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Biomedical subjects

D Chappard

Publications and source records attributed to D Chappard.

At least 109 records · Page 6Linked to original sources

Improved algorithms for automatic bone histomorphometry on a numerized image analysis system.

Quantitative histological methods have proved to be the most effective methods in bone disease research. Faster and more accurate techniques are currently needed. We have developed a simple digitized image analysis system which allows accurate measurements of trabecular bone mass. The algorithm is based on the 'four-connected sets' mathematical theory. Given a numerized image displayed by a CCD camera, the algorithm recognizes all possible four-connected sets and provides area measurements. The first procedure automatically eliminates small, irrelevant profiles (wrinkles, cell nuclei, etc.) while larger profiles are erased interactively. The second procedure similarly erases the artefactual defects within the trabeculae (artefactual cracks or empty osteocytic lacunae). The method was shown to be very accurate and time-saving.

Algorithms↗

Trabecular bone remodeling after seven days of weightlessness exposure (BIOCOSMOS 1667).

Seven male rats were exposed to 7 days of weightlessness in the Soviet mission COSMOS 1667 and compared with seven control rats by bone histomorphometric methods. In proximal tibial metaphysis, the trabecular bone volume was markedly reduced in flight animals. Trabeculae were decreased in number and thickness; this probably leads to alteration of bone mechanical properties. Formation activity (reflected by measurements of osteoid seams) was decreased at trabecular and endosteal levels. Resorption activity (estimated by count of osteoclast number and active resorption surfaces using a histoenzymologic method) remained unchanged. The imbalance between these cellular activities appears to be responsible for the loss of trabecular bone mass. In proximal femoral metaphysis, measurements were performed in an area located under the muscular insertions. The trabecular bone volume, despite a slight decrease in flight rats, was not significantly different from that of control rats. Furthermore, osteoclastic and osteoid parameters were unchanged. Differential responses between these two long bones need additional investigations. In thoracic and lumbar vertebrae no detectable change in bone mass and bone resorption parameters was found.

Animals↗

Spatial distribution of trabeculae in iliac bone from 145 osteoporotic females.

145 women showing clinical and radiological signs of involutional osteoporosis of the spine were biopsed at the ilium for histomorphometric analysis of bone mass including trabecular bone volume and parameters reflecting the spatial distribution of bony elements (mean trabecular plate thickness, density and separation). Results were compared with an age-matched population of 22 healthy females. Postmenopausal osteoporotics (i.e. younger than 75 years) were characterized by a significant reduction in trabecular bone volume, plate density and thickness, while senile osteoporotics (i.e. older than 75 years) did not exhibit any difference with controls. 51% of the osteoporotic patients had a trabecular bone volume higher than the spontaneous vertebral crush threshold defined by Meunier. Osteoporotic patients with trabecular bone volume under the vertebral crush threshold had a significant decrease in all trabecular parameters. On the opposite, patients with trabecular bone volume above the vertebral crush threshold had only a significant decrease in the number of trabeculae. A negative correlation was found between age and plate density in both osteoporotic patients and controls. A linear correlation was found between trabecular bone volume and plate density, but thickness and density of trabecular plates were not correlated. This study confirms that involutional osteoporosis is not only a decreased bone mass disorder. A modified spatial distribution of trabeculae or a mechanically less resistant bone matrix could be additional factors.

Aged↗

[Effects of weightlessness on phospho-calcium metabolism and its hormonal regulation in man during the 51 G Franco-American space flight].

During the 51 G franco-american mission in weightlessness, calcium metabolism and hormonal regulation were analyzed in two astronauts (a male, a female) before (30,7 and 4 days) and after (0,2 and 5 days) the 7-day spaceflight. Calcium, phosphate, alkaline phosphatase, GLA protein, 25 hydro-vitamin D, 1-25 dihydroxyvitamin D, parathyroid hormone, 24 h urinary calcium, total, dialysable and nondialysable hydroxyproline and glycosaminoglycans (GAG) were measured in blood and urine. Only urinary parameters are increased after space flight. Blood parameters, in particular hormone measurements, are unchanged. The data indicate stimulation of resorptive activity which could result in bone matrix atrophy and demineralisation. On the contrary, no bone formation impairment is noted since alkaline phosphatase and GLA protein are unchanged. These changes are not dependent on hormonal variations. They could only reflect the mechanical bone adaptation to weightlessness.

Adult↗

[Relations between trabecular bone mass and the configuration in the space of the trabeculae of bone. Histomorphometric study of a population of 145 osteoporotic women].

145 women with involution osteoporosis and at least one vertebral compression noted on X-Rays, underwent a biopsy of the iliac wing and measurement of the mean cortical thickness, trabecular bony volume and parameters concerning the spatial configuration of bony trabecules (MTPT = Mean Trabecular Plate Thickness, MTPD = Mean Trabercular Plate Density, MTPS = Mean Trabecular Plate Separation), reflecting respectively the thickness, the density and the separation of the trabecules. The results were compared with those of a series of 22 control women of the same age. The post-menopausal osteoporotic patients present a decreased VTO, MTPD and MTPS with normal MTPT. Patients with senile osteoporosis show no difference as compared to the control patients of the same age. Half of the patients had a VTO higher than the spontaneous vertebral fracture threshold. Involution osteoporosis where the VTO exceeds the threshold present a significant decrease of the number of bony trabecules without decreased thickness. There is a negative correlation between age and the number of trabecules; a linear correlation between VTO and thickness, between VTO and trabecular density confirming the close relationship between mass and spatial configuration of the bone. There is a geometric configuration between density and separation of the trabecules, indicating a great sensitivity of the microstructural framework of disorganized trabecular bone. These data show that a decreased bony mass is not the only physiopathological mechanism observed during involution osteoporosis. Alterations of the spatial configuration of the trabecules or the mechanical resistance of the bony matrix are additional factors, the role of which might have been underestimated.

Aged↗

B cell malignancies presenting with unusual bone involvement and mimicking multiple myeloma. Study of nine cases.

Nine unusual cases of B cell malignancies presenting with lytic bone lesions on radiography and/or hypercalcemia, and mimicking multiple myeloma are reported in this study. These cases included acute lymphoblastic leukemia (one patient), non-Hodgkin's lymphoma (five patients), chronic lymphocytic leukemia (two patients), and Waldenström's disease (one patient). Quantitative bone biopsy performed in two patients (one with non-Hodgkin's lymphoma and one with Waldenström's disease) demonstrated a dramatic increase in osteoclastic resorption close to tumor cells in both cases, similar to or even more marked than that observed in multiple myeloma. Finally, significant production of bone resorbing activity was found in short-term liquid culture specimens of tumor cells in eight of the nine cases. These observations clearly indicate that in B cell disorders other than multiple myeloma, the malignant B cells can produce significant bone resorbing activity in vitro and result in excessive osteoclastic resorption and lytic bone lesions, frequent hypercalcemia, and an overall disease picture clinically mimicking multiple myeloma.

Adolescent↗

Effects of weightlessness on bone mass and osteoclast number in pregnant rats after a five-day spaceflight (COSMOS 1514).

Five pregnant growing rats were orbited for 5 days aboard the Soviet COSMOS 1514 biologic satellite. The bone effects of weightlessness were studied and compared to those of five pregnant rats kept in vivarium and five pregnant conditioned rats. Bone histomorphometric studies were performed to investigate the early effects of weightlessness in loaded (tibia-femur) and unloaded (thoracic and lumbar vertebrae) bones. A short exposure to weightlessness does not induce any change in bone mass and inner structure in either type of bone. In unloaded bones, the number of osteoclasts per square millimeter of the trabecular surface significantly increased when measured after histochemical demonstration of tartrate-resistant acid phosphatase. It is likely that a stimulation of bone resorption activity occurs in the trabeculae of unloaded bones during the early phase of a spaceflight. In tibia, osteoid seam thickness and total osteoclastic resorption surfaces at the endosteal level were not modified.

Acid Phosphatase↗

Bone embedding in pure methyl methacrylate at low temperature preserves enzyme activities.

Pure Methyl Methacrylate (MMA), a widely used embedding medium for undecalcified bone studies, was polymerized at low temperature (4 degrees C). MMA was prepared by a new purification procedure yielding a absolutely anhydrous and catalysed resin. The redox system benzoyle peroxide/NN-Dimethylanilin was used as the catalyzer-initiator system providing free radicals for the MMA chemical polymerization. Since the reaction is inhibited at -20 degrees C, complete infiltration of blocks is achieved within 3 d. Polymerization took place at +4 degrees C. The method provides undecalcified bone sections suitable for histomorphometric analysis of osteoid tissue, tetracycline bone labeling and Tartrate Resistant Acid Phosphatase. Enzyme histochemistry was shown to be possible in pure MMA embedded bones, when this low temperature embedding was used.

Alkaline Phosphatase↗

Effects of a 120 day period of bed-rest on bone mass and bone cell activities in man: attempts at countermeasure.

The effects of immobilization on bone mass and bone remodeling in patients with spinal cord injuries are known to simulate weightlessness-induced bone changes in astronauts. Nevertheless, immobilization has never been investigated using histomorphometric studies in healthy volunteers. Twenty healthy male volunteers participated in a '120 day bed-rest' experiment in the USSR. Bone biopsy cores of iliac crest were taken before and at the end of the period of bed-rest. The subjects were divided into five groups. Five subjects underwent a normal ambulatory life (control subjects); three subjects were placed on continuous bed-rest for 120 days (complete immobilization); four subjects were immobilized and underwent a controlled training program; four subjects were immobilized and received treatment with potassium diphosphonate (ethane-1,hydroxy-1,diphosphonate 900 mg/day, per os); and four subjects were immobilized and received diphosphonate plus physical exercise. Parameters of bone mass and bone cellular activities (osteoblastic formation and osteoclastic resorption) were measured using automatic and semi-automatic image analysis systems. Bone mass remained constant in each group. Cellular activity measurements showed that in completely immobilized men, the mineralization rate was lower than in controls without change in osteoid parameters; in contrast, osteoclastic parameters were increased. In immobilized men given the training program, bone formation was normal and bone resorption was increased. In immobilized men treated with diphosphonate, osteoid parameters and resorption activity were decreased. In immobilized men with diphosphonate plus training, the osteoid parameters and the resorption activity were reduced but to a lesser degree than in immobilized men with diphosphonate alone. Failure of bone loss in normal immobilized subjects differed from results found in paraplegic patients. However, a decrease in mineralization rate and an increase in bone resorption activity were found in both studies. Exercise stimulated bone resorption and diphosphonate inhibited the osteoclastic activity. These data emphasize the difficulties in finding good models to stimulate spaceflight conditions on earth. Comparative studies must be done using bone biopsies to determine more precisely the effects of weightlessness on the human skeleton.

Adult↗

[Contribution of bone biopsy with histomorphometry in the evaluation of monoclonal gammopathy].

Histomorphometric analysis of bone specimen from the iliac crest was performed to determine total trabecular resorption surface (TTRS) in 46 patients presenting with a monoclonal gammopathy, which was benign in 17 cases and malignant in 29 others. In benign gammopathies, 14 patients had normal TTRS; no adverse progression, representing a malignant B-cell lymphoproliferative disorder occurred after 40 months of follow-up. The three other patients presented increased areas of resorption in contact with lymphoplasmocyte nodules or pure plasmocytes and had an unfavorable course: multiple myeloma, Waldenström's disease, and chronic lymphocytic leukemia. In myeloma, an increase in TTRS was observed when the bone marrow is invaded confirming the dominant local activity of an "osteoclast activating factor". TTRS is all the more extensive according to bone marrow invasion or in IgA or light chain myeloma, all of which are especially destructive of bone. It thus appears that determination of TTRS by bone histomorphometry is of diagnostic importance to recognize the benign or malignant nature of the disorder and has prognostic value in case of multiple myeloma. Bone histomorphometry thus appears as a useful tool in the thorough assessment of monoclonal gammopathies.

Biopsy↗

[Effects of weightlessness on osseous tissue of the rat after a space flight of 5 days (Cosmos 1514)].

Five pregnant female growing rats have been orbited for five days aboard the Cosmos 1514 soviet biological satellite. They were compared to five female rats kept in vivarium and five female conditioned rats in synchronised way. Histomorphometric studies were performed in order to investigate: 1. The early effects of weightlessness on the bone mass in loading (tibiae and femur) and unloading bones (thoracic and lumbar vertebrae). 2. The changes of osteoblastic and osteoclastic activities. A short exposure does not induce changes in the bone mass and the inner structure of loading and unloading bones. These results fit in well with human data available in the literature: they show that weightlessness doesn't change bone mass in the early phase of a spaceflight. However extrapolation of animal results to men is discussed. In unloading bones (vertebrae) osteoblastic activity was not measurable. Osteoclasts detected by histoenzymologic method don't change as far as their number per mm3 of trabecular bone is concerned. However the number per mm2 of trabecular area increases. It seems likely that an increase of the osteoclastic population occurs in trabecular bone. In loading bones, formation activity (appreciated by the measurement of osteoid seam thickness) and total osteoclastic resorption surfaces were not modified. These results are different from those of longer flights.

Animals↗

[The bone in non-corticoid-treated sarcoidosis. A histomorphometric study].

The sarcoidosis granuloma secretes 1-25 dihydroxyvitamin D (1-25(OH)2 D) at the origin of the hypercalcemia found in about 20 p. cent of sarcoidoses. Its mechanism was considered exclusively of a digestive nature, but the activity of osteoclastic stimulation of 1-25(OH)2 D must consider an associated bony origin. Nine patients with bouts of non-steroid sarcoidosis were studied by osseous histomorphometric analysis. In a patient with hypercalcemia and radiographic manifestations, a high degree of bone remodelling is demonstrated, with stimulation of osteoclastic and osteoblastic activity. In eight patients with normocalcemia, two subgroups are individualized: two patients present intramedullary sarcoidosis granulomas with increased contact resorption; six patients present a normal marrow: the total areas of resorption remain increased. It is concluded that the activity of osteoclastic resorption is increased in sarcoidosis, which may explain partially the hypercalcemia. The absence of titration of 1-25(OH)2 D in our study does not permit to precise the role of this enzyme in the genesis of hyperosteoclastosis, although this seems quite plausible.

Adult↗

[Quantitative histological study of the bone mass and cellular activity after 120-days decubitus. Trial of preventive protocols].

Bone biopsies of iliac crests, performed at the beginning and the end of a decubitus period of 120 days, were studied in 20 healthy volunteers men. The purpose of this experiment is to simulate the bony alterations of astronauts. The bony mass of all bed-ridden patients remained constant. In strictly immobilized patients, the rate of bony mineralization is decreased and the osteoclastic resorption activity is stimulated. In bed-ridden patients subjected to preventive physical exercises, the bony formation and resorption are increased. When prevention is administered with the use of a diphosphonate, the osteoid and osteoclastic parameters are decreased. When both types of prevention are administered, the osteoid parameters and the resorption activity are decreased but less markedly than in patients treated with diphosphonate alone.

Adult↗

[Effects of weightlessness on phosphorus and calcium metabolism and bone remodeling].

Weightlessness results in negative calcium balance which can only reflect a redistribution of calcium in the body: the loss of calcium in the faeces and/or urine is constant, but an increase in urinary hydroxyproline indicating bone collagen destruction is not always detectable; moreover, a slowing down of collagen maturation may be suspected. Bone analysis by histomorphometry in animals and by indirect, non-invasive methods in man shows a decrease in bone mass. However, this bone tissue atrophy might only reflect excessive ageing of the bone during weightlessness, as suggested by slow bone formation and lack of variation in bone resorption. Since the experimental results obtained in men and animals during simulated weightlessness on earth are not strictly identical with those observed in space- flights, their validity may be questioned. Additional studies (notably histomorphometric studies) are therefore required for a better knowledge, as well as prevention, of the problems raised by human life in space.

Animals↗

Uncalcified cartilage resorption in human fetal cartilage canals.

In the human fetus, epiphyses appear as a solid avascular cartilaginous mass until the eleventh week of development. Around the third fetal month of development, vascular canals coming from the perichondrium are recognized in the mineralized epiphyseal cartilage. Whether cartilage canals develop by passive inclusion or active chondrolysis is still a matter of controversy. We studied the relationships between the intracanalar cells and the surrounding matrix on human fetal epiphyses embedded in glycol methacrylate. At the blind end of canals both stellate fibroblast-like cells and vacuolated macrophages are observed. These cellular foci show all characteristics of active chondrolysis (loss of metachromasia, lacunae containing cells intimately associated with matrix, and presence of granular debris). Similar resorptive foci have been observed in the pannus of rheumatoid joints and in the embryonic chick growth plate composed of uncalcified cartilage. A cellular cooperation (fibroblast/macrophage) is necessary for uncalcified cartilage breakdown. In the human fetus, monocytes/macrophages have been recognized in the peripheral blood as early as the twelfth week of gestation. Our observations support the view that chondrolysis due to both fibroblasts (of mesenchymal origin) and macrophages is the basic mechanism for cartilage canal development.

Abortion, Spontaneous↗

Importance of quantitative histology of bone changes in monoclonal gammopathy.

Quantitative histology of bone changes, using undecalcified transiliac bone biopsies (UTBB), was performed blindly in 46 individuals with monoclonal gammopathy (MG), including 17 with MG of undetermined significance (MGUS) and 29 with overt multiple myeloma (MM). Three MGUS presented an excess of osteoclastic resorption (OR) in the vicinity of clusters of tumour cells and developed overt B cell malignancies, chronic lymphocytic leukaemia, Waldenström's disease and MM respectively. On the other hand, MGUS with normal OR remained stable (median follow-up = 28 months), with one exception who developed a systemic amyloidosis. In MM, excessive OR was only observed in areas invaded by myeloma cells. OR was frequently normal in active MM lacking myeloma cells in UTBB. Active MM without lesions on radiography had excessive OR. IgA and pure Bence Jones MM appeared more osteoclastic than IgG cases (P less than 0.05). Of major interest was the finding that one third of MM presented histological bone changes similar to osteoporosis, osteosclerosis or osteoblastic metastasis. Two major findings must be emphasized from the current data: UTBB could be of major interest for the early detection of a B cell malignancy; heterogeneity of myeloma bone condition is unexpected. If some changes appear directly related to the tumour (i.e. excessive OR or osteoblastic dysfunction), some others are probably accidentally associated with it (i.e. osteoporosis), both needing treatment other than chemotherapy.

Aged↗