Abstract
Objective Acids, such as those used in adhesive dentistry, have been shown to solubilize bioactive molecules from dentin. These dentin matrix components (DMC) may promote cell proliferation and differentiation, and ultimately contribute to dentin regeneration. The objective of this study was to evaluate the potential for varying concentrations of DMC extracted from human dentin by phosphoric acid of a range of pHs to stimulate proliferation and mineralization of two different cultured pulp cell populations. Methods DMC were solubilized from powdered human dentin (7 days - 4 °C) by phosphoric acid of pH 1, 3, and 5 and also, EDTA. Extracts were dialyzed for 7 days against distilled water and lyophilized. Undifferentiated mouse dental pulp cells (OD-21) and cells of the odontoblast-like cell line (MDPC-23) were seeded in six-well plates (1 × 105) and cultured for 24 h in DMEM (Dulbecco's modified Eagle's medium) containing 10% (v/v) FBS (fetal bovine serum). The cells were washed with serum-free medium and then treated with different concentrations of DMC (0.01, 0.1, 1.0 and 10.0 μg/ml) daily in serum free medium for 7 days. After 3, 5 (MDPC-23 only), and 7 days of treatment, cell proliferation was measured using 10 vol% Alamar blue solution, which was added to each well for 1 h. Cell numbers were first measured by cell counting (Trypan blue; n = 5) and Alamar blue fluorescence to validate the assay, which was then used for the subsequent assessments of proliferation. Mineralization was assessed by Alizarin Red S assay after 12 days exposure to DMC (n = 5). Controls were media-only (DMEM) and dexamethasone (DEX; positive control). Results were analysed by ANOVA/Tukey's (p ≤ 0.05). Results There was a linear correlation between cell counts and Alamar blue fluorescence (R2 > 0.96 for both cell types), verifying the validity of the Alamar blue assay for these cell types. In general, there was a dose-dependent trend for enhanced cell proliferation with higher concentration of DMC for both cell lines, especially at 10.0 μg/ml. DEX exposure resulted in significantly higher mineralization, but did not affect cell proliferation. DMC exposure demonstrated significantly greater mineralization than media-only control for 10 μg/ml for all extracts, and at lower concentrations for EDTA and pH 5 extracts. Significance Human dentin matrix components solubilized by acids at pH levels found in commercial dentin adhesives enhanced cell proliferation and mineralization of mouse and rat undifferentiated dental pulp cells when presented in adequate concentration.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 334-342 |
| Number of pages | 9 |
| Journal | Dental Materials |
| Volume | 32 |
| Issue number | 3 |
| DOIs | |
| State | Published - Mar 1 2016 |
Fingerprint
Keywords
- Cell mineralization
- Cell proliferation
- Dentin matrix components (DMC)
- MDPC-23
- OD-21 cells
- Odontoblast cells
- Phosphoric acid
- Undifferentiated mouse dental pulp cells
ASJC Scopus subject areas
- Dentistry(all)
- Materials Science(all)
- Mechanics of Materials
Cite this
Dentin matrix components extracted with phosphoric acid enhance cell proliferation and mineralization. / Salehi, Satin; Cooper, Paul; Smith, Anthony; Ferracane, Jack.
In: Dental Materials, Vol. 32, No. 3, 01.03.2016, p. 334-342.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Dentin matrix components extracted with phosphoric acid enhance cell proliferation and mineralization
AU - Salehi, Satin
AU - Cooper, Paul
AU - Smith, Anthony
AU - Ferracane, Jack
PY - 2016/3/1
Y1 - 2016/3/1
N2 - Objective Acids, such as those used in adhesive dentistry, have been shown to solubilize bioactive molecules from dentin. These dentin matrix components (DMC) may promote cell proliferation and differentiation, and ultimately contribute to dentin regeneration. The objective of this study was to evaluate the potential for varying concentrations of DMC extracted from human dentin by phosphoric acid of a range of pHs to stimulate proliferation and mineralization of two different cultured pulp cell populations. Methods DMC were solubilized from powdered human dentin (7 days - 4 °C) by phosphoric acid of pH 1, 3, and 5 and also, EDTA. Extracts were dialyzed for 7 days against distilled water and lyophilized. Undifferentiated mouse dental pulp cells (OD-21) and cells of the odontoblast-like cell line (MDPC-23) were seeded in six-well plates (1 × 105) and cultured for 24 h in DMEM (Dulbecco's modified Eagle's medium) containing 10% (v/v) FBS (fetal bovine serum). The cells were washed with serum-free medium and then treated with different concentrations of DMC (0.01, 0.1, 1.0 and 10.0 μg/ml) daily in serum free medium for 7 days. After 3, 5 (MDPC-23 only), and 7 days of treatment, cell proliferation was measured using 10 vol% Alamar blue solution, which was added to each well for 1 h. Cell numbers were first measured by cell counting (Trypan blue; n = 5) and Alamar blue fluorescence to validate the assay, which was then used for the subsequent assessments of proliferation. Mineralization was assessed by Alizarin Red S assay after 12 days exposure to DMC (n = 5). Controls were media-only (DMEM) and dexamethasone (DEX; positive control). Results were analysed by ANOVA/Tukey's (p ≤ 0.05). Results There was a linear correlation between cell counts and Alamar blue fluorescence (R2 > 0.96 for both cell types), verifying the validity of the Alamar blue assay for these cell types. In general, there was a dose-dependent trend for enhanced cell proliferation with higher concentration of DMC for both cell lines, especially at 10.0 μg/ml. DEX exposure resulted in significantly higher mineralization, but did not affect cell proliferation. DMC exposure demonstrated significantly greater mineralization than media-only control for 10 μg/ml for all extracts, and at lower concentrations for EDTA and pH 5 extracts. Significance Human dentin matrix components solubilized by acids at pH levels found in commercial dentin adhesives enhanced cell proliferation and mineralization of mouse and rat undifferentiated dental pulp cells when presented in adequate concentration.
AB - Objective Acids, such as those used in adhesive dentistry, have been shown to solubilize bioactive molecules from dentin. These dentin matrix components (DMC) may promote cell proliferation and differentiation, and ultimately contribute to dentin regeneration. The objective of this study was to evaluate the potential for varying concentrations of DMC extracted from human dentin by phosphoric acid of a range of pHs to stimulate proliferation and mineralization of two different cultured pulp cell populations. Methods DMC were solubilized from powdered human dentin (7 days - 4 °C) by phosphoric acid of pH 1, 3, and 5 and also, EDTA. Extracts were dialyzed for 7 days against distilled water and lyophilized. Undifferentiated mouse dental pulp cells (OD-21) and cells of the odontoblast-like cell line (MDPC-23) were seeded in six-well plates (1 × 105) and cultured for 24 h in DMEM (Dulbecco's modified Eagle's medium) containing 10% (v/v) FBS (fetal bovine serum). The cells were washed with serum-free medium and then treated with different concentrations of DMC (0.01, 0.1, 1.0 and 10.0 μg/ml) daily in serum free medium for 7 days. After 3, 5 (MDPC-23 only), and 7 days of treatment, cell proliferation was measured using 10 vol% Alamar blue solution, which was added to each well for 1 h. Cell numbers were first measured by cell counting (Trypan blue; n = 5) and Alamar blue fluorescence to validate the assay, which was then used for the subsequent assessments of proliferation. Mineralization was assessed by Alizarin Red S assay after 12 days exposure to DMC (n = 5). Controls were media-only (DMEM) and dexamethasone (DEX; positive control). Results were analysed by ANOVA/Tukey's (p ≤ 0.05). Results There was a linear correlation between cell counts and Alamar blue fluorescence (R2 > 0.96 for both cell types), verifying the validity of the Alamar blue assay for these cell types. In general, there was a dose-dependent trend for enhanced cell proliferation with higher concentration of DMC for both cell lines, especially at 10.0 μg/ml. DEX exposure resulted in significantly higher mineralization, but did not affect cell proliferation. DMC exposure demonstrated significantly greater mineralization than media-only control for 10 μg/ml for all extracts, and at lower concentrations for EDTA and pH 5 extracts. Significance Human dentin matrix components solubilized by acids at pH levels found in commercial dentin adhesives enhanced cell proliferation and mineralization of mouse and rat undifferentiated dental pulp cells when presented in adequate concentration.
KW - Cell mineralization
KW - Cell proliferation
KW - Dentin matrix components (DMC)
KW - MDPC-23
KW - OD-21 cells
KW - Odontoblast cells
KW - Phosphoric acid
KW - Undifferentiated mouse dental pulp cells
UR - http://www.scopus.com/inward/record.url?scp=84958753414&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84958753414&partnerID=8YFLogxK
U2 - 10.1016/j.dental.2015.11.004
DO - 10.1016/j.dental.2015.11.004
M3 - Article
C2 - 26777093
AN - SCOPUS:84958753414
VL - 32
SP - 334
EP - 342
JO - Dental Materials
JF - Dental Materials
SN - 0109-5641
IS - 3
ER -