Bone specimens used for evaluating the mechanical properties of bone may not have been necessarily preserved by the same method before they become available. To this end, the mechanical properties of bone may be affected by freezing, a common preservation method, as well as by formalin and other preservation solutions. In this study, bone specimens were preserved by different methods (i.e., freezing, preservation in saline, preservation in ethanol, and preservation in formalin) to examine the effects of each preservation method on the fracture characteristics of bovine femoral cortical bone. Regarding the crack extension behavior in the fracture toughness test, microcracks accumulated at the top of the slit in the low-load region before the maximum load was reached. When such accumulated microcracks grew to a visible size, a crack was formed from the top of the slit, and then grew with subsequent expansion of the processing zone. The fracture toughness values of the bone groups preserved in formalin and neutral buffered formalin were significantly lower than those of the bone groups preserved by other methods. As the fracture surface by scanning electron microscopic observations was smoother with a loading rate of 20 mm/min than with 1 mm/min, cracks were considered to develop easily, resulting in a lower fracture toughness value. Scanning electron microscopic observations of a test section subjected to a low loading rate showed that the fracture surface of the bone groups preserved in formalin or neutral buffered formalin was flat and smooth. In the other preservation groups, the lamellae adjacent to the top of the slit had undulating contours with plastic deformation. [
Bone fracture toughness has been well studied, however, it is also important to investigate the effect of preservative treatment on the mechanical properties of bones. It is necessary to evaluate crack initiation and propagation after fracture because this process may be different in the case of injured bone tissues. In this study, we attempted to analyze the strain distribution on bone tissue surface by using image correlation techniques in order to elucidate the relationship between microscopic bone damage and strain distribution. Bovine femoral cortical bone was employed as the bone specimen and the three-point bend test method was used to determine the fracture toughness, in accordance with the ASTM E399 guidelines. An Instron type machine was used in the fracture toughness test and the loading rate was set to 1 mm/min. Black and white spray paint was applied in a random pattern to the surface of the specimens, and the specimens were loaded until they were ruptured. Bone surface strain analysis was performed using image correlation techniques and the changes were recorded in a digital image. In order to evaluate the effects of preservative treatment on the mechanical properties of bone, we categorized the specimens into 4 groups: the control group included the specimens that were submitted for testing immediately after machining and the preservation group comprised specimens that were analyzed after preservative treatment with different method (formalin, ethanol and physiological saline solution). A strain analysis performed using image correlation techniques allowed the visualization of the increased strain at the forward end of the slit of the specimens. The strain value at the forward end of the slit (the longitudinal direction of specimens) measured at the time of rupture in the control group was approximately 4 times larger than that in the formalin preservation group, thereby suggesting the embrittlement of bone organic constituents due to preservative treatment.
Microphysiological systems (MPS) are an emerging technology for next-generation drug screening in non-clinical tests. Microphysiological systems are microfluidic devices that reconstitute the physiological functions of a human organ using a three-dimensional in vivo-mimicking microenvironment. In the future, MPSs are expected to reduce the number of animal experiments, improve prediction methods for drug efficacy in clinical settings, and reduce the costs of drug discovery. However, drug adsorption onto the polymers used in an MPS is a critical issue for assessment because it changes the concentration of the drug. Polydimethylsiloxane (PDMS), a basic material used for the fabrication of MPS, strongly adsorbs hydrophobic drugs. As a substitute for PDMS, cyclo-olefin polymer (COP) has emerged as an attractive material for low-adsorption MPS. However, it has difficulty bonding with different materials and, therefore, is not commonly used. In this study, we assessed the drug adsorption properties of each material constituting an MPS and subsequent changes in drug toxicity for the development of a low-adsorption MPSs using COP. The hydrophobic drug cyclosporine A showed an affinity for PDMS and induced lower cytotoxicity in PDMS-MPS but not in COP-MPS, whereas adhesive tapes used for bonding adsorbed a significant quantity of drugs, lowering their availability, and was cytotoxic. Therefore, easily-adsorbed hydrophobic drugs and bonding materials having lower cytotoxicity should be used with a low-adsorption polymer such as COP.
Bone fracture toughness has been well studied, however, it is also important to investigate the effect of preservative treatment on the mechanical properties of bones. It is necessary to evaluate crack initiation and propagation after fracture because this process may be different in the case of injured bone tissues. In this study, we attempted to analyze the strain distribution on bone tissue surface by using image correlation techniques in order to elucidate the relationship between microscopic bone damage and strain distribution. Bovine femoral cortical bone was employed as the bone specimen and the three point bend test method was used to determine the fracture toughness, in accordance with the ASTM E399 guidelines. An Instron type machine was used in the fracture toughness test and the loading rate was set to 1 mm/min. Black and white spray paint was applied in a random pattern to the surface of the specimens, and the specimens were loaded until they were ruptured. Bone surface strain analysis was performed using image correlation techniques and the changes were recorded in a digital image. In order to evaluate the effects of preservative treatment on the mechanical properties of bone, we categorized the specimens into 4 groups: the control group included the specimens that were submitted for testing immediately after machining and the preservation group comprised specimens that were analyzed after preservative treatment with different method (formalin, ethanol and physiological saline solution). A strain analysis performed using image correlation techniques allowed the visualization of the increased strain at the forward end of the slit of the specimens. The strain value at the forward end of the slit (the longitudinal direction of specimens) measured at the time of rupture in the control group was approximately 4 times larger than that in the formalin preservation group, thereby suggesting the embrittlement of bone organic constituents due to preservative treatment.
Bone specimens used for evaluating the mechanical properties of bone have not necessarily been preserved by the same method before they become available. However, the mechanical properties of bone may be influenced by freezing, a common preservation method, as well as by formalin and other preservation solutions. In this study, bone specimens were preserved by different methods (freezing, physiological saline solution, ethanol, formalin and neutral buffered formalin) to examine the effects of preservation method on the fracture characteristics of bovine femoral cortical bone. Regarding the crack extension behavior in the fracture toughness test, microcracks accumulated at the top of the slit in the low load region before the maximum load was reached. When such accumulated microcracks grew to a visible size, a crack was formed from the top of the slit, and then grew with subsequent expansion of the processing zone. The fracture toughness values of the groups preserved in formalin and neutral buffered formalin were significantly lower than those of other preservation groups. As the fracture surface by SEM observations was smoother with a loading rate of 20 mm/min than 1 mm/min, cracks were considered to develop easily, resulting in a lower fracture toughness value. SEM observations of a test section subjected to a low loading rate showed that the fracture surface of the groups preserved in formalin or neutral buffered formalin was flat and smooth. In the other preservation groups, the lamellae adjacent to the top of the slit had undulating fracture surface, with plastic deformation produced.
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