Active Vitamin D prevents anti-osteogenic effect of AGEs on human osteoblasts

Summary

Objective: To investigate whether 1,25-dihydroxyvitamin D₃ (1,25D₃) is able to prevent advanced glycation end products (AGE)-induced alterations of osteoblasts (OB).

Methods: Human OB were isolated and cultured from bone tissue of ten patients with knee osteoarthritis and joint replacement. Cells from passages three to seven were treated with control bovine serum albumin (Co-BSA), AGE-BSA or AGE-BSA supplemented with 1,25D₃ in two different concentrations (medium concentrations: 5 mg/ml AGE-BSA and Co-BSA, respectively; 100 pmol/l and 500 pmol/l 1,25D₃). mRNA and protein expression of bone alkaline phosphatase (bALP), collagen type 1 (Col1) and osteocalcin (OC) were investigated by real-time PCR and Western Blot-analysis, respectively.

Results: AGE-BSA reduced mRNA expression of bALP, Col1 and OC in comparison to Co-BSA significantly. Addition of 1,25D₃ in both concentrations completely prevented the AGEBSA-induced suppression and resulted in a further increase of mRNA and protein expression of bALP, Col1 and OC.

Discussion: 1,25D₃ in a physiological concentration prevents anti-osteogenic effects of AGEs on human OB. Therefore, 1,25D₃ treatment should be beneficial in diseases associated with AGE accumulation, impaired bone formation and vitamin D deficiency such as senile osteoporosis and rheumatoid arthritis.

Zusammenfassung

Ziel unserer Studie war es, zu evaluieren, ob 1,25-Dihydroxycholecalciferol (1,25D₃) durch Advanced Glycation End Products (AGEs) induzierte Alterationen von Osteoblasten (OB) verhindern kann.

Methodik: Humane OB wurden aus Knochengewebe von zehn Patienten mit Gonarthrose und Kniegelenkersatzoperation isoliert und kultiviert. Zellen aus den Passagen drei bis sieben erhielten eine Behandlung mit Rinderserumalbumin (Kontroll-BSA), AGEBSA bzw. AGE-BSA + 1,25D₃ (Mediumkonzentrationen: 5 mg/ml AGE-BSA bzw. Kontroll-BSA; 100 pmol/l und 500 pmol/l 1,25D₃). MRNA- und Protein-Expression von knochenspezifischer alkalischer Phosphatase (bALP), Kollagen I (Col1) und Osteokalzin (OC) wurden mittels quantitativer RT-PCR bzw. Western Blot-Analyse untersucht.

Ergebnisse: AGE-BSA reduzierte die mRNAExpression of bALP, Col1 und OC im Vergleich zu Kontroll-BSA signifikant. Die zusätzliche Gabe von 1,25D₃ in beiden Konzentrationen verhinderte die AGE-BSA-induzierte Suppression vollständig und führte zu einem weiteren Anstieg der mRNA und Protein-Expression von bALP, Col1 und OC.

Diskussion: 1,25D₃ in physiologischer Konzentration antagonisiert antiosteogene Effekte von AGEs auf humane OB. Eine Behandlung mit 1,25D₃ dürfte daher besonders bei Erkrankungen nützlich sein, die mit AGE-Akkumulation, gestörter Knochenformation und Vitamin-D-Defizienz assoziiert sind, wie senile Osteoporose und rheumatoide Arthritis.

Keywords

AGEs - vitamin D - osteoblasts - mRNA expression

Schlüsselwörter

AGEs - Vitamin D - Osteoblasten - mRNAExpression

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