Bacterial cellulose, a random assembled, ribbon-shaped, and nanosized fibrils is produced extracellularly by Gram negative strains of Acetobacter Xylinum. Due to its superior mechanical property and 3-d microstructure which mimics the conformation of extracellular matrix, bacterial cellulose becomes one of the potential biomaterials for designing the scaffold in tissue engineering. However, beside the morphological motifs, more surface-tailoring factors such as the surface properties of materials are also essential in tissue engineering approaches.
In this study, two surface modifications were performed to achieve the goals on (i) promoting the biocompatibility of BC membranes, and (ii) the creation of anti-fouling BC membranes.
In terms of promoting the biocompatibility of BC membranes, surface modifications of bacterial cellulose were performed by plasma treatment carried out by using oxygen (O2), nitrogen (N2) and tetrafluorocarbon (CF4) plasmas in order to discover the optimized cell-surface interactions. The pristine and plasma-treated surfaces were characterized by water contact angle measurement, atomic force microscope (AFM), Fourier transformed infrared (FTIR), and electron spectroscopy for chemical analysis (ESCA). By using oxygen and nitrogen plasmas, the wettability of bacterial cellulose increased while the tetrafluorocarbon plasma decreased the wettability of materials. By using ESCA, the presence of oxygen, nitrogen and fluorine was detected on the O2, N2 and CF4 plasma-treated bacterial cellulose and further deconvolution of C1s peak helped to discover the specific functional groups such as O-C-O and C=O, CO-NH2 and Cx-Fx bindings, incorporated by using O2, N2 and CF4 plasmas, respectively.
The biocompatibility of plasma treated BCs was evaluated by directly cultivating different kinds of cells, L-929 mouse fibroblast, Chinese hamster ovary (CHO) and human embryonic skin fibroblast, under the condition with or without serum in cell culture medium.
Under serum containing medium, for 6h of cell incubation time, cell attachment on the BC-CF4 was about 40-50% higher than that on BC, BC-O2 and BC-N2 and nearly identical to that on TCPS. The cells show lower density on all samples under the serum free cell culture condition. The effect of serum was explained by the proteins, originated from cell culture media, which adsorbed in larger amount onto CF4 plasma-treated BC revealed by both proteins adsorption and ESCA analyses.
For long term cell cultivation time (24 and 48 hr), both L929 and CHO cells revealed elongated shape (over than 3 times of the initial cell length) and higher cell density on BC-CF4 comparing with on the pristine BC, O2 and N2 modified BCs. On the other hand, by cultivating human embryonic skin fibroblast cells on the BC membranes, the improvement of biocompatibility was shown on all the plasma treated surfaces compared to the pristine surface.
This study proposed to apply kinetic parameter to evaluate the initial cell adhesion behavior by pseudo first order Lagergren model. For both L929 and CHO cells, the cell density increased significantly on CF4 plasma treated BC than that on the pristine BC and on other plasmas treated BCs under the cell culture condition with serum. The increment of the adhered cell number can be quantified by the kinetic parameter which increased 2-3 folds for both cell types.
The second goal of this thesis, the incorporation of antifouling property onto BC membranes was facilitated by two methods to graft PEG: (i) plasma activation of PEG to graft on BC (PEG-g-BC), and (ii) chemically grafting PEG onto BC by using IPDI (mPEG-BC). The surface characterization by using SEM and FTIR indicated that the PEG was successfully incorporated onto BC for both PEG-g-BC and mPEG-BC and showed the decreasing of both protein and L-929 cell adhesion. Moreover, mPEG-BC showed particular anti-fouling effect compared to the PEG-g-BC by examining the cell density which was probably due to the brush conformation of PEG-g-BC.

Bacterial cellulose, a random assembled, ribbon-shaped, and nanosized fibrils is produced extracellularly by Gram negative strains of Acetobacter Xylinum. Due to its superior mechanical property and 3-d microstructure which mimics the conformation of extracellular matrix, bacterial cellulose becomes one of the potential biomaterials for designing the scaffold in tissue engineering. However, beside the morphological motifs, more surface-tailoring factors such as the surface properties of materials are also essential in tissue engineering approaches.
In this study, two surface modifications were performed to achieve the goals on (i) promoting the biocompatibility of BC membranes, and (ii) the creation of anti-fouling BC membranes.
In terms of promoting the biocompatibility of BC membranes, surface modifications of bacterial cellulose were performed by plasma treatment carried out by using oxygen (O2), nitrogen (N2) and tetrafluorocarbon (CF4) plasmas in order to discover the optimized cell-surface interactions. The pristine and plasma-treated surfaces were characterized by water contact angle measurement, atomic force microscope (AFM), Fourier transformed infrared (FTIR), and electron spectroscopy for chemical analysis (ESCA). By using oxygen and nitrogen plasmas, the wettability of bacterial cellulose increased while the tetrafluorocarbon plasma decreased the wettability of materials. By using ESCA, the presence of oxygen, nitrogen and fluorine was detected on the O2, N2 and CF4 plasma-treated bacterial cellulose and further deconvolution of C1s peak helped to discover the specific functional groups such as O-C-O and C=O, CO-NH2 and Cx-Fx bindings, incorporated by using O2, N2 and CF4 plasmas, respectively.
The biocompatibility of plasma treated BCs was evaluated by directly cultivating different kinds of cells, L-929 mouse fibroblast, Chinese hamster ovary (CHO) and human embryonic skin fibroblast, under the condition with or without serum in cell culture medium.
Under serum containing medium, for 6h of cell incubation time, cell attachment on the BC-CF4 was about 40-50% higher than that on BC, BC-O2 and BC-N2 and nearly identical to that on TCPS. The cells show lower density on all samples under the serum free cell culture condition. The effect of serum was explained by the proteins, originated from cell culture media, which adsorbed in larger amount onto CF4 plasma-treated BC revealed by both proteins adsorption and ESCA analyses.
For long term cell cultivation time (24 and 48 hr), both L929 and CHO cells revealed elongated shape (over than 3 times of the initial cell length) and higher cell density on BC-CF4 comparing with on the pristine BC, O2 and N2 modified BCs. On the other hand, by cultivating human embryonic skin fibroblast cells on the BC membranes, the improvement of biocompatibility was shown on all the plasma treated surfaces compared to the pristine surface.
This study proposed to apply kinetic parameter to evaluate the initial cell adhesion behavior by pseudo first order Lagergren model. For both L929 and CHO cells, the cell density increased significantly on CF4 plasma treated BC than that on the pristine BC and on other plasmas treated BCs under the cell culture condition with serum. The increment of the adhered cell number can be quantified by the kinetic parameter which increased 2-3 folds for both cell types.
The second goal of this thesis, the incorporation of antifouling property onto BC membranes was facilitated by two methods to graft PEG: (i) plasma activation of PEG to graft on BC (PEG-g-BC), and (ii) chemically grafting PEG onto BC by using IPDI (mPEG-BC). The surface characterization by using SEM and FTIR indicated that the PEG was successfully incorporated onto BC for both PEG-g-BC and mPEG-BC and showed the decreasing of both protein and L-929 cell adhesion. Moreover, mPEG-BC showed particular anti-fouling effect compared to the PEG-g-BC by examining the cell density which was probably due to the brush conformation of PEG-g-BC.

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