The promotion of bone regeneration by nanofibrous hydroxyapatite/chitosan scaffolds by effects on integrin-BMP/Smad signaling pathway in BMSCs

Liu, Huanhuan; Peng, Hongju; Wu, Yan; Zhang, Can; Cai, Youzhi; Xu, Guangyu; Li, Qin; Chen, Xiao; Ji, Junfeng; Zhang, Yanzhong; Ouyang, Hong

doi:10.1016/j.biomaterials.2013.02.048

Cited by 294 publications

(175 citation statements)

References 47 publications

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“…Originating in the mesoderm layer, MSCs are non-hematopoietic stem cells that do not express blood cell surface antigens, but only the surface markers of stromal and interstitial cells, such as CD44, CD73, CD90, and CD105 (Baksh et al, 2004;Liu et al, 2013). This study revealed that 94.38% of the MSCs were CD90-positive, while only 3.99% were CD34-positive and only 1.71% were CD45-positive, in agreement with a previous study (Tyndall, 2012).…”

Section: Discussionsupporting

confidence: 91%

Genetic expression and functional characterization of the RUNX2 gene in human adult bone marrow mesenchymal stem cells

Zhang¹,

Li²

2015

Genet. Mol. Res.

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ABSTRACT. Past studies have revealed the critical role of runt-related transcription factor 2 (RUNX2) in the proliferation and differentiation of mesenchymal stem cells (MSCs). This study therefore aimed to investigate the expression profile of the RUNX2 gene in human bone marrow MSCs and its biological characteristics. Bone marrow MSCs were separated from 12 patients who had received hip joint replacement surgery. After purification and culture, the MSCs were subjected to the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay, flow cytometry, the alkaline phosphatase assay, reverse transcription polymerase chain reaction, and RUNX2 protein quantification. The cell growth curve, staining images, and information on the membrane antigens and the levels of RUNX2 mRNA and protein were obtained based on the results. The growth curve showed, after a 2-day lag period, cultured MSCs entered into the log phase between d3 (Day 3) and d6, when they reached a plateau. Flow cytometry data suggested 94.38% of MSCs were CD90-positive, while only 3.99 and 1.71% of total cells were positive for CD35 and CD45, respectively. With the elongated induction period, cultured MSCs were polygonal in shape. After a 14-day induction, cell fusion occurred in the center of the cell nodule accompanied by the disappearance of cellular structure to form the calcium nodule, which was stained red. There was also a statistically significant increase in the level of RUNX2 protein at d7 and d14. An osteogenic medium is required for the differentiation of adult MSCs, which is also under RUNX2 regulation. These findings are potentially valuable for clinical practice.

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Section: Discussionsupporting

confidence: 91%

Genetic expression and functional characterization of the RUNX2 gene in human adult bone marrow mesenchymal stem cells

Zhang¹,

Li²

2015

Genet. Mol. Res.

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“…For example, spontaneous lineage commitment has been induced in mesenchymal stromal cells (MSCs), by culturing them on collagen-coated polystyrene microbeads [4], or on hydroxyapatite/chitosan nanofibrous scaffolds [5]. Altering cell culture conditions from 2D to 3D results in alteration of cell adhesion, causing major remodeling of the cellular cytoskeleton.…”

Section: Introductionmentioning

confidence: 99%

Optimization of Femtosecond Laser Polymerized Structural Niches to Control Mesenchymal Stromal Cell Fate in Culture

et al. 2014

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We applied two-photon polymerization to fabricate 3D synthetic niches arranged in complex patterns to study the effect of mechano-topological parameters on morphology, renewal and differentiation of rat mesenchymal stromal cells. Niches were formed in a photoresist with low auto-fluorescence, which enabled the clear visualization of the fluorescence emission of the markers used for biological diagnostics within the internal niche structure. The niches were structurally stable in culture up to three weeks. At three weeks of expansion in the niches, cell density increased by almost 10-fold and was 67% greater than in monolayer culture. Evidence of lineage commitment was observed in monolayer culture surrounding the structural niches, and within cell aggregates, but not inside the niches. Thus, structural niches were able not only to direct stem cell homing and colony formation, but also to guide aggregate formation, providing increased surface-to-volume ratios and space for stem cells to adhere and renew, respectively. OPEN ACCESSMicromachines 2014, 5 342

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“…23 In addition, Liu et al demonstrated the effects of nanofibrous hydroxyapatite/chitosan scaffolds on promoting bone marrow mesenchymal stem cell activities. 24 In this study, we developed chitosan nanofibrous products and examined their effects on osteoblast maturation as well as the possible action mechanisms.…”

Section: Introductionmentioning

confidence: 99%

Improving effects of chitosan nanofiber scaffolds on osteoblast proliferation and maturation

Liao²,

Lin³

et al. 2014

IJN

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Osteoblast maturation plays a key role in regulating osteogenesis. Electrospun nanofibrous products were reported to possess a high surface area and porosity. In this study, we developed chitosan nanofibers and examined the effects of nanofibrous scaffolds on osteoblast maturation and the possible mechanisms. Macro- and micro observations of the chitosan nanofibers revealed that these nanoproducts had a flat surface and well-distributed fibers with nanoscale diameters. Mouse osteoblasts were able to attach onto the chitosan nanofiber scaffolds, and the scaffolds degraded in a time-dependent manner. Analysis by scanning electron microscopy further showed mouse osteoblasts adhered onto the scaffolds along the nanofibers, and cell–cell communication was also detected. Mouse osteoblasts grew much better on chitosan nanofiber scaffolds than on chitosan films. In addition, human osteoblasts were able to adhere and grow on the chitosan nanofiber scaffolds. Interestingly, culturing human osteoblasts on chitosan nanofiber scaffolds time-dependently increased DNA replication and cell proliferation. In parallel, administration of human osteoblasts onto chitosan nanofibers significantly induced osteopontin, osteocalcin, and alkaline phosphatase (ALP) messenger (m)RNA expression. As to the mechanism, chitosan nanofibers triggered runt-related transcription factor 2 mRNA and protein syntheses. Consequently, results of ALP-, alizarin red-, and von Kossa-staining analyses showed that chitosan nanofibers improved osteoblast mineralization. Taken together, results of this study demonstrate that chitosan nanofibers can stimulate osteoblast proliferation and maturation via runt-related transcription factor 2-mediated regulation of osteoblast-associated osteopontin, osteocalcin, and ALP gene expression.

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The promotion of bone regeneration by nanofibrous hydroxyapatite/chitosan scaffolds by effects on integrin-BMP/Smad signaling pathway in BMSCs

Cited by 294 publications

References 47 publications

Genetic expression and functional characterization of the RUNX2 gene in human adult bone marrow mesenchymal stem cells

Genetic expression and functional characterization of the RUNX2 gene in human adult bone marrow mesenchymal stem cells

Optimization of Femtosecond Laser Polymerized Structural Niches to Control Mesenchymal Stromal Cell Fate in Culture

Improving effects of chitosan nanofiber scaffolds on osteoblast proliferation and maturation

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