Cell attachment activity of cementum: bone sialoprotein _ identified in cementum

Abstract

Considerable research effort has been directed at preparing root surfaces in a fashion that would promote cell attachment leading to periodontal regeneration; however, no methods have proven to be clinically predictable. Identification of attachment protein(s) associated with the root surface matrix of cementum may prove valuable for developing effective clinical treatments. In this study cementum proteins were extracted from bovine and human teeth by sequential chaotropic extraction using guanidine followed by guanidine/EDTA. The guanidine/EDTA extract, but not guanidine extract, was found to promote attachment of fibroblasts. This attachment activity was inhibitable with synthetic peptide containing the attachment sequence arginine-glycine-aspartic acid (RGD). Fractionation of the guanidine/EDTA extract revealed several fractions with attachment activity. Immunoblot analysis demonstrated that two of these fractions contain the bone-associated RGD containing attachment protein, bone sialoprotein-II (BSP-II). In addition, attachment activity was also noted in other fractions that could not be attributed to BSP-II or fibronectin. These studies indicate that a component of the attachment activity of cementum is likely to be due to BSP-II and that cementum contains additional, as yet undetermined, attachment proteins.

Cell Attachment Activity of Cementum Proteins and Mechanism of Endotoxin Inhibition

“…Western blots revealed the presence of three broad bands cross-reacting with an antibody to bonesialoprotein II. This result confirmed the recent observations of Somerman et al (1991), who reported the presence of bonesialoprotein II in guanidine-EDTA extracts. The p68 was in the region of one of these bands, although additional experiments are necessary to determine whether it is bone sialoprotein.…”

“…However, the time courses of attachment are similar for both proteins (McAllister et al, 1990). Both fibronectin and cementum proteins mediate cell attachment through the arg-gly-asp peptide sequence (McAllister et al, 1990;Somerman et al, 1991), and whether the same cell-surface receptors are involved is not clear. Interestingly, epithelial cells did not attach significantly to plates treated with cementum proteins under the conditions of assay, indicating that healthy root surfaces favor the attachment of fibroblasts over epithelial cells.…”

“…5). Somerman et al (1991) have reported that bone sialoproteins contribute to fibroblast attachment activity of cementumguanidine-EDTA extracts. As confirmation of whether DE-G and HS-0.2 fractions contain bone sialoprotein-HI, Western blots were performed with use of an antibody to bone sialoprotein II.…”

Differences in Composition of Cell-attachment Sialoproteins between Dentin and Bone

“…BSP has been detected in dentin extracts (Fisher et al, 1983) and BSP mRNA expression shown in odontoblasts (Chen et al, 1991). BSP has also been identified in cementum (Somerman et al, 1991). Dentin contains a 53-kDa sialoprotein termed DSP (dentin sialoprotein), which is rich in sialic acid and appears to be specific to dentin D'Souza et al, 1992).…”

Histological assessment of root cementum at periodontally healthy and diseased human teeth.

“…The cementum, which is continuously deposited and rarely remodeled, is an important component of periodontium and also contains various proteins which facilitate cell migration, attachment, differentiation and proliferation (1)(2)(3). Progression of plaque-induced chronic inflammatory periodontal disease leads to continuous loss of periodontal attachment and exposure of the root cementum to the oral environment.…”