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Microstructure, local viscoelasticity and cell culture suitability of 3D hybrid HA/collagen scaffolds.

ABSTRACT: As mechanical properties of cell culture substrates matter, methods for mechanical characterization of scaffolds on a relevant length scale are required. We used multiple particle tracking microrheology to close the gap between elasticity determined from bulk measurements and elastic properties sensed by cells. Structure and elasticity of macroporous, three-dimensional cryogel scaffolds from mixtures of hyaluronic acid (HA) and collagen (Coll) were characterized. Both one-component gels formed homogeneous networks, whereas hybrid gels were heterogeneous in terms of elasticity. Most strikingly, local elastic moduli were significantly lower than bulk moduli presumably due to non-equilibrium chain conformations between crosslinks. This was more pronounced in Coll and hybrid gels than in pure HA gels. Local elastic moduli were similar for all gels, irrespective of their different swelling ratio and bulk moduli. Fibroblast cell culture proved the biocompatibility of all investigated compositions. Coll containing gels enabled cell migration, adhesion and proliferation inside the gels.

SUBMITTER: Roether J

PROVIDER: S-EPMC6300200 | biostudies-literature | 2018

REPOSITORIES: biostudies-literature

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Microstructure, local viscoelasticity and cell culture suitability of 3D hybrid HA/collagen scaffolds.

Roether Johanna J Bertels Sarah S Oelschlaeger Claude C Bastmeyer Martin M Willenbacher Norbert N

PloS one 20181219 12

As mechanical properties of cell culture substrates matter, methods for mechanical characterization of scaffolds on a relevant length scale are required. We used multiple particle tracking microrheology to close the gap between elasticity determined from bulk measurements and elastic properties sensed by cells. Structure and elasticity of macroporous, three-dimensional cryogel scaffolds from mixtures of hyaluronic acid (HA) and collagen (Coll) were characterized. Both one-component gels formed h ...[more]

PMID: 30566463

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Project description:Bioengineering scaffolds have been improved to achieve efficient regeneration of various damaged tissues. In this study, we attempted to fabricate mechanically and biologically activated 3D printed scaffold in which porous gelatin/hydroxyapatite (G/H) as a matrix material provided outstanding mechanical properties with recoverable behavior, and human placental extracts (hPE) embedded in the scaffold were used as bioactive components. Methods: Various cell types (human adipose-derived stem cells; hASCs, pre-osteoblast; MC3T3-E1, human endothelial cell line; EA.hy926, and human dermal fibroblast; hDFs) were used to assess the effect of the hPE on cellular responses. High weight fraction (~ 70 wt%) of hydroxyapatite (HA) in a gelatin solution supplemented with glycerol was used for the G/H scaffold fabrication, and the scaffolds were immersed in hPE for the embedding (G/H/hPE scaffold). The osteogenic abilities of the scaffolds were investigated in cultured cells (hASCs) assaying for ALP activity and expression of osteogenic genes. For the in vivo test, the G/H and G/H/hPE scaffolds were implanted in the rat mastoid obliteration model. Results: The G/H/hPE scaffold presented unique elastic recoverable properties, which are important for efficient usage of implantable scaffolds. The effects of G/H and G/H/hPE scaffold on various in vitro cell-activities including non-toxicity, biocompatibility, and cell proliferation were investigated. The in vitro results indicated that proliferation (G/H = 351.1 ± 13.3%, G/H/hPE = 430.9 ± 8.7% at day 14) and expression of osteogenic markers (ALP: 3.4-fold, Runx2: 3.9-fold, BMP2: 1.7-fold, OPN: 2.4-fold, and OCN: 4.8-fold at day 21) of hASCs grown in the G/H/hPE scaffold were significantly enhanced compared with that in cells grown in the G/H scaffold. In addition, bone formation was also observed in an in vivo model using rat mastoid obliteration. Conclusion: In vitro and in vivo results suggested that the G/H/hPE scaffold is a potential candidate for use in bone tissue engineering.

Dataset Information

Microstructure, local viscoelasticity and cell culture suitability of 3D hybrid HA/collagen scaffolds.

Publications

Microstructure, local viscoelasticity and cell culture suitability of 3D hybrid HA/collagen scaffolds.

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