Wound repair and regeneration : official publication of the Wound Healing Society [and] the European Tissue Repair Society, 2015
Currently, pressure ulcer preventive strategies focus mainly on pressure redistribution. Little a... more Currently, pressure ulcer preventive strategies focus mainly on pressure redistribution. Little attention is paid to reduce the harmful effects of shear- force, because little is known about pathophysiological aspects of shear-force. Even today, no method to measure the effects of shear-force on the skin is available. Therefore, the aim of this study was to investigate the response to shear-forces in terms of analysing a non-invasive biomarker and reactive hyperaemic parameter measured at the skin of healthy participants. A physical model was developed to produce a combination of pressure and shear or pressure alone on the skin. Ten healthy male participants were included and pressure (3.9 kPa) and a combined loading of pressure and shear (2.4 kPa + 14.5 N) was applied at the volar aspect of the forearms for 15 and 30 minutes. A Sebutape sample was used to collect IL-1α and total protein non- invasively. The reactive hyperaemic parameter was derived from a Laser Doppler flowmeter. T...
Discrete element models have often been the primary tool in investigating and characterising the ... more Discrete element models have often been the primary tool in investigating and characterising the viscoelastic behaviour of soft tissues. However, studies have employed varied configurations of these models, based on the choice of the number of elements and the utilised formation, for different subject tissues. This approach has yielded a diverse array of viscoelastic models in the literature, each seemingly resulting in different descriptions of viscoelastic constitutive behaviour and/or stress-relaxation and creep functions. Moreover, most studies do not apply a single discrete element model to characterise both stress-relaxation and creep behaviours of tissues. The underlying assumption for this disparity is the implicit perception that the viscoelasticity of soft tissues cannot be described by a universal behaviour or law, resulting in the lack of a unified approach in the literature based on discrete element representations. This paper derives the constitutive equation for different viscoelastic models applicable to soft tissues with two characteristic times. It demonstrates that all possible configurations exhibit a unified and universal behaviour, captured by a single constitutive relationship between stress, strain and time as: σ+Aσ̇+Bσ¨=Pε̇+Qε¨. The ensuing stress-relaxation G(t) and creep J(t) functions are also unified and universal, derived as [Formula: see text] and J(t)=c2+(ε0-c2)e(-PQt)+σ0Pt, respectively. Application of these relationships to experimental data is illustrated for various tissues including the aortic valve, ligament and cerebral artery. The unified model presented in this paper may be applied to all tissues with two characteristic times, obviating the need for employing varied configurations of discrete element models in preliminary investigation of the viscoelastic behaviour of soft tissues.
Dynamic loading is known to be essential in the biosynthesis and maintenance of healthy matabolis... more Dynamic loading is known to be essential in the biosynthesis and maintenance of healthy matabolism in articular cartilage and chondrocyte-seeded hydrogels. This is thought to be a result of a range of factors, including cell deformation. However, the nature of this deformation remains elusive. This study utilises the well-characterized chondrocyte seeded agarose model and finite element analysis based on a viscoelastic model to demonstrate the cell deformation duringboth static and dynamic compression in the stage of initial culture. The prony series viscoelastic models were adopted to model agarose and chondrocyte which have been implemented in finite element analysis. The findings suggest that cell strain during loaded phase and unloaded phase strongly depends on the frequency and number of cycles, both of which have been implicated in various mechanotransduction pathway. Due to the mechanical properties mismatch between the cell and agarose construct, the cell strain is amplified through-out the cyclic compression.
Medical science monitor : international medical journal of experimental and clinical research
Recent developments in Computer-Integrated and Robot-Aided Surgery (in particular, the emergence ... more Recent developments in Computer-Integrated and Robot-Aided Surgery (in particular, the emergence of automatic surgical tools and robots (as well as advances in Virtual Reality techniques, call for closer examination of the mechanical properties of very soft tissues (such as brain, liver, kidney, etc.). Moreover, internal organs are very susceptible to trauma. In order to protect them properly against car crash and other impact consequences we need to be able to predict the organ deformation. Such prediction can be achieved by proper mathematical modelling followed by a computer simulation. The ultimate goal of our research into the biomechanics of these tissues is development of corresponding, realistic mathematical models. This paper contains experimental results of in vitro, uniaxial, unconfined compression of swine brain tissue obtained by the author in Mechanical Engineering Laboratory, Japan, and discusses liver and kidney in vivo compression experiments conducted in Highway Sa...
Articular cartilage is subjected to dynamic compressive loading during normal activity which infl... more Articular cartilage is subjected to dynamic compressive loading during normal activity which influences chondrocyte metabolism through various mechanotransduction pathways. A well characterised and reproducible model system, involving chondrocytes embedded in agarose gel, has been used to investigate the effects of mechanical compression on chondrocytes, isolated from full depth cartilage or separately from the superficial and deep zone tissue. The role of nitric oxide as a mediator of mechanical-induced effects has also been studied. Chondrocytes were isolated, separately, from full depth, superficial and deep zone cartilage and seeded in 3% agarose constructs. Dynamic compressive strain was applied to the constructs using a range of frequencies (0.3, 1 and 3 Hz). Glycosaminoglycan synthesis, cell proliferation and nitrite production were assessed. In further experiments, constructs were compressed in the presence of 1 mM L-NAME or 10 microM dexamethasone. Glycosaminoglycan synthes...
This study reports the cytoskeletal organisation within chondrocytes, isolated from the superfici... more This study reports the cytoskeletal organisation within chondrocytes, isolated from the superficial and deep zones of articular cartilage and seeded into agarose constructs. At day 0, marked organisation of actin microfilaments was not observed in cells from both zones. Partial or clearly organised microtubules and vimentin intermediate filaments cytoskeletal components were present, however, in a proportion of cells. Staining for microtubules and vimentin intermediate filaments was less marked after 1 day in culture however than on initial seeding. For all three cytoskeletal components there was a dramatic increase in organisation between days 3 and 14 and, in general, organisation was greater within deep zone cells. Clear organisation for actin microfilaments was characterised by a cortical network and punctate staining around the periphery of the cell, while microtubules and vimentin intermediate filaments formed an extensive fibrous network. Cytoskeletal organisation within chon...
We report the Laser Induced Forward Transfer (LIFT) of antibodies from a liquid donor film onto p... more We report the Laser Induced Forward Transfer (LIFT) of antibodies from a liquid donor film onto paper receivers for application as point-of-care diagnostic sensors. To minimise the loss of functionality of the active biomolecules during transfer, a dynamic release layer was employed to shield the biomaterial from direct exposure to the pulsed laser source. Cellulose paper was chosen as the ideal receiver because of its inherent bio-compatibility, liquid transport properties, wide availability and low cost, all of which make it an efficient and suitable platform for point-of-care diagnostic sensors. Both enzyme-tagged and untagged IgG antibodies were LIFT-printed and their functionality was confirmed via a colorimetric enzyme-linked immunosorbent assay. Localisation of the printed antibodies was exhibited, which can allow the creation of complex 2-d patterns such as QR codes or letters for use in a final working device. Finally, a calibration curve was determined that related the intensity of the colour obtained to the concentration of active antibodies to enable quantitative assessment of the device performance. The motivation for this work was to implement a laser-based procedure for manufacturing low-cost, point-of-care diagnostic devices on paper.
This study investigates the influence of cyclic tensile strain, applied to fully contracted fibro... more This study investigates the influence of cyclic tensile strain, applied to fully contracted fibroblast-seeded collagen constructs. The constructs were preloaded to either 2 or 10 mN. The preloaded constructs were subsequently subjected to a further 10% cyclic strain (0-10%) at 1 Hz, using a triangular waveform, or were cultured in the preloaded state. In all cases cellular viability was maintained during the conditioning period. Cell proliferation was enhanced by the application of cyclic strain within constructs preloaded to both 2 and 10 mN. Collagen synthesis was enhanced by cyclic strain within constructs preloaded at 2 mN only. The profile of matrix metalloproteinase (MMP) expression, determined by zymography, was broadly similar in constructs preloaded at 2 mN with or without the application of cyclic strain. By contrast, constructs preloaded at 10 mN and subjected to cyclic strain expressed enhanced levels of staining for latent MMP-1, latent MMP-9, and both latent and active MMP-2, when compared with the other conditioning regimens. The structural stiffness of constructs preloaded at 2 mN and subjected to cyclic strain was enhanced compared with control specimens, reflecting the increase in collagen synthesis. By contrast, the initial failure loads for cyclically strained constructs preloaded at 10 mN were reduced, potentially because of enhanced catabolic activity.
Increasing the thickness of tissue-engineered cartilage is associated with loss of chondrocyte vi... more Increasing the thickness of tissue-engineered cartilage is associated with loss of chondrocyte viability and biosynthetic activity at the tissue center. Exceptionally high volumes of culture medium, however, can maintain cellularity and glycosaminoglycan synthesis throughout 4-mm-thick constructs. We hypothesized that glucose supplementation could replicate the augmentation of tissue formation achieved by medium volume. Chondrocyte-alginate constructs (40x10(6) cells/mL) were cultured for 14 days in 0.4-6.4 mL/10(-6) cells of either low- (5.1 mM) or high- (20.4 mM) glucose medium. Glucose was critical to chondrocyte viability, and glucose uptake increased significantly (P < .001) with both medium volume and glucose supplementation. After 14 days, constructs cultured in 0.4 mL/10(-6) cells of low-glucose medium had a mass of 172 +/- 6.1 mg and glycosaminoglycan (GAG) content of 0.32 +/- 0.03 mg (mean +/- standard deviation). A 4-fold increase in medium volume increased the final construct mass by 44% and GAG content by 207%. An equivalent increase in glucose supply in the absence of volume change increased these parameters by just 10% and 73%, respectively. A similar trend was observed from 0.8 to 3.2 mL/10(-6) cells, when maximal values of construct GAG content and mass were obtained. Therefore, medium volume remains an important consideration for the optimal culture of tissue-engineered cartilage.
Wound repair and regeneration : official publication of the Wound Healing Society [and] the European Tissue Repair Society, 2015
Currently, pressure ulcer preventive strategies focus mainly on pressure redistribution. Little a... more Currently, pressure ulcer preventive strategies focus mainly on pressure redistribution. Little attention is paid to reduce the harmful effects of shear- force, because little is known about pathophysiological aspects of shear-force. Even today, no method to measure the effects of shear-force on the skin is available. Therefore, the aim of this study was to investigate the response to shear-forces in terms of analysing a non-invasive biomarker and reactive hyperaemic parameter measured at the skin of healthy participants. A physical model was developed to produce a combination of pressure and shear or pressure alone on the skin. Ten healthy male participants were included and pressure (3.9 kPa) and a combined loading of pressure and shear (2.4 kPa + 14.5 N) was applied at the volar aspect of the forearms for 15 and 30 minutes. A Sebutape sample was used to collect IL-1α and total protein non- invasively. The reactive hyperaemic parameter was derived from a Laser Doppler flowmeter. T...
Discrete element models have often been the primary tool in investigating and characterising the ... more Discrete element models have often been the primary tool in investigating and characterising the viscoelastic behaviour of soft tissues. However, studies have employed varied configurations of these models, based on the choice of the number of elements and the utilised formation, for different subject tissues. This approach has yielded a diverse array of viscoelastic models in the literature, each seemingly resulting in different descriptions of viscoelastic constitutive behaviour and/or stress-relaxation and creep functions. Moreover, most studies do not apply a single discrete element model to characterise both stress-relaxation and creep behaviours of tissues. The underlying assumption for this disparity is the implicit perception that the viscoelasticity of soft tissues cannot be described by a universal behaviour or law, resulting in the lack of a unified approach in the literature based on discrete element representations. This paper derives the constitutive equation for different viscoelastic models applicable to soft tissues with two characteristic times. It demonstrates that all possible configurations exhibit a unified and universal behaviour, captured by a single constitutive relationship between stress, strain and time as: σ+Aσ̇+Bσ¨=Pε̇+Qε¨. The ensuing stress-relaxation G(t) and creep J(t) functions are also unified and universal, derived as [Formula: see text] and J(t)=c2+(ε0-c2)e(-PQt)+σ0Pt, respectively. Application of these relationships to experimental data is illustrated for various tissues including the aortic valve, ligament and cerebral artery. The unified model presented in this paper may be applied to all tissues with two characteristic times, obviating the need for employing varied configurations of discrete element models in preliminary investigation of the viscoelastic behaviour of soft tissues.
Dynamic loading is known to be essential in the biosynthesis and maintenance of healthy matabolis... more Dynamic loading is known to be essential in the biosynthesis and maintenance of healthy matabolism in articular cartilage and chondrocyte-seeded hydrogels. This is thought to be a result of a range of factors, including cell deformation. However, the nature of this deformation remains elusive. This study utilises the well-characterized chondrocyte seeded agarose model and finite element analysis based on a viscoelastic model to demonstrate the cell deformation duringboth static and dynamic compression in the stage of initial culture. The prony series viscoelastic models were adopted to model agarose and chondrocyte which have been implemented in finite element analysis. The findings suggest that cell strain during loaded phase and unloaded phase strongly depends on the frequency and number of cycles, both of which have been implicated in various mechanotransduction pathway. Due to the mechanical properties mismatch between the cell and agarose construct, the cell strain is amplified through-out the cyclic compression.
Medical science monitor : international medical journal of experimental and clinical research
Recent developments in Computer-Integrated and Robot-Aided Surgery (in particular, the emergence ... more Recent developments in Computer-Integrated and Robot-Aided Surgery (in particular, the emergence of automatic surgical tools and robots (as well as advances in Virtual Reality techniques, call for closer examination of the mechanical properties of very soft tissues (such as brain, liver, kidney, etc.). Moreover, internal organs are very susceptible to trauma. In order to protect them properly against car crash and other impact consequences we need to be able to predict the organ deformation. Such prediction can be achieved by proper mathematical modelling followed by a computer simulation. The ultimate goal of our research into the biomechanics of these tissues is development of corresponding, realistic mathematical models. This paper contains experimental results of in vitro, uniaxial, unconfined compression of swine brain tissue obtained by the author in Mechanical Engineering Laboratory, Japan, and discusses liver and kidney in vivo compression experiments conducted in Highway Sa...
Articular cartilage is subjected to dynamic compressive loading during normal activity which infl... more Articular cartilage is subjected to dynamic compressive loading during normal activity which influences chondrocyte metabolism through various mechanotransduction pathways. A well characterised and reproducible model system, involving chondrocytes embedded in agarose gel, has been used to investigate the effects of mechanical compression on chondrocytes, isolated from full depth cartilage or separately from the superficial and deep zone tissue. The role of nitric oxide as a mediator of mechanical-induced effects has also been studied. Chondrocytes were isolated, separately, from full depth, superficial and deep zone cartilage and seeded in 3% agarose constructs. Dynamic compressive strain was applied to the constructs using a range of frequencies (0.3, 1 and 3 Hz). Glycosaminoglycan synthesis, cell proliferation and nitrite production were assessed. In further experiments, constructs were compressed in the presence of 1 mM L-NAME or 10 microM dexamethasone. Glycosaminoglycan synthes...
This study reports the cytoskeletal organisation within chondrocytes, isolated from the superfici... more This study reports the cytoskeletal organisation within chondrocytes, isolated from the superficial and deep zones of articular cartilage and seeded into agarose constructs. At day 0, marked organisation of actin microfilaments was not observed in cells from both zones. Partial or clearly organised microtubules and vimentin intermediate filaments cytoskeletal components were present, however, in a proportion of cells. Staining for microtubules and vimentin intermediate filaments was less marked after 1 day in culture however than on initial seeding. For all three cytoskeletal components there was a dramatic increase in organisation between days 3 and 14 and, in general, organisation was greater within deep zone cells. Clear organisation for actin microfilaments was characterised by a cortical network and punctate staining around the periphery of the cell, while microtubules and vimentin intermediate filaments formed an extensive fibrous network. Cytoskeletal organisation within chon...
We report the Laser Induced Forward Transfer (LIFT) of antibodies from a liquid donor film onto p... more We report the Laser Induced Forward Transfer (LIFT) of antibodies from a liquid donor film onto paper receivers for application as point-of-care diagnostic sensors. To minimise the loss of functionality of the active biomolecules during transfer, a dynamic release layer was employed to shield the biomaterial from direct exposure to the pulsed laser source. Cellulose paper was chosen as the ideal receiver because of its inherent bio-compatibility, liquid transport properties, wide availability and low cost, all of which make it an efficient and suitable platform for point-of-care diagnostic sensors. Both enzyme-tagged and untagged IgG antibodies were LIFT-printed and their functionality was confirmed via a colorimetric enzyme-linked immunosorbent assay. Localisation of the printed antibodies was exhibited, which can allow the creation of complex 2-d patterns such as QR codes or letters for use in a final working device. Finally, a calibration curve was determined that related the intensity of the colour obtained to the concentration of active antibodies to enable quantitative assessment of the device performance. The motivation for this work was to implement a laser-based procedure for manufacturing low-cost, point-of-care diagnostic devices on paper.
This study investigates the influence of cyclic tensile strain, applied to fully contracted fibro... more This study investigates the influence of cyclic tensile strain, applied to fully contracted fibroblast-seeded collagen constructs. The constructs were preloaded to either 2 or 10 mN. The preloaded constructs were subsequently subjected to a further 10% cyclic strain (0-10%) at 1 Hz, using a triangular waveform, or were cultured in the preloaded state. In all cases cellular viability was maintained during the conditioning period. Cell proliferation was enhanced by the application of cyclic strain within constructs preloaded to both 2 and 10 mN. Collagen synthesis was enhanced by cyclic strain within constructs preloaded at 2 mN only. The profile of matrix metalloproteinase (MMP) expression, determined by zymography, was broadly similar in constructs preloaded at 2 mN with or without the application of cyclic strain. By contrast, constructs preloaded at 10 mN and subjected to cyclic strain expressed enhanced levels of staining for latent MMP-1, latent MMP-9, and both latent and active MMP-2, when compared with the other conditioning regimens. The structural stiffness of constructs preloaded at 2 mN and subjected to cyclic strain was enhanced compared with control specimens, reflecting the increase in collagen synthesis. By contrast, the initial failure loads for cyclically strained constructs preloaded at 10 mN were reduced, potentially because of enhanced catabolic activity.
Increasing the thickness of tissue-engineered cartilage is associated with loss of chondrocyte vi... more Increasing the thickness of tissue-engineered cartilage is associated with loss of chondrocyte viability and biosynthetic activity at the tissue center. Exceptionally high volumes of culture medium, however, can maintain cellularity and glycosaminoglycan synthesis throughout 4-mm-thick constructs. We hypothesized that glucose supplementation could replicate the augmentation of tissue formation achieved by medium volume. Chondrocyte-alginate constructs (40x10(6) cells/mL) were cultured for 14 days in 0.4-6.4 mL/10(-6) cells of either low- (5.1 mM) or high- (20.4 mM) glucose medium. Glucose was critical to chondrocyte viability, and glucose uptake increased significantly (P < .001) with both medium volume and glucose supplementation. After 14 days, constructs cultured in 0.4 mL/10(-6) cells of low-glucose medium had a mass of 172 +/- 6.1 mg and glycosaminoglycan (GAG) content of 0.32 +/- 0.03 mg (mean +/- standard deviation). A 4-fold increase in medium volume increased the final construct mass by 44% and GAG content by 207%. An equivalent increase in glucose supply in the absence of volume change increased these parameters by just 10% and 73%, respectively. A similar trend was observed from 0.8 to 3.2 mL/10(-6) cells, when maximal values of construct GAG content and mass were obtained. Therefore, medium volume remains an important consideration for the optimal culture of tissue-engineered cartilage.
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