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Biochemical Markers of Bone Turnover

Dr Jemil Makadia PG 3rd year Biochemistry dept. LHMC Click to edit Master subtitle style

19th August, 2011

4/19/12

Outline

Structure and function of bone Bone turnover What is metabolic bone diseases? Structure of Collagen-I Bone turnover markers Uses
4/19/12

Structure and function of bone Structure Cortical (80%) fun: mechanical protective Trabecular (20%) metabolic 4/19/12 .

Composition Organic matrix Type-I collagen (90%) Non collagenous protein (osteocalcin) Cellular 4/19/12 Minerals Calcium. Magnesium Na+. Phosphate Carbonate. K+ osteoblast .

Bone turnover Osteoclast precursors Proliferate and fuse to Form Osteoclast Hydrogen Ion Remove mineralisation 4/19/12 Lysosomal enzyme digest metrix matrix bone lining cell Osteoblast .

RANK ligand and Osteoprotegerin Osteoblast RANK ligand Osteoprotegerin(OPG) (Receptor activator of Nuclear factor kB) for bind receptor on 4/19/12 Decoy receptor RANK ligand .

Metabolic bone diseases Result of a partial uncoupling or imbalance between resorption and bone formation • • • • Osteoporosis Osteomalacia / Rickets Renal osteodystrophy Paget’s disease 4/19/12 .

Two α1 and One α2 chains -.Structure of collagen-I Triple helix – three polypeptide chains (1000 AA) -.covalent cross links intra strand and inter strand by Pyridinoline (PYD) and Deoxypyridinoline (DPD) 4/19/12 .amino terminal and carboxy terminal not triple helical (telopeptides) -.

NTELOPEPTID E REGION HELICAL REGION CTELOPEPTID E REGION Click to edit Master subtitle style NT x P yr Dp d CT x 4/19/12 .

Free pyridinoline (PYD) .Free deoxypyridinoline (DPD) .Telopeptides .C-telopeptide (CTx) .N-telopeptide (NTx) .Total DPD and PYD 4/19/12 .Tartrate-resistant acid phosphatase (TRAP) .Pyridinium cross-links .Bone turnover markers • Markers of Bone Resorption .

Osteocalcin (Bone Gla protein.N-terminal (PINP) .Bone ALP .Procollagen type I propeptide (collagen peptides ) .C-terminal (PICP) 4/19/12 . BGP) .• Markers of Bone Formation .

Telopeptides • • • • NTx and CTx Serum and Urine ELISA (immunoassay) It does not recognize precursor that is not cross linked DPD cross link with α2 chain (not α1.also present in skin) 2/3 DPD binds to NTx and 1/3 DPD binds to CTx So NTx is more specific • • • 4/19/12 .

form during collagen maturation . ligament and Aorta whereas PYD is widespread DPD. Dentine.Pyridinium • • PYD and DPD DPD more sensitive and specific because present only in Bone.not metabolize before excreted in urine .mainly from bone .not from diet • 4/19/12 .

HPLC Peak at 5am.• • • • • • Only in Urine Free and Total Free – ELISA Total.8am Second morning void sample (10 am) 4/19/12 .

CTx.hyperthyroidism .metastatic bone disease .postmenopausal women .hyperparathyroidism .Clinical significance (NTx.osteoporosis . PYD and DPD) • Increase in .Paget’s d’s .infancy and adolescence 4/19/12 .

Tartrate-resistant acid phosphatase (TRAP) • • • Two isoform 5a and 5b Osteoclast produce 5b isoform Enzyme is instable and associated with β2 macroglobulin complicated development of methods for detection Monoclonal antibody against TRAP 5b Kinetic method with fluoride inhibition or heparin inhibition 4/19/12 • • .

Hydroxyproline • • Mainly found in collagen 10% of hydroxyproline released during collagen catabolism is excreted in urine Assays involved oxidation of hydroxyproline to pyrrole Not specific because – – • • Other tissues like muscle and skin contain it Collagen is degraded during synthesis and maturation Incresed in paget’s ds. psoriasis. Hyperparathyroidsm. 4/19/12 acromegaly. burns and – .

liver. intestine. kidney and placenta ALP from liver bone and kidney are isoforms of the same gene product It increases in – – – – – • • Osteoporosis Osteomalacia and rickets Hyperparathyroidsm Renal osteodystrophy Thyrotoxicosis 4/19/12 .Bone Alkaline phosphatase • ALP is found in many tissues like bone.

15-41 u/l .• Measurement of bone ALP has several advantages – – – – Long half life Unaffected by diurnal variation More stable in vitro Useful in individuals with impaired renal function • Immunoassay is more sensitive and specific for bone ALP – – Two monoclonal antibodies Immunoadsorption assay using single monoclonal antibody • 4/19/12 Ref value : men.

Osteocalcin • • • Non collagenous protein 1% of total protein in human bone During bone formation 10-30% of OC synthesized from osteoblast is released in circulation Synthesis stimulated by vit-D Excreted by kidney Half life is 5 min. • • • • Immunoassay targeting N-terminal/ midregion 4/19/12 fragment is used.– specimen should be .

• Clinical significance Increase in – – – – – – Osteoporosis Osteomalacia and rickets Hyperparathyroidsm Renal osteodystrophy Thyrotoxicosis Acromegaly Hypoparathyroidism Decrease in – – Hypothyroidism 4/19/12 .

Procollagen peptides • • • N and C terminal of procollagen type-I Also from several other tissue so non specific Helpful in pt on vit-D as ALP and OC are not reliable in that condition 4/19/12 .

Preanalytical and Analytical variables • With in individual variability is 15-60% in urine sample and 5-10% in serum sample Diurnal variation (except ALP) • 4/19/12 .

Uses • • • • Monitoring the therapy (mc and only use) Selection of patient for therapy Prediction of bone loss Prediction of fracture risk After bone resorption therapy significant reduction in bone resorption markers with in few weeks and reach to plateau with in 3-6 months. And bone formation markers take 6-12 months 4/19/12 .

Limitation • • • • Can not detect site of disease Can not diagnose the disease Methods are not specific cost 4/19/12 .

Thank you 4/19/12 .

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