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  • Wet-electrospun PHBV nanofiber reinforced carboxymethyl chitosan-silk hydrogel composite scaffolds for articular cartilage repair.

Wet-electrospun PHBV nanofiber reinforced carboxymethyl chitosan-silk hydrogel composite scaffolds for articular cartilage repair.

Journal of biomaterials applications (2020-07-01)
Oylum Colpankan Gunes, Aylin Ziylan Albayrak, Seyma Tasdemir, Aylin Sendemir
ABSTRACT

The objective of the study was to produce three-dimensional and porous nanofiber reinforced hydrogel scaffolds that can mimic the hydrated composite structure of the cartilage extracellular matrix. In this regard, wet-electrospun poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) nanofiber reinforced carboxymethyl chitosan-silk fibroin (PNFs/CMCht-SF) hydrogel composite scaffolds that were chemically cross-linked by poly(ethylene glycol) diglycidyl ether (PEGDE) were produced. To the best of our knowledge, this is the first study in cartilage regeneration where a three dimensional porous spongy composite scaffold was obtained by the dispersion of wet-electrospun nanofibers within a polymer matrix. All of the produced hydrogel composite scaffolds had an interconnected microporous structure with well-integrated PHBV nanofibers on the pore walls. The scaffold comprising an equal amount of PEGDE and polymer (PNFs/CMCht-SF1:PEGDE1) demonstrated comparable water content (91.4 ± 0.7%), tan δ (0.183 at 1 Hz) and compressive strength (457 ± 85 kPa) values to that of articular cartilage. Besides, based on the histological analysis, this hydrogel composite scaffold supported the chondrogenic differentiation of bone marrow mesenchymal stem cells. Consequently, this hydrogel composite scaffold presented a great promise for cartilage tissue regeneration.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Poly(ethylene glycol) diglycidyl ether, average Mn 500
Sigma-Aldrich
L-Ascorbic acid 2-phosphate sesquimagnesium salt hydrate, ≥95%
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Osmium tetroxide, ReagentPlus®, 99.8%
Sigma-Aldrich
Chloroform, contains ethanol as stabilizer, meets analytical specification of BP, 99-99.4% (GC)
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Trypsin-EDTA solution, 1 ×, sterile; sterile-filtered, BioReagent, suitable for cell culture, 0.5 g porcine trypsin and 0.2 g EDTA, 4Na per liter of Hanks′ Balanced Salt Solution with phenol red
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Transforming Growth Factor-β1 human, TGF-β1, Xeno-free, recombinant, expressed in HEK 293 cells, suitable for cell culture
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ITS Liquid Media Supplement (100×), liquid, sterile-filtered, BioReagent, suitable for cell culture
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Dialysis tubing cellulose membrane, avg. flat width 25 mm (1.0 in.)
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Dexamethasone-Water Soluble, suitable for cell culture, BioReagent
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Alcian Blue 8GX, powder