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Stability of cross-linked collagen
Součková, Gabriela ; Vojtová, Lucy (oponent) ; Jančář, Josef (vedoucí práce)
Theoretical part of this bachelor thesis is based on general information about collagen, most common fibrous protein in body, collagen cross-linking; leading to its better mechanical and chemical properties; and widely used crosslinking agents, mainly based on 1-ethyl-3-(3 dimethylaminopropyl) carbodiimide (EDC). Furthermore it deals with several methods of quantifying EDC in aqueous solutions. Experimental part is concerned with the determination of EDC remained in water used for washing samples in order to purify them after the cross-linking. The goal is to find out if used washing method is adequate enough for certain application or if it needs some adjusting. Even if EDC is practically non-toxic, it provides side effects on colour of the collagenous material after the sterilization by irradiation. Since it has not been found in the literature what specific amount of EDC can stay in a sample or if it is necessary to be completely washed out. According the state-of-the-art, it is difficult to assessed if the washing method is sufficient or not. Moreover, there are also other side products of cross-linking collagen in the final collageneous products that have to be taken into account. However, the aim of this bachelor thesis is to evaluate washing EDC from the collagenous sample only. As for cross-linking method, three concentrations of EDC followed by washing samples for different periods of time were used. Following this method, 48 samples were prepared and three times measured using UV-VIS colorimetric assay. As a result it was found out that for commonly used concentration of cross-linking agent (50 mmol.l-1 of EDC) it is sufficient to wash samples for 30 min in solution of disodium phospnonate (Na2HPO3) followed by washing with water 4 times for 15 min with changing the water at each step. After these steps, the concentration of EDC in washing water was reduced up to 1 mol.l-1. For lower concentrations of cross-linking EDC solution it is sufficient even shorter washing time. Considering the preliminary results of this bachelor thesis, further investigation dealing with this topic should follow.
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Stability of cross-linked collagen
Součková, Gabriela ; Vojtová, Lucy (oponent) ; Jančář, Josef (vedoucí práce)
Theoretical part of this bachelor thesis is based on general information about collagen, most common fibrous protein in body, collagen cross-linking; leading to its better mechanical and chemical properties; and widely used crosslinking agents, mainly based on 1-ethyl-3-(3 dimethylaminopropyl) carbodiimide (EDC). Furthermore it deals with several methods of quantifying EDC in aqueous solutions. Experimental part is concerned with the determination of EDC remained in water used for washing samples in order to purify them after the cross-linking. The goal is to find out if used washing method is adequate enough for certain application or if it needs some adjusting. Even if EDC is practically non-toxic, it provides side effects on colour of the collagenous material after the sterilization by irradiation. Since it has not been found in the literature what specific amount of EDC can stay in a sample or if it is necessary to be completely washed out. According the state-of-the-art, it is difficult to assessed if the washing method is sufficient or not. Moreover, there are also other side products of cross-linking collagen in the final collageneous products that have to be taken into account. However, the aim of this bachelor thesis is to evaluate washing EDC from the collagenous sample only. As for cross-linking method, three concentrations of EDC followed by washing samples for different periods of time were used. Following this method, 48 samples were prepared and three times measured using UV-VIS colorimetric assay. As a result it was found out that for commonly used concentration of cross-linking agent (50 mmol.l-1 of EDC) it is sufficient to wash samples for 30 min in solution of disodium phospnonate (Na2HPO3) followed by washing with water 4 times for 15 min with changing the water at each step. After these steps, the concentration of EDC in washing water was reduced up to 1 mol.l-1. For lower concentrations of cross-linking EDC solution it is sufficient even shorter washing time. Considering the preliminary results of this bachelor thesis, further investigation dealing with this topic should follow.
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