Formation of Octacalcium Phosphate in vitro

Author(s):  
M. Iijima
Crystals ◽  
2020 ◽  
Vol 10 (12) ◽  
pp. 1131
Author(s):  
Maricela Santana ◽  
Gonzalo Montoya ◽  
Raúl Herrera ◽  
Lía Hoz ◽  
Enrique Romo ◽  
...  

Dental cementum contains unique molecules that regulate the mineralization process in vitro and in vivo, such as cementum protein 1 (CEMP1). This protein possesses amino acid sequence motifs like the human recombinant CEMP1 with biological activity. This novel cementum protein 1-derived peptide (CEMP1-p3, from the CEMP1’s N-terminal domain: (QPLPKGCAAVKAEVGIPAPH), consists of 20 amino acids. Hydroxyapatite (HA) crystals could be obtained through the combination of the amorphous precursor phase and macromolecules such as proteins and peptides. We used a simple method to synthesize peptide/hydroxyapatite nanocomposites using OCP and CEMP1-p3. The characterization of the crystals through scanning electron microscopy (SEM), powder X-ray diffraction (XRD), high--resolution transmission electron microscopy (HRTEM), and Raman spectroscopy revealed that CEMP1-p3 transformed OCP into hydroxyapatite (HA) under constant ionic strength and in a buffered solution. CEMP1-p3 binds and highly adsorbs to OCP and is a potent growth stimulator of OCP crystals. CEMP1-p3 fosters the transformation of OCP into HA crystals with crystalline planes (300) and (004) that correspond to the cell of hexagonal HA. Octacalcium phosphate crystals treated with CEMP1-p3 grown in simulated physiological buffer acquired hexagonal arrangement corresponding to HA. These findings provide new insights into the potential application of CEMP1-p3 on possible biomimetic approaches to generate materials for the repair and regeneration of mineralized tissues, or restorative materials in the orthopedic field.


2015 ◽  
Vol 11 ◽  
pp. 520-530 ◽  
Author(s):  
Sachiko Hiromoto ◽  
Motoki Inoue ◽  
Tetsushi Taguchi ◽  
Misao Yamane ◽  
Naofumi Ohtsu

1992 ◽  
Vol 11 (2) ◽  
pp. 130-140,217 ◽  
Author(s):  
Seiji BAN ◽  
Toshikage JINDE ◽  
Jiro HASEGAWA

2021 ◽  
Vol 2021 ◽  
pp. 1-9
Author(s):  
Jianghao Gong ◽  
Shangjun Fu ◽  
Zhenghao Zhou

Objective. To explore the effects of silicone gel nanoparticles modified with octacalcium phosphate on the surface (silica/OCP) polymer drugs on the proliferation of osteoblasts and autophagy. Method. Silica/OCP was prepared in vitro, and the quality of the sample preparation was tested through characterization experiments. The osteoblast cell line (hFOB1.19) was treated with silica/OCP, autophagy inhibitor (3-methyladenine (3-MA)), and silica/OCP+3-MA, respectively. The proliferation of hFOB1.19 cells was detected through the methylthiazolyldiphenyl-tetrazolium bromide (MTT) kit. Flow cytometry was used to detect the cell apoptosis. The change in protein beclin1 and P62 expression in hFOB1.19 cells was observed in Western blot. An ROS detection kit was used to detect the content of reactive oxygen species in hFOB1.19 cells. Results. Silica/OCP was a sphere with a particle size of 50 nm to 130 nm and had an OCP phase in electron projection microscopy and X-ray diffraction techniques. The results indicated that OCP successfully modified silica and the material was successfully prepared. An MTT kit and flow cytometry test showed that the cell viability of the cells treated with silica/OCP increased significantly ( P < 0.05 ), and the intracellular apoptosis phenomenon was significantly decreased ( P < 0.05 ) compared to the control group. Moreover, the inhibition of cell viability and promotion of apoptosis caused by the autophagy inhibitor 3-MA can be rescued. Western blotting demonstrated that the protein level of beclin1 in osteoblasts reached the highest after six hours of treatment with silica/OCP, and the protein level of p62, the substrate protein of autophagy, reached the lowest. At the same time, treatment of cells with the autophagy inhibitor 3-MA and silica/OCP+3-MA found that the protein levels of beclin1 and p62 in the silica/OCP+3-MA group were adjusted back compared to the 3-MA group. After adding the autophagy inhibitor, the reactive oxygen content in the cell was significantly increased ( P < 0.05 ) in the silica/OCP group. In the presence of intracellular reactive oxygen inhibitors catalase and silica/OCP, the cell viability of osteoblasts was significantly lower than that of the silica/OCP group but significantly higher than that of the silica/OCP+3-MA group. The apoptosis level of the silica/OCP+catalase group was also significantly lower than that of the silica/OCP+3-MA group ( P < 0.05 ) but was significantly higher than that of the silica/OCP group ( P < 0.05 ). Conclusion. Silica/OCP nanoparticles can upregulate the level of autophagy in osteoblasts and promote the proliferation of osteoblasts.


2005 ◽  
Vol 284-286 ◽  
pp. 11-14 ◽  
Author(s):  
Yang Leng ◽  
Ren Long Xin ◽  
Ji Yong Chen

Bioactive calcium phosphate (Ca-P) formation in bioceramics surfaces in simulated body fluid (SBF) and in rabbit muscle sites was investigated. The examined bioceamics included most commonly used bioglass®, A-W glass-ceramics and calcium phosphates in orthopedic and dental applications. The Ca-P cyrstal structures were examined with single crystal diffraction patterns in transmission electron microscopy, which reduced possibility of misidentifying Ca-P phases. The experimental results show that capability of Ca-P formation considerably varied among bioceramics, particularly in vivo. Octacalcium phosphate (OCP) was revealed on the all types of bioceramics in vitro and in vivo experiments. This work leads us to rethink how to evaluate bioactivity of bioceramics and other orthopedic materials which exhibit capability of osteoconduction by forming direct bonding with bone.


2019 ◽  
Vol 10 (4) ◽  
pp. 54 ◽  
Author(s):  
Joseph Lazraq Bystrom ◽  
Michael Pujari-Palmer

Phosphoserine modified cements (PMC) exhibit unique properties, including strong adhesion to tissues and biomaterials. While TTCP-PMCs remodel into bone in vivo, little is known regarding the bioactivity and physiochemical changes that occur during resorption. In the present study, changes in the mechanical strength and composition were evaluated for 28 days, for three formulations of αTCP based PMCs. PMCs were significantly stronger than unmodified cement (38–49 MPa vs. 10 MPa). Inclusion of wollastonite in PMCs appeared to accelerate the conversion to hydroxyapatite, coincident with slight decrease in strength. In non-wollastonite PMCs the initial compressive strength did not change after 28 days in PBS (p > 0.99). Dissolution/degradation of PMC was evaluated in acidic (pH 2.7, pH 4.0), and supersaturated fluids (simulated body fluid (SBF)). PMCs exhibited comparable mass loss (<15%) after 14 days, regardless of pH and ionic concentration. Electron microscopy, infrared spectroscopy, and X-ray analysis revealed that significant amounts of brushite, octacalcium phosphate, and hydroxyapatite reprecipitated, following dissolution in acidic conditions (pH 2.7), while amorphous calcium phosphate formed in SBF. In conclusion, PMC surfaces remodel into metastable precursors to hydroxyapatite, in both acidic and neutral environments. By tuning the composition of PMCs, durable strength in fluids, and rapid transformation can be obtained.


2008 ◽  
Vol 396-398 ◽  
pp. 187-190 ◽  
Author(s):  
Raquel Farias Weska ◽  
Grínia M. Nogueira ◽  
Wellington C. Vieira ◽  
Marisa Masumi Beppu

The requirements for scaffolds for bone tissue engineering include appropriate chemistry, morphology and structure to promote cell adhesion and synthesis of new bone matrix. Silk fibroin (SF) represents an important biomaterial for biomedical application, due to its suitable mechanical properties, biodegradability, biocompatibility, and versatility in processing. Our group has developed a new method to obtain a porous SF membrane, and the study of its potential for use as a scaffold for bone regeneration was the aim of this study. Porous membranes were obtained from SF solution, through the compression of a material generated by phase separation. For in vitro calcification experiments, porous SF membrane samples were immersed in SBF at pH 7.4 placed in polyethylene flasks. The experiments were carried out for seven days, at 36.5±0.5 °C. After 48 and 96h, the solutions were changed for fresh SBF with the ion concentration 1.5-fold higher than that of the standard one, to accelerate the calcification process. The characterization of morphology and composition of samples was performed by using scanning electron microscopy (SEM) and energy dispersive X-ray spectroscopy (EDS), respectively. The SEM micrographs indicated that the porous SF membranes presented calcium phosphate deposits after undergoing in vitro calcification. These results were confirmed by EDS spectra, which showed a stoichiometric molar Ca/P ratio ranging from 1.27 to 1.52. This fact may suggest that calcification deposits consisted of mixtures of HAP (Ca/P ratio = 1.67) and transient HAP precursor phases, such as octacalcium phosphate (Ca/P = 1.33) and dicalcium phosphate dehydrate (Ca/P = 1), indicating early stage mineralization. The porous silk fibroin membrane analysed in the current study is a promising material to be used as scaffolds for bone regeneration.


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