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Fabrication of a Biomimetic Nano-Matrix with Janus Base Nanotubes and Fibronectin for Stem Cell Adhesion.

Journal of visualized experiments : JoVE (2020-05-26)
Libo Zhou, Anne Yau, Wuxia Zhang, Yupeng Chen
ABSTRACT

A biomimetic NM was developed to serve as a tissue-engineering biological scaffold, which can enhance stem cell anchorage. The biomimetic NM is formed from JBNTs and FN through self-assembly in an aqueous solution. JBNTs measure 200-300 µm in length with inner hydrophobic hollow channels and outer hydrophilic surfaces. JBNTs are positively charged and FNs are negatively charged. Therefore, when injected into a neutral aqueous solution, they are bonded together via noncovalent bonding to form the NM bundles. The self-assembly process is completed within a few seconds without any chemical initiators, heat source, or UV light. When the pH of the NM solution is lower than the isoelectric point of FNs (pI 5.5-6.0), the NM bundles will self-release due to the presence of positively charged FN. NM is known to mimic the extracellular matrix (ECM) morphologically and hence, can be used as an injectable scaffold, which provides an excellent platform to enhance hMSC adhesion. Cell density analysis and fluorescence imaging experiments indicated that the NMs significantly increased the anchorage of hMSCs compared to the negative control.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Acetonitrile, suitable for HPLC, gradient grade, ≥99.9%
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Methanol, suitable for HPLC, ≥99.9%
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N,N-Dimethylformamide, anhydrous, 99.8%
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Dichloromethane, anhydrous, ≥99.8%, contains 40-150 ppm amylene as stabilizer
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Ethyl acetate, ACS reagent, ≥99.5%
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Citric acid, ACS reagent, ≥99.5%
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Sodium bicarbonate, ACS reagent, ≥99.7%
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Phosphorus(V) oxychloride, ReagentPlus®, 99%
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Hydrochloric acid, 36.5-38.0%, BioReagent, for molecular biology
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Urethane, ≥99%