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Inflammatory response to nano- and microstructured hydroxyapatite.

Mestres G, Espanol M, Xia W, Persson C, Ginebra MP, Ott MK - PLoS ONE (2015)

Bottom Line: Additionally, the effect of supplementing the extracts with calcium ions and/or proteins was investigated.Macrophage activation on the substrates was evaluated by quantifying the release of reactive oxygen species and by morphological observations.However, the difference in macrophage proliferation was ascribed to different ionic exchanges and protein adsorption/retention from the substrates rather than to the texture of materials.

View Article: PubMed Central - PubMed

Affiliation: Materials in Medicine, Div. of Applied Materials Science, Dpt. Engineering Sciences, Uppsala University, Uppsala, Sweden.

ABSTRACT
The proliferation and activation of leukocytes upon contact with a biomaterial play a crucial role in the degree of inflammatory response, which may then determine the clinical failure or success of an implanted biomaterial. The aim of this study was to evaluate whether nano- and microstructured biomimetic hydroxyapatite substrates can influence the growth and activation of macrophage-like cells. Hydroxyapatite substrates with different crystal morphologies consisting of an entangled network of plate-like and needle-like crystals were evaluated. Macrophage proliferation was evaluated on the material surface (direct contact) and also in extracts i.e. media modified by the material (indirect contact). Additionally, the effect of supplementing the extracts with calcium ions and/or proteins was investigated. Macrophage activation on the substrates was evaluated by quantifying the release of reactive oxygen species and by morphological observations. The results showed that differences in the substrate's microstructure play a major role in the activation of macrophages as there was a higher release of reactive oxygen species after culturing the macrophages on plate-like crystals substrates compared to the almost non-existent release on needle-like substrates. However, the difference in macrophage proliferation was ascribed to different ionic exchanges and protein adsorption/retention from the substrates rather than to the texture of materials.

No MeSH data available.


Related in: MedlinePlus

Schema of the preparation of extracts.Schema of the preparation of substrate extracts, which were either used as obtained or supplemented with Ca and/or fetal bovine serum (FBS). “M” stands for medium only supplemented with 1% penicillin/streptomycin. “cM” stands for complete media (DMEM medium supplemented with 10% FBS and 1% penicillin/streptomycin).
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pone.0120381.g001: Schema of the preparation of extracts.Schema of the preparation of substrate extracts, which were either used as obtained or supplemented with Ca and/or fetal bovine serum (FBS). “M” stands for medium only supplemented with 1% penicillin/streptomycin. “cM” stands for complete media (DMEM medium supplemented with 10% FBS and 1% penicillin/streptomycin).

Mentions: b) Indirect cell culture using extracts supplemented with Ca and FBS. HA is known to interact with the surrounding media by taking up proteins [18] and ions such as Ca [19]. To determine the effect of medium impoverishment on cell growth, extracts (medium conditioned with substrates for 24 h) were evaluated as obtained or supplemented with Ca, FBS or both. Cell proliferation/activation studies were essentially carried out as described above, however in this case, only 6.5·103 cells were plated in each well (96-well plate; 2.0·104 cells/cm2). Moreover, the culture medium used to soak the substrates contained either 10% FBS and 1% penicillin/streptomycin (complete medium, cM) or only 1% penicillin/streptomycin (medium, M). The extracts were collected every 24 h and either added as obtained or supplemented with 10% FBS and/or 1 mM calcium chloride anhydrous (stocks of 100% FBS and 100 mM CaCl2) to the cells. As control, fresh medium supplemented with 10% FBS and 1% penicillin/streptomycin was used. All media combinations are schematically explained in Fig. 1. Cells were grown in the established conditions with daily changing of the extracts. Cell number was quantified by LDH after 1, 3 and 7 days, as described previously. The complete study was repeated three times using quadruplicates.


Inflammatory response to nano- and microstructured hydroxyapatite.

Mestres G, Espanol M, Xia W, Persson C, Ginebra MP, Ott MK - PLoS ONE (2015)

Schema of the preparation of extracts.Schema of the preparation of substrate extracts, which were either used as obtained or supplemented with Ca and/or fetal bovine serum (FBS). “M” stands for medium only supplemented with 1% penicillin/streptomycin. “cM” stands for complete media (DMEM medium supplemented with 10% FBS and 1% penicillin/streptomycin).
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4383585&req=5

pone.0120381.g001: Schema of the preparation of extracts.Schema of the preparation of substrate extracts, which were either used as obtained or supplemented with Ca and/or fetal bovine serum (FBS). “M” stands for medium only supplemented with 1% penicillin/streptomycin. “cM” stands for complete media (DMEM medium supplemented with 10% FBS and 1% penicillin/streptomycin).
Mentions: b) Indirect cell culture using extracts supplemented with Ca and FBS. HA is known to interact with the surrounding media by taking up proteins [18] and ions such as Ca [19]. To determine the effect of medium impoverishment on cell growth, extracts (medium conditioned with substrates for 24 h) were evaluated as obtained or supplemented with Ca, FBS or both. Cell proliferation/activation studies were essentially carried out as described above, however in this case, only 6.5·103 cells were plated in each well (96-well plate; 2.0·104 cells/cm2). Moreover, the culture medium used to soak the substrates contained either 10% FBS and 1% penicillin/streptomycin (complete medium, cM) or only 1% penicillin/streptomycin (medium, M). The extracts were collected every 24 h and either added as obtained or supplemented with 10% FBS and/or 1 mM calcium chloride anhydrous (stocks of 100% FBS and 100 mM CaCl2) to the cells. As control, fresh medium supplemented with 10% FBS and 1% penicillin/streptomycin was used. All media combinations are schematically explained in Fig. 1. Cells were grown in the established conditions with daily changing of the extracts. Cell number was quantified by LDH after 1, 3 and 7 days, as described previously. The complete study was repeated three times using quadruplicates.

Bottom Line: Additionally, the effect of supplementing the extracts with calcium ions and/or proteins was investigated.Macrophage activation on the substrates was evaluated by quantifying the release of reactive oxygen species and by morphological observations.However, the difference in macrophage proliferation was ascribed to different ionic exchanges and protein adsorption/retention from the substrates rather than to the texture of materials.

View Article: PubMed Central - PubMed

Affiliation: Materials in Medicine, Div. of Applied Materials Science, Dpt. Engineering Sciences, Uppsala University, Uppsala, Sweden.

ABSTRACT
The proliferation and activation of leukocytes upon contact with a biomaterial play a crucial role in the degree of inflammatory response, which may then determine the clinical failure or success of an implanted biomaterial. The aim of this study was to evaluate whether nano- and microstructured biomimetic hydroxyapatite substrates can influence the growth and activation of macrophage-like cells. Hydroxyapatite substrates with different crystal morphologies consisting of an entangled network of plate-like and needle-like crystals were evaluated. Macrophage proliferation was evaluated on the material surface (direct contact) and also in extracts i.e. media modified by the material (indirect contact). Additionally, the effect of supplementing the extracts with calcium ions and/or proteins was investigated. Macrophage activation on the substrates was evaluated by quantifying the release of reactive oxygen species and by morphological observations. The results showed that differences in the substrate's microstructure play a major role in the activation of macrophages as there was a higher release of reactive oxygen species after culturing the macrophages on plate-like crystals substrates compared to the almost non-existent release on needle-like substrates. However, the difference in macrophage proliferation was ascribed to different ionic exchanges and protein adsorption/retention from the substrates rather than to the texture of materials.

No MeSH data available.


Related in: MedlinePlus