Biomedical subjects

C V C Bouten

Publications and source records attributed to C V C Bouten.

10 recordsLinked to original sources

PubMed2005

Design of scaffolds for blood vessel tissue engineering using a multi-layering electrospinning technique.

Aiming to develop a scaffold architecture mimicking morphological and mechanically that of a blood vessel, a sequential multi-layering electrospinning (ME) was performed on a rotating mandrel-type collector. A bi-layered tubular scaffold composed of a stiff and oriented PLA outside fibrous layer and a pliable and randomly oriented PCL fibrous inner layer (PLA/PCL) was fabricated. Control over the level of fibre orientation of the different layers was achieved through the rotation speed of the collector. The structural and mechanical properties of the scaffolds were examined using scanning electron microscopy (SEM) and tensile testing. To assess their capability to support cell attachment, proliferation and migration, 3T3 mouse fibroblasts and later human venous myofibroblasts (HVS) were cultured, expanded and seeded on the scaffolds. In both cases, the cell-polymer constructs were cultured under static conditions for up to 4 weeks. Environmental-scanning electron microscopy (SEM), confocal laser scanning microscopy (CLSM), histological examination and biochemical assays for cell proliferation (DNA) and extracellular matrix production (collagen and glycosaminoglycans) were performed. The findings suggest the feasibility of ME to design scaffolds with a hierarchical organization through a layer-by-layer process and control over fibre orientation. The resulting scaffolds achieved the desirable levels of pliability (elastic up to 10% strain) and proved to be capable to promote cell growth and proliferation. The electrospun PLA/PCL bi-layered tube presents appropriate characteristics to be considered a candidate scaffold for blood vessel tissue engineering.

3T3 Cells↗

PubMed2005

Mechanical and failure properties of single attached cells under compression.

Eukaryotic cells are continuously subjected to mechanical forces under normal physiological conditions. These forces and associated cellular deformations induce a variety of biological processes. The degree of deformation depends on the mechanical properties of the cell. As most cells are anchorage dependent for normal functioning, it is important to study the mechanical properties of cells in their attached configuration. The goal of the present study was to obtain the mechanical and failure properties of attached cells. Individual, attached C2C12 mouse myoblasts were subjected to unconfined compression experiments using a recently developed loading device. The device allows global compression of the cell until cell rupture and simultaneously measures the associated forces. Cell bursting was characterized by a typical reduction in the force, referred to as the bursting force. Mean bursting forces were calculated as 8.7+/-2.5 microN at an axial strain of 72+/-4%. Visualization of the cell using confocal microscopy revealed that cell bursting was preceded by the formation of bulges at the cell membrane, which eventually led to rupturing of the cell membrane. Finite element calculations were performed to simulate the obtained force-deformation curves. A finite element mesh was built for each cell to account for its specific geometrical features. Using an axisymmetric approximation of the cell geometry, and a Neo-Hookean constitutive model, excellent agreement between predicted and measured force-deformation curves was obtained, yielding an average Young's modulus of 1.14+/-0.32 kPa.

Animals↗

PubMed2005

Compression-induced damage in a muscle cell model in vitro.

Soft tissue breakdown can be initiated at the muscle layer associated with bony prominences, leading to the development of pressure ulcers. Both the magnitude and duration of pressure are important factors in this breakdown process. The present study utilizes a physical model, incorporating C2C12 mouse myoblasts in a homogeneous agarose gel, to examine the damaging effects of prolonged applied pressure. Identical cylindrical cores cut from the agarose/cell suspension were subjected to two separate compressive strains, of 10 and 20 per cent. The strain was applied for time periods ranging from 0.5 to 12 hours, using a specially designed loading apparatus. After each compression period, sections taken from the central horizontal plane of the individual constructs were stained using either haematoxylin and eosin or with the fluorescent probes, Calcein AM and ethidium homodimer-1, and assessed for cell damage. It was found that constructs subjected to the higher strain values demonstrated significantly higher values of non-viable cells for equivalent time points compared to the unstrained constructs. Further analysis on sections using the DNA nick-translation method suggested that this increase was primarily due to apoptosis. These findings imply a relationship between the duration of applied compression and damage to muscle cells seeded in the gel, which was particularly apparent at the strain level of 20 per cent, equivalent to a clinically relevant pressure of 32 mmHg (4.3 kPa). Such an approach might be useful in establishing damage threshold levels at a cellular level.

Animals↗

PubMed2004

A computational model for collagen fibre remodelling in the arterial wall.

As the interaction between tissue adaptation and the mechanical condition within tissues is complex, mathematical models are desired to study this interrelation. In this study, a mathematical model is presented to investigate the interplay between collagen architecture and mechanical loading conditions in the arterial wall. It is assumed that the collagen fibres align along preferred directions, situated in between the principal stretch directions. The predicted fibre directions represent symmetrically arranged helices and agree qualitatively with morphometric data from literature. At the luminal side of the arterial wall, the fibres are oriented more circumferentially than at the outer side. The discrete transition of the fibre orientation at the media-adventitia interface can be explained by accounting for the different reference configurations of both layers. The predicted pressure-radius relations resemble experimentally measured sigma-shaped curves. As there is a strong coupling between the collagen architecture and the mechanical loading condition within the tissue, we expect that the presented model for collagen remodelling is useful to gain further insight into the processes involved in vascular adaptation, such as growth and smooth muscle tone adaptation.

Adaptation, Physiological↗

PubMed2003

Quantifying pressure sore-related muscle damage using high-resolution MRI.

To obtain insight into the etiology of deep pressure sores, understanding of the relationship between prolonged transverse loading and local muscle damage is required. To date, the amount and location of muscle damage have been determined by histological examination. In the present study, we determined whether T2-weighted high-resolution magnetic resonance imaging (MRI) can also be applied to evaluate muscle tissue after prolonged transverse loading. The tibialis anterior muscle and overlying skin in the right hindlimbs of five rats were compressed between an indenter and the tibia. The in vivo magnetic resonance images of the loaded and contralateral hindlimbs were obtained 24 h after load application. The tibialis anterior muscles were then processed for histological examination. In the magnetic resonance images of all five loaded hindlimbs, signal intensity appeared higher in the loaded regions of the muscle compared with the unloaded regions. The location of the higher signal intensity coincided with the location of damage assessed from histology. Also the amount of damage determined with MRI was in good agreement with the amount of damage assessed from histological examination. Because MRI is nondestructive, it is a promising alternative for histology in research on pressure sore etiology, especially in follow-up studies to evaluate the development of muscle damage in time and in clinical studies.

Animals↗

PubMed2003

Monitoring the biomechanical response of individual cells under compression: a new compression device.

Skeletal muscle cells are sensitive to sustained compression, which can lead to the development of pressure sores. Although it is known that this type of tissue breakdown depends on the magnitude and duration of the applied load, the exact relationship between cell deformation and damage remains unclear. To gain more insight into this process, a method has been developed, that incorporates the use of a new loading device and confocal microscopy. The loading device is able to compress individual cells, either statically or dynamically, while measuring the resulting forces. Experiments can be performed under ideal environmental conditions, comparable with those of a CO2 incubator. First compression experiments on C2C12 mouse myoblasts showed the shape changes that cells undergo during static compression by the loading device. Calculations using the three-dimensional confocal images showed no change in volume and an increase in the surface area of the cell as a result of compression. The device presented here provides a useful way to monitor the biomechanical response of skeletal muscle cells during long-term compression experiments. Therefore it will contribute to the knowledge about strain-induced cell damage, as seen in pressure sores and other mechanically induced clinical conditions.

Animals↗

PubMed2003

Remodelling of continuously distributed collagen fibres in soft connective tissues.

Extracellular matrix remodelling plays an essential role in tissue engineering of load-bearing structures. The goal of this study is to model changes in collagen fibre content and orientation in soft connective tissues due to mechanical stimuli. A theory is presented describing the mechanical condition within the tissue and accounting for the effects of collagen fibre alignment and changes in fibre content. A fibre orientation tensor is defined to represent the continuous distribution of collagen fibre directions. A constitutive model is introduced to relate the fibre configuration to the macroscopic stress within the material. The constitutive model is extended with a structural parameter, the fibre volume fraction, to account for the amount of fibres present within the material. It is hypothesised that collagen fibre reorientation is induced by macroscopic deformations and the amount of collagen fibres is assumed to increase with the mean fibre stretch. The capabilities of the model are demonstrated by considering remodelling within a biaxially stretched cube. The model is then applied to analyse remodelling within a closed stented aortic heart valve. The computed preferred fibre orientation runs from commissure to commissure and resembles the fibre directions in the native aortic valve.

Adaptation, Physiological↗

PubMed2003

The relevance of large strains in functional tissue engineering of heart valves.

BACKGROUND: Exposing the developing tissue to flow and pressure in a bioreactor has been shown to enhance tissue formation in tissue-engineered heart valves. Animal studies showed excellent functionality in these valves in the pulmonary position. However, they lack the mechanical strength for implantation in the high-pressure aortic position. Improving the in vitro conditioning protocol is an important step towards the use of these valves as aortic heart valve replacements. In this study, the relevance of large strains to improve the mechanical conditioning protocol was investigated. METHODS: Using a newly developed device, engineered heart valve tissue was exposed to increasing cyclic strain in vitro. Tissue formation and mechanical properties were analyzed and compared to unstrained controls. RESULTS: Straining resulted in more pronounced and organized tissue formation with superior mechanical properties over unstrained controls. Overall tissue properties improved with increasing strain levels. CONCLUSIONS: The results demonstrate the significance of large strains in promoting tissue formation. This study may provide a methodological basis for tissue engineering of heart valves appropriate for systemic pressure applications.

Absorbable Implants↗

PubMed2003

Can loaded interface characteristics influence strain distributions in muscle adjacent to bony prominences?

Pressure distributions at the interface between skin and supporting tissues are used in design of supporting surfaces like beds, wheel chairs, prostheses and in sales brochures to support commercial products. The reasoning behind this is, that equal pressure distributions in the absence of high pressure gradients is assumed to minimise the risk of developing pressure sores. Notwithstanding the difficulty in performing reproducible and accurate pressure measurements, the question arises if the interface pressure distribution is representative of the internal mechanical state of the soft tissues involved. The paper describes a study of the mechanical condition of a supported buttock contact, depending on cushion properties, relative properties of tissue layers and friction. Numerical, mechanical simulations of a buttock on a supporting cushion are described. The ischial tuberosity is modelled as a rigid body, whereas the overlying muscle, fat and skin layers are modelled as a non-linear Ogden material. Material parameters and thickness of the fat layer are varied. Coulomb friction between buttock and cushion is modelled with different values of the friction coefficient. Moreover, the thickness and properties of the cushion are varied. High shear strains are found in the muscle near the bony prominence and the fat layer near the symmetry line. The performed parameter variations lead to large differences in shear strain in the fat layer but relatively small variations in the skeletal muscle. Even with a soft cushion, leading to a high reduction of the interface pressure the deformation of the skeletal muscle near the bone is high enough to form a risk, which is a clear argument that interface pressures alone are not sufficient to evaluate supporting surfaces.

Adipose Tissue↗

PubMed2003

Compression induced cell damage in engineered muscle tissue: an in vitro model to study pressure ulcer aetiology.

The aetiology of pressure ulcers is poorly understood. The complexity of the problem, involving mechanical, biochemical, and physiological factors demands the need for simpler model systems that can be used to investigate the relative contribution of these factors, while controlling others. Therefore, an in vitro model system of engineered skeletal muscle tissue constructs was developed. With this model system, the relationship between compressive tissue straining and cell damage initiation was investigated under well-defined environmental conditions. Compression of the engineered muscle tissue constructs revealed that cell death occurs within 1-2 h at clinically relevant straining percentages and that higher strains led to earlier damage initiation. In addition, the uniform distribution of dead cells throughout the constructs suggested that sustained deformation of the cells was the principle cause of cell death. Therefore, it is hypothetised that sustained cell deformation is an additional mechanism that plays a role in the development of pressure ulcers.

Animals↗