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      Poly(acrylic acid)-regulated Synthesis of Rod-Like Calcium Carbonate Nanoparticles for Inducing the Osteogenic Differentiation of MC3T3-E1 Cells

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          Abstract

          Calcium carbonate, especially with nanostructure, has been considered as a good candidate material for bone regeneration due to its excellent biodegradability and osteoconductivity. In this study, rod-like calcium carbonate nanoparticles (Rod-CC NPs) with desired water dispersibility were achieved with the regulation of poly (acrylic acid). Characterization results revealed that the Rod-CC NPs had an average length of 240 nm, a width of 90 nm with an average aspect ratio of 2.60 and a negative ζ-potential of −22.25 ± 0.35 mV. The degradation study illustrated the nanoparticles degraded 23% at pH 7.4 and 45% at pH 5.6 in phosphate-buffered saline (PBS) solution within three months. When cultured with MC3T3-E1 cells, the Rod-CC NPs exhibited a positive effect on the proliferation of osteoblast cells. Alkaline phosphatase (ALP) activity assays together with the osteocalcin (OCN) and bone sialoprotein (BSP) expression observations demonstrated the nanoparticles could induce the differentiation of MC3T3-E1 cells. Our study developed well-dispersed rod-like calcium carbonate nanoparticles which have great potential to be used in bone regeneration.

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          Most cited references35

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          The effect of calcium ion concentration on osteoblast viability, proliferation and differentiation in monolayer and 3D culture.

          Our research group aims to develop an osteochondral composite using type II collagen gel with hydroxyapatite (HAp) deposited on one side. Soaking gels in Ca2+ and phosphate solution is indispensable to HAp deposition, so relationships between cell behavior and Ca2+ concentration were examined in two- and three-dimensional cultures. The present results indicate that 2-4 mM Ca2+ is suitable for proliferation and survival of osteoblasts, whereas slightly higher concentrations (6-8 mM) favor osteoblast differentiation and matrix mineralization in both 2- and 3-dimensional cultures. Higher concentrations (>10 mM) are cytotoxic. Purely from the perspective of calcium deposition, higher concentrations lead to increased accumulation of Ca2+. Culturing cells in phosphate-containing gel in media with Ca2+ also leads to time-dependent formation of HAp in the gel. Considering the viability of embedded cells, culturing scaffolds in media with Ca2+ concentrations around 5mM is useful for both HAp deposition and osteoblast behavior.
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            Guided bone regeneration membrane made of polycaprolactone/calcium carbonate composite nano-fibers.

            In this study, new type of guided bone regeneration (GBR) membranes were fabricated by polycaprolactone (PCL)/CaCO3 composite nano-fibers with two different PCL to calcium carbonate (CaCO3) ratios (PCL:CaCO3=75:25 wt% and 25:75 wt%). The composite nano-fibers were successfully fabricated by electrospinning method and CaCO3 nano-particles on the surface of nano-fibers were confirmed by energy disperse X-ray (EDX) analysis. In order to achieve mechanical stability of GBR membranes, composite nano-fibers were spun on PCL nano-fibrous membranes which has high tensile strength, i.e., the membranes consist of two layers of functional layer (PCL/CaCO3) and mechanical support layer (PCL). Two different GBR membranes were prepared, i.e., GBR membrane (A)=PCL:CaCO3=75:25 wt%+PCL, GBR membrane (B)=PCL:CaCO3=25:75 wt%+PCL. Osteoblast attachment and proliferation of GBR membrane (A) and (B) were discussed by MTS assay and scanning electron microscope (SEM) observation. As a result, absorbance intensity of GBR membrane (A) and tissue culture polystyrene (TCPS) increased during 5 days seeding time. In contrast, although absorbance intensity of GBR membrane (B) also increased, its value was lower than membrane (A). SEM observation showed that no significant difference in osteoblast attachment manner was seen on GBR membrane (A) and (B). Because of good cell attachment manner, there is a potential to utilize PCL/CaCO3 composite nano-fibers to GBR membranes.
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              Synthesis of micro and nano-sized calcium carbonate particles and their applications

              This paper is an authoritative review of the synthesis of nano and micro-sized calcium carbonate particles and their applications. Calcium carbonate nano and micro-particles have a large number of industrial applications due to their beneficial properties such as high porosity, high surface area to volume ratio, non-toxicity and biocompatibility towards bodily fluids. Consequently, there has been significant research to deliver easy ways of synthesising nano and micro-sized calcium carbonate particles at specific sizes, polymorphs and morphologies. A majority of their synthesis approaches are based on either the biomimetic or the CO 2 bubbling methods. This review paper describes these methods, and the effects of experimental parameters such as additive types and concentration, pH, temperature, [Ca 2+ ] : [CO 3 2− ] ratio, solvent ratio, mixing mode and agitation time on the properties of the particles produced. The current and potential uses of calcium carbonate particles in areas such as material filling, biomedical, environmental and the food industry have also been discussed.
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                Author and article information

                Contributors
                Role: Academic Editor
                Journal
                Int J Mol Sci
                Int J Mol Sci
                ijms
                International Journal of Molecular Sciences
                MDPI
                1422-0067
                06 May 2016
                May 2016
                : 17
                : 5
                : 639
                Affiliations
                [1 ]Bio-X Center, College of Life Sciences, Zhejiang Sci-Tech University, Hangzhou 310018, China; vivi-yangwei@ 123456hotmail.com (W.Y.); yaochenxue092@ 123456163.com (C.Y.); cuizy2015@ 123456126.com (Z.C.); dandanlaw@ 123456126.com (D.L.); chencen313@ 123456gmail.com (C.C.)
                [2 ]Institute of Natural Sciences, Yonsei University, Seoul 120-749, Korea; inseop@ 123456yonsei.ac.kr
                [3 ]College of Materials and Textiles, Zhejiang Sci-Tech University, Hangzhou 310018, China; yaoj@ 123456zstu.edu.cn
                Author notes
                [* ]Correspondence: kongxiangdong@ 123456gmail.com ; Tel.: +86-571-8684-3196
                [†]

                These authors contributed equally to this work.

                Article
                ijms-17-00639
                10.3390/ijms17050639
                4881465
                27164090
                d919b2b9-25f3-46ea-b6e9-f9d2767b89da
                © 2016 by the authors; licensee MDPI, Basel, Switzerland.

                This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC-BY) license ( http://creativecommons.org/licenses/by/4.0/).

                History
                : 20 March 2016
                : 22 April 2016
                Categories
                Article

                Molecular biology
                calcium carbonate,nanoparticles,rod-like,bone regeneration
                Molecular biology
                calcium carbonate, nanoparticles, rod-like, bone regeneration

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