Manufacturing of Zinc Oxide Nanoparticle (ZnO NP)-Loaded Polyvinyl Alcohol (PVA) Nanostructured Mats Using Ginger Extract for Tissue Engineering Applications.

In this research, as an alternative to chemical and physical methods, environmentally and cost-effective antimicrobial zinc oxide nanoparticles (ZnO NP) were produced by the green synthesis method. The current study focuses on the production of ZnO NP starting from adequate precursor and Zingiber officinale aqueous root extracts (ginger). The produced ZnO NP was loaded into electrospun nanofibers at different concentrations for various tissue engineering applications such as wound dressings. The produced ZnO NPs and ZnO NP-loaded nanofibers were examined by Scanning Electron Microscopy (SEM) for morphological assessments and Fourier-transform infrared spectrum (FT-IR) for chemical assessments. The disc diffusion method was used to test the antimicrobial activity of ZnO NP and ZnO NP-loaded nanofibers against three representatives strains, Escherichia coli (Gram-negative bacteria), Staphylococcus aureus (Gram-positive bacteria), and Candida albicans (fungi) microorganisms. The strength and stretching of the produced fibers were assessed using tensile tests. Since water absorption and weight loss behaviors are very important in tissue engineering applications, swelling and degradation analyses were applied to the produced nanofibers. Finally, the MTT test was applied to analyze biocompatibility. According to the findings, ZnO NP-loaded nanofibers were successfully synthesized using a green precipitation approach and can be employed in tissue engineering applications such as wound dressing.

Atmospheric Pressure Plasma Surface Treatment of Polymers and Influence on Cell Cultivation.

Atmospheric plasma treatment is an effective and economical surface treatment technique. The main advantage of this technique is that the bulk properties of the material remain unchanged while the surface properties and biocompatibility are enhanced. Polymers are used in many biomedical applications; such as implants, because of their variable bulk properties. On the other hand, their surface properties are inadequate which demands certain surface treatments including atmospheric pressure plasma treatment. In biomedical applications, surface treatment is important to promote good cell adhesion, proliferation, and growth. This article aim is to give an overview of different atmospheric pressure plasma treatments of polymer surface, and their influence on cell-material interaction with different cell lines.

A novel treatment strategy for preterm birth: Intra-vaginal progesterone-loaded fibrous patches.

Progesterone-loaded poly(lactic) acid fibrous polymeric patches were produced using electrospinning and pressurized gyration for intra-vaginal application to prevent preterm birth. The patches were intravaginally inserted into rats in the final week of their pregnancy, equivalent to the third trimester of human pregnancy. Maintenance tocolysis with progesterone-loaded patches was elucidated by recording the contractile response of uterine smooth muscle to noradrenaline in pregnant rats. Both progesterone-loaded patches indicated similar results from release and thermal studies, however, patches obtained by electrospinning had smaller average diameters and more uniform dispersion compared to pressurized gyration. Patches obtained by pressurized gyration had better results in production yield and tensile strength than electrospinning; thereby pressurized gyration is better suited for scaled-up production. The patches did not affect cell attachment, viability, and proliferation on Vero cells negatively. Consequently, progesterone-loaded patches are a novel and successful treatment strategy for preventing preterm birth.

Preparation of electrospun PCL-based scaffolds by mono/multi-functionalized GO.

In the present study, sythetic biodegradable polymer poly(ε-caprolactone) (PCL) and graphene oxide (GO) were combined together to prepare 3D, composite tissue scaffolds (PCL/GO scaffolds) by using electrospinning technique. Also, the influence of Gly-Arg-Gly-Asp-Ser-Pro (GRGDSP) and/or thiophene (Th) modified GO on the composite PCL/GO mats (PCL/GO, PCL/GO-GRGDSP, PCL/GO-Th, PCL/GO-GRGDSP-Th) was further investigated. Characteristic examinations of the scaffolds were carried out by scanning electron microscope (SEM), contact angle (CA) measurements, x-ray photoelectron spectroscopy, TGA, electrical conductivity tests, phosphate buffer saline absorption and shrinkage tests and mechanical tests. All of the scaffolds were exhibited suitable bead-free and uniform morphology according to SEM images. With the addition of GO, better hydrophilicity and a slight CA decrease (∼5°) for the PCL/GO scaffolds were observed. Mechanical properties were reinforced drastically with the addition and well-dispersion of GO into PCL matrix. The incorporation of PCL and GO exhibited enhanced electrical conductivity and the highest value was found for PCL/GO-GRGDSP-Th (2%) as 15.06 µS cm-1. The MG-63 osteoblast cell culture studies (MTT assay, ALP activity, Alizarin-Red staining, fluorescence and SEM analyses) showed that PCL/GO-GRGDSP-Th (1%) scaffolds exhibited the highest biocompatibility performance (1.87 fold MTT absorbance value comparing with neat PCL) due to the advanced properties of GO and the biological interfaces.

Development of Amoxicillin-Loaded Electrospun Polyurethane/Chitosan/ $\beta$ -Tricalcium Phosphate Scaffold for Bone Tissue Regeneration.

Biocompatible nanocomposite electrospun fibers containing Polyurethane/Chitosan/ $\beta $ -Tri calcium phosphate with diverse concentrations were designed and produced through the electrospinning process for bone tissue engineering applications. After the production process, density measurement, viscosity, electrical conductivity, and tensile strength measurement tests were carried out as physical analyses of blended solutions. The chemical structural characterization was scrutinized using Fourier transform infrared spectrometer (FTIR), and scanning electron microscopy (SEM) was used to observe the morphological details of developed electrospun scaffolds. Cell viability, attachment, and proliferation were performed using a L929 fibroblast cell line. Based on the physical, SEM, FTIR analysis, and cell culture studies, preferable nanofiber composition was selected for further studies. Amoxicillin (AMX) was loaded to that selected nanofiber composition for examination of the drug release. In comparison with other studies on similar AMX controlled products, higher drug loading and encapsulation efficiencies were obtained. It has been clearly found that the developed nanofiber composites have potential for bone tissue engineering applications.

Dielectric barrier discharge and jet type plasma surface modifications of hybrid polymeric poly (ε-caprolactone)/chitosan scaffolds.

In this study, dry air plasma jet and dielectric barrier discharge Ar + O2 or Ar + N2 plasma modifications and their effects on wettability, topography, functionality and biological efficiency of the hybrid polymeric poly (ε-caprolactone)/chitosan scaffolds were reported. The samples treated with Ar + O2 dielectric barrier discharge plasma (80 sccm O2 flow rate, 3-min treatment) or with dry air plasma jet (15-cm nozzle-sample distance, 13-min treatment) had the closest wettability (49.11 ± 1.83 and 53.60 ± 0.95, respectively) to the commercial tissue culture polystyrene used for cell cultivation. Scanning electron microscopy images and X-ray photoelectron spectrometry analysis showed increase in topographical roughness and OH/NH2 functionality, respectively. Increased fluid uptake capacity for the scaffolds treated with Ar + O2 dielectric barrier discharge plasma (73.60% ± 1.78) and dry air plasma jet (72.48% ± 0.75) were also noted. Finally, initial cell attachment as well as seven-day cell viability, growth and proliferation performances were found to be significantly better for both plasma treated scaffolds than for untreated scaffolds.

Development of core-shell coaxially electrospun composite PCL/chitosan scaffolds.

This study was related to combining of synthetic Poly (ε-caprolactone) (PCL) and natural chitosan polymers to develop three dimensional (3D) PCL/chitosan core-shell scaffolds for tissue engineering applications. The scaffolds were fabricated with coaxial electrospinning technique and the characterizations of the samples were done by thickness and contact angle (CA) measurements, scanning electron microscopy (SEM), transmission electron microscopy (TEM), X-Ray Photoelectron Spectroscopy (XPS) analyses, mechanical and PBS absorption and shrinkage tests. The average inter-fiber diameter values were calculated for PCL (0.717±0.001µm), chitosan (0.660±0.007µm) and PCL/chitosan core-shell scaffolds (0.412±0.003µm), also the average inter-fiber pore size values exhibited decreases of 66.91% and 61.90% for the PCL and chitosan scaffolds respectively, compared to PCL/chitosan core-shell ones. XPS analysis of the PCL/chitosan core-shell structures exhibited the characteristic peaks of PCL and chitosan polymers. The cell culture studies (MTT assay, Confocal Laser Scanning Microscope (CLSM) and SEM analyses) carried out with L929 ATCC CCL-1 mouse fibroblast cell line proved that the biocompatibility performance of the scaffolds. The obtained results showed that the created micro/nano fibrous structure of the PCL/chitosan core-shell scaffolds in this study increased the cell viability and proliferation on/within scaffolds.

Effects of nozzle type atmospheric dry air plasma on L929 fibroblast cells hybrid poly (ε-caprolactone)/chitosan/poly (ε-caprolactone) scaffolds interactions.

In the study presented here, in order to improve the surface functionality and topography of poly (ε-caprolactone) (PCL)/chitosan/PCL hybrid tissue scaffolds fabricated layer by layer with electrospinning technique, an atmospheric pressure nozzle type plasma surface modification was utilized. The optimization of the plasma process parameters was carried out by monitoring the changes in surface hydrophilicity by using contact angle measurements. SEM, AFM and XPS analyses were utilized to observe the changes in topographical and chemical properties of the modified surfaces. The results showed that applied plasma modification altered the nanotopography and the functionality of the surfaces of the scaffolds. The modification applied for 9 min from a distance of 17 cm was found to provide the possible contact angle value (75.163±0.083) closest to the target value which is the value of tissue culture polystyrene (TCPS) petri dishes (∼49.7°), compared to the unmodified samples (84.46±3.86). In vitro cell culture was carried out by L929 mouse fibroblast cell line in order to examine the effects of plasma surface modification on cell-material interactions. Standard MTT assay showed improved cell viability on/within modified scaffolds confirmed with the observations of the cell attachment and the morphology by means of SEM, fluorescence and confocal imaging. The experiments performed in the study proved the enhanced biocompatibility of the nozzle type dry air plasma modified scaffolds.

DBD atmospheric plasma-modified, electrospun, layer-by-layer polymeric scaffolds for L929 fibroblast cell cultivation.

This paper reported a study related to atmospheric pressure dielectric barrier discharge (DBD) Ar + O2 and Ar + N2 plasma modifications to alter surface properties of 3D PCL/Chitosan/PCL layer-by-layer hybrid scaffolds and to improve mouse fibroblast (L929 ATCC CCL-1) cell attachment, proliferation, and growth. The scaffolds were fabricated using electrospinning technique and each layer was electrospun sequentially on top of the other. The surface modifications were performed with an atmospheric pressure DBD plasma under different gas flow rates (50, 60, 70, 80, 90, and 100 sccm) and for different modification times (0.5-7 min), and then the chemical and topographical characterizations of the modified samples were done by contact angle (CA) measurements, scanning electron microscopy (SEM), atomic force microscopy, and X-ray photoelectron spectroscopy. The samples modified with Ar + O2 plasma for 1 min under 70 cm(3)/min O2 flow rate (71.077° ± 3.578) showed a 18.83% decrease compare to unmodified samples' CA value (84.463° ± 3.864). Comparing with unmodified samples, the average fiber diameter values for plasma-modified samples by Ar + O2 (1 min 70 sccm) and Ar + N2 (40 s 70 sccm) increased 40.756 and 54.295%, respectively. Additionally, the average inter-fiber pore size values exhibited decrease of 37.699 and 48.463% for the same Ar + O2 and Ar + N2 plasma-modified samples, respectively, compare to unmodified samples. Biocompatibility performance was determined with MTT assay, fluorescence, Giemsa, and confocal imaging as well as SEM. The results showed that Ar + O2-based plasma modification increased the hydrophilicity and oxygen functionality of the surface, thus affecting the cell viability and proliferation on/within scaffolds.

Influence of water/O2 plasma treatment on cellular responses of PCL and PET surfaces.

In this study, low pressure water/O2 plasma treatment was performed in order to obtain COOH functionalities on the surface of poly-ε-caprolactone (PCL) membranes as well as non-woven polyester fabric (NWPF) discs. The plasma treatments were performed in a cylindrical, capacitively coupled RF-plasma-reactor and then following steps were performed: in situ (oxalyl chloride vapors) gas/solid reaction to convert -OH functionalities into -COCl groups; and hydrolysis under open laboratory conditions using air moisture for final-COOH functionalities. COOH and OH functionalities on modified surfaces were detected quantitatively by using fluorescent labeling technique and an UVX 300G sensor. Electron spectroscopy for chemical analysis (ESCA) was used to evaluate the relative surface atomic compositions and the carbon and oxygen linkages located in non-equivalent atomic positions of untreated and modified surfaces. Atomic force microscope (AFM) analysis showed that nanoscale features of the PCL surfaces are dramatically changed during the surface treatments. Scanning electron microscopy (SEM) results indicated the changes in the relatively smooth appearance of the untreated NWPF discs after the plasma treatment. Periodontal ligament (PDL) fibroblasts were used in cell culture studies. Cell culture results showed that plasma treated PCL membranes and NWPF discs were favorable for the PDL cell spreading, growth and viability due to the presence of functional groups and/or nanotopographies on their surfaces.

Insulin and heparin co-immobilized 3D polyester fabrics for the cultivation of fibroblasts in low-serum media.

Insulin and/or heparin immobilized/co-immobilized non-woven polyester fabric (NWPF) discs were developed for the cultivation of L929 mouse fibroblasts in low-serum media. At first, NWPF discs were hydrolyzed to obtain a carboxylic acid group-introduced matrix (NWPF-hydrolyzed). Insulin and heparin co-immobilized NWPF (NWPF-insulin-heparin) was prepared by the grafting of PEO onto NWPF-hydrolyzed disc (NWPF-PEO), followed by the reaction first with insulin and then heparin. In the presence of spacer arm, PEO, the amount of immobilized insulin molecules significantly increased from 6.96 to 84.45 microg/cm(2). The amount of heparin bound to the NWPF-PEO (5.93 microg/cm(2)) was higher than that of the insulin immobilized surface (4.59 microg/cm(2)). Insulin and heparin immobilized NWPF discs were observed with fluorescence microscopy by labeling the insulin and heparin with 8-anilino-1-naphthalene sulfonic acid (ANS) or fluorescein isothiocyanate (FITC), respectively. L929 fibroblasts were used to check the cell adhesion and cell growth capabilities of modified NWPF discs in low-serum media (containing 5% fetal bovine serum). Optical photographs showed that after 2nd day of the culture, fibroblastic cells spread along the length of modified fibers, eventually filling the interfiber space. At the end of 6-day growth period, cell yield in the presence of immobilized heparin was a little bit higher than that of the immobilized insulin. Co-immobilized (insulin/heparin) NWPF discs did not accelerate the cell growth as well as insulin or heparin immobilized discs.