Ammonia plasma-treated electrospun polyacrylonitryle nanofibrous membrane: the robust substrate for protein immobilization through glutaraldhyde coupling chemistry for biosensor application.

The surface of polyacrylonitrile electrospun nanofibrous membrane (PAN NFM) was aminated by the ammonia plasma treatment. The content of amine groups has been estimated for different time of plasma treatment. The newly generated amine groups were successfully activated by glutaraldehyde (Ga) for the covalent attachment of the protein molecules on the NFM surface. Bio-functionalization of ammonia plasma treated PAN NFM was carried out by the primary antibodies (Ab) immobilization as a protein model through Ga coupling chemistry. For comparison, the immobilization of Ab was also performed through physical interactions. Attenuated total reflection-fourier transform infrared spectroscopy (ATR-FTIR) was used for the characterization of surface functional groups of PAN NFM after different modifications. The surface morphology of the NFM after immobilization was characterized using scanning electron microscope (SEM). The efficacy of Ab immobilization was estimated by enzyme-linked immuno sorbent assay (ELISA) method. X- Ray photoelectron spectroscopy (XPS) was performed to confirm the covalent immobilization of Ab on the modified PAN NFM. Results show that ammonia plasma treatment effectively increased the amount of Ab immobilization through Ga coupling chemistry. Our findings suggest that this is a versatile model for the preparation of stable bio-functionalized NFM which is applicable in different field of biomedical science.

Study of epithelial differentiation and protein expression of keratinocyte-mesenchyme stem cell co-cultivation on electrospun nylon/B. vulgaris extract composite scaffold.

Employing of the composite electrospun scaffold containing herbal extract in conjugation with co-culturing of cells can open up new window to the design of efficient biomaterials for skin tissue regeneration. Here, we introduce the synergistic effect of composite electrospun nanofibrous scaffold of nylon66 loaded with Beta vulgaris (B. vulgaris) (extract of beet roots, a plants whose widely used in Iranian folk medicine as wound healing medicine) and co-culture of mesenchymal stem-cells (MSCs)-human keratinocyte (H-keratino) differentiation towards epithelial lineage. In vitro biocompatibility was examined through MTT assay and epithelial differentiation checked by real-time PCR and immunocytochemistry (ICC) assay after co-culturing of MSCs and H-keratino on proposed scaffold. Significant enhancement in cell proliferation was detected after cell culturing on the composite type of electrospun scaffold containing B. vulgaris. Moreover, after 14days of co-culturing process, gene expression results revealed that both composite and non-composite nylon66 electrospun scaffold promote epithelial differentiation compared to mono-cell culturing of H-keratino in terms of several markers as Cytokeratin 10, Cytokeratin 14 and Involucrin and ICC of some dermal proteins like Cytokeratin 14 and Loricrin. To the best of our knowledge, findings of this study will introduce new way for the generation of novel biomaterials for the development of current skin tissue engineering.