented in the territorial matrix, and these components were longitudinally arranged within the trabeculae of the interterritorial matrix. Furthermore, the territorial matrix was a more densely packed structure than the interterritorial matrix. Our results revealed the following major differences between the two cartilages: The degree of orientation of sulfated glycosaminoglycans was higher in the femoral cartilage matrix areas as compared to the identical structures of the tibial cartilage; the collagen structure was more densely packed in the interterritorial matrix of the superficial and mineralization zones of the femoral cartilage than in the tibial cartilage, and except for the zone of mineralization, the degree of collagen orientation was higher in the territorial matrix of the femoral than the tibial cartilage. These findings suggest that the extracellular matrix of femoral condylar cartilage has a more densely packed molecular structure than the softer tibial cartilage matrix. This structural difference may have an influence on the pathogenesis of diseases involving articular cartilage.

Animals

Indentation instrument for the measurement of cartilage stiffness under arthroscopic control.

Changes in the biomechanical properties of articular cartilage are one of the first signs of the tissue degeneration. We have developed a small size indentation instrument for the quantification of cartilage stiffness under arthroscopic control. During measurement, the indenter imposes a constant deformation on the cartilage and the maximal indenter force, by which the cartilage resists the deformation, is used as a measure for cartilage stiffness. The instrument has been tested in laboratory conditions with elastomer and cadaver knee joint cartilage samples. A linear relationship was found between indenter force and elastomer stiffness (r = 0.990, n = 14) as well as between indenter force and cartilage shear modulus obtained by a reference device (r = 0.879, n = 22). Also, the correlation between two repeated measurements at the measurement sites, used to evaluate the reproducibility, was linear (r = 0.953, n = 16). Quantitative detection of cartilage stiffness is possible with the instrument.

Arthroscopes

Decreased torque and electromyographic activity in the extensor thigh muscles in chondromalacia patellae.

The alterations in thigh muscle properties of chondromalacia patellae patients during isometric and dynamic endurance tests were studied using a variokinetic knee testing system linked to surface EMG. A total of 41 patients (chondromalacia group) with arthroscopically certified chondromalacia of the patella were studied. The control group consisted of 31 healthy adult volunteers with no history of knee pain or trauma. Peak torque values were 21% (p < 0.01) and force output values 25% (p < 0.05) lower on the symptomatic side of the chondromalacia group than in the control group. The decrease in the ratio between integrated EMG (IEMG) and measured force were found in all parts of the quadriceps femoris muscle in patients with chondromalacia of the patella in isometric extension. No change in the normalized IEMG levels of the thigh muscles were found between chondromalacia patients and controls in dynamic endurance test. The severity of the chondromalacia of the patella did not affect the level of electromyographic activation in thigh muscles. The ratio of normalized EMG levels of vastus medialis and vastus lateralis did not differ between the groups. The present study showed that chondromalacia patellae patients have reduced force and electromyographic activation levels of quadriceps femoris muscle. Especially, the explosive strength of the quadriceps femoris muscle is reduced.

Adolescent

Proteoglycan depletion and size reduction in lesions of early grade chondromalacia of the patella.

OBJECTIVE: To determine the content and molecular size of proteoglycans (PGs) in patellar chondromalacia (CM) and control cartilages as a first step in investigating the role of matrix alterations in the pathogenesis of this disease. METHODS: Chondromalacia tissue from 10 patients was removed with a surgical knife. Using identical techniques, apparently healthy cartilage of the same site was obtained from 10 age matched cadavers (mean age 31 years in both groups). Additional pathological cartilage was collected from 67 patients with grades II-IV CM (classified according to Outerbridge) using a motorised shaver under arthroscopic control. The shaved cartilage chips were collected with a dense net from the irrigation fluid of the shaver. The content of tissue PGs was determined by Safranin O precipitation or uronic acid content, and the molecular size by mobility on agarose gel electrophoresis. RESULTS: The mean PG content of the CM tissue samples with a knife was dramatically reduced, being only 15% of that in controls. The cartilage chips collected from shaving operations of grades II, III, and IV CM showed a decreasing PG content: 9%, 5%, and 1% of controls, respectively. Electrophoretic analysis of PGs extracted with guanidium chloride from the shaved tissue samples suggested a significantly reduced size of aggrecans in the mild (grade II) lesions. CONCLUSION: These data show that there is already a dramatic and progressive depletion of PGs in CM grade II lesions. This explains the softening of cartilage, a typical finding in the arthroscopic examination of CM. The PG size reduction observed in grade II implicates proteolytic attack as a factor in the pathogenesis of CM.

Adult

Articular cartilage thickness and glycosaminoglycan distribution in the young canine knee joint after remobilization of the immobilized limb.

The recovery of articular cartilage from atrophy induced by joint immobilization was investigated in immature dogs. In a previous study, we showed that 11 weeks of immobilization of the knee (stifle) joint of young dogs reduced the concentration of articular cartilage glycosaminoglycans (GAGs) by 13-47%. In the present study, right hindlimbs from six female beagles were immobilized for 11 weeks, as in the previous study, and then were remobilized for 15 weeks. Cartilage from the knee joint was compared with cartilage from nonimmobilized knees of eight age-matched control beagles. Histological samples taken from 11 different locations of the knee joint were stained with safranin O, and microspectrophotometry was used to demonstrate distribution of GAGs in the tissue. After remobilization, GAG concentration was restored in the patellofemoral region and tibial condyles. On the summits of the femoral condyles, and especially at the periphery of the femoral condyles, GAG concentration remained 8-26% less than the control values. On the summits, the thickness of the uncalcified cartilage was as much as 15% less than in the age-matched controls. Consequently, the changes induced by unloading were reversible to a great extent, but a full restoration of articular cartilage was not obtained at all sites of the knee joint within the 15 weeks of remobilization. Immobilization of the skeletally immature joint therefore may affect the development of articular cartilage in such a way that very slow recovery or permanent alterations are induced.

Animals

Lateral release in chondromalacia patellae using clinical, radiologic, electromyographic, and muscle force testing evaluation.

Lateral release (division of the lateral retinaculum of the patella) is a common operative procedure to improve alignment of the patella in patients with chondromalacia patellae. We investigated the effects of lateral release using clinical follow-up and studied the changes in the radiologic measurements of the patellar alignment and electromyographic function of the stabilizing muscles of the knee, as well as using muscle-force tests of the thigh. The study included 75 patients. Diagnostic arthroscopy was done for all patients. In 42 patients with obvious incongruency of the patellofemoral joint observed at arthroscopy, open division of the lateral patellar retinaculum was performed in addition to arthroscopy. Four years after surgery, the results showed that although division of the lateral retinaculum of the patella reduced lateral patellar tilting and lateral patellar displacement, the postoperative physical capacity of the patients was not improved. Lateral release did not impair the electrical activity of thigh muscles. The Lysholm scores of the lateral release group and the control group were equal after four years.

Adult