Circadian Variation of Urinary Type I Collagen Crosslinked C-Telopeptide and Free and Peptide-Bound Forms of Pyridinium Crosslinks

doi:10.1016/S8756-3282(97)00225-1

Bone

Volume 22, Issue 1, January 1998, Pages 73-78

https://doi.org/10.1016/S8756-3282(97)00225-1 Get rights and content

Abstract

This study was performed to investigate the circadian variation of urinary CrossLaps (CTx), which was the type I collagen peptide released during bone matrix degradation, and peptide-bound and free forms of urinary pyridinium crosslinks. Urine was obtained during the 24 h of the study in seven separate collections as follows: from 23:00 h to the first void (FV) followed by FV at 11:00, 11:00–14:00, 14:00–17:00, 17:00–20:00, 20:00–23:00, and 23:00 h to FV the next morning. Total, free, and peptide-bound pyridinoline (Pyr) and deoxypyridinoline (Dpyr) excretion measured by high-performance liquid chromatography (HPLC) and CTx measured by enzyme-linked immunosorbent assay in nine premenopausal women aged 22–40 years and nine osteoporotic women aged 65–83 years was analyzed. Among three parameters of Pyr measured by HPLC, a significant day and night difference was found only in total Pyr (21.9% higher at night than during the day in premenopausal women and 24.0% in osteoporotic women, whereas no significant day and night variation was found in free and peptide-bound Pyr in either group. In contrast, total and peptide-bound Dpyr were significantly (37.9% and 66.9%) higher at night than those during the day in premenopausal women (38.0%) and osteoporotic women (48.8%). For free Dpyr, there were no day and night differences in the two groups. The day and night variances were significantly greater in peptide-bound Dpyr than with total Dpyr in both groups. In urinary CTx, a significant circadian variation with a peak at night and a nadir at 17:00 h was found (p < 0.0001) (premenopausal was 54.0% higher at night than during the day; osteoporotic was 38.4%). In conclusion, urinary CTx represented remarkable circadian variation compared with urinary pyridinium crosslinks measured by HPLC. Furthermore, free pyridinium crosslinks did not undergo a circadian variation. Peptide-bound crosslinks might contribute mostly to the circadian variation of total excretion of pyridinium crosslinks.

Introduction

Diurnal rhythm in bone turnover has been reported, as assessed by several bone metabolic markers. So far, urinary pyridinoline (Pyr) and deoxypyridinoline (Dpyr), urinary hydroxyproline, serum osteocalcin, the carboxyterminal propeptide of type I procollagen (PICP), the carboxyterminal pyridinoline crosslinked telopeptide of type I collagen (ICTP), and urinary excretion of crosslinked N-telopeptides of type I collagen (NTx) were reported to undergo circadian periodicities with higher values at night.2, 4, 13, 21, 24, 25, 26, 29, 34

Urinary Pyr and Dpyr are two mature crosslinking amino acids that form covalent crosslinks between adjacent collagen chains in extracellular matrix.3, 15They have been reported to be sensitive markers in assessing bone metabolic diseases.11, 22, 23, 27, 28, 33They are released from mature collagen during bone resorption and excreted in the urine in the free (∼40%–50%) or peptide-bound (50%–60%) form.16, 31, 35Using high-performance liquid chromatography (HPLC), total and free pyridinium crosslinks can be measured.[16]Several recent clinical studies disclosed that changes in the free fraction of Pyr or Dpyr might not reflect changes in the excretion of total Pyr or Dpyr.6, 17, 19Therefore, the variation of each form of urinary pyridinium crosslinks needs to be evaluated.

A direct immunoassay, using antibodies that recognize urinary type I C-telopeptide breakdown products [CrossLaps (CTx)] is now available.[8]CTx showed greater response to bisphosphonate therapy and discriminated postmenopausal status or bone metabolic diseases from premenopausal status or normals, respectively.1, 7, 18Urinary CTx was reported to undergo circadian variation in early postmenopausal women and in men.[36]

The aims of this study were: (1) to compare the circadian rhythm of bone collagen degradation as determined by urinary excretion of Pyr or Dpyr using HPLC and CTx in premenopausal women and osteoporotic women, and (2) to assess the circadian variation of free or peptide-bound forms of pyridinium crosslinks.

Section snippets

Subjects

Nine healthy women aged 22–40 years [mean ± standard deviation (SD) 30.6 ± 6.7 years] and nine osteoporotic women aged 65–83 years (75.5 ± 7.5 years) were recruited. None had been treated with calcitonin or bisphosphonates in the past 6 months. All osteoporotic women had a femoral neck fracture and/or vertebral fractures, but none of the vertebral fractures had occurred during the 6 months prior to this study. Since we found that the values of the urinary biochemical markers do not change

Pyridinium Crosslinks and CTx in Premenopausal Women and Osteoporotic Women

The study population and mean assay results are shown in Table 1. Mean urinary concentrations of all pyridinium crosslinks measured by HPLC and CTx measured by ELISA were significantly higher in osteoporotic than in premenopausal women (p < 0.05, unpaired t test) (Table 1). The values of all parameters of urinary Pyr and Dpyr in osteoporotic women were approximately two- and threefold higher, respectively, than those in premenopausal women. When the values of free crosslinks measured by HPLC

Discussion

In this study, urinary CTx showed significant circadian variation with a peak value at night and a nadir during daytime. The pattern of the circadian variation of urinary CTx was congruent with those in other metabolic bone markers.2, 4, 13, 21, 24, 25, 26, 29, 34Alexandersen et al.[1]reported that urinary CTx underwent circadian variation in postmenopausal women and in men. In this study, urinary CTx underwent circadian variation in both premenopausal and osteoporotic women. Furthermore, we

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Original ArticlesCircadian Variation of Urinary Type I Collagen Crosslinked C-Telopeptide and Free and Peptide-Bound Forms of Pyridinium Crosslinks

Abstract

Introduction

Section snippets

Subjects

Pyridinium Crosslinks and CTx in Premenopausal Women and Osteoporotic Women

Discussion

Methods Enzymol

Bone

Anal Biochem

Circadian variation in CrossLapsA new marker of bone resorption

J Bone Miner Res

Circadian rhythm of urinary crosslinks, pyridinoline and deoxypyridinoline in normal human and involutional osteoporosis

J Bone Miner Metab

Quantitative analysis of pyridinium crosslinks of collagen in urine using ion-paired reversed-phase high performance chromatography

Anal Biochem

Circadian variation in urinary excretion of bone collagen cross-links

J Bone Miner Res

The effect of calcium supplementation on the circadian rhythm of bone resorption

J Clin Endocrinol Metab

Different responses of biochemical markers of bone resorption to bisphosphonate therapy in Paget disease

Clin Chem

Application of an enzyme immunoassay for a new marker of bone resorption (CrossLaps)Follow-up on hormone replacement therapy and osteoporosis risk assessment

J Clin Endocrinol Metab

Immunoassay for quantifying of type I collagen degradation products in urine evaluated

Clin Chem

Comparison of immuno- and HPLC-assays for the measurement of urinary collagen cross-links

J Endocrinol Invest

The renal clearance of free and conjugated pyridinium cross-links of collagen

J Bone Miner Res

Biochemical markers of bone turnover in patients with metastatic bone disease

Clin Chem

The effect of day-to-day and diurnal variation on two immunoassats for pyridinium crosslinks in urine

J Bone Miner Res

Abnormalities in circadian patterns of bone resorption and renal calcium conservation in type I osteoporosis

J Clin Endocrinol Metab

Evaluation of bone turnover in type I osteoporosis using biochemical markers specific for both bone formation and bone resorption

Osteopor Int

Analysis of pyridinoline, a crosslinking compound of collagen fibers in human urine

J Biochem

Original Articles
Circadian Variation of Urinary Type I Collagen Crosslinked C-Telopeptide and Free and Peptide-Bound Forms of Pyridinium Crosslinks