Recovery and determination of 3H-tetracycline from whole bones

Jackman, Kenneth Van; Klein, L.; Lacey, Stephen H.

doi:10.1016/0006-2944(73)90015-x

Cited by 15 publications

(7 citation statements)

References 10 publications

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“…Because 3 HTC is removed from skeletal sites during bone resorption [8], and is not reincorporated during bone turnover, urinary excretion of 3 HTC closely reflects bone resorption [3].…”

mentioning

confidence: 99%

Urinary 3 H-Tetracycline and Pyridinium Crosslinks Differ in Their Response to Calcium Restriction in Mature and Aged Rats

Talbott

Chowdhury

Shapses

1999

Calcified Tissue International

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The aim of this study was to evaluate bone resorption (BR) in rats by two methods: chronic 3H-tetracycline labeling (3HTC) and pyridinium crosslink excretion (PYDX), and compare the sensitivity of these markers in two age groups. Female Sprague-Dawley rats at 12-29 weeks of age ("mature", n = 12) and at 40-57 weeks of age ("aged", n = 22) were examined. Skeletal incorporation of 3HTC in aged rats was 43 +/- 8% of that in mature animals (P < 0.01), indicating an age-related decrease in bone turnover. BR was modulated over 9 weeks by calcium restriction (CR), measured by urinary excretion of both 3HTC and PYDX, and compared with age-matched, calcium-adequate controls. At baseline, urinary excretion of 3HTC was not significantly different between age groups, whereas urinary PYDX was 14-20% higher in mature compared with aged rats (P < 0.01). CR produced a 32-39% peak increase in BR (P < 0.01) compared with controls that did not differ significantly between marker or age group. Urinary 3HTC was elevated at weeks 1-3 (P < 0. 01) and reached maximal values at week 2 (32 +/- 17%). Urinary PYDX, however, was not elevated until week 2, reached maximal levels at week 3 (39 +/- 15%), and remained elevated until week 6 (P < 0.01). These data indicate that although both markers are elevated by CR, marker response differs with age, and variability exists for acute and chronic responses.

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“…Because 3 HTC is removed from skeletal sites during bone resorption [8], and is not reincorporated during bone turnover, urinary excretion of 3 HTC closely reflects bone resorption [3].…”

mentioning

confidence: 99%

Urinary 3 H-Tetracycline and Pyridinium Crosslinks Differ in Their Response to Calcium Restriction in Mature and Aged Rats

Talbott

Chowdhury

Shapses

1999

Calcified Tissue International

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“…We have shown that all the animals were equally radioactive with respect to each isotope per whole bone at the end of the labelling period [24,27]. Previous results ( [24] and unpublished observation) have shown by chemical analysis and autoradiography that this multiple-labeling procedure resulted in a uniform distribution of isotope in bone.…”

Section: Chemical Attd Isotopic Analyses Of Plasma and Bonementioning

confidence: 73%

“…dry weights were obtained. The bone mineral was extracted repeatedly with 0.5 N HC1 at 4~ [24,27] until greater than 95% of the radioactivity (~H-tetracycline and 45Ca) was removed.…”

Section: Physical Measurements Attd Extraction Of Mitt Eralmentioning

confidence: 99%

Effects of ethane-1-hydroxy-1,1-diphosphonate (EHDP) upon the kinetics of bone resorption and bone formation at the whole bone level in prelabelled chicks

Klein

Wong

1983

Calcif Tissue Int

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Chicks chronically prelabelled with 45Ca, 3H-tetracycline, and 3H-proline were used to measure the weekly effect of EHDP (5 mg P/kg for 28 days) on bone turnover at the whole bone level in vivo. Direct measurements were made of cortical bone resorption (loss of 3H-tetracycline and 3H-collagen from whole femur, blood-bone ratio of 45Ca), skeletal collagen formation and bone mineralization (collagen and calcium mass per whole femur, respectively). Chicks were sacrificed after 5, 14, 21 and 28 days of EHDP administration. By five days of treatment, EHDP caused a greater inhibition of bone mineralization (86%) than bone resorption (30-39%) without affecting skeletal collagen mass. The blood-bone ratio of 45Ca decreased 34%, which was similar in degree to the inhibition of 3H-tetracycline loss (30%) and 3H-collagen loss (39%). Almost complete inhibition of bone resorption and bone mineralization occurred by 14 days of treatment without effects on skeletal collagen mass. No additional effect was seen at 21 and 28 days of EHDP treatment. In chicks, EHDP inhibits almost completely bone mineralization (bone formation) and cortical bone resorption without affecting skeletal collagen mass.

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“…It was previously shown that all the animals were equally radioactive with respect to each isotope at the end of the labeling period (17,18). Calcium conservation can be estimated by comparing the relative losses of [3H]tetracycline and [3H]collagen to that of 45Ca in terms of total radioactivity per whole bone.…”

Section: Methodsmentioning

confidence: 99%

“…After the bones were defatted twice with CHCI3/ MeOH, 2:1 (vol/vol), and dried in vacuo over NaOH pellets, dry weights were obtained. The bone mineral was extracted repeatedly with 0.5 M HC1 at 4VC (17,18) until no significant amount of radioactivity was removed. Whole bones from the opposite side were fixed in 100% ethanol and viewed under ultraviolet light.…”

Section: Methodsmentioning

confidence: 99%

Direct measurement of bone resorption and calcium conservation during vitamin D deficiency or hypervitaminosis D

Klein

1980

Proc. Natl. Acad. Sci. U.S.A.

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When bone is remodeled during the growth of a given size bone to a larger size, some bone is resorbed and some is deposited. Much of the resorbed bone mineral, calcium, can be reutilized during bone formation. The net and absolute effects of normal growth, vitamin D deficiency, or (4,5) pointed out that conservation of calcium could not be measured by the classical methods of histology and kinetics. From autoradiographic data in immature dogs, he concluded that incorporated calcium was conserved systemically rather than locally. Similar conclusions were drawn from kinetic studies in which the specific radioactivity of 45Ca approached equality in various parts of the rat skeleton (6, 7) and in redistribution of 90Sr in uniformly labeled beagles (8). However, the rapid ionic exchange of 45Ca between bone and body fluids after pulse labeling has complicated the transient kinetics of isotopes in young rats (9, 10). Due to this complication, various workers (11, 12) have expressed the need for a more direct measurement of bone turnover. In addition, tracer methods for calcium kinetics involve only plasma tracer concentrations and focus on net pool turnover rather than on more absolute measurements of bone turnover (13).Bone resorptiont has been observed directly by using histological measurements of the increase in diameter of the bone marrow space in rapidly growing rats (14). Resorption has been measured kinetically by either 45Ca under steady-state conditions in adult rats (15) or 40Ca dilution of the natural isotope 48Ca in human subjects (16). Yet none of these approaches quantified the conservation of bone calcium, although it was recognized that such conservation was occurring (15).The purpose of the present study is to determine whether D deficiency or hypervitaminosis D has a disproportional effect on bone formation or bone resorption up to and above what would be expected to occur due to the growth process alone. The major effect of the two experimental conditions is to inhibit bone mineralization indirectly or bone formation directly without increasing bone resorption. The constancy of bone resorption under these experimental conditions suggests that bone mineralization is the major regulator of bone mass. MATERIALS AND METHODS Animals

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Recovery and determination of 3H-tetracycline from whole bones

Cited by 15 publications

References 10 publications

Urinary 3 H-Tetracycline and Pyridinium Crosslinks Differ in Their Response to Calcium Restriction in Mature and Aged Rats

Urinary 3 H-Tetracycline and Pyridinium Crosslinks Differ in Their Response to Calcium Restriction in Mature and Aged Rats

Effects of ethane-1-hydroxy-1,1-diphosphonate (EHDP) upon the kinetics of bone resorption and bone formation at the whole bone level in prelabelled chicks

Direct measurement of bone resorption and calcium conservation during vitamin D deficiency or hypervitaminosis D

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