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  • Effect of incorporation of nanoscale bioactive glass and hydroxyapatite in PCL/chitosan nanofibers for bone and periodontal tissue engineering.

Effect of incorporation of nanoscale bioactive glass and hydroxyapatite in PCL/chitosan nanofibers for bone and periodontal tissue engineering.

Journal of biomedical nanotechnology (2013-04-30)
K T Shalumon, S Sowmya, D Sathish, K P Chennazhi, Shantikumar V Nair, R Jayakumar
ABSTRACT

A biomimetic scaffold which can very closely mimic the extracellular matrix of the bone was fabricated by incorporating nano-bioceramic particles such as nano bioglass (nBG) and nano hydroxyapatite (nHAp) within electrospun nanofibrous scaffold. A comparative study between nHAp incorporated poly(caprolactone) (PCL)-chitosan (CS) and nBG incorporated PCL-CS nanofibrous scaffolds was carried out and their feasibility in tissue engineering was investigated. All the samples were optimized to obtain fibers of similar diameter from 100-200 nm for the ease of comparison between the samples. Protein adsorption studies showed that PCL-CS incorporated with 3 wt% nHAp and 3 wt% nBG adsorbed more proteins on their surface than other samples. Cell attachment and proliferation studies using human periodontal ligament fibroblast cells (hPLFs) and osteoblast like cells (MG-63 cell lines) showed that nBG incorporated samples are slightly superior to nHAp incorporated counterparts. Cell viability test using alamar blue assay and live/dead staining confirms that the scaffolds are cytocompatible. ALP activity confirmed the osteoblastic behavior of hPDLFs. Also the presence of nHAp and nBG enhanced the ALP activity of hPDLF on the PCH3 and PCB3 scaffolds. These studies indicate that nBG incorporated electrospun scaffolds are comparatively better candidates for orthopedic and periodontal tissue engineering applications.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Chitosan, high molecular weight
Sigma-Aldrich
Phosphatase, Alkaline bovine, recombinant, expressed in Pichia pastoris, ≥4000 units/mg protein
Sigma-Aldrich
Polycaprolactone, average Mn 45,000
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Chitosan, from shrimp shells, practical grade
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Chitosan, low molecular weight
Sigma-Aldrich
Chitosan, from shrimp shells, ≥75% (deacetylated)
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Chitosan, medium molecular weight
Sigma-Aldrich
Polycaprolactone, average Mw ~14,000, average Mn ~10,000 by GPC
Sigma-Aldrich
Polycaprolactone, average Mn 80,000
Sigma-Aldrich
Hydroxyapatite, purum p.a., ≥90% (as Ca3(PO4)2, KT)
Sigma-Aldrich
Hydroxyapatite, puriss., meets analytical specification of Ph. Eur., BP, FCC, E341, ≥90% (calculated on glowed substance)
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Phosphatase, Alkaline from bovine intestinal mucosa, buffered aqueous glycerol solution, ≥4,000 DEA units/mg protein
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Phosphatase, Alkaline from Escherichia coli, buffered aqueous glycerol solution, 20-50 units/mg protein (in glycine buffer)
Sigma-Aldrich
Phosphatase, Alkaline from Escherichia coli, ammonium sulfate suspension, 30-90 units/mg protein (modified Warburg-Christian, in glycine buffer)
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Phosphatase, Alkaline from bovine intestinal mucosa, lyophilized powder, ≥10 DEA units/mg solid
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Phosphatase, Alkaline from bovine intestinal mucosa, buffered aqueous solution, ≥2,000 DEA units/mg protein
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Phosphatase, Alkaline from porcine kidney, lyophilized powder, ≥100 DEA units/mg protein
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Phosphatase, Alkaline from Escherichia coli, lyophilized powder, 30-60 units/mg protein (in glycine buffer)
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Phosphatase, Alkaline from bovine intestinal mucosa, ≥5,500 DEA units/mg protein
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Phosphatase, Alkaline from bovine intestinal mucosa, BioUltra, ≥5,700 DEA units/mg protein
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Phosphatase, Alkaline from calf intestinal mucosa, suitable for enzyme immunoassay, solution (clear, colorless), ~2500 U/mg protein (~10 mg/ml)
Sigma-Aldrich
Chitosan from shrimp shells, low-viscous
Sigma-Aldrich
Phosphatase, Alkaline shrimp, ≥900 DEA units/mL, buffered aqueous glycerol solution, recombinant, expressed in proprietary host