An Improved Cryosection Method for Polyethylene Glycol Hydrogels Used in Tissue Engineering

Ruan, Jia‐Ling; Tulloch, Nathaniel L.; Muskheli, Veronica; Genova, E. Erin; Mariner, Peter D.; Anseth, Kristi S.; Murry, Charles E.

doi:10.1089/ten.tec.2012.0460

Cited by 43 publications

(46 citation statements)

References 31 publications

(33 reference statements)

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“…duPont Hospital for Children using a protocol adapted from Ruan et al [32]. Briefly, PEG hydrogels were removed from culture medium and fixed with 4% paraformaldehyde for 30 minutes, rinsed, and then infiltrated with Tissu-Tek® Optimal Cutting Temperature embedding medium (OCT; Sakura-Finetek, Torrance, CA) overnight at 4°C.…”

Section: Methodsmentioning

confidence: 99%

Decreasing matrix modulus of PEG hydrogels induces a vascular phenotype in human cord blood stem cells

et al. 2015

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Adult and congenital cardiovascular diseases are significant health problems that are often managed using surgery. Bypass grafting is a principal therapy, but grafts fail at high rates due to hyperplasia, fibrosis, and atherosclerosis. Biocompatible, cellularized materials that attenuate these complications and encourage healthy microvascularization could reduce graft failure, but an improved understanding of biomaterial effects on human stem cells is needed to reach clinical utility. Our group investigates stem-cell-loaded biomaterials for placement along the adventitia of at-risk vessels and grafts. Here, the effects of substrate modulus on human CD34+ stem cells from umbilical cord blood were evaluated. Cells were isolated by immunomagnetic separation and encapsulated in 3, 4, and 6 weight% PEG hydrogels containing 0.032% gelatin and 0.0044% fibronectin. Gels reached moduli of 0.34, 4.5, and 9.1 kPa. Cell viability approached 100%. Cell morphologies appeared similar across gels, but proliferation was significantly lower in 6 wt% gels. Expression profiling using stem cell signaling arrays indicated enhanced self-renewal and differentiation into vascular endothelium among cells in the lower weight percent gels. Thus, modulus was associated with cell proliferation and function. Gels with moduli in the low kilopascal range may be useful in stimulating cell engraftment and microvascularization of graft adventitia.

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Section: Methodsmentioning

confidence: 99%

Decreasing matrix modulus of PEG hydrogels induces a vascular phenotype in human cord blood stem cells

et al. 2015

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“…27 The sections of scaffolds were dried by cold air before hematoxylin and eosin (H&E) staining. Then the slices were immersed into hematoxylin solution for 6 min and washed in water for 3 times.…”

Section: Cellular Morphology Studymentioning

confidence: 99%

Preparation, characterization, and evaluation of genipin crosslinked chitosan/gelatin three-dimensional scaffolds for liver tissue engineering applications

Zhang

Wang

Yan

et al. 2016

J. Biomed. Mater. Res.

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In liver tissue engineering, scaffolds with porous structure desgined to supply nutrient and oxygen exchange for three-dimensional (3-D) cells culture, and maintain liver functions. Meanwhile, genipin, as a natural crosslinker, is widely used to crosslink biomaterials in tissue engineering, with lower cytotoxicity and better biocompatibility. In present study, chitosan/gelatin 3-D scaffolds crosslinked by genipin, glutaraldehyde or 1-(3-dimethylaminopropyl)-3-ethyl-carbodimide hydrochloride (EDC) were prepared and characterized by Fourier-transform infrared (FT-IR) and scanning electron microscopy (SEM). The biocompatibility of chitosan/gelatin scaffolds corsslinked with different crosslinkers was investigated by cell viability, morphology and liver specific functions. The result showed that the 1% and 2% genipin crosslinked chitosan/gelatin scaffolds possess ideal porosity. The genipin crosslinked 3-D scaffolds possessed the best biocompatibility than that of the others, and maintained liver specific functions when HepG2 cells seeded on scaffolds. The cellular morphology of HepG2 cells seeded on scaffolds showed that cells could penetrate into the scaffolds and proliferate significantly. Therefore, genipin crosslinked chitosan/gelatin scaffolds could be a promising biomaterial used in liver tissue engineering. © 2016 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 104A: 1863-1870, 2016.

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“…Subsequently, samples were fixed in 10% buffered formalin (Fisher Scientific, Pittsburgh, PA, USA) overnight and partially demineralized in 10% formic acid (VWR, Radnor, PA, USA) for 48 to 72 h to allow for sectioning of the tissue. To preserve the hydrogel during cryosectioning, samples were soaked in HistoPrep (Fisherbrand, Fair Lawn, NJ, USA) overnight at 4 °C (Ruan et al 2013). The following day, samples were rinsed and embedded in OCT Compound (Tissue-Tek, Torrance, CA, USA) and flash-frozen in dry ice/ethanol slurry.…”

Section: Histologymentioning

confidence: 99%

Ex Vivo Modeling of Multidomain Peptide Hydrogels with Intact Dental Pulp

et al. 2015

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Preservation of a vital dental pulp is a central goal of restorative dentistry. Currently, there is significant interest in the development of tissue engineering scaffolds that can serve as biocompatible and bioactive pulp-capping materials, driving dentin bridge formation without causing cytotoxic effects. Our earlier in vitro studies described the biocompatibility of multidomain peptide (MDP) hydrogel scaffolds with dental pulp-derived cells but were limited in their ability to model contact with intact 3-dimensional pulp tissues. Here, we utilize an established ex vivo mandible organ culture model to model these complex interactions. MDP hydrogel scaffolds were injected either at the interface of the odontoblasts and the dentin or into the pulp core of mandible slices and subsequently cultured for up to 10 d. Histology reveals minimal disruption of tissue architecture adjacent to MDP scaffolds injected into the pulp core or odontoblast space. Additionally, the odontoblast layer is structurally preserved in apposition to the MDP scaffold, despite being separated from the dentin. Alizarin red staining suggests mineralization at the periphery of MDP scaffolds injected into the odontoblast space. Immunohistochemistry reveals deposition of dentin sialophosphoprotein by odontoblasts into the adjacent MDP hydrogel, indicating continued functionality. In contrast, no mineralization or dentin sialophosphoprotein deposition is evident around MDP scaffolds injected into the pulp core. Collagen III expression is seen in apposition to gels at all experimental time points. Matrix metalloproteinase 2 expression is observed associated with centrally injected MDP scaffolds at early time points, indicating proteolytic digestion of scaffolds. Thus, MDP scaffolds delivered centrally and peripherally within whole dental pulp tissue are shown to be biocompatible, preserving local tissue architecture. Additionally, odontoblast function and pulp vitality are sustained when MDP scaffolds are intercalated between dentin and the odontoblast region, a finding that has significant implications when considering these materials as pulp-capping agents.

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An Improved Cryosection Method for Polyethylene Glycol Hydrogels Used in Tissue Engineering

Cited by 43 publications

References 31 publications

Decreasing matrix modulus of PEG hydrogels induces a vascular phenotype in human cord blood stem cells

Decreasing matrix modulus of PEG hydrogels induces a vascular phenotype in human cord blood stem cells

Preparation, characterization, and evaluation of genipin crosslinked chitosan/gelatin three-dimensional scaffolds for liver tissue engineering applications

Ex Vivo Modeling of Multidomain Peptide Hydrogels with Intact Dental Pulp

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