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EDX-microanalysis for the determination of calcium in trabecular bone samples
Abstracts from the Bone Histomorphometry
415
Workshop
osteons, cMWT was derived from the measurement of the shortest osteon diameter (D) and of the corresponding haversian canal diameter (d): cMWT = (D-d)/2. In the whole group, there was a significant correlation between tMWT and cMWT (Spearman’s R=0.72, P< ,001) as well as within each sex (males R=.56, P< .05; females R= 1 .O, P= .Ol). cMWT was 57.2+7.6 pm and decreased significantly with age in the group of 19(R= -0.46, P< .05) but not significantly in each sex (males R= -0.33, females I?= -0.77). tMWT was 49.2+5.9pm and decreased significantly with age in the whole group(R= -0.52, P< .05) and in males (R= -0.58, P< .05) but not in females (R= -0.77, N=6). These data show that at the BMU level bone formation -in trabecular bone parallels the one in cortical bone.
BONE HISTOMORPHOMETRY IN RHEUMATOID ARTHRITIS P. Lips, R. van Soesbergen, A. Bloat and J.C. Netelenbos Academisch Ziekenhuis Vrqe Universiteit and Slotervaarfziekenhuis, Amsterdam Iliac crest biopsies and vitamin D status were investigated in 17 patients with long standing rheumatoid arthritis, 16 women and 1 man, mean age+SD 70.1+5.9 years. Histomorphometric parameters were as follows: trabecular bone volume (mean+SD) 13.0f4.4 %, relative osteoid volume 3.4k2.1 %, osteoid surface 24.3+13.6%, thickness index of osteoid seams 14.3k4.7, resorption surface 4.2k2.1 %, thickness index of osteoid seams 14.3k4.7, resorption surface 4.2k2.1 %, mineralization rate O.SS+ .l 1 pm/day. The serum concentration of 25-hydroxyvitamin D (25(OH)D) was 16.9+11.1 nmol/L compared with 32.9k13.6 nmol/L in 74 aged controls. Osteomalacia was not observed. Five biopsies showed evidence of high turnover, i.e., an increased resorption surface (2 6 %) and extended osteoid surface (2 20%). In the latter group serum 25(OH)D was 11.4k4.4 nmol/L. In rheumatoid arthritis low 25(OH)D levels may be caused by decreased sunshine exposure and by increased hepatic conversion due to antiinflammatory drugs. The vitamin D deficiency may provoke high bone turnover, which may accelerate bone loss.
cation lag time of 20 days. Resorption was estimated from % crenated surface (%RS). Results: The normal female data fitted an exponential model of bone loss after the age of 49. The same model was applied for forearm mineral density measurements in 388 postmenopausal women and yielded a similar exponential function with a very comparable rate of bone turnover. Conclusions: Age-related bone loss in normal women is entirely due to a resetting of fractional resorption rate and in normal men to a reduced BFR. In male osteoporosis, the accelerated bone loss is entirely due to increased bone resorption.
EDX-MICROANALYSIS FOR THE DETERMINATION OF CALCIUM IN TRABECULAR BONE SAMPLES K. Obrant and R. Odselium Ma/m6
Sweden
Histochemical techniques permit only comparative estimation of the calcium content in trabecular bone samples. By means of EDX-microanalysis the concentration can be determined in any part of the samples, but this technique has to our knowledge never been used on human trabecular bone samples. There is a greater applicability of this technique, if, regardless of the impaired morphological resolution, conventional fixation for light microscopy is used. A methodological study was thus designed, in which the concentration of calcium was determined in different parts of tibia1 metaphyseal bone trabeculae from five men who had a fracture of the same diaphysis some months earlier. Five previously healthy men who had sustained sudden death served as controls. The results were also related to the corresponding Goldner and von Kossa stainings. The concentration of calcium in central parts of normal tibia1 trabecular bone was fairly constant to 45 weight %, whereas in the post traumatic cases the concentration was considerably less. It was also discovered that the Goldner staining was at least 10 times more sensitive for calcium than the von Kossa staining. In conclusion, this paper describes an accurate method for quantitative determination of single elements in anatomically and histochemically defined parts of trabecular bone samples.
BONE FORMATION AND RESORPTION IN NORMAL AND OSTEOPOROTIC MEN AND WOMEN
CHANGES OF COLLAGEN IMPERFECTA
B.E.C. Nordin,* T. Hubert
H. Oxlund, U. Pedersen, C.C. Danielsen, I. Oxlund and 0. Elbrldnd /nstifufe.of Anatomy C, Aarhus University and E.N.T. Dept., Aarhus Kommunehospital.
J. Aaron,+ D. Carter,+ K. Sagreiya+ and
*De@ of Endocrinology, Royal Adelaide Hospital +MRC Unit, The General Infirmary, Leeds +Division of C/inica/ Chemistry, institute of Medicine & Veterinary Science, South Australia To examine further the relationship between bone formation, bone resorption and trabecular bone volume, we now report iliac crest biopsy results from 25 young normal females (YNF); 26 old normal females (ONF); 51 osteoporotic women (FOP); 22 young normal men (YNM); 35 old normal men (ONM); 18 osteoporotic men (MOP); and 18 agematched controls (MC). Bone formation rate (BFR) was calculated from the product of total forming surface, % calcification fronts, and mean seam width assuming a calcifi-
IN OSTEOGENESIS
Osteogenesis imperfecta is a heterogeneous group of genetic disorders that are characterized primarily by increased fragility of bones. Other tissues, such as skin, scleras and teeth, are affected. Changes in the structure of collagen seem to be a common feature. The benefit of biophysical and biochemical studies of tissues from patients with osteogenesis imperfecta is not confined to these relatively rare disorders but may provide new information about collagen function, collagen metabolism, role of collagen types, etc. The major collagen type of bone is type I collagen. In skin 70-85% is collagen type I and 15-30% is collagen type
415
Workshop
osteons, cMWT was derived from the measurement of the shortest osteon diameter (D) and of the corresponding haversian canal diameter (d): cMWT = (D-d)/2. In the whole group, there was a significant correlation between tMWT and cMWT (Spearman’s R=0.72, P< ,001) as well as within each sex (males R=.56, P< .05; females R= 1 .O, P= .Ol). cMWT was 57.2+7.6 pm and decreased significantly with age in the group of 19(R= -0.46, P< .05) but not significantly in each sex (males R= -0.33, females I?= -0.77). tMWT was 49.2+5.9pm and decreased significantly with age in the whole group(R= -0.52, P< .05) and in males (R= -0.58, P< .05) but not in females (R= -0.77, N=6). These data show that at the BMU level bone formation -in trabecular bone parallels the one in cortical bone.
BONE HISTOMORPHOMETRY IN RHEUMATOID ARTHRITIS P. Lips, R. van Soesbergen, A. Bloat and J.C. Netelenbos Academisch Ziekenhuis Vrqe Universiteit and Slotervaarfziekenhuis, Amsterdam Iliac crest biopsies and vitamin D status were investigated in 17 patients with long standing rheumatoid arthritis, 16 women and 1 man, mean age+SD 70.1+5.9 years. Histomorphometric parameters were as follows: trabecular bone volume (mean+SD) 13.0f4.4 %, relative osteoid volume 3.4k2.1 %, osteoid surface 24.3+13.6%, thickness index of osteoid seams 14.3k4.7, resorption surface 4.2k2.1 %, thickness index of osteoid seams 14.3k4.7, resorption surface 4.2k2.1 %, mineralization rate O.SS+ .l 1 pm/day. The serum concentration of 25-hydroxyvitamin D (25(OH)D) was 16.9+11.1 nmol/L compared with 32.9k13.6 nmol/L in 74 aged controls. Osteomalacia was not observed. Five biopsies showed evidence of high turnover, i.e., an increased resorption surface (2 6 %) and extended osteoid surface (2 20%). In the latter group serum 25(OH)D was 11.4k4.4 nmol/L. In rheumatoid arthritis low 25(OH)D levels may be caused by decreased sunshine exposure and by increased hepatic conversion due to antiinflammatory drugs. The vitamin D deficiency may provoke high bone turnover, which may accelerate bone loss.
cation lag time of 20 days. Resorption was estimated from % crenated surface (%RS). Results: The normal female data fitted an exponential model of bone loss after the age of 49. The same model was applied for forearm mineral density measurements in 388 postmenopausal women and yielded a similar exponential function with a very comparable rate of bone turnover. Conclusions: Age-related bone loss in normal women is entirely due to a resetting of fractional resorption rate and in normal men to a reduced BFR. In male osteoporosis, the accelerated bone loss is entirely due to increased bone resorption.
EDX-MICROANALYSIS FOR THE DETERMINATION OF CALCIUM IN TRABECULAR BONE SAMPLES K. Obrant and R. Odselium Ma/m6
Sweden
Histochemical techniques permit only comparative estimation of the calcium content in trabecular bone samples. By means of EDX-microanalysis the concentration can be determined in any part of the samples, but this technique has to our knowledge never been used on human trabecular bone samples. There is a greater applicability of this technique, if, regardless of the impaired morphological resolution, conventional fixation for light microscopy is used. A methodological study was thus designed, in which the concentration of calcium was determined in different parts of tibia1 metaphyseal bone trabeculae from five men who had a fracture of the same diaphysis some months earlier. Five previously healthy men who had sustained sudden death served as controls. The results were also related to the corresponding Goldner and von Kossa stainings. The concentration of calcium in central parts of normal tibia1 trabecular bone was fairly constant to 45 weight %, whereas in the post traumatic cases the concentration was considerably less. It was also discovered that the Goldner staining was at least 10 times more sensitive for calcium than the von Kossa staining. In conclusion, this paper describes an accurate method for quantitative determination of single elements in anatomically and histochemically defined parts of trabecular bone samples.
BONE FORMATION AND RESORPTION IN NORMAL AND OSTEOPOROTIC MEN AND WOMEN
CHANGES OF COLLAGEN IMPERFECTA
B.E.C. Nordin,* T. Hubert
H. Oxlund, U. Pedersen, C.C. Danielsen, I. Oxlund and 0. Elbrldnd /nstifufe.of Anatomy C, Aarhus University and E.N.T. Dept., Aarhus Kommunehospital.
J. Aaron,+ D. Carter,+ K. Sagreiya+ and
*De@ of Endocrinology, Royal Adelaide Hospital +MRC Unit, The General Infirmary, Leeds +Division of C/inica/ Chemistry, institute of Medicine & Veterinary Science, South Australia To examine further the relationship between bone formation, bone resorption and trabecular bone volume, we now report iliac crest biopsy results from 25 young normal females (YNF); 26 old normal females (ONF); 51 osteoporotic women (FOP); 22 young normal men (YNM); 35 old normal men (ONM); 18 osteoporotic men (MOP); and 18 agematched controls (MC). Bone formation rate (BFR) was calculated from the product of total forming surface, % calcification fronts, and mean seam width assuming a calcifi-
IN OSTEOGENESIS
Osteogenesis imperfecta is a heterogeneous group of genetic disorders that are characterized primarily by increased fragility of bones. Other tissues, such as skin, scleras and teeth, are affected. Changes in the structure of collagen seem to be a common feature. The benefit of biophysical and biochemical studies of tissues from patients with osteogenesis imperfecta is not confined to these relatively rare disorders but may provide new information about collagen function, collagen metabolism, role of collagen types, etc. The major collagen type of bone is type I collagen. In skin 70-85% is collagen type I and 15-30% is collagen type