Collagen type I prevents glyoxal-induced apoptosis in osteoblastic cells cultured on titanium alloy

Cells Tissues Organs. 2004;177(1):29-36. doi: 10.1159/000078425.

Abstract

Advanced glycation end products (AGEs) irreversibly cross-link proteins with sugars and accumulate at a higher age and in diabetes, processes which can interfere with the integration of implants into the tissue. Glyoxal is a highly reactive glycating agent involved in the formation of AGEs and is known to induce apoptosis, as revealed by the upregulation of caspase-3 and fractin (caspase-3 being a key enzyme activated during the late stage of apoptosis and fractin being a caspase-cleaved actin fragment). In this study, we investigated the influence of collagen type I coating on the cytotoxic effect of glyoxal on rat calvarial osteoblastic cells and on human osteosarcoma cells (Saos-2) grown on titanium alloy, Ti6Al4V. Activation of caspase-3 and fractin was measured by counting immunohistochemically stained cells and by flow cytometry with propidium iodide (detection of the apoptosis indicating a sub-G1 peak). Our results showed an increased number of apoptotic osteoblasts after incubation with glyoxal on Ti6Al4V discs. However, the number of apoptotic cells on collagen-coated titanium was significantly smaller than on uncoated titanium after the same treatment. The present findings demonstrate that osteoblasts treated with glyoxal undergo apoptosis, whereas collagen type I coating of titanium alloys (used for implants) has an antiapoptotic function.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Alloys / pharmacology*
  • Animals
  • Apoptosis*
  • Caspase 3
  • Caspases / metabolism
  • Cell Culture Techniques / methods
  • Cell Line
  • Cell Line, Tumor
  • Cells, Cultured
  • Chromatin / metabolism
  • Collagen Type I / physiology*
  • Cross-Linking Reagents / pharmacology
  • Dose-Response Relationship, Drug
  • Glycation End Products, Advanced / metabolism
  • Glyoxal / pharmacology*
  • Humans
  • Immunohistochemistry
  • Microscopy, Fluorescence
  • Osteoblasts / metabolism
  • Osteoblasts / pathology*
  • Osteosarcoma / metabolism
  • Rats
  • Time Factors
  • Titanium / pharmacology*
  • Up-Regulation

Substances

  • Alloys
  • Chromatin
  • Collagen Type I
  • Cross-Linking Reagents
  • Glycation End Products, Advanced
  • Glyoxal
  • Titanium
  • CASP3 protein, human
  • Casp3 protein, rat
  • Caspase 3
  • Caspases