A Synthetic Thermosensitive Hydrogel for Cartilage Bioprinting and Its Biofunctionalization with Polysaccharides

Abbadessa, Anna; Mouser, Vivian H. M.; Blokzijl, Maarten M.; Gawlitta, Debby; Dhert, Wouter J.A.; Hennink, Wim E.; Malda, Jos; Vermonden, Tina

doi:10.1021/acs.biomac.6b00366

Cited by 115 publications

(98 citation statements)

References 74 publications

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“…To visualize the orientation of the collagen fibers, other sections were stained with picrosirius red (1 μg/ml, 60 min) with Weigert's hematoxylin as counterstain. Collagen types I, II, and VI were visualized with immunohistochemistry as previously described (Abbadessa et al, 2016). The stained sections were scanned with a NanoZoomer-XR Digital slide scanner (C12000-02, Hamamatsu, Almere, The Netherlands) while the picrosirius stained sections were evaluated with a light microscope and a polarizing filter (BX51 with a DP70 camera, Olympus, Hamburg, Germany).…”

Section: Histology and Immunohistochemistrymentioning

confidence: 99%

Ex vivo model unravelling cell distribution effect in hydrogels for cartilage repair

Mouser

2018

ALTEX

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Summary The implantation of chondrocyte-laden hydrogels is a promising cartilage repair strategy. Chondrocytes can be spatially positioned in hydrogels and thus in defects, while current clinical cell therapies introduce chondrocytes in the defect depth. The main aim of this study was to evaluate the effect of spatial chondrocyte distribution on the reparative process. To reduce animal experiments, an ex vivo osteochondral plug model was used and evaluated. The role of the delivered and endogenous cells in the repair process was investigated. Full thickness cartilage defects were created in equine osteochondral plugs. Defects were filled with (A) chondrocytes at the bottom of the defect, covered with a cell-free hydrogel, (B) chondrocytes homogeneously encapsulated in a hydrogel, and (C, D) combinations of A and B with different cell densities. Plugs were cultured for up to 57 days, after which the cartilage and repair tissues were characterized and compared to baseline samples. Additionally, at day 21, the origin of cells in the repair tissue was evaluated. Best outcomes were obtained with conditions C and D, which resulted in well-integrated cartilage-like tissue that completely filled the defect, regardless of the initial cell density. A critical role of the spatial chondrocyte distribution in the repair process was observed. Moreover, the osteochondral plugs stimulated cartilage formation in the hydrogels when cultured in the defects. The resulting repair tissue originated from the delivered cells. These findings confirm the potential of the osteochondral plug model for the optimization of the composition of cartilage implants and for studying repair mechanisms.

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Section: Histology and Immunohistochemistrymentioning

confidence: 99%

Ex vivo model unravelling cell distribution effect in hydrogels for cartilage repair

Mouser

2018

ALTEX

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“…HAMA was analyzed by 1 in which an integration value of 1 is given to the signal at chemical shift of 6.2 ppm. The DM of HAMA was also measured according to a High Performance Liquid Chromatography (HPLC)-based method, as reported previously [24,42].…”

Section: Synthesis and Characterization Of Polymersmentioning

confidence: 99%

“…Our more recent studies have shown that by blending these PEG/methacrylated pHPMAlac triblock copolymers with UV cross-linkable polysaccharides, i.e. methacrylated chondroitin sulfate [23] or methacrylated hyaluronic acid [24], hydrogels with improved mechanical characteristics and degradation profiles can be obtained.…”

Section: Introductionmentioning

confidence: 99%

“…Specifically, the more pronounced thermosensitivity, shear thinning and yield stress behavior led to a superior 3D printability of hydrogels based on polysaccharides and PEG/methacrylated pHPMAlac copolymers compared with hydrogels only composed of PEG/methacrylated pHPMAlac copolymers [23,24]. Nevertheless, mechanistic insights behind the beneficial effect on mechanical properties due to the addition of polysaccharides to hydrogels of PEG/methacrylated pHPMAlac are lacking.…”

Section: Introductionmentioning

confidence: 99%

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Two-component thermosensitive hydrogels: Phase separation affecting rheological behavior

Abbadessa

Landín

Blenke

et al. 2017

European Polymer Journal

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Please cite this article as: Abbadessa, A., Landín, M., Oude Blenke, E., Hennink, W.E., Vermonden, T., Twocomponent thermosensitive hydrogels: Phase separation affecting rheological behavior, European Polymer Journal (2017), doi: http://dx. AbstractExtracellular matrices are mainly composed of a mixture of different biopolymers and therefore the use of two or more building blocks for the development of tissue-mimicking hydrogels is nowadays an attractive strategy in tissue-engineering. Multi-component hydrogel systems may undergo phase separation, which in turn can lead to new, unexpected material properties. The aim of this study was to understand the role of phase separation on the mechanical properties and 3D printability of hydrogels composed of triblock copolymers of polyethylene glycol (PEG) and methacrylated poly(N-(2-hydroxypropyl) methacrylamide-mono/dilactate) (pHPMAlac) blended with methacrylated hyaluronic acid (HAMA). To this end, hydrogels composed of different concentrations of PEG/pHPMAlac and HAMA, were analyzed for phase behavior and rheologicalproperties. Subsequently, phase separation and rheological behavior as function of the two polymer concentrations were mathematically processed to generate a predictive model. Results showed that PEG/pHPMAlac/HAMA hydrogels were characterized by hydrophilic, HAMA-richer internal domains dispersed in a more hydrophobic continuous phase, composed of PEG/pHPMAlac, and that the volume fraction of the dispersed phase increased by increasing HAMA concentration.Storage modulus, yield stress and viscosity increased with increasing HAMA concentration for low/medium HAMA contents (• 0.75% w/w), while a further increase of HAMA resulted in a 0.75% w/w), while a further increase of HAMA resulted in a decrease of the mentioned properties. On the other hand, by increasing the concentration of PEG/pHPMAlac these rheological properties were enhanced. The generated models showed a good fitting with experimental data, and were used to identify an exemplary 3D printability window for PEG/pHPMAlac/HAMA hydrogels, which was verified by rheological characterization and preparation of 3D printed scaffolds. In conclusion, a clear relationship between phase separation and rheological behavior in these two-component hydrogels can be described by complex functions of the two polymer concentrations. The predictive model generated in this study can be used as a valid tool for the identification of hydrogel compositions with desired, selected characteristics.3 Keywords: Thermosensitive polymers, hyaluronic acid, rheological properties, confocal laser scanning microscopy, neurofuzzy logic, 3D bioprinting. Highlights-Polymer ratios determine extent of phase separation in PEG/pHPMAlac/HAMA hydrogels.-Phase separation leads to hydrophilic domains dispersed in a more hydrophobic phase.-Phase separation affects rheological properties in a controllable fashion.-Mathematical models enabled a priori identification of hydrogels with desired properties.-Hydrogel 3D printing was achieved with a f...

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“…The OC plugs were embedded in paraffin and tissue sections (5 µm) were cut for the visualization of matrix components (Abbadessa et al, 2016). Safranin-O/Fast green staining was performed to visualize collagens (green/blue) and GAGs (red) (Wall and Board, 2014).…”

Section: Histology and Immunohistochemistrymentioning

confidence: 99%

Ex vivo model unravelling cell distribution effect in hydrogels for cartilage repair_suppl

Mouser

2017

ALTEX

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A Synthetic Thermosensitive Hydrogel for Cartilage Bioprinting and Its Biofunctionalization with Polysaccharides

Cited by 115 publications

References 74 publications

Ex vivo model unravelling cell distribution effect in hydrogels for cartilage repair

Ex vivo model unravelling cell distribution effect in hydrogels for cartilage repair

Two-component thermosensitive hydrogels: Phase separation affecting rheological behavior

Ex vivo model unravelling cell distribution effect in hydrogels for cartilage repair_suppl

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