Centrifugally-spun polyhydroxybutyrate fibres: effect of process solvent on structure, morphology and cell response

The structure and morphology of a novel form of poly(beta-hydroxybutyrate) produced by gel-spinning is described. The entangled fibrous nature of the material, which resembles 'cotton wool' suggests possible functions in wound scaffolding devices. The surface structure and fibre diameter distribution of the fibres have been investigated using phase contrast and scanning electron microscopy. Fibres were found to possess a variety of surface irregularities, such as pores and indentations. with diameters mainly in the range 1-15 microm. Additionally, individual fibres were occasionally found to be fused or forked together with neighbours. The effects of blending with various polysaccharides and of altering the process solvent on fibre morphology were also investigated. Under hydrolytic degradation conditions (pH 10.6, 70 degrees C) the fibres degraded by gradual fragmentation and erosion to fibre fragments, particulate matter and eventually to monomer. Altering the production process influenced both the fibre diameter distributions and surface morphology of the constituent fibres. Mammalian and human epithelial cells were used to study the cellular interaction with the spun fibres. SEM studies show that there is little or no cell adhesion to the unmodified fibres, but surface treatment by means of acid and alkali washes promoted cell proliferation on the materials, probably as a result of the introduction of hydroxyl and carboxyl at the surface. Fabrication of non-woven mats, which were subsequently acid or alkali treated, provided a conventional way of forming a cell-adhesive matrix which may have potential value as a wound scaffold. Neither cell line exhibited any cytotoxic response to these polymers.     

Electrospinning and Evaluation of PHBV-Based Tissue Engineering Scaffolds with Different Fibre Diameters,Surface Topography and Compositions

While electrospinning is an effective technology for producing poly(hydroxybutyrate-co-hydroxyvalerate) (PHBV) micrometre-scale fibrous scaffolds for tissue regeneration, electrospinning of PHBV fibrous scaffolds composed of sub-micrometre fibres, surface-porous fibres or nanocomposite fibres is rarely explored. In this study, the average PHBV fibre diameter was successfully reduced to the sub-micrometre scale by dissolving a conductivity-enhancing salt in the polymer solution for electrospinning. Surface-porous fibres were made using a mixture of solvents, and carbonated hydroxyapatite (CHA) nanoparticles were incorporated into the fibres with the aid of an ultrasonic power source. Water contact angle measurements demonstrated that both fibre diameter reduction and CHA incorporation enhanced the wettability of the fibrous scaffolds. Tensile properties of the scaffolds were not undermined by the reduction of fibre diameter and the presence of surface pores. In vitro biological evaluation using a human osteoblast-like cell line (SaOS-2) demonstrated that all types of fibrous scaffolds supported cell attachment, spreading and proliferation. Analysis of cell morphology revealed similar projected cell areas on all types of scaffolds. However, cells on sub-micrometre fibres possessed a lower cell aspect ratio than cells on microfibres. The reduction of fibre diameter to the sub-micrometre scale enhanced cell proliferation after 14 days cell culture, while the incorporation of CHA nanoparticles in microfibres significantly enhanced the alkaline phosphatase activity of SaOS-2 cells. The control of fibre diameter, surface topography and composition is important in developing electrospun PHBV-based scaffolds for specific tissue-engineering applications.     

A comparison between the physiological and histochemical characterisation of urethral striated muscle in the guinea pig

In this in vitro study comparison has been made between the actomyosin ATPase activity and the contractile properties of the external urethral sphincter in the guinea pig. Histochemical analysis showed the external urethral sphincter to contain a mixture of alkali and acid stable actomyosin ATPase positive fibres in the ratio of 3:1. External urethral sphincter isometric contraction measurements were undertaken using specimens mounted transversely or longitudinally with respect to the urethral lumen. These contraction parameters have shown that the majority of fibres which constitute the external urethral sphincter correspond to the fast twitch type. In order to determine whether the relatively small fibre diameter of urethral striated muscle influenced the contraction results, when compared with controls, corrections for different volume ratios of slow and fast fibres were applied to the results. These modified values revealed close correlation between muscle fibre actomyosin ATPase content and isometric contraction responses.     

Gravity spun polycaprolactone fibres for soft tissue engineering: interaction with fibroblasts and myoblasts in cell culture

Poly(epsilon-caprolactone) (PCL) fibres were produced by wet spinning from solutions in acetone under low shear (gravity flow) conditions. As-spun PCL fibres exhibited a mean strength and stiffness of 7.9 MPa and 0.1 GPa, respectively and a rough, porous surface morphology. Cold drawing to an extension of 500% resulted in increases in fibre strength (43 MPa) and stiffness (0.3 GPa) and development of an oriented, fibrillar surface texture. The proliferation rate of Swiss 3T3 mouse fibroblasts and C2C12 mouse myoblasts on as-spun, 500% cold-drawn and gelatin-modified PCL fibres was determined in cell culture to provide a basic measure of the biocompatibility of the fibres. Proliferation of both cell types was consistently higher on gelatin-coated fibres relative to as-spun fibres at time points below 7 days. Fibroblast growth rates on cold-drawn PCL fibres exceeded those on as-spun fibres but myoblast proliferation was similar on both substrates. After 1 day in culture, both cell types had spread and coalesced on the fibres to form a cell layer, which conformed closely to the underlying topography. The high fibre compliance combined with a potential for modifying the fibre surface chemistry with cell adhesion molecules and the surface architecture by cold drawing to enhance proliferation of fibroblasts and myoblasts, recommends further investigation of gravity-spun PCL fibres for 3-D scaffold production in soft tissue engineering.     

Post-self-assembly experimentation on extruded collagen fibres for tissue engineering applications

Extruded collagen fibres have been shown to constitute a biomimetic three-dimensional scaffold with numerous tissue engineering applications. The multi-step fabrication process of this material provides opportunities for further advancements to improve the properties of the final product. Herein we investigated the influence of the post-self-assembly washing baths on the structural, mechanical and thermal properties of these fibres. The surface morphology and the inter-fibre packing were similar for every treatment. The overnight incubation in isopropanol yielded fibres with the highest temperature and energy of denaturation (p<0.013). Typical s- and j-shape stress-strain curves were obtained for all treatments in the dry and wet state respectively. Rehydration of the fibres resulted in increased fibre diameter (p<0.006) and reduced stress (p<0.001), force (p<0.001) and modulus (p<0.002) values for every treatment. In the dry state, the alcohol-treated fibres were characterized by the highest stress (p<0.002) values; whilst in the wet state the Tris-HCl-treated fibres were the weakest (p<0.006). For every treatment, in both dry and wet state, a strong and inverse relationship between the fibre diameter and the stress at break was observed. Overall, the fibres produced were characterized by properties similar to those of native tissues.     

Dependence of cross-bridge kinetics on myosin light chain isoforms in rabbit and rat skeletal muscle fibres

Cross-bridge kinetics underlying stretch-induced force transients was studied in fibres with different myosin light chain (MLC) isoforms from skeletal muscles of rabbit and rat. The force transients were induced by stepwise stretches (< 0.3% of fibre length) applied on maximally Ca²⁺-activated skinned fibres. Fast fibre types IIB, IID (or IIX) and IIA and the slow fibre type I containing the myosin heavy chain isoforms MHC-IIb, MHC-IId (or MHC-IIx), MHC-IIa and MHC-I, respectively, were investigated. The MLC isoform content varied within fibre types. Fast fibre types contained the fast regulatory MLC isoform MLC2f and different proportions of the fast alkali MLC isoforms MLC1f and MLC3f. Type I fibres contained the slow regulatory MLC isoform MLC2s and the slow alkali MLC isoform MLC1s. Slow MLC isoforms were also present in several type IIA fibres. The kinetics of force transients differed by a factor of about 30 between fibre types (order from fastest to slowest kinetics: IIB > IID > IIA ≫ I). The kinetics of the force transients was not dependent on the relative content of MLC1f and MLC3f. Type IIA fibres containing fast and slow MLC isoforms were about 1.2 times slower than type IIA fibres containing only fast MLC isoforms. We conclude that while the cross-bridge kinetics is mainly determined by the MHC isoforms present, it is affected by fast and slow MLC isoforms but not by the relative content of MLC1f and MLC3f. Thus, the physiological role of fast and slow MLC isoforms in type IIA fibres is a fine-tuning of the cross-bridge kinetics.     

Morphology of diaphragm neuromuscular junctions on different fibre types

Summary We hypothesize that the morphology of the neuromuscular junction on different muscle fibre types varies, reflecting differences in activation history. In the rat diaphragm muscle, we used a three-colour fluorescent immunocytochemical technique to simultaneously visualize (1) innervating axons and presynaptic nerve terminals, (2) motor endplates and (3) myosin heavy chain isoform expression (muscle fibre type). Laser-scanning confocal microscopy was then used to optically section the triple-labelled muscle fibres, and create three-dimensional views of the neuromuscular junction. Type I fibres were innervated by the smallest axons, and type IIa, IIx and IIb fibres by progressively larger axons. Absolute planar areas of nerve terminals and endplates progressively increased from type I, IIa, IIx to IIb fibres. When normalized for fibre diameter planar areas of nerve terminals were largest on type I fibres, with no difference among type II fibres. The normalized planar area of endplates were larger for type I and IIb fibres, compared to type IIa and IIx fibres. The three-dimensional surface area of endplates was largest on type I fibres, with no differences across type II fibres. When normalized for fibre diameter, endplate surface areas increase progressively from type I, IIa, IIx to IIb fibres. The branches increased progressively from type I, IIa, IIx to IIb fibres. Conversely, individual branch length was longest on type I fibres, and shortest on type IIb fibres. The extent of overlap of pre- and postsynaptic elements of the neuromuscular junction decreased progressively on type I, IIa, IIx and IIb fibres. We conclude that these morphological differences at the neuromuscular function of different fibre types reflect differences in activation history and may underlie phenotypic differences in neuromuscular transmission.     

Characterisation of antibacterial copper releasing degradable phosphate glass fibres

Phosphate-based glass fibres (PGF) of the general formula Na(2)O-CaO-P(2)O(5) are degradable in an aqueous environment, and therefore can function as antibacterial delivery systems through the inclusion of ions such as copper. In this study, PGF with varying amounts of copper oxide (CuO) were developed for potential uses in wound healing applications. PGF with 0, 1, 5 and 10 mol% CuO were produced with different diameters and characterised in terms of structural and antibacterial properties. The effect of CuO and fibre pulling speed on the glass properties were investigated using rapid differential scanning calorimetry, differential thermal analysis and X-ray diffraction. The effect of two fibre diameters on short-term (3 h) attachment and killing against Staphylococcus epidermidis were investigated and were related to their rate of degradation in deionised water, as well as copper ion release measured using ion chromatography. Thermal analysis showed that there was a significant increase in the PGF glass transition temperature as the CuO content increased. There was a significant decrease in the rate of degradation with increasing CuO content and an increase in fibre diameter. Over 6 h, both the amount and rate of copper ions released increased with CuO content, as well as a reduction in fibre diameter thus increasing the surface area to volume ratio. There was a decrease in the number of viable staphylococci both attached to the CuO-containing fibres and in the surrounding environment.     

Scanning electron microscopic studies of the myelinated nerve fibres of the mouse sciatic nerve with special reference to the Schwann cell cytoplasmic network external to the myelin sheath

Summary The three-dimensional morphology of the surface of myelinated nerve fibres in the mouse sciatic nerve was studied by scanning electron microscopy after combined potassium hydroxide treatment and collagenase digestion (to remove the surrounding collagen fibrils and basal laminae from nerve fibres) as well as by transmission electron microscopy.The myelinated nerve fibre appeared as a long cylinder with sporadic annular constrictions corresponding to the nodes of Ranvier. Slight swellings of the surface due to Schwann cell nuclei were usually found at the middle of each internode. The surface of the nerve fibre clearly exhibited a network of bulges, which consisted of longitudinal bands extending from the nuclear swelling to the nodes of Ranvier through the internode, and transverse trabeculae bridging between these longitudinal bands. These bulges on the surface of nerve fibres were the site of the retained Schwann cell cytoplasm external to the myelin lamellae. These cytoplasmic networks on myelinated fibres presumably corresponded to the networks described by Cajal following silver impregnation. In addition, other thin elevations and focal round swellings were also found associated with these longitudinal bands and transverse trabeculae. These networks of Schwann cell cytoplasm are considered to be cytoplasmic channels for nutrition.The two apposing paranodal bulbs of nodes of Ranvier were often asymmetrical in their structure. The networks of the paranodal region were more complicated than those in the internode. The networks of Schwann cell cytoplasm converged into a continuous circumferential collar toward the node, which in turn gave rise to finger-like projections into the nodal gap.     

Fibre composition of human intrinsic tongue muscles

The muscle fibre composition of three human intrinsic tongue muscles, the longitudinalis, verticalis and transversus, was investigated in four anterior to posterior regions of the tongue using morphological and enzyme- and immunohistochemical techniques. All three muscles typically contained type I, IIA and IM/IIC fibres. Type I fibres expressed slow myosin heavy chain (MyHC), type II fibres fast MyHC, mainly fast A MyHC, whereas type IM/IIC coexpressed slow and fast MyHCs. Type II fibres were in the majority (60%), but regional differences in proportion and diameter of fibre types were obvious. The anterior region of the tongue contained a predominance of relatively small type II fibres (71%), in contrast to the posterior region which instead showed a majority of larger type I and type IM/IIC fibres (66%). In general, the fibre diameter was larger in the posterior region. This muscle fibre composition of the tongue differs from those of limb, orofacial and masticatory muscles, probably reflecting genotypic as well as phenotypic functional specialization in oral function. The predominance of type II fibres and the regional differences in fibre composition, together with intricate muscle structure, suggest generally fast and flexible actions in positioning and shaping the tongue, during vital tasks such as mastication, swallowing, respiration and speech.     

Expression of Po mRNA in myelinating Schwann cells is related to fibre size

Summary This study examines whether there is a relationship between the abundance of expression for Po mRNA in myelinated Schwann cells and fibre diameter. Individual teased sciatic nerve fibres from young adult rats were hybridized with radiolabelled probe for Po mRNA which is expressed in the perinuclear cytoplasm of the mid-internode. Signal intensity was measured as optical density of the developed autoradiograms. A highly significant positive linear correlation was present between signal intensity and fibre diameter. In a companion study, individual fibres were mounted in Araldite resin and transversely serially sectioned at 4 m for autoradiography. Grain densities were determined for fibres of different diameters. Again, larger diameter fibres were associated with higher grain densities. The results indicate that the abundance of Po mRNA expressed by a myelin-producing Schwann cell is related to fibre diameter with axonal size probably being the critical determinant. Axons may regulate Po expression through the number of signalling molecules exposed on or released from the axolemma.     

An aligned 3D neuronal-glial co-culture model for peripheral nerve studies

In this study we report on the development of a 3D in vitro peripheral nerve model using aligned electrospun polycaprolactone fibre scaffolds manufactured under tightly controlled and reproducible conditions with uniform diameters of 1 μm, 5 μm and 8 μm. Fibres were characterized by SEM for diameter, density and alignment properties and formed in to scaffolds for 3D in vitro culture. Three different approaches were adopted using: i) neuronal or primary Schwann cell cultures alone; ii) neuronal and primary Schwann cells in co-culture and iii) isolated dorsal root ganglion cultures, containing both neuronal and Schwann cells, with immunohistochemical and 3D confocal microscopy analysis. Neurite guidance was evident on all fibres diameters with the longest neurites detected on 8 μm fibres when cultured alone. However, co-culture with primary Schwann cells was found to enable neurite formation on all scaffolds. Dorsal root ganglion explants when grown on scaffolds showed both organised aligned neurite guidance and notably the co-localization of Schwann cells with neurites. Neurite lengths of up to 2.50 mm were routinely observed using 1 μm diameter fibres after 10 days and all cultures demonstrated a migrating Schwann cell 'front' of up to 2.70 mm (1 μm diameter fibres). Thus, a direct relationship was found between fibre diameter, neurite outgrowth and Schwann cell morphology. This work therefore supports the use of aligned electrospun PCL microfibres for the development of 3D peripheral nerve models in vitro. We envisage such models having future value in a number of areas including developmental biology, disease studies and the design of devices and scaffolds for peripheral nerve repair.     

Structure, distribution and innervation of muscle spindles in avian fast and slow skeletal muscle

Muscle spindles in 2 synergistic avian skeletal muscles, the anterior (ALD) and posterior (PLD) latissimus dorsi, were studied by light and electron microscopy to determine whether morphological or quantitative differences existed between these sensory receptors. Differences were found in the density, distribution and location of muscle spindles in the 2 muscles. They also differed with respect to the morphology of their capsules and intracapsular components. The slow ALD possessed muscle spindles which were evenly distributed throughout the muscle, whereas in the fast PLD they were mainly concentrated around the single nerve entry point into the muscle. The muscle spindle index (number of spindles per gram wet muscle weight) in the ALD was more than double that of its fast-twitch PLD counterpart (130.5±2.0 vs 55.4±2.0 respectively, n=6). The number of intrafusal fibres per spindle ranged from 1 to 8 in the ALD and 2 to 9 in the PLD, and their diameters varied from 5.0 to 16.0 μm and 4.5 to 18.5 μm, respectively. Large diameter intrafusal fibres were more frequently encountered in spindles of the PLD. Unique to the ALD was the presence of monofibre muscle spindles (12.7% of total spindles observed in ALD) which contained a solitary intrafusal fibre. In muscle spindles of both the ALD and PLD, sensory nerve endings terminated in a spiral fashion on the intrafusal fibres in their equatorial regions. Motor innervation was restricted to either juxtaequatorial or polar regions of the intrafusal fibres. Outer capsule components were extensive in polar and juxtaequatorial regions of ALD spindles, whereas inner capsule cells of PLD spindles were more numerous in juxtaequatorial and equatorial regions. Overall, muscle spindles of the PLD exhibited greater complexity with respect to the number of intrafusal fibres per spindle, range of intrafusal fibre diameters and development of their inner capsules. It is postulated that the differences in muscle spindle density and structure observed in this study reflect the function of the muscles in which they reside.     

Regional differences in fibre type composition in the human temporalis muscle

Anatomical and electromyographic studies point to regional differences in function in the human temporalis muscle. During chewing and biting the anterior portions of the muscle are in general more intensively activated and they are capable of producing larger forces than the posterior portions. It was hypothetised that this heterogeneity in function is reflected in the fibre type composition of the muscle. The composition and surface area of different fibre types in various anteroposterior portions of the temporalis muscle were investigated in 7 cadavers employing immunohistochemistry with a panel of monoclonal antibodies against different isoforms of myosin heavy chain. Pure slow muscle fibres, type I, differed strongly in number across the muscle. In the most posterior portion of the muscle there were 24% type I fibres, in the intermediate portion 57%, and in the most anterior portion 46%. The mean fibre cross-sectional area (m-fcsa) of type I fibres was 1849 μm², which did not differ significantly across the muscle. The proportion of pure fast muscle fibres, type IIA and IIX, remained more or less constant throughout the muscle at 13% and 11% respectively; their m-fcsa was 1309 μm² and 1206 μm², respectively, which did not differ significantly throughout the muscle. Pure type IIB fibres were not found. The relative proportion of hybrid fibres was 31% and did not differ significantly among the muscle portions. Fibre types I+IIA and cardiac α+I+IIA were the most abundant hybrid fibre types. In addition, 5% of the type I fibres had an additional myosin isoform which has only recently been described by means of electrophoresis and was named Ia. In the present study they were denoted as hybrid type I+Ia muscle fibres. It is concluded that intramuscular differences in type I fibre distribution are in accordance with regional differences in muscle function.     

Gravity spinning of polycaprolactone fibres for applications in tissue engineering

Poly(epsilon-caprolactone) (PCL) fibres have been produced by wet spinning from solutions in acetone under low shear (gravity flow) conditions. The tensile strength and stiffness of as-spun fibres were highly dependent on the concentration of the spinning solution. Use of a 6% w/v solution resulted in fibres having strength and stiffness of 1.8 MPa and 0.01 GPa, respectively, whereas these values increased to 9.9 MPa and 0.1 GPa when fibres were produced from 20% w/v solutions. Cold drawing to an extension of 500% resulted in further increases in fibre strength (up to 50 MPa) and stiffness (0.3 GPa). The surface morphology of as-spun fibres was modified, to yield a directional grooved pattern by drying in contact with a mandrel having a machined topography characterised by a peak-peak separation of 91 microm and a peak height of 30 microm. Limited in vitro studies of cell behaviour in contact with the fibres were performed using cell culture. The number of attached fibroblasts and myoblasts on as-spun PCL fibres after 5 days in cell culture was lower than on tissue culture plastic by a factor 2 and 1.5, respectively, but higher than on Dacron monofilament by a factor of 4 and 11, respectively. The high fibre compliance and the potential for controlling the fibre surface architecture to promote contact guidance effects together with the maintained proliferation of fibroblasts and myoblasts on as-spun PCL fibres in vitro recommends their use for 3-D scaffold production in soft tissue engineering.     

Electron microscopic serial section analysis of nodes of Ranvier in lumbosacral spinal roots of the cat: ultrastructural organization of nodal compartments in fibres of different sizes

The general ultrastructural organization of nodes of Ranvier in peripheral nerve fibres from 2 to 20 microns in diameter (D) was investigated in the adult cat using serially sectioned ventral and dorsal spinal roots. The study was performed in order to collect and systematize information considered necessary for a morphometric analysis of the node of Ranvier. In all cases a node of Ranvier could be divided into a central nodal axon segment and a surrounding nodal Schwann cell compartment. The latter included a nodal gap matrix substance, more or less overlapping nodal Schwann cell collars and, as a rule, also a Schwann cell brush-border emanating from the nodal Schwann cell collars and occupying the nodal gap. The relative size and the organization level of the nodal Schwann cell compartment increased with increasing fibre size up to a fibre diameter of 8-10 microns. At this fibre size the nodal gap was of a fairly even height (1 micron) all around the nodal axon and contained a thick brush-border of densely packed, more or less radially arranged Schwann cell microvilli. In very small fibres (D less than 3 microns) the nodal gap was low (less than 0.1 microns) and contained no or few microvilli. In fibres greater than 10 microns in diameter the relative size and the degree of structural order of the nodal Schwann cell compartment decreased with increasing fibre size. Drastic sectorial variations in nodal gap height and local thinning-out of the brush-border became prominent features in the largest fibres. The possible in vivo organization of the nodal Schwann cell compartment is discussed. Preliminary calculations indicate that the extracellular space directly surrounding the nodal axon might be quite small and that the area open for free communication between this extracellular space and the endoneurial space might be very much restricted, measuring as little as 2% of the area of the nodal axolemma. Algorithms for calculating various nodal structural parameters are discussed.     

Effect of embedded optical fibres on the mechanical properties of cochlear electrode arrays

Incorporating optical fibres in cochlear electrode arrays has been proposed to provide sensors to help minimise insertion trauma and also for the delivery of light in optical nerve stimulation applications. However, embedding an optical fibre into an electrode array may change its stiffness properties, which can affect the level of trauma during insertion. This report uses measurements of buckling and deflection force to compare the stiffness properties of a range of cochlear electrode arrays (Nucleus straight array, rat array, cat array and guinea pig array) with custom arrays containing an embedded optical fibre. The cladding diameters of the optical fibres tested were 125 µm, 80 µm and 50 µm. The results show that the stiffness of the optical-fibre-embedded arrays is related to the diameter of the optical fibre. Comparison with wired arrays suggests optical fibres with a diameter of 50 µm could be embedded into an electrode array without significantly changing the stiffness properties of the array.     

The surface area of sheep cardiac Purkinje fibres

1. Measurements combining the techniques of point counting and line integration were performed on light and electron micrographs of Purkinje fibres from the sheep's heart. The measurements were aimed at determining membrane areas of importance for the cellular electrophysiology of this tissue.2. The mean volume fractions of the cells occupied by various constituents were: myofibrils, 0.234; mitochondria, 0.103; and nuclei, 0.009. The mean volume fraction of the fibres occupied by the interspaces between the tightly packed cells was 0.0023.3. The mean fractions of intercellular surface area occupied by junctional specializations were: nexus, 0.17; desmosome, 0.023; and fascia adherens, 0.014.4. The mean surface to volume ratio of the Purkinje cells and fibres was 0.46 mu(-1) which is 11.5 times the value of the surface to volume ratio of a long right circular cylinder 100 mu in diameter.5. There are two reasons for the increment in the surface to volume ratio of the fibre (when compared to that of a long right circular cylinder 100 mu in diameter): the multicellular composition of the fibres and the extensive folding of the surface of the cells.6. After correction for the intercellular nexal area the surface to volume ratio of a long cylindrical fibre 100 mu in diameter was 0.39 mu(-1), or about 10 times the value for a long right circular cylinder 100 mu in diameter. The surface to volume ratio of the tissue interspaces in the same fibre was 170 mu(-1).7. It was concluded that the total sarcolemmal area in this tissue is great enough so that the specific membrane capacitance could be about 1 muF/cm(2) and the specific membrane resistance 20,000 Omega cm(2).     

The influences of muscle fibre proportions and areas upon EMG during maximal dynamic knee extensions

This study is an investigation of the relationship between muscle morphology and surface electromyographic (EMG) parameters [mean frequency of the power spectrum (MNF), signal amplitude (root mean square, RMS) and the signal amplitude ratio (SAR; i.e. the ratio between the RMS level during the passive part of the contraction cycle and the RMS level during the active part of the contraction cycle)] during 100 maximal dynamic knee extensions at 90° · s⁻¹. Each contraction cycle comprised of 1 s of active knee extension and 1 s of passive knee flexion. The surface EMG was recorded from the vastus lateralis muscle. Twenty clinically healthy subjects participated in the study, and muscle biopsy samples of the vastus lateralis were obtained from 19 of those subjects. The relationships between muscle morphology and EMG were investigated at three stages of the test: initially, during the fatigue phase (initial 40 contractions), and at the endurance level (the final 50 contractions). Major findings on correlations are that SAR and MNF tended to correlate positively with the proportion of type 1 fibres, and RMS correlated positively with the proportion of type 2 muscle fibres. The muscle fibre areas showed little correlation with the EMG variables under investigation. The results of the present study showed that the three EMG variables of a dynamic endurance test that were investigated (RMS, MNF and SAR) were clearly correlated with the proportions of the different fibre types, but only to a small extent with fibre areas. These findings contradict some of the theoretical models of the EMG, especially for parameters in the frequency domain. Accepted: 17 June 1999     

New biotextiles for tissue engineering: Development,characterization and in vitro cellular viability

This work proposes biodegradable textile-based structures for tissue engineering applications. We describe the use of two polymers, polybutylene succinate (PBS) proposed as a viable multifilamentand silk fibroin (SF), to produce fibre-based finely tuned porous architectures by weft knitting. PBS is here proposed as a viable extruded multifilament fibre to be processed by a textile-based technology. A comparative study was undertaken using a SF fibre with a similar linear density. The knitted constructs obtained are described in terms of their morphology, mechanical properties, swelling capability, degradation behaviour and cytotoxicity. The weft knitting technology used offers superior control over the scaffold design (e.g. size, shape, porosity and fibre alignment), manufacturing and reproducibility. The presented fibres allow the processing of a very reproducible intra-architectural scaffold geometry which is fully interconnected, thus providing a high surface area for cell attachment and tissue in-growth. The two types of polymer fibre allow the generation of constructs with distinct characteristics in terms of the surface physico-chemistry, mechanical performance and degradation capability, which has an impact on the resulting cell behaviour at the surface of the respective biotextiles. Preliminary cytotoxicity screening showed that both materials can support cell adhesion and proliferation. These results constitute a first validation of the two biotextiles as viable matrices for tissue engineering prior to the development of more complex systems. Given the processing efficacy and versatility of the knitting technology and the interesting structural and surface properties of the proposed polymer fibres it is foreseen that the developed systems could be attractive for the functional engineering of tissues such as skin, ligament, bone or cartilage.