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The actions of parathyroid hormone on bone: relation to bone remodeling and turnover, calcium homeostasis, and metabolic bone disease. Part I of IV parts: mechanisms of calcium transfer between blood and bone and their cellular basis: morphological and kinetic approaches to bone turnover.

The supracellular organization of living bone enables the study of isolated cellular and subcellular systems to be related to the study of the whole organism. Bone is formed by osteoblasts in successive stages, separated in both time and space, of matrix formation and primary mineralization. Osteoblasts are joined by tight junctions and largely cover the osteoid seam which separates them from mineralized bone. Secondary mineralization is not completed for several months and is not regulated by the osteoblast. Bone is resorbed by osteoclasts which simultaneously accomplish mineral dissolution and matrix digestion. Active osteoblasts occupy about 5% of the free bone surface, osteoid seams with less active osteoblasts about 10%, active osteoclasts about 0.5%, and Howship's lacunae at which bone remodeling is either quiescent or arrested about 5%. The remaining 80% of the free bone surface is covered by a leaky envelope of thin flattened cells, termed surface osteocytes. Some osteoblasts become permanently buried in the bone as deep osteocytes, around which a specialized and metabolically active perilacunar bone is formed. This bone is less highly mineralized and can temporarily lose or gain calcium in accordance with homeostatic needs. Deep osteocytes maintain contact with each other and with the surface osteocytes, their cell processes within canaliculi being joined by gap junctions. Remodeling of cortical bone proceeds with the excavation by osteoclasts of a longitudinal tunnel which is refilled by osteoblasts to form a new osteon. The anatomically discrete longitudinally oriented structure consisting of a cutting cone of osteoclasts in front and a closing cone of osteoblasts behind is termed a cortical remodeling unit. The events of centrifugal resorption and centripetal formation which occur in a single cross section is termed a cortical remodeling cycle. Normally each new cycle is slightly out of phase with its predecessor. The quantities which characterize cortical remodeling are the birth rate of new remodeling cycles or activation frequency (mu), and the durations of the resorptive period (sigma r), the quiescent interval (sigma q) and the formation period (sigma f). The average distances traveled by the osteoclast and osteoblast are indicated respectively by the mean cement line diameter and mean wall thickness of completed osteons. These quantities show little interindividual variation. Because of this constancy the magnitude of bone turnover (the bone formation rate) is almost entirely a function of mu, the activation frequency of new remodeling cycles. Variations in the velocity of advance of osteoclasts (the linear resorption rate) or of osteoblasts (the appositional rate) alter inversely both the extent of surface engaged in resorption or formation and the time taken to replace a particular moiety of bone, but in a steady state do not influence the rate of turnover of the skeleton as a whole...

Animals↗

The actions of parathyroid hormone on bone: relation to bone remodeling and turnover, calcium homeostasis, and metabolic bone diseases. II. PTH and bone cells: bone turnover and plasma calcium regulation.

Kinetic and morphologic studies in patients with parathyroid disease, and a wide variety of studies in experimental animals indicate that one major effect of PTH is to increase the proliferation of osteoprogenitor cells into osteoclasts and so to increase bone turnover. PTH stimulates bone cells by increasing cell membrane permeability to calcium and consequently increasing calcium influx and by activating membrane-bound adenyl-cyclase. It is likely that the former event precedes the latter and that calcium is the second messenger and cyclic AMP the third messenger. PTH increases the production by bone cells of lactate, citric and carbonic acids, lysosomal enzymes, collagenase, and hyaluronic acid, some or all of which are concerned in the mechanism of bone resorption. With the exception of lactate which probably comes mainly from osteocytes, the increase in metabolic activity is largely due to the increase in the number of osteoclasts. There is also ultrastructural, biochemical, and biophysical evidence that PTH stimulates existing osteoclasts, but this most likely represents the transformation of inactive cells into an active state, and is a transient and nonsustainable effect. As yet, there is no evidence that either increased osteoprogenitor cell proliferation or increased osteoclast activity is mediated by adenyl-cyclase activation. PTH also acts on the deep osteocyte to cause rapid mobilization of calcium from the zone of hypomineralized metabolically active perilacunar bone. This effect is mediated by adenyl-cyclase activation and is preceded by a slight fall in plasma calcium probably due to the movement of calcium into bone cells. The function of this rapid hypercalcemic response to PTH is correct errors in the prevailing steady-state level of plasma calcium...

Animals↗

Morphological changes of autoclaved autogenic bone implantation and autoclaved autogenic bone supplemented with allogenic demineralized bone matrix in rat parietal bone.

The healing process of resected, autoclaved (121 degrees C, 20 minutes) and re-implanted bone in the rat parietal bone was compared with that of autoclaved bone that was supplemented with allogenic bone matrix (AAA-bone), using a scanning electron microscope and a light microscope. In the implant without AAA-bone, bone union and replacement of the autoclaved bone was seen at 2 weeks after implantation. There was no evidence of any inflammatory reaction around the autoclaved bone. The implant was gradually replaced by the new bone. In the implant with AAA-bone, the new bone formation around the implanted bone was more abundant than that of the implant without AAA-bone. An inflammatory reaction was also observed after 1 week. The replacement of the implant with AAA-bone was inferior to the nonsupplemented group. The reason for the poor replacement was the disturbance of the blood supply in the implant by abundant new bone formation. In these results, the autoclaved bone re-implantation was an excellent bone substitute with osteoconductive ability and biocompatibility. The implantation with AAA-bone was good for the new bone formation, but the position and the technique of supplement with AAA-bone have to be more deeply investigated.

Animals↗

[Study of bone formation in fetal rat parietal bone using serum-free bone-forming organ culture system: effect of insulin on bone formation].

Serum-free bone-forming organ culture system using twenty-day fetal rat parietal bones was established. The effects of insulin on bone formation were studied in this system. Bones were cultured on grids and placed on a rocking platform under an aerobic condition. This condition increased the bone-forming activities of cultured rat parietal bones. The calcium content, dry weight of bone, and lactate produced by bone tissue in the medium were measured to study the precise bone-forming process of vital bones. Using this improved bone-forming organ culture system, insulin increased calcium content and dry weight of bone. These effects were maximum in bones treated with insulin at 10(-6) M. Histo-morphometrical analysis showed that the areas of mineralized bone and bone matrix increased, and that many osteoblasts and few osteoclasts appeared in bones treated with insulin at 10(-6) M or higher. Insulin inhibited the release of 45Ca into the medium. From these results, it was suggested that insulin has a stimulatory effect on bone formation, enhancing both bone mineralization and bone matrix formation.

Animals↗

The actions of parathyroid hormone on bone: relation to bone remodeling and turnover, calcium homeostasis, and metabolic bone disease. Part IV of IV parts: The state of the bones in uremic hyperaparathyroidism--the mechanisms of skeletal resistance to PTH in renal failure and pseudohypoparathyroidism and the role of PTH in osteoporosis, osteopetrosis, and osteofluorosis.

In early chronic renal failure, the state of the bones resembles that of type II primary hyperparathyroidism. Cortical bone becomes thinner and more porous, and there is increased extent of surface remodeling. These changes are followed in turn by osteomalacia and osteitis fibrosa, although sometimes these may be alternate rather than successive stages. Bone turnover is less than would be expected for the elevation of PTH level, probably because of 1,25 (OH)2D3 deficiency. The resorption velocity and lamellar bone appositional rates are depressed, but woven bone appositional rate may be increased, possibly because of hyperphosphatemia. Bone mass reflects the summation of three independent processes: loss of lamellar bone due to hyperparathyroidism (depending on the extent of insulation by osteoid); accumulation of partly mineralized osteoid because of osteomalacia; accumulation of woven bone because of osteitis fibrosa. Osteosclerosis may be growth-related metaphyseal, subchondral or diffuse axial, and periosteal neostosis may also occur. Some patients on hemodialysis lose bone because of planing rather than lacunar or dissecting resorption, combined with depression of both lamellar and woven bone formation. Hyperparathyroid bone disease tends to improve slowly after renal transplantation. Persistent hypocalcemia reflects a defect in the calcium homeostatic system and cannot be explained solely by the known stimuli to secondary hyperparathyroidism. The increment in plasma calcium in response to PTH infusion is subnormal, both in early chronic and in acute renal failure, probably because of 1,25(OH)2D3 deficiency. This is also the most likely explanation for the depressed level of blood-bone equilibrium. The activity of all three of the PTH responsive cell systems in bone is depressed in renal failure, probably because all three require 1,25(OH)2D3 in order to function normally. In pseudohypoparathyroidism, as in chronic renal failure, hypocalcemia results from a defect in the regulation of the blood-bone equilibrium. The bone-remodeling system shows all gradations of response, from slight depression of bone turnover to overt osteitis fibrosa, but bone turnover is never as low as in PTH deficiency. These differences may reflect the presence or absence of resistance to PTH of the osteoprogenitor cell as well as of the calcium homeostatic system, or may be due to varying degrees of 1,25(OH)2D3 deficiency, as in chronic renal failure. An increase in plasma calcium in response to PTH can occur either in the untreated state or after treatment with vitamin D because either the error-correcting or remodeling system remains responsive to PTH. Pseudohypoparathyroidism may be subdivided into three types, depending on whether the urinary cyclic-AMP response to PTH remains defective despite treatment with vitamin D, improves with treatment, or is normal before treatment. Only the former is associated with the genetic syndrome of Albright's hereditary osteodystrophy...

Bone Diseases↗

Reconstruction of the bone--bone marrow organ by osteogenin, a bone morphogenetic protein, and demineralized bone matrix in calvarial defects of adult primates.

Information concerning the efficacy of osteogenin, a bone morphogenetic protein, and demineralized bone matrix in orthotopic sites in nonhuman primates is a prerequisite for potential clinical application in humans. After exposure of the calvaria, 84 cranial defects, 25 mm in diameter, were prepared in 26 adult male baboons (Papio ursinus). Defects were implanted with insoluble collagenous bone matrix (ICBM, the inactive collagenous residue after dissociative extraction of bone matrix with 4 M guanidine hydrochloride) reconstituted with osteogenin fractions isolated from baboon bone matrix by chromatography on heparin-Sepharose and hydroxyapatite-Ultrogel (Og Hep-HA) or osteogenin further purified using Sephacryl S-200 gel filtration chromatography (Og S-200). Baboon osteogenin with the highest biologic activity in a rodent bioassay, as determined by alkaline phosphatase activity, calcium content, and histologic analysis, was used for orthotopic implantation in baboons. Additional defects were implanted with baboon demineralized bone matrix (DBM) or ICBM without osteogenin as control. Defects also were grafted with corticocancellous bone harvested from the iliac crest or left ungrafted to monitor the spontaneous regeneration potential of the adult baboon calvaria. Undecalcified bone sections at 7 microns were prepared from the harvested specimens 30 and 90 days after surgery. Histomorphometry demonstrated that Og S-200 induced copious amounts of bone and osteoid as early as day 30 (P < 0.01 versus ICBM, autogenous grafts and untreated defects). At day 90, in implants of Og S-200, Og Hep-HA, and DBM, bone and marrow formation was extensive, culminating in complete regeneration of the craniotomies. In implants of DBM, bone formed with an intervening phase of cartilage development. This provides the phenotypic evidence of endochondral bone differentiation by induction in defects of membranous calvarial bone in adult primates. These results establish the potential therapeutic application of osteogenin and demineralized bone matrix for the architectural reconstruction of the bone-bone marrow organ in humans.

Animals↗

[The injection of acrylic bone cement prevents bone collapse in the intercalar bones lacking bony support: an experimental sheep semilunar bone model].

OBJECTIVES: In a sheep semilunar bone model, we investigated whether collapse in the intercalar bones lacking bony support could be prevented by the injection of acrylic bone cement. METHODS: The study included 16 limbs of eight sheep. Preoperatively, anteroposterior and lateral views of the carpal joints in the fore limbs were obtained. The animals were divided into four groups. In group 1 (n=3) no surgical procedure was performed in the right semilunar bones, whereas the periosteum on the contralateral side was elevated (group 2; n=3). The first two groups were left as controls. In Group 3 (n=5) the left semilunar bones were filled with acrylic bone cement following decancellation of the bone, while the right semilunar bones were left decancellated (group 4; n=5). The sheep were monitored for three months. Radiographs of the carpal joints were obtained to evaluate collapse occurrence in the semilunar bones. Thereafter, the animals were sacrificed and the semilunar bones were excised for biomechanical and histological examinations. Osteonecrosis and cartilage damage were sought and resistance to compressive forces was investigated. RESULTS: Radiologically, the extent of collapse was statistically significant in the semilunar bones in group 4 (p<0.05). The use of acrylic bone cement was found to prevent collapse in group 3, with no significant difference being noted between preoperative and postoperative semilunar bone heights (p>0.05). Biomechanically, the least resistance to compressive forces was measured in group 4 (p<0.05). Histologically, cartilage damage and osteonecrosis were only seen in group 4. CONCLUSION: Our data suggest that the use of acrylic bone cement prevents collapse in the semilunar bones, without inducing any cartilage damage or osteonecrosis.

Animals↗

Demineralized bone matrix supplied by bone banks for a carrier of recombinant human bone morphogenetic protein (rhBMP-2): a substitute for autogeneic bone grafts.

Four commercially available preparations of demineralized freeze-dried human bone powders (DFDB) were investigated for endogenous bone morphogenetic protein (BMP) as observed by osteoinductive activity. Composites of DFDB without and with 1 microgram or 10 micrograms of recombinant human bone morphogenetic protein-2 (rhBMP-2) were implanted into the hindquarter muscles of Swiss-Webster mice. The four batches of DFDB without rhBMP-2 were placed also in hindquarter muscles of athymic mice. Three weeks after implantation, the area of induced bone and cartilage formation was measured by radiographic and histomorphometric methods. In normal mice, without rhBMP-2, DFDB implants induced development of dense fibrous connective tissue with minimal, if any, new bone. In athymic mice, DFDB induced development of small patches of appositional new bone. In contrast, in normal mice, composites of DFDB and rhBMP-2 induced development of large areas of heterotopic bone and bone marrow formation. The bone morphogenetic response occurred with a statistically significant difference (p < 0.0001) between implants of 1 microgram and the 10-microgram rhBMP-2 composites. Thus, DFDB from all four bone banks demonstrated comparable capacity to serve as a carrier for rhBMP-2.

Analysis of Variance↗

Comparison of the bone robusticity index and bone weight/bone length index with the results of bone densitometry and bone histomorphometry in experimental studies.

In experimental studies of bone in rats, two morphometric indices reflecting bone density have been proposed, the bone robusticity index and bone weight/bone length index. In rats, the bone mineral content (BMC) and bone mineral density (BMD) of a selected bone can be determined using dual-energy X-ray absorptiometry (DXA); bone volume can be measured by histomorphometry and other techniques. This study was undertaken to compare two morphometric indices (bone robusticity and bone weight/bone length) with the results of DXA and histomorphometry. Forty female Wistar rats (100 days old, mean weight 239+/-12 g) were studied: 20 controls and 20 ovariectomized rats (OVX). The morphometric indices and BMD differed significantly (Friedman test) in the overall group of rats; no differences were observed in the control group, but significant differences were apparent in the OVX group (p<0.0001). The morphometric indices correlated more closely with BMC than with BMD; the femur length/ femur weight index had closer correlations than the robusticity index. Nonetheless, both morphometric indices differed significantly from BMD determined by DXA under abnormal conditions, which makes them unreliable for use in these circumstances.

Absorptiometry, Photon↗

Compressed homologous cancellous bone and bone morphogenetic protein (BMP)-7 or bone marrow accelerate healing of long-bone critical defects.

We studied 18 adult male New Zealand rabbits with a critical right-sided ulnar defect of 15 mm. In six animals the defect was grafted with homologous compressed cancellous bone, in six animals with homologous compressed cancellous bone including 300 micro g bone morphogenetic protein (BMP)-7 and in six animals with homologous compressed cancellous bone including 0.5 ml autologous bone marrow. The defect was studied using radiographs every second week for 10 weeks. At the conclusion of the experiment the animals were killed and the defect studied by histology and histomorphometry. In all animals treated with the addition of autologous bone marrow and in five of six animals treated with the addition of BMP-7, the defect healed. There was no union in animals treated with homologous compressed cancellous bone without additive. The histological picture of the regenerated area was similar in the two experimental groups. Woven bone contained small marrow spaces with fibrous tissue and capillaries. The osteoid seams were on average greater in animals that received autologous bone marrow as compared to animals that received BMP-7.

Animals↗

Requirement of donor-derived stromal cells in the bone marrow for successful allogeneic bone marrow transplantation. Complete prevention of recurrence of autoimmune diseases in MRL/MP-Ipr/Ipr mice by transplantation of bone marrow plus bones (stromal cells) from the same donor.

MRL/MP-Ipr/Ipr (MRL/Ipr) mice possess radioresistant (9.5 Gy) abnormal stem cells and show a recurrence of autoimmune diseases within 5 mo of conventional allogeneic bone marrow transplantation. We recently have found that the MHC preference exists between hemopoietic stem cells and stromal cells; when bones are engrafted, donor-derived stromal cells present in the engrafted bones can migrate into the recipient bone marrows, which are replaced with both donor-derived stromal cells and hematopoietic cells. Based on these findings, we attempted to prevent the recurrence of autoimmune diseases in MRL/Ipr mice by the transplantation of both bone marrow cells and bone (as a source of stromal cells). MRL/Ipr mice were irradiated (8.5 Gy) and then reconstituted with C57BL/6 bone marrow cells plus bone grafts. The mice survived more than 48 wk after this treatment. Immunohistologic studies revealed that the mice were completely free from both lymphadenopathy and autoimmune diseases such as lupus nephritis and rheumatoid arthritis. Sera from these mice showed normal levels of circulating immune complexes and rheumatoid factors. Normal functions of both T cells and B cells were noted. Abnormal T cells such as Thy-1+B220+ cells present in nontreated MRL/Ipr mice could not be seen in the thus-treated mice. In addition, to our surprise, spleen cells from treated mice showed completely normal in vitro primary anti-SRBC responses. These results indicate that stromal cells in allogeneic bone marrow transplantation play a crucial role not only in the prevention of graft failure but also in the successful cooperation among APCs, T cells, and B cells. Although MRL/Ipr mice are radiosensitive and usually die of interstitial pneumonia or fatty liver due to the side effects of radiation, it should be noted that this strategy allows a reduction in the radiation dose (9.5 Gy-->8.5 Gy), and that these mice can survive more than 48 wk without showing any symptoms of autoimmune diseases.

Animals↗

Isolation of bone from muscles prevents the development of experimental callus-like heterotopic bone. A study of the interaction of bone and muscle in new bone formation.

Heterotopic bone (HB) has previously been provoked in the compartment of the profundus part of the vastus intermedius muscle (VIP) by manipulation of the rabbit knee immobilized in extension. The authors isolated the femoral bone of adult rabbits with a permeable or an impermeable polymer membrane to investigate the interactions of bone, periosteum, and muscle in callus-like HB formation, using the HB model. Heterotopic bone developed beside but not over the membrane if isolation was carried out before the immobilization-manipulation period, regardless of the type of membrane used. In cases where the femoral bone was isolated from the muscle by insertion of the tubular membrane one to two weeks after the immobilization-manipulation had begun, there was HB formation over and beside the membrane. No HB developed under the membrane in any of the cases of femoral bone isolation. Thus direct and extensive contact between bone and muscle appears to be essential at the onset of formation of experimental HB. An intertissue exchange of soluble substances derived from bone or muscle, however, does not seem to play an essential role here. After the onset of HB formation, its further development will not be prevented by subsequent isolation of muscle and bone. The study elucidates the interaction between bone and adjacent muscles and the results may be valuable in further investigations on HB and fracture healing.

Animals↗

Cementum-like bone production in solitary bone cysts. (so-called "cementoma" of long bones). Report of three cases. Electron microscopic observations supporting a synovial origin to the simple bone cyst.

Three cases of simple bone cysts (S.B.C) in association with cementum-like bone production are reported. Analysis of our cases and the 2 previously reported in the literature as so-called cementomas has led us to conclude that the "cementoma" of long bone is not a distinct entity, but merely, a form of S.B.C. associated with a peculiar, poorly cellular form of bone which mimics tooth cementum by light microscopy only. Electron microscopic studies of this substance demonstrate collagen fibers and numerous matrix vesicles which form the initial sites of calcification. Matrix vesicles are a product of osteoblastic activity and are never found in the cementum of the tooth or oral cementum producing tumors. We also report the first ultramiscroscopic observations on the wall of the simple bone cyst and its lining. Two cell types constitute the lining, having features identical to those described for type A and type B synovial cells. Because of this new observation, we believe, the most reasonable explanation for the simple bone cyst is that it represents a congenital "rest" of synovial tissue displaced into the thin, cortical metaphyseal region of bones at the synovial-capsularbone reflection. Its benign nature and slow growth would explain its discovery in early childhood and the marked preponderance of its proximal humerofemoral location dependent upon the fact that these 2 bones have the largest area of capsular to metaphyseal bone reflection.

Adolescent↗

Intraoperative bone and bone marrow sampling: a simple method for accurate measurement of uptake of radiopharmaceuticals in bone and bone marrow.

Accurate estimation of bone marrow uptake of radiopharmaceuticals is of crucial importance for accurate whole body dosimetry. In this study, a method for obtaining normal bone marrow and bone during routine surgery without inconvenience to volunteers is suggested and compared to an indirect method. In five volunteers (group 1), 4 MBq 111In-labelled human polyclonal IgG (111In-IgG) was administered 48 h before placement of a total hip prosthesis. After resection of the femoral head and neck, bone marrow was aspirated from the medullary space with a biopsy needle. In five patients, suspected of having infectious disease (group 2), bone marrow uptake was calculated according to a well-accepted method using regions of interest over the lumbar spine, 48 h after injection of 75 MBq 111In-IgG. Bone marrow uptake in group 1 (4.5 +/- 1.3% D kg-1) was significantly lower than that in group 2 (8.5 +/- 2.1% D kg-1) (P < 0.01). Blood and plasma activity did not differ significantly for both groups. This method provides a system for directly and accurately measuring uptake and retention in normal bone marrow and bone of all radiopharmaceuticals at various time points. It is a safe and simple procedure without any discomfort to the patient. Since small amounts of activity are sufficient, the radiation dose to the patient is low.

Aged↗

Relations of bone mineral content, ash weight and bone mass: implication for correction of bone mineral content for bone size.

Bone mineral content (BMC) was measured by photon absorptiometry in fresh bone specimens: 35 iliac crest and 39 forearms were used. BMC was related to ash weight, fat-free weight and the amount of bone determined by quantitative histologic methods. Ninety-five per cent of the variation in bone mineral concentration (BMC') in the iliac crest could be explained by variation in ash weight, and 88% of the variation in BMC in the forearm could be explained by variation in the amount of bone determined histologically. The total cross-sectional bone area of radius and ulna could be calculated from the width of the structures measured on the photon absorption curves. Using this parameter and the relation between BMC and ash weight, the actual in-vivo bone mineral content in the forearm could be expressed in terms of ash-weight in grams per cm3 of total bone.

Anthropometry↗

The penetration of lincomycin into normal human bone. Determination of penetration into compact bone, spongy bone and bone marrow.

The penetration of lincomycin into normal bone was studied in 10 patients with fracture of the neck of the femur, a separate determination being made of the lincomycin concentration in serum, bone marrow, spongy bone and compact bone. The concentration of lincomycin in bone marrow was found to be at the same level as that in the serum. The concentration in spongy bone amounted in most cases to 50 to 75 percent of the concentration in the serum, whereas the concentration in compact bone varied from 0 to 15 percent of that in the serum.

Aged↗

Bone-to-bone, joint-to-bone and joint-to-joint ratios in normal and diseased skeletal states using region-of-interest technique and bone-seeking radiopharmaceuticals.

Bone-to-bone, iliosacral joint-to-os sacrum and joint-to-joint ratios were computed using the region-of-interest technique 2 to 3 hrs. after injection of 99mTc Sn-methylene-diphosphonate or 99mTc Sn-pyrophosphate in 139 patients with skeletal diseases (bone tumours, degenerative changes of the spine and joints, inflammatory changes of joints) as well as in 123 patients with normal skeletal states. In the latter group, iliosacral joint-to-os sacrum ratios decreased with increasing age of the patients. In patients with osseous metastases of the spine ratios of 0.80 to 4.0 occurred ( reference area second vertebra below or above the affected vertebra). In degenerative changes of the spine values of 0.80 to 1.69 were computed. These results show, that 74% of the spine metastases could not be differentiated from benign changes of the spine by determining their relative amounts of bone uptake. In bone tumours of the extremities and in rheumatoid or gouty arthritis of the small joints (hands and feet) the highest ratios, i.e. contrasts, occurred referring to a contralateral reference area. Osteoarthritic and inflammatory alterations of the big joints could not be differentiated because of percentual distribution of the increased joint-to-joint ratios turned out to be nearly identical.

Adult↗