Glycosylation pattern and cell attachment-inhibiting property of human salivary mucins

Matthias O. Gabriel, Thorsten Grünheid, Andrej Zentner

Research output: Contribution to journalArticle

10 Citations (Scopus)

Abstract

Background: Whole human saliva (WHS) and its high molecular weight mucin constituent (Muc) inhibit fibroblast attachment and might influence periodontal and peri-implant wound healing. The aim of this work was to study the potential role of glycosylation of Muc in fibroblast attachment-inhibiting property and to examine in vitro the effect of WHS and Muc on epithelial cell attachment. Methods: Muc was isolated from WHS by CsCl density gradient ultracentrifugation; covalently immobilized on polystyrene; and subjected to enzymic digestion by N-glycanase, O-glycanase, and sialidase, or chemical desulfation and periodate treatment. Wells of tissue culture microtiter plates were incubated with WHS, Muc, or buffer as control; suspensions of normal human oral keratinocytes, spontaneously immortalized human keratinocytes, or human gingival fibroblasts were applied; and cell attachment determined using methylene blue assay. Results: While enzymic cleavage of N-linked carbohydrates showed no effect, selective removal of O-linked residues and sialic acid as well as desulfation and periodate oxidation resulted in statistically significant reduction of cell attachment-inhibiting property of immobilized Muc. Significantly lower numbers of attached cells of each cell type were found in wells pretreated with WHS or Muc. Conclusions: Inhibition of cell attachment may be mediated by the carbohydrate residues suggesting specific interactions between the salivary constituent and the cell surface. Exposure of root and implant surfaces to saliva during early wound healing events might influence healing by inhibiting surface colonization by oral keratinocytes and fibroblasts, and enhancing wound repair in the form of long junctional epithelium rather than regeneration.

Original languageEnglish
Pages (from-to)1175-1181
Number of pages7
JournalJournal of Periodontology
Volume76
Issue number7
DOIs
Publication statusPublished - Jul 2005
Externally publishedYes

Fingerprint

Mucins
Glycosylation
Saliva
Fibroblasts
Keratinocytes
Epithelial Attachment
Wound Healing
Carbohydrates
Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase
Methylene Blue
Ultracentrifugation
Polystyrenes
Neuraminidase
N-Acetylneuraminic Acid
Regeneration
Digestion
Suspensions
Buffers
Cell Count
Molecular Weight

Keywords

  • Cell adhesion
  • Cell attachment
  • Cells, epithelial
  • Glycosylation
  • Mucin/ physiology
  • Saliva
  • Wound healing

ASJC Scopus subject areas

  • Dentistry(all)

Cite this

Glycosylation pattern and cell attachment-inhibiting property of human salivary mucins. / Gabriel, Matthias O.; Grünheid, Thorsten; Zentner, Andrej.

In: Journal of Periodontology, Vol. 76, No. 7, 07.2005, p. 1175-1181.

Research output: Contribution to journalArticle

Gabriel, Matthias O. ; Grünheid, Thorsten ; Zentner, Andrej. / Glycosylation pattern and cell attachment-inhibiting property of human salivary mucins. In: Journal of Periodontology. 2005 ; Vol. 76, No. 7. pp. 1175-1181.
@article{2769da7003dc4501876af191376d536f,
title = "Glycosylation pattern and cell attachment-inhibiting property of human salivary mucins",
abstract = "Background: Whole human saliva (WHS) and its high molecular weight mucin constituent (Muc) inhibit fibroblast attachment and might influence periodontal and peri-implant wound healing. The aim of this work was to study the potential role of glycosylation of Muc in fibroblast attachment-inhibiting property and to examine in vitro the effect of WHS and Muc on epithelial cell attachment. Methods: Muc was isolated from WHS by CsCl density gradient ultracentrifugation; covalently immobilized on polystyrene; and subjected to enzymic digestion by N-glycanase, O-glycanase, and sialidase, or chemical desulfation and periodate treatment. Wells of tissue culture microtiter plates were incubated with WHS, Muc, or buffer as control; suspensions of normal human oral keratinocytes, spontaneously immortalized human keratinocytes, or human gingival fibroblasts were applied; and cell attachment determined using methylene blue assay. Results: While enzymic cleavage of N-linked carbohydrates showed no effect, selective removal of O-linked residues and sialic acid as well as desulfation and periodate oxidation resulted in statistically significant reduction of cell attachment-inhibiting property of immobilized Muc. Significantly lower numbers of attached cells of each cell type were found in wells pretreated with WHS or Muc. Conclusions: Inhibition of cell attachment may be mediated by the carbohydrate residues suggesting specific interactions between the salivary constituent and the cell surface. Exposure of root and implant surfaces to saliva during early wound healing events might influence healing by inhibiting surface colonization by oral keratinocytes and fibroblasts, and enhancing wound repair in the form of long junctional epithelium rather than regeneration.",
keywords = "Cell adhesion, Cell attachment, Cells, epithelial, Glycosylation, Mucin/ physiology, Saliva, Wound healing",
author = "Gabriel, {Matthias O.} and Thorsten Gr{\"u}nheid and Andrej Zentner",
year = "2005",
month = "7",
doi = "10.1902/jop.2005.76.7.1175",
language = "English",
volume = "76",
pages = "1175--1181",
journal = "Journal of Periodontology",
issn = "0022-3492",
publisher = "American Academy of Periodontology",
number = "7",

}

TY - JOUR

T1 - Glycosylation pattern and cell attachment-inhibiting property of human salivary mucins

AU - Gabriel, Matthias O.

AU - Grünheid, Thorsten

AU - Zentner, Andrej

PY - 2005/7

Y1 - 2005/7

N2 - Background: Whole human saliva (WHS) and its high molecular weight mucin constituent (Muc) inhibit fibroblast attachment and might influence periodontal and peri-implant wound healing. The aim of this work was to study the potential role of glycosylation of Muc in fibroblast attachment-inhibiting property and to examine in vitro the effect of WHS and Muc on epithelial cell attachment. Methods: Muc was isolated from WHS by CsCl density gradient ultracentrifugation; covalently immobilized on polystyrene; and subjected to enzymic digestion by N-glycanase, O-glycanase, and sialidase, or chemical desulfation and periodate treatment. Wells of tissue culture microtiter plates were incubated with WHS, Muc, or buffer as control; suspensions of normal human oral keratinocytes, spontaneously immortalized human keratinocytes, or human gingival fibroblasts were applied; and cell attachment determined using methylene blue assay. Results: While enzymic cleavage of N-linked carbohydrates showed no effect, selective removal of O-linked residues and sialic acid as well as desulfation and periodate oxidation resulted in statistically significant reduction of cell attachment-inhibiting property of immobilized Muc. Significantly lower numbers of attached cells of each cell type were found in wells pretreated with WHS or Muc. Conclusions: Inhibition of cell attachment may be mediated by the carbohydrate residues suggesting specific interactions between the salivary constituent and the cell surface. Exposure of root and implant surfaces to saliva during early wound healing events might influence healing by inhibiting surface colonization by oral keratinocytes and fibroblasts, and enhancing wound repair in the form of long junctional epithelium rather than regeneration.

AB - Background: Whole human saliva (WHS) and its high molecular weight mucin constituent (Muc) inhibit fibroblast attachment and might influence periodontal and peri-implant wound healing. The aim of this work was to study the potential role of glycosylation of Muc in fibroblast attachment-inhibiting property and to examine in vitro the effect of WHS and Muc on epithelial cell attachment. Methods: Muc was isolated from WHS by CsCl density gradient ultracentrifugation; covalently immobilized on polystyrene; and subjected to enzymic digestion by N-glycanase, O-glycanase, and sialidase, or chemical desulfation and periodate treatment. Wells of tissue culture microtiter plates were incubated with WHS, Muc, or buffer as control; suspensions of normal human oral keratinocytes, spontaneously immortalized human keratinocytes, or human gingival fibroblasts were applied; and cell attachment determined using methylene blue assay. Results: While enzymic cleavage of N-linked carbohydrates showed no effect, selective removal of O-linked residues and sialic acid as well as desulfation and periodate oxidation resulted in statistically significant reduction of cell attachment-inhibiting property of immobilized Muc. Significantly lower numbers of attached cells of each cell type were found in wells pretreated with WHS or Muc. Conclusions: Inhibition of cell attachment may be mediated by the carbohydrate residues suggesting specific interactions between the salivary constituent and the cell surface. Exposure of root and implant surfaces to saliva during early wound healing events might influence healing by inhibiting surface colonization by oral keratinocytes and fibroblasts, and enhancing wound repair in the form of long junctional epithelium rather than regeneration.

KW - Cell adhesion

KW - Cell attachment

KW - Cells, epithelial

KW - Glycosylation

KW - Mucin/ physiology

KW - Saliva

KW - Wound healing

UR - http://www.scopus.com/inward/record.url?scp=23044458129&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=23044458129&partnerID=8YFLogxK

U2 - 10.1902/jop.2005.76.7.1175

DO - 10.1902/jop.2005.76.7.1175

M3 - Article

VL - 76

SP - 1175

EP - 1181

JO - Journal of Periodontology

JF - Journal of Periodontology

SN - 0022-3492

IS - 7

ER -