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The Crosstalk between Osteoclasts and Osteoblasts Is Dependent upon the Composition and Structure of Biphasic Calcium Phosphates.

Shiwaku Y, Neff L, Nagano K, Takeyama K, de Bruijn J, Dard M, Gori F, Baron R - PLoS ONE (2015)

Bottom Line: We found that OC differentiation is partially impaired by increased HA content, but not by the presence of micropores within BCP scaffolds, as indicated by TRAP staining and gene profile expression.We found that BCPs with low percentage of HA favored the expression of positive coupling factors, including sphingosine-kinase 1 (SPHK1) and collagen triple helix repeat containing 1 (Cthrc1).All together our studies suggest that the chemical composition of biomaterials affects not only the differentiation and activity of OCs but also their potential to locally regulate bone formation.

View Article: PubMed Central - PubMed

Affiliation: Department of Oral Medicine, Infection and Immunity, Harvard School of Dental Medicine, Boston, MA, United States of America.

ABSTRACT
Biphasic calcium phosphates (BCPs), consisting of hydroxyapatite (HA) and β-tricalcium phosphate (β-TCP), exhibit good biocompatibility and osteoconductivity, maintaining a balance between resorption of the biomaterial and formation of new bone. We tested whether the chemical composition and/or the microstructure of BCPs affect osteoclasts (OCs) differentiation and/or their ability to crosstalk with osteoblasts (OBs). To this aim, OCs were cultured on BCPs with HA content of 5, 20 or 60% and their differentiation and activity were assessed. We found that OC differentiation is partially impaired by increased HA content, but not by the presence of micropores within BCP scaffolds, as indicated by TRAP staining and gene profile expression. We then investigated whether the biomaterial-induced changes in OC differentiation also affect their ability to crosstalk with OBs and regulate OB function. We found that BCPs with low percentage of HA favored the expression of positive coupling factors, including sphingosine-kinase 1 (SPHK1) and collagen triple helix repeat containing 1 (Cthrc1). In turn, the increase of these secreted coupling factors promotes OB differentiation and function. All together our studies suggest that the chemical composition of biomaterials affects not only the differentiation and activity of OCs but also their potential to locally regulate bone formation.

No MeSH data available.


Related in: MedlinePlus

Formation of podosome rings in osteoclasts cultured on BCPs.(A) Cells were labeled for nuclei (blue), f-actin (red) and αVβ3 integrin (green). Bars = 10 μm. Right two columns are pictures of podosome rings or filopodia in high magnification. (B) Percentage of covered with podosome rings in perimeter of osteoclasts cultured on BCPs. Data are expressed as the mean ± S.D. n = 6–8. **p<0.01 significant difference from dentin (control). (C) Number of filopodia of osteoclasts cultured on BCPs. Data are expressed as the mean ± S.D. n = 6–8. **p<0.01 significant difference from dentin (control).
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pone.0132903.g004: Formation of podosome rings in osteoclasts cultured on BCPs.(A) Cells were labeled for nuclei (blue), f-actin (red) and αVβ3 integrin (green). Bars = 10 μm. Right two columns are pictures of podosome rings or filopodia in high magnification. (B) Percentage of covered with podosome rings in perimeter of osteoclasts cultured on BCPs. Data are expressed as the mean ± S.D. n = 6–8. **p<0.01 significant difference from dentin (control). (C) Number of filopodia of osteoclasts cultured on BCPs. Data are expressed as the mean ± S.D. n = 6–8. **p<0.01 significant difference from dentin (control).

Mentions: Immunofluorescent labeling of f-actin and αVβ3 integrin show well-organized podosome rings (Fig 4A), in OCs cultured on dentin. Comparable structures were formed on both 5HA BCPs and on 20HA dense BCP. In contrast, podosome rings and belts were absent from cells plated on 60HA dense BCP (Fig 4B). Higher magnification of the cell’s periphery demonstrated that podosome-like structures were absent from cells plated on dentin (all podosomes were concentrated in the rings and belts that formed) whereas some podosomes appeared at the periphery of cells plated on 5HA BCPs. Importantly, podosomes were totally absent from cells plated on 60HA BCP and there was instead a progressive increase in the number of filopodia as the concentration of HA present in the BCP increased (Fig 4B and 4C). These findings indicate that the characteristic functional cytoskeletal organization of OCs with formation of podosome belts is impaired when cells are cultured on BCPs that contain a higher concentration of HA compared to OCs cultured on 5HA dense and 5HA porous BCPs.


The Crosstalk between Osteoclasts and Osteoblasts Is Dependent upon the Composition and Structure of Biphasic Calcium Phosphates.

Shiwaku Y, Neff L, Nagano K, Takeyama K, de Bruijn J, Dard M, Gori F, Baron R - PLoS ONE (2015)

Formation of podosome rings in osteoclasts cultured on BCPs.(A) Cells were labeled for nuclei (blue), f-actin (red) and αVβ3 integrin (green). Bars = 10 μm. Right two columns are pictures of podosome rings or filopodia in high magnification. (B) Percentage of covered with podosome rings in perimeter of osteoclasts cultured on BCPs. Data are expressed as the mean ± S.D. n = 6–8. **p<0.01 significant difference from dentin (control). (C) Number of filopodia of osteoclasts cultured on BCPs. Data are expressed as the mean ± S.D. n = 6–8. **p<0.01 significant difference from dentin (control).
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4507990&req=5

pone.0132903.g004: Formation of podosome rings in osteoclasts cultured on BCPs.(A) Cells were labeled for nuclei (blue), f-actin (red) and αVβ3 integrin (green). Bars = 10 μm. Right two columns are pictures of podosome rings or filopodia in high magnification. (B) Percentage of covered with podosome rings in perimeter of osteoclasts cultured on BCPs. Data are expressed as the mean ± S.D. n = 6–8. **p<0.01 significant difference from dentin (control). (C) Number of filopodia of osteoclasts cultured on BCPs. Data are expressed as the mean ± S.D. n = 6–8. **p<0.01 significant difference from dentin (control).
Mentions: Immunofluorescent labeling of f-actin and αVβ3 integrin show well-organized podosome rings (Fig 4A), in OCs cultured on dentin. Comparable structures were formed on both 5HA BCPs and on 20HA dense BCP. In contrast, podosome rings and belts were absent from cells plated on 60HA dense BCP (Fig 4B). Higher magnification of the cell’s periphery demonstrated that podosome-like structures were absent from cells plated on dentin (all podosomes were concentrated in the rings and belts that formed) whereas some podosomes appeared at the periphery of cells plated on 5HA BCPs. Importantly, podosomes were totally absent from cells plated on 60HA BCP and there was instead a progressive increase in the number of filopodia as the concentration of HA present in the BCP increased (Fig 4B and 4C). These findings indicate that the characteristic functional cytoskeletal organization of OCs with formation of podosome belts is impaired when cells are cultured on BCPs that contain a higher concentration of HA compared to OCs cultured on 5HA dense and 5HA porous BCPs.

Bottom Line: We found that OC differentiation is partially impaired by increased HA content, but not by the presence of micropores within BCP scaffolds, as indicated by TRAP staining and gene profile expression.We found that BCPs with low percentage of HA favored the expression of positive coupling factors, including sphingosine-kinase 1 (SPHK1) and collagen triple helix repeat containing 1 (Cthrc1).All together our studies suggest that the chemical composition of biomaterials affects not only the differentiation and activity of OCs but also their potential to locally regulate bone formation.

View Article: PubMed Central - PubMed

Affiliation: Department of Oral Medicine, Infection and Immunity, Harvard School of Dental Medicine, Boston, MA, United States of America.

ABSTRACT
Biphasic calcium phosphates (BCPs), consisting of hydroxyapatite (HA) and β-tricalcium phosphate (β-TCP), exhibit good biocompatibility and osteoconductivity, maintaining a balance between resorption of the biomaterial and formation of new bone. We tested whether the chemical composition and/or the microstructure of BCPs affect osteoclasts (OCs) differentiation and/or their ability to crosstalk with osteoblasts (OBs). To this aim, OCs were cultured on BCPs with HA content of 5, 20 or 60% and their differentiation and activity were assessed. We found that OC differentiation is partially impaired by increased HA content, but not by the presence of micropores within BCP scaffolds, as indicated by TRAP staining and gene profile expression. We then investigated whether the biomaterial-induced changes in OC differentiation also affect their ability to crosstalk with OBs and regulate OB function. We found that BCPs with low percentage of HA favored the expression of positive coupling factors, including sphingosine-kinase 1 (SPHK1) and collagen triple helix repeat containing 1 (Cthrc1). In turn, the increase of these secreted coupling factors promotes OB differentiation and function. All together our studies suggest that the chemical composition of biomaterials affects not only the differentiation and activity of OCs but also their potential to locally regulate bone formation.

No MeSH data available.


Related in: MedlinePlus