Spreading, proliferation and differentiation of human dental pulp stem cells on chitosan scaffolds immobilized with RGD or fibronectin

Sana, Farzin Asghari; Yurtsever, Merve Çapkın; Bayrak, Gökçe Kaynak; Tunçay, Ekin Ö.; Kiremitçi, Arlin; Gümüşderelı́oğlu, Menemşe

doi:10.1007/s10616-017-0072-9

Cited by 43 publications

(25 citation statements)

References 39 publications

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“…Scanning electron microscopy (SEM) of the prepared ECM revealed a smooth gel surface with nanoscale pores. The gel was subsequently coated with 0.2 mg/cm 2 fibronectin to overcome it, as cell adhesion molecules can bind to the RGD fragment of fibronectin [ 33 ]. SEM analysis of cells plated on the fibronectin-coated substrate showed clear pseudopod extensions that facilitate substrate binding (Fig.…”

Section: Resultsmentioning

confidence: 99%

Extracellular matrix stiffness controls osteogenic differentiation of mesenchymal stem cells mediated by integrin α5

et al. 2018

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BackgroundHuman mesenchymal stem cell (hMSC) differentiation into osteoblasts has important clinical significance in treating bone injury, and the stiffness of the extracellular matrix (ECM) has been shown to be an important regulatory factor for hMSC differentiation. The aim of this study was to further delineate how matrix stiffness affects intracellular signaling through integrin α5/β1, FAK, and Wnt signaling, subsequently regulating the osteogenic phenotype of hMSCs.MethodshMSCs were cultured on tunable polyacrylamide hydrogels coated with fibronectin with stiffness corresponding to a Young’s modulus of 13–16 kPa and 62–68 kPa. After hMSCs were cultured on gels for 1 week, gene expression of alpha-1 type I collagen, BGLAP, and RUNX2 were evaluated by real-time PCR. After hMSCs were cultured on gels for 24 h, signaling molecules relating to integrin α5 (FAK, ERK, p-ERK, Akt, p-Akt, GSK-3β, p-GSK-3β, and β-catenin) were evaluated by western blot analysis.ResultsOsteogenic differentiation was increased on 62–68 kPa ECM, as evidenced by alpha-1 type I collagen, BGLAP, and RUNX2 gene expression, calcium deposition, and ALP staining. In the process of differentiation, gene and protein expression of integrin α5/β1 increased, together with protein expression of the downstream signaling molecules FAK, p-ERK, p-Akt, GSK-3β, p-GSK-3β, and β-catenin, indicating that these molecules can affect the osteogenic differentiation of hMSCs. An antibody blocking integrin α5 suppressed the stiffness-induced expression of all osteoblast markers examined. In particular, alpha-1 type I collagen, RUNX2, and BGLAP were significantly downregulated, indicating that integrin α5 regulates hMSC osteogenic differentiation. Downstream expression of FAK, ERK, p-ERK, and β-catenin protein was unchanged, whereas Akt, p-Akt, GSK-3β, and p-GSK-3β were upregulated. Moreover, expression of Akt and p-Akt was upregulated with anti-integrin α5 antibody, but no difference was observed for FAK, ERK, and p-ERK between the with or without anti-integrin α5 antibody groups. At the same time, expression of GSK-3β and p-GSK-3β was upregulated and β-catenin levels showed no difference between the groups with or without anti-integrin α5 antibody. Since Akt, p-Akt, GSK-3β, and p-GSK-3β displayed the same changes between the groups with or without anti-integrin α5 antibody, we then detected the links among them. Expression of p-Akt and p-GSK-3β was reduced effectively in the presence of the Akt inhibitor Triciribine. However, Akt, GSK-3β, and β-catenin were unchanged. These results suggested that expression of p-GSK-3β was regulated by p-Akt on 62–68 kPa ECM.ConclusionsTaken together, our results provide evidence that matrix stiffness (62–68 kPa) affects the osteogenic outcome of hMSCs through mechanotransduction events that are mediated by integrin α5.Electronic supplementary materialThe online version of this article (10.1186/s13287-018-0798-0) contains supplementary material, which is available to authorized users.

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Section: Resultsmentioning

confidence: 99%

Extracellular matrix stiffness controls osteogenic differentiation of mesenchymal stem cells mediated by integrin α5

et al. 2018

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“…One prominent example of this is fibronectin, which was massively produced early on. Fibronectin plays a role in cell attachment and represents an important molecule for cells to attach firmly to the scaffolds 57 . We detected several laminins, which are part of a family of glycoproteins crucial for basement membrane function 58 , that were not found in the Li et al .…”

Section: Discussionmentioning

confidence: 99%

Quantifying extracellular matrix turnover in human lung scaffold cultures

Rosmark

Åhrman

Müller

et al. 2018

Sci Rep

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Remodelling of the extracellular matrix is accomplished by altering the balance between matrix macromolecule production and degradation. However, it is not well understood how cells balance production of new matrix molecules and degradation of existing ones during tissue remodelling and regeneration. In this study, we used decellularized lung scaffolds repopulated with allogenic lung fibroblasts cultured with stable isotope labelled amino acids to quantify the balance between matrix production and degradation at a proteome-wide scale. Specific temporal dynamics of different matrisome proteins were found to correspond to the proliferative activity of the repopulating cells and the degree of extracellular deposition. The remodeling of the scaffold was characterized by an initial phase with cell proliferation and high production of cell adhesion proteins such as emilin-1 and fibronectin. Extended culture time resulted in increased levels of core matrisome proteins. In a comparison with monolayer cultures on plastic, culture in lung scaffolds lead to a pronounced accumulation of proteoglycans, such as versican and decorin, resulting in regeneration of an extracellular matrix with greater resemblance to native lung tissue compared to standard monolayer cultures. Collectively, the study presents a promising technique for increasing the understanding of cell- extracellular matrix interactions under healthy and diseased conditions.

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“…In the case of fibronectin derived sequences, the PHSRN (Pro-His-Ser-Arg-Asn), REDV (Arg-Glu-Asp-Val), LDV (Leu-Asp-Val), and KQAGDV (Lys-Gln-Ala-Gly-Asp-Val) are usually used in TE. Similar to collagen-derived sequences, they also possess the ability to bind to integrin receptors resulting in enhanced adhesion and proliferation of many cells, such as fibroblasts, MSCs, and endothelial cells [38,[120][121][122][123].…”

Section: Peptidesmentioning

confidence: 99%

“…The main advantages of freeze-drying are the ability to regulate pore size from 20 to approximately 300 µm by the change of freezing procedure as well as the possibility to obviate high temperature. In turn, long timescale, high energy consumption, and the possibility to generate irregular pore size constitute the drawbacks of this method [123,[221][222][223][224]. Electrospinning is more complicated technique in which electrostatic forces are used for formation of thin fibers from polymer solution.…”

Section: Application Of Polymer Scaffolds Modified With Proteins and mentioning

confidence: 99%

Proteins and Peptides as Important Modifiers of the Polymer Scaffolds for Tissue Engineering Applications—A Review

2020

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Polymer scaffolds constitute a very interesting strategy for tissue engineering. Even though they are generally non-toxic, in some cases, they may not provide suitable support for cell adhesion, proliferation, and differentiation, which decelerates tissue regeneration. To improve biological properties, scaffolds are frequently enriched with bioactive molecules, inter alia extracellular matrix proteins, adhesive peptides, growth factors, hormones, and cytokines. Although there are many papers describing synthesis and properties of polymer scaffolds enriched with proteins or peptides, few reviews comprehensively summarize these bioactive molecules. Thus, this review presents the current knowledge about the most important proteins and peptides used for modification of polymer scaffolds for tissue engineering. This paper also describes the influence of addition of proteins and peptides on physicochemical, mechanical, and biological properties of polymer scaffolds. Moreover, this article sums up the major applications of some biodegradable natural and synthetic polymer scaffolds modified with proteins and peptides, which have been developed within the past five years.

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Spreading, proliferation and differentiation of human dental pulp stem cells on chitosan scaffolds immobilized with RGD or fibronectin

Cited by 43 publications

References 39 publications

Extracellular matrix stiffness controls osteogenic differentiation of mesenchymal stem cells mediated by integrin α5

Extracellular matrix stiffness controls osteogenic differentiation of mesenchymal stem cells mediated by integrin α5

Quantifying extracellular matrix turnover in human lung scaffold cultures

Proteins and Peptides as Important Modifiers of the Polymer Scaffolds for Tissue Engineering Applications—A Review

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