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Electrospun hydroxyapatite-containing chitosan nanofibers crosslinked with genipin for bone tissue engineering

Paper ID Volume ID Publish Year Pages File Format Full-Text
6815 519 2012 12 PDF Available
Electrospun hydroxyapatite-containing chitosan nanofibers crosslinked with genipin for bone tissue engineering

Reconstruction of large bone defects remains problematic in orthopedic and craniofacial clinical practice. Autografts are limited in supply and are associated with donor site morbidity while other materials show poor integration with the host's own bone. This lack of integration is often due to the absence of periosteum, the outer layer of bone that contains osteoprogenitor cells and is critical for the growth and remodeling of bone tissue. In this study we developed a one-step platform to electrospin nanofibrous scaffolds from chitosan, which also contain hydroxyapatite nanoparticles and are crosslinked with genipin. We hypothesized that the resulting composite scaffolds represent a microenvironment that emulates the physical, mineralized structure and mechanical properties of non-weight bearing bone extracellular matrix while promoting osteoblast differentiation and maturation similar to the periosteum. The ultrastructure and physicochemical properties of the scaffolds were studied using scanning electron microscopy and spectroscopic techniques. The average fiber diameters of the electrospun scaffolds were 227 ± 154 nm as spun, and increased to 335 ± 119 nm after crosslinking with genipin. Analysis by X-ray diffraction, Fourier transformed infrared spectroscopy and energy dispersive spectroscopy confirmed the presence of characteristic features of hydroxyapatite in the composite chitosan fibers. The Young's modulus of the composite fibrous scaffolds was 142 ± 13 MPa, which is similar to that of the natural periosteum. Both pure chitosan scaffolds and composite hydroxyapatite-containing chitosan scaffolds supported adhesion, proliferation and osteogenic differentiation of mouse 7F2 osteoblast-like cells. Expression and enzymatic activity of alkaline phosphatase, an early osteogenic marker, were higher in cells cultured on the composite scaffolds as compared to pure chitosan scaffolds, reaching a significant, 2.4 fold, difference by day 14 (p < 0.05). Similarly, cells cultured on hydroxyapatite-containing scaffolds had the highest rate of osteonectin mRNA expression over 2 weeks, indicating enhanced osteoinductivity of the composite scaffolds. Our results suggest that crosslinking electrospun hydroxyapatite-containing chitosan with genipin yields bio-composite scaffolds, which combine non-weight-bearing bone mechanical properties with a periosteum-like environment. Such scaffolds will facilitate the proliferation, differentiation and maturation of osteoblast-like cells. We propose that these scaffolds might be useful for the repair and regeneration of maxillofacial defects and injuries.

Chitosan; Hydroxyapatite; Genipin; Electrospinning; Bone tissue engineering; Osteoblast differentiationAlamar blue, (AB); alkaline phosphatase, (ALP); chitosan, (CTS); chitosan fibers crosslinked with genipin, (CTS-GP); chitosan-hydroxyapatite fibers cros
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Electrospun hydroxyapatite-containing chitosan nanofibers crosslinked with genipin for bone tissue engineering
Database: Elsevier - ScienceDirect
Journal: Biomaterials - Volume 33, Issue 36, December 2012, Pages 9167–9178
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Physical Sciences and Engineering Chemical Engineering Bioengineering