scholarly journals Cobalt-containing Calcium Phosphate Induces Resorption of Biomineralized Collagen by Human Osteoclasts

2020 ◽  
Author(s):  
Daniel de Melo Pereira ◽  
Matthias Schumacher ◽  
Pamela Habibović
Keyword(s):  
Cell Culture ◽  
Calcium Phosphate ◽  
Calcium Release ◽  
Type I Collagen ◽  
Cell Culture Supernatant ◽  
Type I ◽  
Cobalt Ions ◽  
Collagen Membranes ◽  
Human Osteoclasts ◽  
Osteoclast Resorption

Abstract Background: Biomineralized collagen, consisting of fibrillary type-I collagen with embedded hydroxyapatite mineral, is a bone-mimicking material with potential application as a bone graft substitute. Despite the chemical and structural similarity with bone extracellular matrix, no evidence exists so far that biomineralized collagen can be resorbed by osteoclasts. The aim of the current study was to induce resorption of biomineralized collagen by osteoclasts by a two-fold modification: increasing the calcium phosphate content and introducing cobalt ions (Co2+), which have been previous shown to stimulate resorptive activity of osteoclasts.Methods: To this end, we produced biomineralized collagen membranes and coated them with a cobalt-containing calcium phosphate (CoCaP). Human osteoclasts, derived from CD14+ monocytes from peripheral blood, were differentiated directly on the membranes. Their morphology was assessed by laser confocal microscopy and their capacity for resorption observed by scanning electron microscopy (SEM), as well as indirectly quantified by calcium release into cell culture supernatant. Results: The CoCaP coating increased the mineral content of the membranes by 4 wt.% and their elastic modulus from 1 to 10 MPa. The coated membranes showed a sustained Co2+ release of about 7 nM per 2 days. In contrast to uncoated membranes, on CoCaP-coated biomineralized collagen membranes, osteoclasts sporadically formed actin rings, and caused resorption lacunae to form, as observed by SEM and confirmed by increase in Ca2+ concentration in cell culture medium. The effect of the CoCaP layer on osteoclast function is thought to be mainly caused by the increase of membrane stiffness, although the effect of Co2+, which was released in very low amounts, cannot be fully excluded.Conclusions: This work shows the potential of this relatively simple approach to induce osteoclast resorption of biomineralized collagen, despite the fact that the extent of osteoclast resorption was limited, and the method needs further optimization,. Moreover, the coating method is suitable for incorporating bioactive ions of interest into biomineralized collagen, which is typically not possible using the common biomineralization methods, such as polymer-induced liquid precursor method.

scholarly journals Cobalt-containing calcium phosphate induces resorption of biomineralized collagen by human osteoclasts

2021 ◽  
Vol 25 (1) ◽  
Author(s):  
Daniel de Melo Pereira ◽  
Matthias Schumacher ◽  
Pamela Habibovic
Keyword(s):  
Cell Culture ◽  
Calcium Phosphate ◽  
Culture Medium ◽  
Type I Collagen ◽  
Type I ◽  
Cell Culture Medium ◽  
Cobalt Ions ◽  
Collagen Membranes ◽  
Human Osteoclasts ◽  
Osteoclast Resorption

Abstract Background Biomineralized collagen, consisting of fibrillar type-I collagen with embedded hydroxyapatite mineral, is a bone-mimicking material with potential application as a bone graft substitute. Despite the chemical and structural similarity with bone extracellular matrix, no evidence exists so far that biomineralized collagen can be resorbed by osteoclasts. The aim of the current study was to induce resorption of biomineralized collagen by osteoclasts by a two-fold modification: increasing the calcium phosphate content and introducing cobalt ions (Co2+), which have been previously shown to stimulate resorptive activity of osteoclasts. Methods To this end, we produced biomineralized collagen membranes and coated them with a cobalt-containing calcium phosphate (CoCaP). Human osteoclasts, derived from CD14+ monocytes from peripheral blood, were differentiated directly on the membranes. Upon fluorescent staining of nuclei, F-actin and tartrate-resistant alkaline phosphatase, the cells were analyzed by laser confocal microscopy. Their resorption capacity was assessed by scanning electron microscopy (SEM), as well as indirectly quantified by measuring the release of calcium ions into cell culture medium. Results The CoCaP coating increased the mineral content of the membranes by 4 wt.% and their elastic modulus from 1 to 10 MPa. The coated membranes showed a sustained Co2+ release in water of about 7 nM per 2 days. In contrast to uncoated membranes, on CoCaP-coated biomineralized collagen membranes, osteoclasts sporadically formed actin rings, and induced formation of resorption lacunae, as observed by SEM and confirmed by increase in Ca2+ concentration in cell culture medium. The effect of the CoCaP layer on osteoclast function is thought to be mainly caused by the increase of membrane stiffness, although the effect of Co2+, which was released in very low amounts, cannot be fully excluded. Conclusions This work shows the potential of this relatively simple approach to induce osteoclast resorption of biomineralized collagen, although the extent of osteoclast resorption was limited, and the method needs further optimization. Moreover, the coating method is suitable for incorporating bioactive ions of interest into biomineralized collagen, which is typically not possible using the common biomineralization methods, such as polymer-induced liquid precursor method.

Valve interstitial cell culture: Production of mature type I collagen and precise detection

2017 ◽  
Vol 80 (8) ◽  
pp. 936-942 ◽  
Author(s):  
Jana Liskova ◽  
Daniel Hadraba ◽  
Elena Filova ◽  
Miroslav Konarik ◽  
Jan Pirk ◽  
...  
Keyword(s):  
Cell Culture ◽  
Type I Collagen ◽  
Interstitial Cell ◽  
Type I ◽  
Mature Type ◽  
Culture Production

Gene Expression of C2C12 Cells on Calcium Phosphate Ceramic In Vitro

2005 ◽  
Vol 288-289 ◽  
pp. 265-268 ◽  
Author(s):  
Yan Fei Tan ◽  
Ling Li Zhang ◽  
Xin Lai He ◽  
Wei Qiang Xiao ◽  
Hong Song Fan ◽  
...  
Keyword(s):  
Calcium Phosphate ◽  
Type I Collagen ◽  
C2c12 Cell ◽  
C2c12 Cells ◽  
Type I ◽  
Calcium Phosphate Ceramic ◽  
Mtt Method ◽  
Difference Expression

The osteoinduction of Calcium Phosphate (CaP) had been proved and generally been investigated by in vivo implantation. However, the mechanism of the osteoinductivity was not clear and it was difficult to judge the osteoinductivity in vitro. In this study, Mouse C2C12 cell line, a kind of myoblast precursor cell, was employed to co-culture with CaP. The induction of cell differentiation by materials was tested by MTT method, fluorescence observation, especially the mRNA expression of Osteocalcin, Type I collagen and Fibronectin by RT-PCR. It was founded that C2C12 cells could be induced to expression osteocalcin when growth on the surface of the HA/TCP ceramics. At the same time, the ceramics with different composition and sintering temperature seemed to induce difference expression level of the related genes. The results proved that phase composition was one of the most important factors in the regulation of bone-related genes. This study provided a potential model to evaluate the osteoinductivity of CaP ceramics in vitro.

Coprecipitation of Cell Adhesion Molecule with Calcium Phosphate on Hydroxyapatite Ceramic

2006 ◽  
Vol 309-311 ◽  
pp. 767-770 ◽  
Author(s):  
Yu Sogo ◽  
Yuusuke Ishikawa ◽  
Nao Kondo ◽  
Eiji Uchimura ◽  
Ayako Oyane ◽  
...  
Keyword(s):  
Calcium Phosphate ◽  
Cell Adhesion Molecule ◽  
Type I Collagen ◽  
Alveolar Bone ◽  
Composite Layer ◽  
Phosphate Solution ◽  
Type I ◽  
Hydroxyapatite Ceramic ◽  
Tooth Roots ◽  
Calcified Tissue

Fibronectin (FN) and type I collagen (Col), which are kinds of extracelluar matrices, were coprecipitated with calcium phosphate to form a composite layer on a hydroxyapatite (HAP) ceramic using a supersaturated calcium phosphate solution (CP solution). The amounts of protein immobilized in the layers were determined to be 20.97±3.04 µg·cm-2 for FN, 5.26±0.19 µg·cm-2 for Col and 21.72±2.30 µg·cm-2 for simultaneously immobilized FN and Col. When osteoblastic MC3T3-E1 cells were cultured on the HAP ceramics with the composite layer containing FN and/or Col, calcified tissue was formed through the activity of the cells. The result showed that the composite layer accelerated the differentiation of MC3T3-E1 to bone-forming cells. It is assumed that osteoblastic cells in alveolar bone migrated and differentiated on the surface of the tooth roots when the artificial tooth roots were covered with the composite layer.

EFFECT OF COLLAGEN ON THE MORPHOLOGY AND STRUCTURE OF CALCIUM PHOSPHATE NANOPARTICLES

2014 ◽  
Vol 26 (05) ◽  
pp. 1450061
Author(s):  
Hoda Salemi ◽  
Aliasghar Behnamghader ◽  
Mohamadreza Baghaban Eslaminejad ◽  
Mohammad Ataei
Keyword(s):  
Calcium Phosphate ◽  
Electrostatic Interactions ◽  
Type I Collagen ◽  
Bone Matrix ◽  
Control Sample ◽  
Molar Ratio ◽  
Type I ◽  
Sem Images ◽  
Collagen Concentration ◽  
Calcium Phosphate Nanoparticles

Collagen and noncollagenous proteins have an important role in the formation of mineral constituent of bone matrix. In this research, the morphology and phase characteristics of calcium phosphate nanoparticles in presence of collagen were investigated. The synthesis reaction was initiated by mixing H 3 PO 4 as phosphorous source and CaCl 2 as calcium source and type I collagen. Collagen concentration in suspension and Ca to P ratio was 1% and 1.67, respectively. The samples (with collagen and without collagen), were heat treated at 600°C and characterized by X-Ray diffraction (XRD), Fourier transformation infrared (FTIR) and scanning electron microscopy (SEM). More smaller and flake-like shape particles were observed in the SEM images of sample synthesized in the presence of collagen compared to the control sample which was constituted of larger granular particles. The XRD results revealed that the synthesized mineral powders with collagen were composed of hydroxyapatite and octacalcium phosphate. P – O and OH characteristic peaks were identified in FTIR spectra. In hybrid sample, the shift of amides band, revealed the electrostatic interactions between calcium phosphate ions and carboxyl or amino groups of collagen fibrils. The Ca to P molar ratio for sample with collagen was 1.9. It was found that the sample synthesized in the presence of collagen has a similar microstructure to natural bone.

scholarly journals Effects of ECM Proteins and Cationic Polymers on the Adhesion and Proliferation of Rat Islet Cells

2008 ◽  
Vol 2 (1) ◽  
pp. 133-137 ◽  
Author(s):  
Guoping Chen ◽  
Naoki Kawazoe ◽  
Tetsuya Tateishi
Keyword(s):  
Cell Culture ◽  
Type Iv Collagen ◽  
Type I Collagen ◽  
Islet Cells ◽  
Type Ii Collagen ◽  
Type I ◽  
Cationic Polymers ◽  
Type Iv ◽  
Ecm Proteins ◽  
Cationic Polyelectrolytes

The effects of extracellular matrix (ECM) proteins and cationic polymers on the adhesion and proliferation of rat islet cells, RIN-5F cells, were investigated. ECM proteins of laminin, fibronectin, vitronectin, type I collagen, type II collagen, and type IV collagen, and cationic polyelectrolytes of poly(L-lysine) and poly(allylamine) were coated on the wells of polystyrene cell culture plates. Their effects on the adhesion and proliferation of RIN-5F in serum-free and serum mediums were compared. The cell number on the laminin-coated surface was the highest among the coated surfaces. Laminin promoted cell adhesion more strongly than did the other ECM proteins and cationic polyelectrolytes. Vitronectin, type IV collagen, and poly(L-lysine) showed moderate effects, but type I collagen and type II collagen did not have any effects on adhesion and proliferation compared with the uncoated polystyrene cell culture plate. Fibronectin promoted cell adhesion but not cell proliferation. Cationic poly(allylamine) had an inhibitory effect in serum-free medium and for longterm culture in serum medium. The ECM proteins of laminin, vitronectin, and type IV collagen, and cationic poly(Llysine) will be useful for the surface modification and construction of biomaterials and scaffolds for islet cell culture and tissue engineering.

scholarly journals Regulation and enzymatic basis of bone resorption by human osteoclasts

2007 ◽  
Vol 112 (11) ◽  
pp. 567-575 ◽  
Author(s):  
Karen Fuller ◽  
Barrie Kirstein ◽  
Timothy J. Chambers
Keyword(s):  
Bone Resorption ◽  
Type I Collagen ◽  
Control Period ◽  
Cysteine Protease Inhibitor ◽  
Type I ◽  
Small Contribution ◽  
Short Period ◽  
Osteoclastic Differentiation ◽  
Human Osteoclasts ◽  
Resorptive Activity

Although much has been learned recently of the mechanisms that regulate osteoclastic differentiation, much less is known of the means through which their resorptive activity is controlled. This is especially so for human osteoclasts. We have recently developed an assay that allows us to measure resorptive activity while minimizing confounding effects on differentiation by optimizing osteoclastogenesis, so that measurable resorption occurs over a short period, and by relating resorption in each culture during the test period to the resorption that had occurred in the same culture in a prior control period. In the present study, we found that RANKL (receptor activator of nuclear factor κB ligand) strongly stimulated the release of CTX-I (C-terminal telopeptide degradation product of type I collagen) by osteoclasts over a similar range to that over which it induces osteoclastic differentiation, consistent with a distinct action on osteoclastic function. CT (calcitonin) dose-dependently inhibited bone resorption, whereas PTH (parathyroid hormone), IL (interleukin)-1, TNF-α (tumour necrosis factor-α), IL-6, IL-8, VEGF (vascular endothelial growth factor), MCP-1 (monocyte chemoattractant protein-1), MIP-1γ (macrophage inflammatory protein-1γ), IFN (interferon)-γ and dibutyryl cGMP had no significant effect. Ca2+, cyclosporin A, IFN-β and dibutyryl cAMP all strongly suppressed resorption. Bone resorption was also strongly suppressed by alendronate, the cysteine protease inhibitor E64 and the cathepsin K inhibitor MV061194. Inhibitors of MMPs (matrix metalloproteinases) had no effect on CTX-I release. Moreover, the release of the MMP-derived collagen fragment ICTP (C-terminal cross-linked telopeptide of type I collagen) represented less that 0.01% of the quantity of CTX-I released in our cultures. This suggests that MMPs make, at most, a very small contribution to the bone-resorptive activity of osteoclasts.

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