Development of a morphogenetically active scaffold for three-dimensional growth of bone cells: biosilica-alginate hydrogel for SaOS-2 cell cultivation

Müller, Wernér E.G.; Schröder, Heinz C.; Feng, Qingling; Schloßmacher, Ute; Link, Thorben; Wang, Xiaohong

doi:10.1002/term.1745

Cited by 30 publications

(25 citation statements)

References 76 publications

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“…Green Tracker staining shows living cells in green, while propidium iodide staining shows the nucleus of the dead cells in red. While cells proliferate evenly on the coverslip surface (Figures 10(a)-10(e)), they form clusters of different sizes on the hydrogel as it was demonstrated earlier on 3D scaffolds [41][42][43]. These clusters grow in size as SAOS-2 cells proliferate (Figures 10(f)-10(j)).…”

Section: Alamar Blue Viability Test and Proliferation Imagingsupporting

confidence: 52%

Poly-γ-Glutamic Acid Nanoparticles Based Visible Light-Curable Hydrogel for Biomedical Application

Bakó

Kerényi

Hrubi

et al. 2016

Journal of Nanomaterials

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Nanoparticles and hydrogels have gained notable attention as promising potential for fabrication of scaffolds and delivering materials. Visible light-curable systems can allow for the possibility ofin situfabrication and have the advantage of optimal applicability. In this study nanogel was created from methacrylated poly-gamma-glutamic acid nanoparticles by visible (dental blue) light photopolymerization. The average size of the particles was 80 nm by DLS, and the NMR spectra showed that the methacrylation rate was 10%. Polymerization time was 3 minutes, and a stable nanogel with a swelling rate of 110% was formed. The mechanical parameters of the prepared structure (compression stress 0.73 MPa, and Young’s modulus 0.93 MPa) can be as strong as necessary in a real situation, for example, in the mouth. A retaining effect of the nanogel was found for ampicillin, and the biocompatibility of this system was tested by Alamar Blue proliferation assay, while the cell morphology was examined by fluorescence and laser scanning confocal microscopy. In conclusion, the nanogel can be used for drug delivery, or it can be suitable for a control factor in different systems.

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Section: Alamar Blue Viability Test and Proliferation Imagingsupporting

confidence: 52%

Poly-γ-Glutamic Acid Nanoparticles Based Visible Light-Curable Hydrogel for Biomedical Application

Bakó

Kerényi

Hrubi

et al. 2016

Journal of Nanomaterials

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“…Recently, we developed a morphogenetically active scaffold, based on biosilica-alginate hydrogel [8,15]. Alginates, polysaccharides, allow the fabrication of a variety of biomaterials suitable for tissue engineering, e.g., gels and fibers, and are suitable vehicles for injectable solutions as pastes [16].…”

Section: Introductionmentioning

confidence: 99%

The Marine Sponge-Derived Inorganic Polymers, Biosilica and Polyphosphate, as Morphogenetically Active Matrices/Scaffolds for the Differentiation of Human Multipotent Stromal Cells: Potential Application in 3D Printing and Distraction Osteogenesis

et al. 2014

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The two marine inorganic polymers, biosilica (BS), enzymatically synthesized from ortho-silicate, and polyphosphate (polyP), a likewise enzymatically synthesized polymer consisting of 10 to >100 phosphate residues linked by high-energy phosphoanhydride bonds, have previously been shown to display a morphogenetic effect on osteoblasts. In the present study, the effect of these polymers on the differential differentiation of human multipotent stromal cells (hMSC), mesenchymal stem cells, that had been encapsulated into beads of the biocompatible plant polymer alginate, was studied. The differentiation of the hMSCs in the alginate beads was directed either to the osteogenic cell lineage by exposure to an osteogenic medium (mineralization activation cocktail; differentiation into osteoblasts) or to the chondrogenic cell lineage by incubating in chondrocyte differentiation medium (triggering chondrocyte maturation). Both biosilica and polyP, applied as Ca2+ salts, were found to induce an increased mineralization in osteogenic cells; these inorganic polymers display also morphogenetic potential. The effects were substantiated by gene expression studies, which revealed that biosilica and polyP strongly and significantly increase the expression of bone morphogenetic protein 2 (BMP-2) and alkaline phosphatase (ALP) in osteogenic cells, which was significantly more pronounced in osteogenic versus chondrogenic cells. A differential effect of the two polymers was seen on the expression of the two collagen types, I and II. While collagen Type I is highly expressed in osteogenic cells, but not in chondrogenic cells after exposure to biosilica or polyP, the upregulation of the steady-state level of collagen Type II transcripts in chondrogenic cells is comparably stronger than in osteogenic cells. It is concluded that the two polymers, biosilica and polyP, are morphogenetically active additives for the otherwise biologically inert alginate polymer. It is proposed that alginate, supplemented with polyP and/or biosilica, is a suitable biomaterial that promotes the growth and differentiation of hMSCs and might be beneficial for application in 3D tissue printing of hMSCs and for the delivery of hMSCs in fractures, surgically created during distraction osteogenesis.

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“…The established method was applied [ 68 ]. Normal human citrated plasma (P9523 Sigma) was mixed with 100 μM of prewarmed polyP, complexed with Ca 2+ in a stoichiometric molar ratio of 2 (with respect to phosphate monomer) to 1 Ca 2+ [ 41 ], or 10 μg/ml of beads formed of N , O -CMC, and Na-polyP (as outlined above) and hardened with 2.5% [w/v] aqueous CaCl 2 solution [ 49 ] were added at 37°C in coagulometer cuvettes; the measurements had been performed in a Thrombotrack 4 coagulometer (Diagnostica Skalpeli, Zagreb; Croatia). After addition of CaCl 2 [ 69 ] the clotting time was determined.…”

Section: Methodsmentioning

confidence: 99%

“…Likewise anabolically influencing the cell’s metabolism is biosilica, a natural polymer that causes, especially in bone-(related cells), an inducing effect on cytokines, e.g. BMP-2 [ 49 ].…”

Section: Introductionmentioning

confidence: 99%

Modular Small Diameter Vascular Grafts with Bioactive Functionalities

et al. 2015

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We report the fabrication of a novel type of artificial small diameter blood vessels, termed biomimetic tissue-engineered blood vessels (bTEBV), with a modular composition. They are composed of a hydrogel scaffold consisting of two negatively charged natural polymers, alginate and a modified chitosan, N,O-carboxymethyl chitosan (N,O-CMC). Into this biologically inert scaffold two biofunctionally active biopolymers are embedded, inorganic polyphosphate (polyP) and silica, as well as gelatin which exposes the cell recognition signal, Arg-Gly-Asp (RGD). These materials can be hardened by exposure to Ca2+ through formation of Ca2+ bridges between the polyanions, alginate, N,O-CMC, and polyP (alginate-Ca2+-N,O-CMC-polyP). The bTEBV are formed by pressing the hydrogel through an extruder into a hardening solution, containing Ca2+. In this universal scaffold of the bTEBV biomaterial, polycations such as poly(l-Lys), poly(d-Lys) or a His/Gly-tagged RGD peptide (three RGD units) were incorporated, which promote the adhesion of endothelial cells to the vessel surface. The mechanical properties of the biopolymer material (alginate-Ca2+-N,O-CMC-polyP-silica) revealed a hardness (elastic modulus) of 475 kPa even after a short incubation period in CaCl2 solution. The material of the artificial vascular grafts (bTEBVs with an outer size 6 mm and 1.8 mm, and an inner diameter 4 mm and 0.8 mm, respectively) turned out to be durable in 4-week pulsatile flow experiments at an alternating pressure between 25 and 100 mbar (18.7 and 75.0 mm Hg). The burst pressure of the larger (smaller) vessels was 850 mbar (145 mbar). Incorporation of polycationic poly(l-Lys), poly(d-Lys), and especially the His/Gly-tagged RGD peptide, markedly increased the adhesion of human, umbilical vein/vascular endothelial cells, EA.HY926 cells, to the surface of the hydrogel. No significant effect of the polyP samples on the clotting of human plasma is measured. We propose that the metabolically degradable polymeric scaffold bTEBV is a promising biomaterial for future prosthetic vascular grafts.

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Development of a morphogenetically active scaffold for three-dimensional growth of bone cells: biosilica-alginate hydrogel for SaOS-2 cell cultivation

Cited by 30 publications

References 76 publications

Poly-γ-Glutamic Acid Nanoparticles Based Visible Light-Curable Hydrogel for Biomedical Application

Poly-γ-Glutamic Acid Nanoparticles Based Visible Light-Curable Hydrogel for Biomedical Application

The Marine Sponge-Derived Inorganic Polymers, Biosilica and Polyphosphate, as Morphogenetically Active Matrices/Scaffolds for the Differentiation of Human Multipotent Stromal Cells: Potential Application in 3D Printing and Distraction Osteogenesis

Modular Small Diameter Vascular Grafts with Bioactive Functionalities

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