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Biocomposite material for 3D print in the field of regenerative medicine
Chaloupková, Kateřina ; Obruča, Stanislav (referee) ; Přikryl, Radek (advisor)
The presented thesis deals with preparation of material for use in regenerative medicine based on poly(3-hydroxybutyrate) and its characterization. In addition to poly (3-hydroxybutyrate), there were used other materials lactic acid (PLA), tricalcium phosphate (TCP) and two types of plasticizers Citroflex®B-6 (CB6) and Syncroflex3114 (S3114). These materials were selected based on their biocompatibility and, in the case of TCP, also bioactivity. TCP allows new bone to grow on the surface of the scaffold. PLA was used to improve the mechanical properties of the material. Both plasticizers have been used to improve the processability of the material. Theoretical part of this work contains a literature review describing basic information about used materials. Aim of the experimental part is to prepare the material, characterization of properties and determination of printability on a 3D printer. The material is examined for thermal properties by thermogravimetric analysis and differential scanning calorimetry. This work also deals with the matter of 3D printing, especially FDM technology. It has been found that materials containing the syncroflex plasticizer are better processed and therefore printed on a 3D printer. The printability tests performed are temperature towers and filling studies. Printed samples were subjected to mechanical tests of tensile and bending tests. Experiments of cytotoxicity and biocompatibility of the material were also performed. Within the work, TCP particles were characterized using a particle size analyzer. The average TCP particle size is 10,76 µm. Using SEM-EDX, the distribution of TCP in sample filaments was subsequently observed, where it was found that by mixing TCP particles with the remaining components of materials, TCP particles agglomerate into formations up to 20 µm in size. Roughness of materials was determined by confocal microscopy. Cytotoxicity was also tested in the extracts of samples on mouse fibroblasts. Cytotoxicity was determined by metabolic activity assay and light microscopy. The metabolic activity test proved the biocompatibility of the observed materials; therefore, it was possible to perform cell proliferation and biocompatibility tests directly on the samples. Assays were performed using human mesenchymal stem cells. DNA quantification was used to determine cell proliferation. Shape of cells was subsequently observed by confocal microscopy. Tests confirmed growth of cells and their appropriate shape. Stem cell differentiation into bone was performed by measuring alkaline phosphatase activity.
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Optimization of poly(3-hydroxybutyrate) based biocomposite with respect to its printability and mechanical properties
Chaloupková, Kateřina ; Obruča, Stanislav (referee) ; Přikryl, Radek (advisor)
The presented theses deals with preparation and optimalization of biocompatible material based on poly(3-hydroxybutyrate). Other components of prepared samples are polylactid acid, hydroxyapatite and commercially available plasticizer Syncroflex3114. These components were chosen based on their biocompatibility and properties that can be possibly used in tissue engineering. Theoretical part of this theses contains general overview of bone tissue and review of materials used in bone tissue regeneration. Part of this thesis also deals with the problematics of scaffolds. Aim of the experimental part is a planned experiment, which is used to optimize the mixture with respect to printability and mechanical properties. The first step is the preparation of samples based on the proposed conditions and their subsequent processing into a filament with an exact diameter of 1,75 mm for 3D printing using the fused deposition modeling method. From the prepared filaments, test specimens were printed for the following experiments: temperature tower, warping coefficient measurement, bending and pressure test. Data from these experiments were processed using a mathematical model in the form of graphs and equations which show the effect of material components on the measured quantity. It was found that the amount of plasticizer in the sample affects the properties the most. This effect is negative in all cases and worsens the properties of the material. The result of the planned experiment is also a mixture optimized for the best possible printability and mechanical properties (bending modulus 3,3 GPa and pressure modulus 2,3 GPa). With regard to the potential application of the material in bone tissue engineering, the first accelerated biodegradation screening tests were performed for selected samples. The results of accelerated degradation tests are ambiguous and further optimization is needed. Simultaneously with the diploma thesis, biological testing of scaffolds printed on a 3D printer from prepared samples took place. All tested samples were found to be biocompatible.
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Optimization of poly(3-hydroxybutyrate) based biocomposite with respect to its printability and mechanical properties
Chaloupková, Kateřina ; Obruča, Stanislav (referee) ; Přikryl, Radek (advisor)
The presented theses deals with preparation and optimalization of biocompatible material based on poly(3-hydroxybutyrate). Other components of prepared samples are polylactid acid, hydroxyapatite and commercially available plasticizer Syncroflex3114. These components were chosen based on their biocompatibility and properties that can be possibly used in tissue engineering. Theoretical part of this theses contains general overview of bone tissue and review of materials used in bone tissue regeneration. Part of this thesis also deals with the problematics of scaffolds. Aim of the experimental part is a planned experiment, which is used to optimize the mixture with respect to printability and mechanical properties. The first step is the preparation of samples based on the proposed conditions and their subsequent processing into a filament with an exact diameter of 1,75 mm for 3D printing using the fused deposition modeling method. From the prepared filaments, test specimens were printed for the following experiments: temperature tower, warping coefficient measurement, bending and pressure test. Data from these experiments were processed using a mathematical model in the form of graphs and equations which show the effect of material components on the measured quantity. It was found that the amount of plasticizer in the sample affects the properties the most. This effect is negative in all cases and worsens the properties of the material. The result of the planned experiment is also a mixture optimized for the best possible printability and mechanical properties (bending modulus 3,3 GPa and pressure modulus 2,3 GPa). With regard to the potential application of the material in bone tissue engineering, the first accelerated biodegradation screening tests were performed for selected samples. The results of accelerated degradation tests are ambiguous and further optimization is needed. Simultaneously with the diploma thesis, biological testing of scaffolds printed on a 3D printer from prepared samples took place. All tested samples were found to be biocompatible.
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Biocomposite material for 3D print in the field of regenerative medicine
Chaloupková, Kateřina ; Obruča, Stanislav (referee) ; Přikryl, Radek (advisor)
The presented thesis deals with preparation of material for use in regenerative medicine based on poly(3-hydroxybutyrate) and its characterization. In addition to poly (3-hydroxybutyrate), there were used other materials lactic acid (PLA), tricalcium phosphate (TCP) and two types of plasticizers Citroflex®B-6 (CB6) and Syncroflex3114 (S3114). These materials were selected based on their biocompatibility and, in the case of TCP, also bioactivity. TCP allows new bone to grow on the surface of the scaffold. PLA was used to improve the mechanical properties of the material. Both plasticizers have been used to improve the processability of the material. Theoretical part of this work contains a literature review describing basic information about used materials. Aim of the experimental part is to prepare the material, characterization of properties and determination of printability on a 3D printer. The material is examined for thermal properties by thermogravimetric analysis and differential scanning calorimetry. This work also deals with the matter of 3D printing, especially FDM technology. It has been found that materials containing the syncroflex plasticizer are better processed and therefore printed on a 3D printer. The printability tests performed are temperature towers and filling studies. Printed samples were subjected to mechanical tests of tensile and bending tests. Experiments of cytotoxicity and biocompatibility of the material were also performed. Within the work, TCP particles were characterized using a particle size analyzer. The average TCP particle size is 10,76 µm. Using SEM-EDX, the distribution of TCP in sample filaments was subsequently observed, where it was found that by mixing TCP particles with the remaining components of materials, TCP particles agglomerate into formations up to 20 µm in size. Roughness of materials was determined by confocal microscopy. Cytotoxicity was also tested in the extracts of samples on mouse fibroblasts. Cytotoxicity was determined by metabolic activity assay and light microscopy. The metabolic activity test proved the biocompatibility of the observed materials; therefore, it was possible to perform cell proliferation and biocompatibility tests directly on the samples. Assays were performed using human mesenchymal stem cells. DNA quantification was used to determine cell proliferation. Shape of cells was subsequently observed by confocal microscopy. Tests confirmed growth of cells and their appropriate shape. Stem cell differentiation into bone was performed by measuring alkaline phosphatase activity.
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