Osteoblast proliferation and differentiation on dentin slices are modulated by pretreatment of the surface with tetracycline or osteoclasts

Zvi Schwartz, C. H. Lohmann, M. Wieland, D. L. Cochran, D. D. Dean, M. Textor, Lynda Bonewald, B. D. Boyan

Research output: Contribution to journalArticle

39 Citations (Scopus)

Abstract

Background: Implant surface roughness and chemical composition, as well as other factors, affect the ability of osteogenic cells to form bone adjacent to an implant. The same principles may also apply to the tooth root and some reports have shown that surface modification of the root may lead to improved restoration of the periodontal apparatus. The most common of these surface modification techniques involves demineralization with citric acid or treatment with tetracycline to expose collagen fibrils. In addition, during normal bone remodeling, osteoclasts demineralize the extracellular matrix, leaving resorption pits and exposed collagen fibrils. In this study, the effect of different dentin surface-preparation techniques on osteoblasts were compared. Methods: Slices of sperm whale dentin were mechanically polished and surfaces were treated with tetracycline-HCl (TCN) or were cultured with mouse bone marrow cells to create a surface with osteoclast (OC) resorption pits or left untreated. Profilometry, x-ray photoelectron spectroscopy (XPS), and scanning electron microscopy (SEM) were used to evaluate the 3 different dentin surfaces. MG63 osteoblast-like cells were cultured on the 3 different surfaces and the effect of dentin surface preparation technique on MG63 cell proliferation (cell number), differentiation (alkaline phosphatase specific activity of isolated cells and cell layer lysates; osteocalcin production), and local factor production (transforming growth factor (TGF)-β1 and prostaglandin E2 (PGE2) compared. Results: Profilometry showed the polished and TCN surfaces were smooth with comparable R(a) values, whereas the OC surfaces were slightly rougher due to resorption pits which covered 3.7% of the surface. XPS measurements showed that TCN treatment reduced the Ca and P content of the surface, indicating that it had dissolved the mineral. Osteoclast-resorption also reduced the Ca and P content, but to a lesser extent. MG63 cell proliferation on polished dentin and tissue culture polystyrene was equivalent. In contrast, cells grown on the TCN- and OC- treated surfaces exhibited increased proliferation. No effect of surface treatment on cell alkaline phosphatase activity was observed, but activity in the cell layer lysates was increased on the TCN- and OC-treated surfaces. Osteocalcin production was reduced on all dentin surfaces, but the greatest reduction was found on the TCN-treated surface. Production of both TGF-β1 and PGE2 was increased on the treated surfaces. All effects were greatest in cultures grown on the TCN-treated dentin. Conclusions: These data indicate that demineralization of the dentin surface promotes proliferation of osteoblasts and early differentiation events like production of alkaline phosphatase and autocrine mediators such as PGE2 and TGF-β1. However, later differentiation events like osteocalcin production are decreased. Osteoclast- mediated bone resorption elicits similar responses; less than 4% of the dentin surface resulted in approximately 75% of the response caused by TCN treatment. These observations suggest that greater attention should be paid to the effects of osteoclastic resorption in designing methods for enhancing bone and cementum formation adjacent to root surfaces.

Original languageEnglish (US)
Pages (from-to)586-597
Number of pages12
JournalJournal of Periodontology
Volume71
Issue number4
StatePublished - Apr 2000
Externally publishedYes

Fingerprint

Osteoclasts
Dentin
Tetracycline
Osteoblasts
Osteocalcin
Transforming Growth Factors
Dinoprostone
Alkaline Phosphatase
Photoelectron Spectroscopy
Collagen
Cell Proliferation
Sperm Whale
X-Rays
Tooth Root
Dental Cementum
Bone Remodeling
Polystyrenes
Bone Resorption
Osteogenesis
Citric Acid

Keywords

  • Bone regeneration
  • Cell culture
  • Dental implants
  • Dentin/cytology
  • Dentin/surgery
  • Osteoblasts
  • Osteoclasts
  • Tooth root

ASJC Scopus subject areas

  • Dentistry(all)

Cite this

Schwartz, Z., Lohmann, C. H., Wieland, M., Cochran, D. L., Dean, D. D., Textor, M., ... Boyan, B. D. (2000). Osteoblast proliferation and differentiation on dentin slices are modulated by pretreatment of the surface with tetracycline or osteoclasts. Journal of Periodontology, 71(4), 586-597.

Osteoblast proliferation and differentiation on dentin slices are modulated by pretreatment of the surface with tetracycline or osteoclasts. / Schwartz, Zvi; Lohmann, C. H.; Wieland, M.; Cochran, D. L.; Dean, D. D.; Textor, M.; Bonewald, Lynda; Boyan, B. D.

In: Journal of Periodontology, Vol. 71, No. 4, 04.2000, p. 586-597.

Research output: Contribution to journalArticle

Schwartz, Z, Lohmann, CH, Wieland, M, Cochran, DL, Dean, DD, Textor, M, Bonewald, L & Boyan, BD 2000, 'Osteoblast proliferation and differentiation on dentin slices are modulated by pretreatment of the surface with tetracycline or osteoclasts', Journal of Periodontology, vol. 71, no. 4, pp. 586-597.
Schwartz, Zvi ; Lohmann, C. H. ; Wieland, M. ; Cochran, D. L. ; Dean, D. D. ; Textor, M. ; Bonewald, Lynda ; Boyan, B. D. / Osteoblast proliferation and differentiation on dentin slices are modulated by pretreatment of the surface with tetracycline or osteoclasts. In: Journal of Periodontology. 2000 ; Vol. 71, No. 4. pp. 586-597.
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TY - JOUR

T1 - Osteoblast proliferation and differentiation on dentin slices are modulated by pretreatment of the surface with tetracycline or osteoclasts

AU - Schwartz, Zvi

AU - Lohmann, C. H.

AU - Wieland, M.

AU - Cochran, D. L.

AU - Dean, D. D.

AU - Textor, M.

AU - Bonewald, Lynda

AU - Boyan, B. D.

PY - 2000/4

Y1 - 2000/4

N2 - Background: Implant surface roughness and chemical composition, as well as other factors, affect the ability of osteogenic cells to form bone adjacent to an implant. The same principles may also apply to the tooth root and some reports have shown that surface modification of the root may lead to improved restoration of the periodontal apparatus. The most common of these surface modification techniques involves demineralization with citric acid or treatment with tetracycline to expose collagen fibrils. In addition, during normal bone remodeling, osteoclasts demineralize the extracellular matrix, leaving resorption pits and exposed collagen fibrils. In this study, the effect of different dentin surface-preparation techniques on osteoblasts were compared. Methods: Slices of sperm whale dentin were mechanically polished and surfaces were treated with tetracycline-HCl (TCN) or were cultured with mouse bone marrow cells to create a surface with osteoclast (OC) resorption pits or left untreated. Profilometry, x-ray photoelectron spectroscopy (XPS), and scanning electron microscopy (SEM) were used to evaluate the 3 different dentin surfaces. MG63 osteoblast-like cells were cultured on the 3 different surfaces and the effect of dentin surface preparation technique on MG63 cell proliferation (cell number), differentiation (alkaline phosphatase specific activity of isolated cells and cell layer lysates; osteocalcin production), and local factor production (transforming growth factor (TGF)-β1 and prostaglandin E2 (PGE2) compared. Results: Profilometry showed the polished and TCN surfaces were smooth with comparable R(a) values, whereas the OC surfaces were slightly rougher due to resorption pits which covered 3.7% of the surface. XPS measurements showed that TCN treatment reduced the Ca and P content of the surface, indicating that it had dissolved the mineral. Osteoclast-resorption also reduced the Ca and P content, but to a lesser extent. MG63 cell proliferation on polished dentin and tissue culture polystyrene was equivalent. In contrast, cells grown on the TCN- and OC- treated surfaces exhibited increased proliferation. No effect of surface treatment on cell alkaline phosphatase activity was observed, but activity in the cell layer lysates was increased on the TCN- and OC-treated surfaces. Osteocalcin production was reduced on all dentin surfaces, but the greatest reduction was found on the TCN-treated surface. Production of both TGF-β1 and PGE2 was increased on the treated surfaces. All effects were greatest in cultures grown on the TCN-treated dentin. Conclusions: These data indicate that demineralization of the dentin surface promotes proliferation of osteoblasts and early differentiation events like production of alkaline phosphatase and autocrine mediators such as PGE2 and TGF-β1. However, later differentiation events like osteocalcin production are decreased. Osteoclast- mediated bone resorption elicits similar responses; less than 4% of the dentin surface resulted in approximately 75% of the response caused by TCN treatment. These observations suggest that greater attention should be paid to the effects of osteoclastic resorption in designing methods for enhancing bone and cementum formation adjacent to root surfaces.

AB - Background: Implant surface roughness and chemical composition, as well as other factors, affect the ability of osteogenic cells to form bone adjacent to an implant. The same principles may also apply to the tooth root and some reports have shown that surface modification of the root may lead to improved restoration of the periodontal apparatus. The most common of these surface modification techniques involves demineralization with citric acid or treatment with tetracycline to expose collagen fibrils. In addition, during normal bone remodeling, osteoclasts demineralize the extracellular matrix, leaving resorption pits and exposed collagen fibrils. In this study, the effect of different dentin surface-preparation techniques on osteoblasts were compared. Methods: Slices of sperm whale dentin were mechanically polished and surfaces were treated with tetracycline-HCl (TCN) or were cultured with mouse bone marrow cells to create a surface with osteoclast (OC) resorption pits or left untreated. Profilometry, x-ray photoelectron spectroscopy (XPS), and scanning electron microscopy (SEM) were used to evaluate the 3 different dentin surfaces. MG63 osteoblast-like cells were cultured on the 3 different surfaces and the effect of dentin surface preparation technique on MG63 cell proliferation (cell number), differentiation (alkaline phosphatase specific activity of isolated cells and cell layer lysates; osteocalcin production), and local factor production (transforming growth factor (TGF)-β1 and prostaglandin E2 (PGE2) compared. Results: Profilometry showed the polished and TCN surfaces were smooth with comparable R(a) values, whereas the OC surfaces were slightly rougher due to resorption pits which covered 3.7% of the surface. XPS measurements showed that TCN treatment reduced the Ca and P content of the surface, indicating that it had dissolved the mineral. Osteoclast-resorption also reduced the Ca and P content, but to a lesser extent. MG63 cell proliferation on polished dentin and tissue culture polystyrene was equivalent. In contrast, cells grown on the TCN- and OC- treated surfaces exhibited increased proliferation. No effect of surface treatment on cell alkaline phosphatase activity was observed, but activity in the cell layer lysates was increased on the TCN- and OC-treated surfaces. Osteocalcin production was reduced on all dentin surfaces, but the greatest reduction was found on the TCN-treated surface. Production of both TGF-β1 and PGE2 was increased on the treated surfaces. All effects were greatest in cultures grown on the TCN-treated dentin. Conclusions: These data indicate that demineralization of the dentin surface promotes proliferation of osteoblasts and early differentiation events like production of alkaline phosphatase and autocrine mediators such as PGE2 and TGF-β1. However, later differentiation events like osteocalcin production are decreased. Osteoclast- mediated bone resorption elicits similar responses; less than 4% of the dentin surface resulted in approximately 75% of the response caused by TCN treatment. These observations suggest that greater attention should be paid to the effects of osteoclastic resorption in designing methods for enhancing bone and cementum formation adjacent to root surfaces.

KW - Bone regeneration

KW - Cell culture

KW - Dental implants

KW - Dentin/cytology

KW - Dentin/surgery

KW - Osteoblasts

KW - Osteoclasts

KW - Tooth root

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