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  • From design of bio-based biocomposite electrospun scaffolds to osteogenic differentiation of human mesenchymal stromal cells.

From design of bio-based biocomposite electrospun scaffolds to osteogenic differentiation of human mesenchymal stromal cells.

Journal of materials science. Materials in medicine (2014-03-04)
Julien Ramier, Daniel Grande, Thibault Bouderlique, Olya Stoilova, Nevena Manolova, Iliya Rashkov, Valérie Langlois, Patricia Albanese, Estelle Renard
ABSTRACT

Electrospinning coupled with electrospraying provides a straightforward and robust route toward promising electrospun biocomposite scaffolds for bone tissue engineering. In this comparative investigation, four types of poly(3-hydroxybutyrate) (PHB)-based nanofibrous scaffolds were produced by electrospinning a PHB solution, a PHB/gelatin (GEL) mixture or a PHB/GEL/nHAs (hydroxyapatite nanoparticles) mixed solution, and by electrospinning a PHB/GEL solution and electrospraying a nHA dispersion simultaneously. SEM and TEM analyses demonstrated that the electrospun nHA-blended framework contained a majority of nHAs trapped within the constitutive fibers, whereas the electrospinning-electrospraying combination afforded fibers with a rough surface largely covered by the bioceramic. Structural and morphological characterizations were completed by FTIR, mercury intrusion porosimetry, and contact angle measurements. Furthermore, an in vitro investigation of human mesenchymal stromal cell (hMSC) adhesion and proliferation properties showed a faster cell development on gelatin-containing scaffolds. More interestingly, a long-term investigation of hMSC osteoblastic differentiation over 21 days indicate that hMSCs seeded onto the nHA-sprayed scaffold developed a significantly higher level of alkaline phosphatase activity, as well as a higher matrix biomineralization rate through the staining of the generated calcium deposits: the fiber surface deposition of nHAs by electrospraying enabled their direct exposure to hMSCs for an efficient transmission of the bioceramic osteoinductive and osteoconductive properties, producing a suitable biocomposite scaffold for bone tissue regeneration.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Hydroxyapatite, purum p.a., ≥90% (as Ca3(PO4)2, KT)
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Glutaraldehyde solution, technical, ~50% in H2O (5.6 M)
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Hydroxyapatite, synthetic, 99.8% trace metals basis (excludes Mg)
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Glutaraldehyde solution, 50 wt. % in H2O
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Glutaraldehyde solution, Grade II, 25% in H2O
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Cetylpyridinium chloride, meets USP testing specifications
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Glutaraldehyde solution, Grade I, 70% in H2O, specially purified for use as an electron microscopy fixative or other sophisticated use
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Glutaraldehyde solution, Grade I, 25% in H2O, specially purified for use as an electron microscopy fixative
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Glutaraldehyde solution, Grade I, 50% in H2O, specially purified for use as an electron microscopy fixative or other sophisticated use
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Calcium phosphate tribasic, 34.0-40.0% Ca basis
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Hydroxyapatite, puriss., meets analytical specification of Ph. Eur., BP, FCC, E341, ≥90% (calculated on glowed substance)
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Hydroxyapatite, nanoparticles, dispersion, 10 wt. % in H2O, <200 nm particle size (BET)
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Glutaraldehyde solution, Grade I, 8% in H2O, specially purified for use as an electron microscopy fixative or other sophisticated use
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Glutaraldehyde solution, 50% in H2O, suitable for photographic applications
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Hydroxyapatite, nanopowder, <200 nm particle size (BET), ≥97%, synthetic
Cetylpyridinium chloride, European Pharmacopoeia (EP) Reference Standard
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USP
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