A M Säämänen

University of Kuopio, Kuopio, Eastern Finland Province, Finland

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Publications (24)55.24 Total impact

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    ABSTRACT: 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.
    Journal of Orthopaedic Research 04/1994; 12(2):161-7. DOI:10.1002/jor.1100120203 · 2.97 Impact Factor
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    ABSTRACT: The effects of long distance running training on blood parameters, hormone responses and bone growth were studied in young growing dogs. A genetically uniform group of female beagles matched with respect to age and body mass were used. The runner dogs (n = 10) underwent gradually increased running exercise up to 40 km.day-1 on a treadmill with 15 degrees uphill gradient 5 days each week during a period of 1 year, while the littermate control dogs (n = 10) were kept in their cages throughout the study. Low plasma lactate concentrations of the runners measured immediately after the running training indicated the aerobic metabolism of the dogs while running. Significant decreases of blood haemoglobin concentrations (11%), blood erythrocyte number (10%), and erythrocyte packed cell volume (12%) were found in the runner group. Throughout the experiment, the value of thyroxine was slightly lower (13%) in the runners but no changes were found in tri-iodothyronine, free thyroxine, or cortisol serum concentrations. Serum oestradiol concentration at 56 weeks was significantly lower (42%) in the runner group than in the control group but was not as low (27%) at 70 weeks. Somatomedin-C concentration had decreased significantly by 37% at the age of 56 weeks in the runner group but was again at the level of the control dogs at the end of experiment (at 70 weeks). Ulna and radius bone mass as a ratio to the body mass had significantly increased in the runners. It would seem from our study that long distance running has a positive effect on bone growth.(ABSTRACT TRUNCATED AT 250 WORDS)
    European Journal of Applied Physiology and Occupational Physiology 02/1993; 67(4):321-9. DOI:10.1007/BF00357630
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    ABSTRACT: Collagen and proteoglycans in the intervertebral disc (LI-II) of young beagle dogs (age 55 weeks) were analyzed following a 15 weeks' daily 20 km running training on a treadmill with 15 degree uphill inclination. In nucleus pulposus no statistically significant alterations were found in the content of proteoglycans or collagen. In annulus fibrosus the total tissue wet weight and total amount of collagen (hydroxyproline) increased by 34-36% in the runners as compared to age-matched, untrained controls. Since the total amount of proteoglycans did not increase, the annulus fibrosus became relatively depleted of proteoglycans, as indicated by the 27% reduction in uronic acid concentration, expressed either per wet weight or hydroxyproline. The average molecular size of the remaining nonaggregating proteoglycans was larger, and there was also a trend towards increased proportion of proteoglycans aggregating with hyaluronan. Most of the chondroitin sulfate side chains were 6-sulfated (65-66%). Running did not alter the sulfation or length of the chondroitin sulfate chains. The decreased proteoglycan/collagen ratio in annulus fibrosus may result in altered mechanical properties of the tissue and reflects its adaptation to enhanced motion and stress.
    International Journal of Sports Medicine 02/1993; 14(1):48-51. DOI:10.1055/s-2007-1021145 · 2.37 Impact Factor
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    ABSTRACT: The influences of the strenuous running training program on the knee joint articular cartilage was studied in six female beagle dogs. At the age of 15 weeks, the dogs started running on a treadmill inclined 15 degrees uphill. Thereafter, the dogs were trained for 40 weeks, five times a week. For the final 15 weeks, the dogs ran 20 km/day. Six age-matched female beagles served as controls. The cartilage surfaces were intact after the running exercise. The training reduced the thickness of the uncalcified cartilage by 6% in the medial femoral condyle. The glycosaminoglycan concentration was reduced an average of 11% on the summits of the femoral condyles. The reduction was most pronounced (41%) in the superficial 50-micron cartilage zone. In other regions of the knee, such a decrease of glycosaminoglycans was not observed. A shift to strenuous running voided the increase in cartilage thickness and proteoglycan content previously observed after moderate running. Strenuous running induced marked depletion of proteoglycans from the superficial layer of the femoral condyles at sites subjected to highest impact loads.
    Clinical Orthopaedics and Related Research 11/1992; DOI:10.1097/00003086-199210000-00046 · 2.88 Impact Factor
  • Duodecim; lääketieteellinen aikakauskirja 02/1992; 108(12):1097-107.
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    ABSTRACT: Human whole skin was labeled for 24 h with [6-3H]-glucosamine in organ culture and epidermis, dermis and culture medium were separately analyzed for the molecular mass and content of the [3H]-labeled hyaluronan (HA). Gel filtration on Sephacryl S-1000 of HA purified by HPLC showed a large proportion of the newly synthesized HA to be of a very high molecular mass (greater than 2 X 10(6) Da) in both epidermis and dermis, whereas HA in the medium was of a smaller size. After 24 h chase, most of the high molecular mass HA, and 42-48% of total labeled HA disappeared from both tissue compartments. The size of labeled HA recovered in the chase media was further reduced but the content roughly corresponded to that lost from tissue. The amount of unlabeled HA was not significantly altered in epidermis, whereas in dermis it was reduced to about 10% of the initial values during 5-d culture. The results demonstrate that HA of both epidermis and dermis is synthesized as a very high molecular mass compound but rapidly undergoes a limited degradation into large fragments. The fragmentation of HA is suggested to enhance its diffusion from the tissues, particularly dermis.
    Journal of Investigative Dermatology 08/1991; 97(1):126-30. DOI:10.1111/1523-1747.ep12478553 · 6.37 Impact Factor
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    ABSTRACT: The distribution of proteoglycans (PGs) at 11 sites on the knee (stifle joint) cartilage of young female beagle dogs was studied following cast immobilization for 11 weeks in 90 degrees flexion and after a subsequent remobilization for 15 weeks. Immobilization induced a reduction in PG uronic acid at all sites (mean of -38%), but the greatest depletion (-64%) occurred at the anterior and posterior extremes of the femoral condyles, i.e., at locations where the immobilized cartilage lost contact to the opposing cartilage. Following remobilization, the content of uronic acid remained lower than in the age-matched controls (-18% on average), particularly at the minimum contact sites most affected by immobilization (-33%). The chondroitin-6-sulfate to chondroitin-4-sulfate ratio was reduced by immobilization in most locations (average of -14%) and returned to control values after remobilization. There was no consistent change in the percentage of aggregating PGs observed in Sephacryl S-1000 gel filtration after immobilization or remobilization. However, following remobilization, the aggregating PGs showed an enhanced proportion of the slower mobility band in agarose gel electrophoresis, indicative of a larger monomer size. In the contralateral, load-bearing knee joint, both the uronic acid content and PG monomer type distribution were identical to those observed in the experimental joint, suggesting that the state reached after the remobilization period was due to factor(s) influencing both sides. The results suggest that contact forces between articulating surfaces are required to maintain normal PG content and that the control mechanism works locally at each cartilage site. Restriction of joint mobility and loading in young animals is concluded to cause persistent changes in cartilage matrix. Furthermore, the use of the contralateral joint as the sole control in this kind of studies, although experimentally convenient, seems not to be appropriate.
    Journal of Orthopaedic Research 12/1990; 8(6):863-73. DOI:10.1002/jor.1100080612 · 2.97 Impact Factor
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    ABSTRACT: The indentation stiffness of knee articular cartilage subjected to strenuous physical training (SPT: treadmill running 20 km day-1 for 15 weeks, n = 6) of young Beagles was tested and compared to that obtained from age-matched (55 weeks, n = 9) controls. The mathematical solution for the shear modulus, as determined from indentation of an elastic layer bonded to a rigid half space, was extended to small Poisson's ratios and applied to the analysis of cartilage response after a step stress (0.39 MPa) application. In these measurements with an impervious, plane-ended indenter, the equilibrium deformation was systematically greater than values predicted from the instant response by the linear biphasic theory. Therefore, the accurate determination of Poisson's ratio from the creep curves was not possible. The mean shear modulus (calculated by using the deformation at 900 s after load application and assuming a constant Poisson's ratio of 0.40 for the matrix) of canine knee articular cartilage was 0.37 MPa. While the cartilage thickness was not affected by SPT, the cartilage of the lateral tibial plateau was stiffer (13.3%, p less than 0.05) than that in controls. However, in the femoral condyles, the stiffness was at the control level or even below. Our results on cartilage structure and properties suggest that SPT, in contrast to our previous findings with moderate training, does not necessarily improve the biological properties of articular cartilage in young animals.
    Journal of Biomechanics 02/1990; 23(12):1239-46. DOI:10.1016/0021-9290(90)90381-C · 2.66 Impact Factor
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    ABSTRACT: The levels and types of proteoglycans in articular cartilage of the knees of young beagle dogs were studied after 15 weeks of running exercise, at 4 km/day. Running increased the levels of proteoglycans in the cartilage of the patella, the superior patellofemoral groove, and the summit of the medial condyle of the femur, all of which are considered contact sites subject to enhanced loading caused by running. The elevated content of uronic acid at the femoral sites proved to be due to proteoglycans that were unable to aggregate with hyaluronic acid. There was no change in the content of aggregating proteoglycans. Analysis of chondroitinase AC-derived disaccharides at the same sites showed an increase in chondroitin-6-sulfate content as compared with chondroitin-4-sulfate levels. We believe that this modulation of the proteoglycan matrix reflects enhanced tissue maturation and physiologic adjustment to higher local contact pressures.
    Arthritis & Rheumatology 10/1989; 32(10):1282-92. DOI:10.1002/anr.1780321014 · 7.87 Impact Factor
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    ABSTRACT: The restoration of the biomechanical properties of articular cartilage was studied after 15 weeks of remobilization of the knee joint in beagles previously immobilized with a cast for 11 weeks. The shear moduli were determined with an indentation creep test immediately after load application and at equilibrium at six predefined test points of femoral, tibial, and patellar cartilages. Permeability of the cartilage was estimated from the creep measurements. The values were compared with nontreated, age-matched (55 weeks) controls and with cartilage collected immediately after immobilization. Remobilization reduced the high creep rates created by immobilization and shifted the depressed equilibrium shear moduli towards those of the controls. However, in the femoral condylar cartilage, the equilibrium shear modulus remained at lower level (p less than 0.05) and permeability at higher level (p less than 0.05) as compared with the controls. We conclude that articular cartilage, showing signs of atrophy after long-term immobilization, was capable of restoring its biomechanical properties during remobilization. This repair was not, however, completed in all parts of the knee joint by the end of the observation period.
    Journal of Orthopaedic Research 02/1989; 7(3):352-8. DOI:10.1002/jor.1100070307 · 2.97 Impact Factor
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    ABSTRACT: The instant, creep and equilibrium responses of canine knee articular cartilages were determined after a constant load application with an in situ indentation creep test and related to the chemical composition of the tissue. Instantly, the cartilage stiffness correlated inversely with the proportion of proteoglycans (PGs) extractable with guanidium chloride. The tibial cartilage, rich in PGs but relatively poor in collagen, showed a low resistance to instant rearrangement of the solid matrix after load application. However, the resistance of the tibial cartilage to water flow during creep deformation was similar or even higher than in the femur. The rate of creep correlated inversely with the PG content. The equilibrium modulus of the femoral cartilage (0.40 MPa), 29 per cent higher than in the tibia (0.31 MPa), was related to the content of PGs, while in the tibia the direct correlation between PGs and modulus was not observed. Our results suggest that while PGs control the fluid flow in articular cartilage, a high PG content alone does not guarantee high stiffness of the cartilage. Instead, the properties of the collagen network are suggested to control particularly the instant shape alterations of the articular cartilage under compression.
    ARCHIVE Engineering in Medicine 1971-1988 (vols 1-17) 11/1988; 17(4):157-62. DOI:10.1243/EMED_JOUR_1988_017_042_02
  • A M Säämänen, M Tammi, I Kiviranta, H J Helminen
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    ABSTRACT: Proteoglycans (PGs) and collagen were quantified in the knee articular cartilages of femoral and tibial medial condyles following 1-8 weeks of moderate running exercise of 4- to 6-month-old rabbits. The total content of PGs extractable with 4 mol/l guanidium chloride was elevated in the weight-bearing cartilage of the tibial medial condyle, while their concentration, expressed as uronic acid per wet weight, and collagen remained unchanged. The content of glucosamine (GlcN) and its ratio to galactosamine (GalN) was elevated in femoral cartilage PGs purified by centrifugation in dissociative CsCl gradients, indicating an increase in keratan sulfate. After 8 weeks of running, the chondroitin sulfate chains of PGs from tibial medial condyle contained less unsulfated disaccharide units. The content of chondroitin sulfate was elevated in the nonextractable residue of the tibial medial condyle as indicated by uronic acid and GaN assays. The content of nonextractable GlcN was increased even more, both in tibial medial and femoral cartilages. Moderate running thus increased (1) keratan sulfate-rich PGs, (2) the degree of sulfation of the chondroitin sulfate chains, and (3) nonextractable PGs; these modulatory alterations probably enhance the stability and elastic stiffness of the PG matrix.
    International Journal of Sports Medicine 05/1988; 9(2):127-33. DOI:10.1055/s-2007-1024993 · 2.37 Impact Factor
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    ABSTRACT: The local influences of physical exercise on thickness and glycosaminoglycan (GAG) content of canine articular cartilage were measured by microspectrophotometry of Safranin O- and periodic acid-Schiff (PAS)-stained tissue sections. Female Beagle dogs were housed in individual cages (bottom 0.9 x 1.2 m) and divided into runner (n = 6) and control (n = 8) groups. The training program started at the age of 15 weeks. During the subsequent 10 weeks, the dogs were accustomed to running on a treadmill inclined 15 degrees uphill. Thereafter, the dogs ran 1 h daily, 5 days a week, at a speed of 4 km/h for 15 weeks. At the age of 40 weeks, the dogs were killed, and the samples for histology were taken from 11 different anatomical locations of the right knee (stifle) joint. The thickness of the uncalcified cartilage increased 19-23% on the lateral condyle and patellar surface of the femur, whereas the enhancement was smaller in other parts of the trained cartilage. The calcified cartilage did not show thickness alterations. Total GAGs were augmented by 28% in the summits on the femoral condyles, more on the medial than lateral side. The increased GAGs appeared to be predominantly chondroitin sulphates and were localized in the intermediate, deep, and even in the calcified zones, whereas the superficial zone did not show changes. There was a concomitant increase of non-GAG oligosaccharides in the intermediate and deep zones, but not in the calcified cartilage.(ABSTRACT TRUNCATED AT 250 WORDS)
    Journal of Orthopaedic Research 02/1988; 6(2):188-95. DOI:10.1002/jor.1100060205 · 2.97 Impact Factor
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    ABSTRACT: Casting of the right knee (stifle) joints of young beagle dogs for 11 weeks caused up to 48% reduction in the glycosaminoglycan (GAG) concentration of the uncalcified articular cartilage, as assessed by a new microspectrophotometric method. The GAGs were depleted mainly in the superficial zone of the cartilage. Although the thickness of the uncalcified cartilage was not decreased, the calcified cartilage under the tidemark was thinned by 6-25% at the femoral condyles. The increased weight-bearing in the limb opposite the one in the splint caused uncalcified cartilage thickness to be augmented by 19% and GAG concentration by 25-35% in the intermediate, deep, and calcified zones of the summits of the femoral condyles; the changes were smaller in other, less loaded parts of the joint. It is concluded that in young dogs, increased weight-bearing augments local proteoglycan content of the articular cartilage matrix, while unloading reduces it.
    Arthritis & Rheumatology 08/1987; 30(7):801-9. DOI:10.1097/01241398-198801000-00032 · 7.87 Impact Factor
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    ABSTRACT: The right knees of 4-month-old NZW rabbits were splinted in extension for 1 to 8 weeks. Biochemical changes of the knee articular cartilage were noted after decreased (splinted leg) and increased loading (created by the shift of body weight onto the left, contralateral limb). Increased loading accelerated changes associated with maturation of articular cartilage, which include accumulation of hyaluronic acid (HA) and keratan sulfate-rich proteoglycans (KS, PG) that are tightly bound to the tissue. After 8-weeks of splinting the content of extractable PGs in the tibial medial condyle decreased. The lost material was apparently replaced by PGs with a higher degree of sulfation of the chondroitin sulfate (Ch-S) chains. Reduced loading disturbed normal maturation as evidenced by inhibition of the accumulation of KS-rich, non-extractable PGs. Collagen content increased in all samples of different joint sites and groups during the 8-week experiment. The content of extractable PGs decreased slightly, while the content of non-extractable, especially KS-rich PGs increased. The greatest changes occurred in the tibial medial condyle, where the KS content was highest.
    Connective Tissue Research 02/1987; 16(2):163-75. DOI:10.3109/03008208709002004 · 1.98 Impact Factor
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    ABSTRACT: Ethanol replacement by CO2 of glutaraldehyde-fixed and ethanol-dehydrated rabbit articular cartilage specimens was monitored with both gas chromatograph and alcometer prior to critical point drying (CPD). The surface structure of the patellar specimens was also systematically registered with a semiquantitative scanning electron microscopic method. After a 2 h interval, when about 28 microliters of ethanol/15 min CO2 extract was removed, the articular surface was smooth, although small areas of striated surface and superficial splits were present. A long-term CO2 treatment (16 h) removed ethanol completely, but increased superficial splitting of the articular surface after CPD. Air-drying of the specimens gave rise to inferior preservation of the cartilage: large areas with pitted and leafy surface qualities, but no superficial splits, were present on the surface. It was evident that prolonged ethanol replacement by CO2, prior to CPD, degraded surface structure of the articular cartilage which should be taken into consideration in the planning and design of experiments. Ethanol removal by CO2 could conveniently be monitored by an alcometer.
    Journal of Microscopy 04/1985; 137(Pt 3):305-12. DOI:10.1111/j.1365-2818.1985.tb02587.x · 2.15 Impact Factor
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    ABSTRACT: The influences of joint immobilization and running exercise on the articular cartilage surfaces of the patella and lateral tibial condyle of young rabbits were investigated by the semiquantitative stereomicroscopic and scanning electron microscopic methods during a period of 8 weeks. The smoothness of the articular surfaces was disturbed already 1 week after the onset of immobilization. Leafy, slightly rough and rough surface qualities associated with superficial splits were observed. The changes were of the same nature after a longer period of immobilization. Running exercise on the treadmill (150-300 m twice a day, 12-24 min per turn, 5 times a week) led to an increase of the striated surface quality at 1 to-2-week intervals as compared with the controls. It was apparent that during immobilization remarkable changes of the articular cartilage took place within the first week, while running exercise up to 8 weeks elicited only transient or minor alterations of the articular surface.
    Acta Anatomica 02/1985; 122(1):62-8.
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    ABSTRACT: Staining of articular cartilage by the periodic acid-Schiff (PAS) method was measured using microspectrophotometry. Standard PAS technique with 2 h oxidation produced a distinct Schiff reaction in the cartilage sections. The staining increased with depth of the articular cartilage demonstrating distribution of the glycoproteins. The modified PAS method included a second, longer periodic acid treatment, which made the uronic acid of glycosaminoglycans PAS-positive. The modified PAS method proved to be highly specific for chondroitin sulphate, which was determined from the samples with gas chromatography. A statistically significant correlation between the Schiff reactivity and galactosamine content of the sections was observed. It is concluded that for articular cartilage standard and modified PAS methods are useful procedures for demonstrating local changes of glycoproteins and chondroitin sulphate, respectively.
    Histochemistry 02/1985; 83(4):303-6. DOI:10.1007/BF00684375
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    ABSTRACT: A new microspectrophotometric method was developed for quantitation of glycosaminoglycans with Safranin O dye in articular cartilage matrix. From histological sections molar extinction coefficient of Safranin O was determined and used to measure the dye content of the sections. The amount of glycosaminoglycans was determined with depth of bovine articular cartilage by both gas chromatography and thin layer chromatography to calculate the fixed negative charge content. Comparison between the results revealed that binding of Safranin O to glycosaminoglycan polyanions was stoichiometric and showed minimal nonspecific staining. The method provides an accurate technique for quantitation and localization of fixed negative charge content of glycosaminoglycans in the articular cartilage matrix. Specific enzyme digestions enable detection of separate glycosaminoglycans.
    Histochemistry 02/1985; 82(3):249-55. DOI:10.1007/BF00501401
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    ABSTRACT: The influences of joint immobilization and running exercise on the articular cartilage surfaces of the patella and lateral tibial condyle of young rabbits were investigated by the semiquantitative stereomicroscopic and scanning electron microscopic methods during a period of 8 weeks. The smoothness of the articular surfaces was disturbed already 1 week after the onset of immobilization. Leafy, slightly rough and rough surface qualities associated with superficial splits were observed. The changes were of the same nature after a longer period of immobilization. Running exercise on the treadmill (150–300 m twice a day, 12–24 min per turn, 5 times a week) led to an increase of the striated surface quality at 1 to-2-week intervals as compared with the controls. It was apparent that during immobilization remarkable changes of the articular cartilage took place within the first week, while running exercise up to 8 weeks elicited only transient or minor alterations of the articular surface.
    Cells Tissues Organs 01/1985; 122(1):62-68. DOI:10.1159/000145984 · 1.96 Impact Factor