Dulbecco s modified eagle s medium

The mouse embryonic fibroblast continuous cell line (NIH/3T3, ATCC^® CRL-1658™, Teddington, UK) was used for the cytocompatibility test, according to the EN ISO 10993-5 standard, with modification. As a culture medium, the ATCC-formulated Dulbecco’s Modified Eagle’s Medium (BioSera, Nuaille, France), containing 10% calf serum (BioSera, Nuaille, France) and Penicillin/Streptomycin at 100 U mL ${}^{-1}$ (PAA Laboratories GmbH, Pasching, Austria) was used. The tested samples were prepared with a dimension of 10 mm × 10 mm and sterilized by UV radiation (wavelength of 258 nm) for 30 min and placed into the 24 well-plate. The cells were seeded onto the samples in the concentration of $1\times {10}^4$ for an hour for adhesion of the cells. Tissue polystyrene was used as a reference. After the pre-cultivation, a sufficient amount of the medium was added and incubated for 48 h at 37 ${}^{°}$ C. The changes in cell morphology were observed with an inverted fluorescent microscope (Olympus, IX 81; Hamburg, Germany). To assess the cytotoxic effect, an MTT assay (Duchefa, Biochemie, Haarlem, The Netherlands) was performed. The absorbance was measured by an Infinite M200 Pro NanoQuant absorbance reader (Tecan, Männedorf, Switzerland). All tests were performed in triplicate.

A total of 106 human primary pancreatic adenocarcinoma tissues were collected between 2010 and 2012 at Fudan University Shanghai Cancer Center. The samples were immediately snap-frozen in liquid nitrogen and histologically examined in a timely manner. All samples were obtained with informed consent, and the project was approved by the Clinical Research Ethics Committee of Fudan University Shanghai Cancer Center.
The PCI35 and PCI55 human pancreatic carcinoma cell lines were a gift from Professor Mukaida Naofumi (Kanazawa University, Kanazawa, Japan). The SW1990, MiaPaca-2, PANC-1, BxPC-3, and Capan-1 human pancreatic carcinoma cell lines were purchased from the American Type Culture Collection (ATCC). SW1990 and Capan-1 cells were cultured in Dulbecco's Modified Eagle's Medium (Biosera), and all other cells were maintained in RPMI 1640 (Biosera). All media were supplemented with 10% FBS (Biosera), and the cells were cultured at 37°C in an incubator containing 5% CO₂. The cell lines were authenticated by DNA profiling of short tandem repeats and amelogenin (Beijing Microread Genetics Co. Ltd.).

The cell compatibility was detected according to previously described procedures [37 (link)]. The test samples (films Col/Chi, Col/Chi-Ta and DC-Col/Chi) were sterilized by UV radiation prior to testing. Mouse embryonic fibroblast cell line (ATCC CRL-1658 NIH/3T3; Marlboro, MA, USA) was used to test the adhesion and proliferation of cells on the surfaces. ATCC-formulated Dulbecco’s modified Eagle’s medium (Biosera, Nuaille, France) containing 10% calf serum (Biosera) and 100 U·mL⁻¹ penicillin/streptomycin (PAA, Trasadingen, Switzerland) was used as the culture medium.
In the case of adhesion, the cells were seeded on the samples in the concentration of 1.106 cells mL⁻¹. After one hour, the cells were stained with Hoechst 33258 (Molecular Probes, Carlsbad, CA, USA). To determine the ability of cells to proliferate on the surfaces, the cells were seeded at an initial concentration of 1.105 cells mL⁻¹ and cultivated for 48 h. After 48 h the cells were fixed and stained with Hoechst 33258 and ActinRed 555 (Thermo Fisher Scientific, Waltham, MA, USA). Micrographs were taken using the fluorescence microscope Olympus IX 81 (Olympus, Tokyo, Japan).

Bombyx mori (B. mori, silkworm) cocoons were
purchased from Kozabirlik (Bursa, Turkey). CNFs (<100 ppm iron
content), sodium carbonate (Na₂CO₃), lithium
bromide (LiBr), 1,1,1,3,3,3-hexafluoro-2-propanol (HFIP), protease
type XIV (>3.5 units/mg), and methanol (CH₃OH) were
purchased
from Sigma-Aldrich. NaCl was purchased from Isolab. Dulbecco’s
modified Eagle’s medium, penicillin–streptomycin, fetal
bovine serum (FBS), and trypsin–EDTA were purchased from Biosera.
3-(4,5-Dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium
bromide (MTT), dimethyl sulfoxide (DMSO), glutaraldehyde, and hexamethyldisilazane
(HMDS) were purchased from Glentham, Genaxxon Bioscience, Merck, and
Sigma, respectively.

The mouse embryonic fibroblast continuous cell line (NIH/3T3, ATCC^® CRL-1658™, Teddington, UK) was used for cytocompatibility test according to the EN ISO 10993-5 standard, with modification. As a culture medium, the ATCC-formulated Dulbecco’s Modified Eagle’s Medium (BioSera, Nuaille, France), containing 10% calf serum (BioSera, France) and Penicillin/Streptomycin at 100 U mL ${}^{-1}$ (PAA Laboratories GmbH, Pasching, Austria) was used. The tested samples were prepared with a dimension of 10 mm × 10 mm and sterilized by UV–radiation (wavelength of 258 nm) for 30 min and placed into the 24 well-plate. The cells were seeded onto the samples in the concentration of $1\times {10}^4$ for an hour for adhesion of the cells. After the pre-cultivation, a sufficient amount of the medium was added and incubated for 48 h at 37 ${}^{\circ }$ C. The changes in cell morphology were observed with an inverted fluorescent microscope (Olympus, IX 81). To assess the cytotoxic effect, an MTT assay (Duchefa, Biochemie, Haarlem, The Netherlands) was performed. The absorbance was measured by an Infinite M200 Pro NanoQuant absorbance reader (Tecan, Männedorf, Switzerland). All tests were performed in triplicates.

In vitro cytocompatibility was studied using primary mouse embryonic fibroblast cells (NIH/3T3, ATCC^® CRL-1658^TM, USA). The samples were prepared with dimensions of 10 × 10 mm foil and sterilized under UV-radiation (wavelength of 253.7 nm) for 30 min. As a culture medium, the ATCC-formulated Dulbecco’s modified Eagle’s medium (BioSera, France) containing 10% calf serum (BioSera, France) and 100 U mL⁻¹ penicillin/streptomycin (BioSera, France). The cells were seeded onto the samples in a concentration of 2 × 10⁴ cells per mm² and placed in an incubator for 24 h at 37 °C.
After the incubation period, the cell viability was determined using the 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay (Duchefa Biochemie, Amsterdam, the Netherlands). First, the cells were washed with PBS (BioSera, France) and a fresh medium containing MTT in the concentration of 0.5 mg per mL was added. After 4 h, formed formazan crystals were dissolved in DMSO and the absorbance was measured by an Infinite M200 Pro NanoQuant absorbance reader (Tecan, Switzerland) at 570 nm and the reference wavelength was adjusted to 690 nm. The results are presented as a reduction of cell viability in percentage when compared to cells cultivated on pure PLA. All tests were performed three times.

The mouse embryonic fibroblast continuous cell line (NIH/3T3, ATCC^® CRL-1658™, Teddington, UK) was used for cytocompatibility test, according to the EN ISO 10993-5 standard, with modification. As a culture medium, the ATCC-formulated Dulbecco’s Modified Eagle’s Medium (BioSera, Nuaille, France), containing 10% calf serum (BioSera, France) and Penicillin/Streptomycin at 100 U mL ${}^{-1}$ (PAA Laboratories GmbH, Pasching, Austria) was used. The tested samples were prepared with a dimension of 10 mm × 10 mm and sterilized by UV–radiation (wavelength of 258 nm) for 30 min and placed into the 24 well-plate. The cells were seeded onto the samples in the concentration of $1\times {10}^4$ for an hour for adhesion of the cells. After the precultivation, a sufficient amount of the medium was added and incubated for 72 h at 37 ${}^{\circ }$ C. The changes in cell morphology were observed with an inverted fluorescent microscope (Olympus, IX 81). In order to assess the cytotoxic effect, an MTT assay (Duchefa, Biochemie, Haarlem, The Netherlands) was performed. The absorbance was measured by an Infinite M200 Pro NanoQuant absorbance reader (Tecan, Männedorf, Switzerland). All tests were performed three times.