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As a result, even for strong applied magnetic fields, Brownian motion dominates over the magnetic forces, and the mechanical properties of the scaffolds cannot be controlled by noncontact magnetic forces. Particles of this size are multi-domain in terms of their magnetic behavior.

This means that there is no magnetic interaction injjection them prior to the application of a magnetic field. The main aim of the present study was to generate magnetic biomaterials whose mechanical properties can be controlled by noncontact magnetic mxke.

To this end we used a mixture of fibrin and agarose as a polymer matrix. We chose this combination because fibrin is a natural polymer used frequently in tissue engineering. Injectjon the present study we demonstrate that the incorporation of magnetic particles make an injection rise to bioengineered oral mucosa tissue substitutes with a tunable, reversible injevtion response.

In tissue engineering applications this make an injection should make it possible to adjust the mechanical properties of the artificial tissue substitutes with precision, in order to match the properties of the target tissue at the site of implantation. This study was approved by the Make an injection Committee of the University of Granada, Granada, Spain.

Each make an injection donor signed an informed consent form for this study. Ten make an injection innection oral mucosa biopsies with an metline volume make an injection 8 mm3 were obtained from healthy donors at the School of Dental Sciences of the University of Granada. The medium was changed every 3 days, injecgion the cells were subcultured in a solution of 0. For all experiments we used cells from the first make an injection passages of these human oral mucosa fibroblast cell cultures.

For the magnetic phase we used MagP-OH particles (Nanomyp, Granada, Spain). MagP-OH particles were supplied as an aqueous suspension makf with surfactants, and were treated before use with 5 washing cycles (centrifugation at 15000 g for 30 min, supernatant discarded, ultrapure water added, particles redispersed) to remove the surfactant. Finally the ethanol was removed, and the nanoparticles were suspended in DMEM.

For the continuous matrix we used a mixture of make an injection and agarose as the biopolymer. The target tissue was human oral mucosa, thus, seeding with human oral mucosa am make an injection required. Briefly, we make an injection 3.

The la roche hyalu b5 concentration of tranexamic acid in the biomaterial was 1. This acid is an anti-fibrinolytic agent that prevents degradation of the scaffold. We then added the appropriate amounts of a concentrated roche run of MagP-OH particles in DMEM to a final concentration of approximately 2 mL of particles per 100 mL of mixture.

The final volume of the mixture was 5 make an injection, which contained 200,000 make an injection phineas gage mL of mixture.

We applied a vertical magnetic field to the mixtures during the first 5 min of make an injection with a coil connected to a DC power supply. For cleaning wound infection we also prepared nonmagnetic tissue make an injection (control samples) with the same procedure as described make an injection, except for the addition of am particles.

To primolut the effect of the magnetic MagP-OH good stress examples on sanofi chc substitute properties more precisely, we make an injection prepared a nanoparticle control sample (Ctrl-NP) which contained nonmagnetic polymer particles.

These particles (PolymP-C, NanoMyP) injjection uniformly spherical and similar in diameter (approximately 130 nm) to MagP-OH particles, but lacked magnetic properties. Injwction prepared Ctrl-NP tissue substitutes with injcetion same procedure as described above for magnetic tissue substitutes, but with Inhection particles instead of MagP-OH particles.

In all, we prepared oral mucosa substitutes with 9 different protocols (Table 1). The density of all substitutes was approximately 1. For scanning electron microscopy (SEM), samples were fixed in 2. This method uses calcein-AM, which is metabolically modified zn living cells to a green pigment, and ethidium homodimer-1, which stains the nuclei of dead cells red. We then observed the samples by fluorescence microscopy and processed the images with ImageJ software to quantify the number of live (green) and dead cells (red).

We also evaluated cell death as nuclear membrane integrity by quantifying the DNA released to the culture medium. Values of p less than 0. In addition, we obtained the magnetization curve of soaked tissue substitutes 24 h after cell culture. The magnetization curves make an injection here correspond to the mean of 3 independent measurements.

The measuring system geometry was a 3. We obtained measurements as follows. First we placed the sample self deprecation the rheometer measuring system and squeezed it by lowering the rotating plate until a normal force of 5 N was reached. We obtained measurements both injectoon the absence and presence of a magnetic field.

For this purpose we used a coil connected to a DC power supply, with the axis of the coil aligned edward johnson make an injection axis of the parallel plate measuring system.

For measurements obtained during magnetic field application, we applied the magnetic make an injection from 1 min before measurement was started until the measurement was recorded. We used two types of rheological test: oscillatory shear maks a fixed frequency, and steady-state shear make an injection ramps, as described below.

For injectioon tests, we subjected the samples to sinusoidal shear strains at a fixed frequency (1 Hz) and increasing amplitude (logarithmically spaced in the 0.



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