Cell attachment activity of cementum proteins and mechanism of endotoxin inhibition

J Dent Res. 1991 Sep;70(9):1272-7. doi: 10.1177/00220345910700090801.

Abstract

Cementum occupies a unique anatomical location where soft connective tissues of the periodontium are attached to root surfaces. Cell attachment properties of proteins present in cementum were studied. Human and bovine cementum were extracted with 0.5 mol/L CH3COOH followed by 4 mol/L guanidine, and proteins were separated by ion-exchange chromatography and SDS-polyacrylamide gel electrophoresis. Cells were labeled with radioactive amino acids and added to tissue-culture plastic plates incubated with cementum proteins, and attachment was measured. Results showed that cementum proteins promoted the attachment of smooth muscle cells, endothelial cells, and fibroblasts, but not epithelial cells. Fibroblasts attached more efficiently than other cell types, and they manifested spreading with re-organization of actin filaments. No attachment occurred to plates incubated with endotoxin from A. actinomycetemcomitans. Fewer fibroblasts attached to plates treated with cementum proteins in the presence of endotoxin, but cells pre-treated with endotoxin attached normally. Attachment was not inhibited when plates were incubated first with attachment proteins and then with endotoxin; however, it was decreased when endotoxin or bovine serum albumin preceded cementum proteins. Cementum proteins with Mr 68,000, 61,000, 55,000, and 36,000 (p68, p61, p55, and p36, respectively) manifested attachment activity, while protein(s) with Mr 23,000-24,000 did not. Western blots revealed that guanidine extracts contained three bands cross-reacting with anti-bovine sialoprotein-II antibody, but the p61, p55, and p36 were negative. We conclude that cementum contains bovine sialoprotein-II and at least four other fibroblast attachment proteins, and that they do not support epithelial cell attachment.(ABSTRACT TRUNCATED AT 250 WORDS)

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Actinobacillus
  • Animals
  • Annexins
  • Calcium-Binding Proteins / isolation & purification
  • Calcium-Binding Proteins / physiology
  • Cattle
  • Cell Adhesion / physiology*
  • Cell Adhesion Molecules / antagonists & inhibitors
  • Cell Adhesion Molecules / isolation & purification
  • Cell Adhesion Molecules / physiology*
  • DEAD-box RNA Helicases
  • Dental Cementum / chemistry*
  • Dental Cementum / physiology
  • Electrophoresis, Polyacrylamide Gel
  • Endothelium / cytology
  • Endotoxins / pharmacology*
  • Epithelial Cells
  • Fibroblasts / chemistry*
  • Fibroblasts / physiology
  • Fibronectins / physiology
  • Humans
  • Integrin-Binding Sialoprotein
  • Muscle, Smooth / cytology
  • Nuclear Proteins / isolation & purification
  • Nuclear Proteins / physiology
  • Protein Kinases*
  • RNA Helicases*
  • Sialoglycoproteins / isolation & purification
  • Sialoglycoproteins / physiology

Substances

  • Annexins
  • Calcium-Binding Proteins
  • Cell Adhesion Molecules
  • Endotoxins
  • Fibronectins
  • IBSP protein, human
  • Integrin-Binding Sialoprotein
  • Nuclear Proteins
  • Sialoglycoproteins
  • Protein Kinases
  • Ddx5 protein, human
  • DEAD-box RNA Helicases
  • RNA Helicases