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Biomedical subjects

M D Grynpas

Publications and source records attributed to M D Grynpas.

At least 73 records · Page 4Linked to original sources

Effect of age and osteoarthritis on bone mineral in rhesus monkey vertebrae.

We have used vertebrae of free-ranging rhesus macaques to study the effect of age and osteoarthritis on bone mineralization and bone density and to relate these findings to weight, sex, parity, and mineral chemistry. Bone mineralization was determined using the density fractionation technique and bone density using dual-photon absorptiometry. Arthritis was determined osteologically. We found a relationship between mineralization, age, and osteophytes, such that mineralization rose with age in nonarthritics and decreased with age in arthritics. This could also be seen when the females were examined separately. In males, only an increase in mineralization with age could be seen. In females mineralization decreases with parity. Also in females, DPA density decreases with age and increases with parity. No relationships with DPA density could be seen using males and females together or males alone. In conclusion, we have shown that normal skeletal aging in rhesus monkeys is accompanied by an increase in mineralization similar to that in other species, but this is not true in the presence of osteoarthritis. In the females parity has an important effect because it seems to build up bone mass even though the bone present may be undermineralized.

Absorptiometry, Photon↗

The effects of diet, age, and sex on the mineral content of primate bones.

The effect of diet, age, and sex on the mineral content of primate bones was determined for free-ranging rhesus monkeys (Macaca mulatta) from the Caribbean Primate Research Center. Monkeys in this study were of known age and sex and had been provided with either a low protein (15%) or a high protein (25%) diet for most of their lives. Instrumental neutron activation analysis was used to assess bone mineral content. Results showed that diet had no significant effect on the bulk mineral composition of Ca, Mg, Br, and Cl in the bones. Of the minerals analyzed, only Na and Mn showed significant diet-related effects. The bone Ca content was found to be lower in females than in males when controlled for age. Finally, Ca content was found to be higher in young adults, lower at middle age, and higher in old age in both male and female monkeys. In conclusion, this study has shown that increasing protein content in the diet does not change the bulk mineral content of primate bones. The nondietary effect that Ca content of monkey bones is lower during middle age has not been previously reported.

Aging↗

Decreased bone carbonate content in response to metabolic, but not respiratory, acidosis.

In vitro cultured neonatal mouse calvariae release calcium and buffer the medium proton concentration in response to a decrease in the medium pH caused by a reduction in bicarbonate concentration ([HCO3-]), a model of metabolic acidosis, but not to an equivalent decrease in pH caused by an increase in the partial pressure of carbon dioxide (PCO2), a model of respiratory acidosis. We have postulated that the medium is in equilibrium with the carbonated apatite in bone. To determine whether bone carbonate is depleted during models of acidosis, we cultured calvariae in control medium (pH approximately 7.4, PCO2 approximately 43, [HCO3-] approximately 26) or in medium in which the pH was equivalently reduced by either a decrease in [HCO3-] (metabolic acidosis, pH approximately 7.1, [HCO3-] approximately 13) or an increase in PCO2 (respiratory acidosis, pH approximately 7.1, PCO2 approximately 86) and determined net calcium flux (JCa) and bone carbonate content. We found that compared with control, after 3, 24, and 48 h there was a decrease in bone carbonate content during metabolic but not during respiratory acidosis. Compared with control, at 3 h JCa increased with both respiratory and metabolic acidosis; however, at 24 and 48 h JCa increased only with metabolic acidosis. JCa was correlated inversely with percent bone carbonate content in control and metabolic acidosis at all time periods studied (r = -0.809, n = 23, P < 0.001). Thus a model of metabolic acidosis appears to increase JCa from bone, perhaps due to the low [HCO3-] inducing bone carbonate dissolution.(ABSTRACT TRUNCATED AT 250 WORDS)

Acidosis↗

Changes in bone mineralization, architecture and mechanical properties due to long-term (1 year) administration of pamidronate (APD) to adult dogs.

The goal of this study was to evaluate the effect of 1 year of APD administration on bone mineralization and bone mineral chemistry in the dog. Disodium pamidronate (APD) was given orally by gavage to mature beagle dogs at doses of 0, 2.5, 12.5 and 25.0 mg/kg per day (0.1% concentration for 12 months) as part of a long-term toxicity study. The os ilium and a vertebra were used to determine the mineralization profile and, subsequently, each density fraction was analysed chemically. The ribs were used to determine lattice parameters of the apatite crystal size using X-ray diffraction. The sternum was used to determine selected morphometric parameters using image analysis of specimen X-ray films and subsequently to determine mechanical properties using velocity-of-sound techniques. We found that for both the ilium and the vertebrae there was a significant shift of the mineralization profile towards greater density in a dose-related manner. This effect levelled off with the highest dose because the shift in mineralization profile correlated better with the square root of the dose than with the dose. Together with data on crystal size, which show an increase in lattice parameters and a decrease in crystal size with dose, our data lead us to believe that long-term administration of APD leads to an increase in bone mineralization without major changes in bone chemistry of Ca, Mg, and P and with a decrease in bone apatite crystal size. The image analysis shows a dose-related increase in trabecular bone volume and thickness.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

The effect of fluoride treatment on bone mineral crystals in the rat.

In order to investigate the effect of fluoride on bone mineral crystals, we gave groups of female rats 8 mM NaF/L water and distilled water to control groups. The rats were sacrificed at six weeks, three months, and six months. The fluor content of the bone was determined by neutron activation. X-ray diffraction showed no difference in bone crystal size/strain with fluoride treatment. Fourier transform Infrared Spectroscopy (FTIR) showed an increased crystallinity in fluoride-fed animals, which seems to be associated with a decrease of labile phosphate environment. Three carbonate bands have been found in fluoridated and normal bone samples. The distribution of carbonate ions on type A and B sites is strongly affected by fluoride. Type A carbonate is always present in bone, but decreases with increasing bone fluoride content. A carbonate band found at 866 cm-1 may correspond to a fluoride interaction with type B carbonate ions. Lastly, phosphate bands have been found to be shifted towards high wave number, which is probably related to the change in unit cell size induced by the fluoride ion. All these changes induced by fluoride reduce the solubility of bone crystals by direct incorporation of fluoride ions in the apatite lattice and by decreasing the labile phosphate environments.

Animals↗

Age-related changes in the human femoral cortex.

Bone undergoes structural changes with aging, but the nature of qualitative changes remains to be established. Blocks of midshaft femur were taken at autopsy from men of four different age groups: 20-25 years, 40-45 years, 60-65 years, and 80-85 years. Each femoral specimen was analyzed by density fractionation, a technique that allows the separation of bone by extent of mineralization and maturity. In the 20-25 group, lower density bone predominates. The 40-45 group is characterized by more highly mineralized bone with an increase in the 2.1-2.2 g/cc fraction. At 60-65 years, an increase in the lower density fraction was found, indicating an increase in new bone formation. At 80-85 years, there is an increase in the highest density bone (2.2-2.3 g/cc), which may represent regions of interstitial bone not properly removed through remodeling processes. Chemical studies did not reveal any change in Ca, P, Ca + PO4, or Ca/P molar ratio with respect to age. X-ray diffraction studies show no changes in apatite crystal size with respect to age or degree of mineralization. Morphological studies documented increased remodeling activity and endosteal trabecularization in the older age groups, as well as increased intracortical porosity. An increase in the highest density fraction with aging may represent a pool of bone mineral that is less accessible to remodeling, which may be the interstitial bone.

Adult↗

Subchondral bone in osteoarthritis.

To determine whether subchondral bone in osteoarthritis differs from that seen in normal human aging, osteoarthritic femoral heads removed for total hip arthroplasty were compared with normal age-matched and young autopsy controls. Standardized, 1-cm deep, weight-bearing and nonweight-bearing subchondral bone blocks, as well as cancellous core bone, 2-4 cm deep on the articular surface, were examined in each femoral head. Mineralization was assessed using density fractionation and chemical analysis, and compared to histomorphometry. In osteoarthritis, both weight-bearing and nonweight-bearing surface subchondral bone showed a lower degree of mineralization than age-matched and young controls. Histomorphometric analysis showed that subchondral bone thickness, as well as all osteoid parameters and eroded surfaces, were increased in osteoarthritic samples versus controls. Mineralization in the deep cancellous core bone increased with normal aging but underwent less change with osteoarthritis. Histomorphometry of the cancellous core showed that osteoid parameters, but not bone volume, were increased in osteoarthritis versus controls. In conclusion, osteoarthritis is associated with a thickening of the subchondral bone with an abnormally low mineralization pattern.

Aged↗

Fluoride effects on bone crystals.

Fluoride is known to have biological effects on bone cells as well as physicochemical effects on bone crystals. This review concentrates on the latter. Fluoride increases the stability of the apatite lattice and decreases the solubility of the apatite crystals. In bone mineral, this ion has been shown to affect bone crystal structure by increasing crystallinity and reducing specific surface area. These changes in turn lead to changes in the chemistry of bone mineral. Bone mineral deposition is delayed by fluoride. This ion does not diffuse into bone already formed, but is incorporated during mineralization. Subsequently fluoride tends to accumulate in the most highly mineralized bone. Bone treated with fluoride has been shown to be more resistant to acid dissolution than normal bone, which would explain the reduced rate of resorption of fluoridated bone. The distribution of fluoride in bone is not uniform, but its net effect is to increase bone mineral density probably by an increased packing of bone crystals. Finally, there is a debate as to whether fluoride produces a bone of different quality. Whether these changes in the quality of bone will prove to be helpful or harmful remain to be determined.

Animals↗

Effects of low doses of strontium on bone quality and quantity in rats.

Strontium (Sr) has been shown to increase bone mass when given at low doses. In this study, the diets of rats containing 0.50% calcium were supplemented with Sr (0.19 and 0.40% of SrCl2 orally) for periods of four and eight weeks. Long bones and vertebrae were studied by density fractionation and each fraction was analyzed chemically. X-ray diffraction was used to determine crystal size. Static and dynamic histomorphometric parameters of bone formation were also measured. We found a shift towards lower density in the mineralization profiles of Sr supplemented rats (0.40%), as well as a decrease in bone crystal size at the larger dose of Sr. The CO3 content and the Ca/Sr ratio of the bone decreased with increasing Sr content. We found an increase in the vertebral trabecular bone volume together with an increase in osteoid volume in Sr supplemented rats. This study shows that Sr at the larger dose induces bone hypomineralization as well as an increase in bone mass in rats fed a relatively low calcium diet.

Animals↗

Maturation of bone and dentin matrices in rats flown on the Soviet biosatellite Cosmos 1887.

We have studied the chemistry, hydroxyapatite crystal size, and maturational changes in bone and dentin from rats exposed to microgravity for 12 days in a Soviet biosatellite (Cosmos 1887). Bone ash was reduced in vertebrae (L5) but not in the non-weight-bearing calvaria or mandibles. All tissues had a relatively normal percentage composition of Ca, P, and Mg. Nevertheless, flight rat calvaria and vertebral tissues tended to exhibit lower Ca/P and higher Ca/Mg ratios that any of their weight-matched controls groups, and gradient density analysis (calvaria) indicated a strong shift to the fractions lower specific gravity that was commensurate with impaired rates of matrix-mineral maturation. X-ray diffraction data were confirmatory. Bone hydroxyapatite crystal growth in the mandibles of flight rats was preferentially altered in such a way as to reduce their size (C-axis dimension). But in the mandibular diastemal region devoid of muscle attachments, flight rat bone and dentin were normal with respect to the Ca, P, Mg, and Zn concentrations and Ca/P and Ca/Mg ratios of age-matched controls. These observations affirm the concept that while microgravity most adversely affects the maturation of newly formed matrix and mineral moieties in weight-bearing bone, such effects occur throughout the skeleton.

Animals↗

Spontaneous osteoarthritis in rhesus macaques. II. Characterization of disease and morphometric studies.

From examination of the articular tissue of 35 animals from the Caribbean Primate Research Centre, we identified the epidemiological and histomorphometric features of the spontaneous osteoarthritis (OA) that affects the free-ranging rhesus macaques. The frequency of this disease increases with aging, and in females, with increased parity. Histological and morphological studies demonstrate that as in humans, the disease is characterized by persistence of the chondrocyte density typified by the cartilage of young animals. Owing to its epidemiologic and histologic resemblance to the disease in man, we conclude that degenerative arthritis affecting rhesus macaques provides a useful model for the study of factors contributing to the pathogenesis of OA.

Animals↗

The emergence and maturation of the first apatite crystals in an in vitro bone formation system.

Using chick periosteal bone forming cultures we have followed the appearance of the first crystals during de Novo bone formation. The first detectable mineral deposits are crystalline. By x-ray diffraction and dark field imaging we found crystal length to grow from around 100 angstroms on day 4 to (135-160) Angstroms by day 8. The Ca/P ratio rose from 1.46 at day 4 to 1.60 at day 8.

Animals↗

Spontaneous osteoarthritis in rhesus macaques. I. Chemical and biochemical studies.

We analyzed the articular cartilage of 41 rhesus macaques for elemental concentrations including [Ca] calcium, [P] phosphorus, [Mg] magnesium, [S] sulfur, [K] potassium and glycosaminoglycan (GAG) and collagen content. Our findings revealed an increase in all elements with aging. Within the osteoarthritic (OA) groups, young OA had higher levels of all elements than old OA, and significantly greater levels compared to young normals. In addition, GAG content was dramatically increased in young OA. Old OA did not differ from old normals for any elements although GAG levels decreased. Collagen was shown to be stable with increasing age in both normal and OA cartilage, but decreased in OA cartilage. This macaque model of spontaneous OA is valuable because it demonstrates early OA changes typical of experimental models and late changes similar to human OA.

Aging↗

Bone mineral and osteoporosis in aging rhesus monkeys.

55 femora of free ranging rhesus monkeys from the Caribbean Primate Research Center ranging in age from 2-25 years were analyzed using histology, density fractionation and chemical analysis and correlated with morphometric data. The vertebral density was determined by photon absorptiometry. 119 caudal vertebrae from the skeletal collection were analyzed chemically. Cortical mineral density and porosity increased with age, and vertebral density and cortical area increased with weight. There was significant sexual dimorphism in many histological variables reflecting size differences and cortical porosity was greater in males. The chemical analysis showed no influence of the diet on bone mineral but that Ca increase with age in females but not in males. Finally a subgroup of aged, animals had hypermineralized, and more porous bone. This subpopulation may prove to be a valuable model for senile osteoporosis.

Academies and Institutes↗

Rhesus macaques as an experimental model for degenerative arthritis.

To understand the pathogenesis of degenerative arthritis, an experimental model of the disease in which systemic factors can be investigated is required. This study reviews the evidence that the spontaneous degenerative arthritis in free ranging rhesus macaques at the Caribbean Primate Research Center meets the criteria for such a model. Two forms of degenerative arthritis in rhesus macaques have been identified: osteoarthritis and calcium pyrophosphate dihydrate crystal deposition disease. These diseases resemble spontaneous human arthritis with respect to age, sex, joint histology and cartilage composition. The availability of large numbers of affected primates as well as the availability of age/sex matched controls free of disease are additional factors that make spontaneous degenerative arthritis in rhesus macaques a suitable experimental model for the study of the human disease.

Academies and Institutes↗

Bone mineral and glycosaminoglycans in newborn and mature rabbits.

The relation between bone mineralization and glycosaminoglycans (GAG) was investigated in newborn and mature rabbit diaphyseal bone. Using the density fractionation technique, the bone was separated into fractions of increasing density from 1.4 to 2.3 grams/ml. Each fraction was analyzed by X-ray diffraction to determine the average crystal size. The GAG content of each fraction and of the unfractionated bone was determined by direct extraction and on a few fractions by sequential extraction in guanidine hydrochloride and guanidine/EDTA. There was a decrease in the GAG content with animal age and with increasing fraction density in the newborn rabbit. In one overlapping fraction (2.0-2.1 grams/ml), the GAG content was twice as high in the newborn as in the mature animal. Finally, the crystal size substantially increased from newborn to mature rabbits. Therefore, calcification and maturation of bone is associated with a decrease in the proteoglycan content of the organic matrix.

Animals↗

Crystals in calcified epiphyseal cartilage and cortical bone of the rat.

The size of hydroxyapatite (HAP) crystals in calcified cartilage and cortical bone of the rat has been studied and compared with that of synthetic poorly crystalline hydroxyapatite (PCHA). Crystal size was determined by X-ray diffraction and selected-area dark field imaging, and their elemental compositions were determined by emission spectroscopy. The crystal size of cartilage, bone, and PCHA were found to be between 120 and 170 A in length by 50 A in width as determined by both X-ray diffraction and dark field imaging. Cartilage had a lower ash weight than bone but both have the same Ca/P ratio of 1.6. These findings, though in agreement with the X-ray diffraction literature, differ from observations made by conventional bright field electron microscopy. We conclude that mineral sites in cartilage and bone, which exist as platelike structures, are in fact aggregates of small 120-170 X 50 A HAP crystals. The consequences of these findings was discussed in view of crystal relationships with collagen and other macromolecules.

Animals↗