RESUMEN
The various connective tissues of the body have different functions, which result from their specific structure and composition. The identification of this structure-function relationship is of great importance for various disciplines such as medicine, biology or tissue engineering. Connective tissue consists mainly of an extracellular matrix (ECM) and a limited number of cells. It is extremely adaptable because the activity of the cells remodels the composition and structure of the ECM in order to adapt the mechanical properties (functions) to the new demands (e.g. an increased mechanical stimulus).
Asunto(s)
Tejido Conectivo , Matriz Extracelular , Tejido Conectivo/fisiología , Humanos , Matriz Extracelular/fisiología , Matriz Extracelular/química , Fenómenos Biomecánicos/fisiología , Modelos Biológicos , Animales , Ingeniería de Tejidos/métodosRESUMEN
Connective tissues can be recognized as an important structural support element in muscles. Recent studies have also highlighted its importance in active force generation and transmission between muscles, particularly through the epimysium. In the present study, we aimed to investigate the impact of the endomysium, the connective tissue surrounding muscle fibers, on both passive and active force production. Pairs of skeletal muscle fibers were extracted from the extensor digitorum longus muscles of rats and, after chemical skinning, their passive and active force-length relationships were measured under two conditions: (i) with the endomysium between muscle fibers intact, and (ii) after its dissection. We found that the dissection of the endomysium caused force to significantly decrease in both active (by 22.2 % when normalized to the maximum isometric force; p < 0.001) and passive conditions (by 25.9 % when normalized to the maximum isometric force; p = 0.034). These findings indicate that the absence of endomysium compromises muscle fiber's not only passive but also active force production. This effect may be attributed to increased heterogeneity in sarcomere lengths, enhanced lattice spacing between myofilaments, or a diminished role of trans-sarcolemmal proteins due to dissecting the endomysium. Future investigations into the underlying mechanisms and their implications for various extracellular matrix-related diseases are warranted.
Asunto(s)
Fibras Musculares Esqueléticas , Animales , Ratas , Fibras Musculares Esqueléticas/fisiología , Ratas Wistar , Tejido Conectivo/fisiología , Sarcómeros/fisiología , Masculino , Músculo Esquelético/fisiología , Fenómenos Biomecánicos , Contracción Isométrica/fisiología , Contracción Muscular/fisiologíaRESUMEN
PURPOSE: Muscle mechanics is paramount in our understanding of motor performance. However, little is known regarding the sensitivity of fascicle dynamics and connective tissues stiffness to exercise duration and ambient temperature during running, both increasing muscle temperature. This study aimed to determine gastrocnemius medialis (GM) fascicle dynamics in vivo during running in temperate and hot conditions, as well as muscle-tendon unit responses. METHODS: Using ultrafast ultrasound, 15 participants (8 men, 7 women; 26 ± 3 yr) were tested before, during (2 and 40 min), and after a running task (40 min at 10 km·h -1 ) in temperate (TEMP; ~23°C) and hot (HOT: ~38°C) conditions. RESULTS: Although core, skin temperatures, and heart rate increased from the beginning to the end of the exercise and in a larger extent in HOT than TEMP ( P < 0.001), the physiological stress elicited did not alter running temporal parameters and GM fascicle operating lengths, with similar behavior of the fascicles on their force-length relationship, over time (2 vs 40 min) or across condition (TEMP vs HOT; P ≥ 0.248). Maximal voluntary force production did not reported statistical changes after exercise ( P = 0.060), and the connective tissues stiffness measured (i.e., passive muscle and stiffness of the series-elastic elements) did not show neither time ( P ≥ 0.281), condition ( P ≥ 0.256) nor time-condition interaction ( P ≥ 0.465) effect. CONCLUSIONS: This study revealed that prolonged running exercise does not alter muscle-tendon unit properties and interplay, which are not influenced by ambient temperature. These findings may rule out potential detrimental effects of heat on muscle properties and encourage further investigations on longer and more intense running exercise.
Asunto(s)
Calor , Músculo Esquelético , Carrera , Ultrasonografía , Humanos , Masculino , Carrera/fisiología , Femenino , Adulto , Músculo Esquelético/fisiología , Músculo Esquelético/diagnóstico por imagen , Frecuencia Cardíaca/fisiología , Tendones/fisiología , Tendones/diagnóstico por imagen , Adulto Joven , Temperatura Cutánea/fisiología , Factores de Tiempo , Temperatura Corporal/fisiología , Tejido Conectivo/fisiologíaRESUMEN
Oculomotricity is a multidimensional domain characterised by a delicate interplay of anatomical structures and physiological processes. This manuscript meticulously dissects the nuances of this interplay, bringing to the fore the integral role of the extraocular muscles (EOMs) and their intricate relationship with the myriad orbital connective tissues as it harmoniously orchestrates binocular movements, ensuring synchronised and fluid visual tracking. Historically, the peripheral oculomotor apparatus was conceptualised as a rudimentary system predominantly driven by neural directives. While widely accepted, this perspective offered a limited view of the complexities inherent in ocular movement mechanics. The twentieth century heralded a paradigm shift in this understanding. With advances in anatomical research and imaging techniques, a much clearer picture of the gross anatomy of the EOMs emerged. This clarity challenged traditional viewpoints, suggesting that the inherent biomechanical properties of the EOMs, coupled with their associated tissue pulleys, play a pivotal role in dictating eye movement dynamics. Central to this revised understanding is the "arc of contact" paradigm. This concept delves deep into the mechanics of eye rotation, elucidating the significance of the point of contact between the EOMs and the eyeball. The arc of contact is not just a static anatomical feature; its length and orientation play a crucial role in determining the effective torque generated by a muscle, thereby influencing the amplitude and direction of eye rotation. The dynamic nature of this arc, influenced by the position and tension of the muscle pulleys, offers a more comprehensive model for understanding ocular kinematics. Previously overlooked in traditional models, muscle pulleys have now emerged as central players in the biomechanics of eye movement. These anatomical structures, formed by dense connective tissues, guide the paths of the EOMs, ensuring that their pulling angles remain optimal across a range of gaze directions. The non-linear paths resulting from these pulleys provide a more dynamic and intricate understanding of eye movement, challenging two-dimensional, linear models of orbital anatomy. The implications of these revelations extend beyond mere theoretical knowledge. The insights garnered from this research promise transformative potential in the realm of strabismus surgery. Recognising the pivotal role of muscle pulleys and the "arc of contact" paradigm allows for more precise surgical interventions, ensuring better post-operative outcomes and minimising the risk of complications. Surgical procedures that previously relied on basic mechanical principles now stand to benefit from a more nuanced understanding of the underlying anatomical and physiological dynamics. In conclusion, this manuscript serves as a testament to the ever-evolving nature of scientific knowledge. Challenging established norms and introducing fresh perspectives pave the way for more effective and informed clinical interventions in strabismus surgery.
Asunto(s)
Tejido Conectivo , Músculos Oculomotores , Órbita , Estrabismo , Humanos , Músculos Oculomotores/anatomía & histología , Músculos Oculomotores/fisiología , Estrabismo/cirugía , Tejido Conectivo/anatomía & histología , Tejido Conectivo/fisiología , Órbita/anatomía & histología , Movimientos Oculares/fisiología , Fenómenos Biomecánicos/fisiologíaRESUMEN
The classical view of muscles as independent motors has been challenged over the past decades. An alternative view has emerged in which muscles are not isolated but embedded in a three-dimensional connective tissue network that links them to adjacent muscles and other non-muscular structures in the body. Animal studies showing that the forces measured at the distal and proximal ends of a muscle are not equal have provided undisputable evidence that these connective tissue linkages are strong enough to serve as an extra pathway for muscular force transmission. In this historical review, we first introduce the terminology and anatomy related to these pathways of muscle force transmission and provide a definition for the term epimuscular force transmission. We then focus on important experimental evidence indicating mechanical interactions between synergistic muscles that may affect force transmission and/or influence the muscles' force generating capacity. We illustrate that there may exist different expressions of the highly relevant force-length properties depending on whether the force is measured at the proximal or distal tendon and depending on the dynamics of surrounding structures. Changes in length, activation level or disruption of the connective tissue of neighboring muscles, can affect how muscles interact and produce force on the skeleton. While most direct evidence is from animal experiments, studies on humans also suggest functional implications of the connective tissues surrounding muscles. These implications may explain how distant segments, which are not part of the same joint system, affect force generation at a given joint, and, in clinical conditions, explain observations from tendon transfer surgeries, where a muscle transferred to act as an antagonist continues to produce agonistic moments.
Asunto(s)
Tejido Conectivo , Fuerza Muscular , Tono Muscular , Músculo Esquelético , Humanos , Músculo Esquelético/fisiología , Animales , Tejido Conectivo/fisiologíaRESUMEN
The "motor unit" or the "muscle" has long been considered the quantal element in the control of movement. However, in recent years new research has proved the strong interaction between muscle fibers and intramuscular connective tissue, and between muscles and fasciae, suggesting that the muscles can no longer be considered the only elements that organize movement. In addition, innervation and vascularization of muscle is strongly connected with intramuscular connective tissue. This awareness induced Luigi Stecco, in 2002, to create a new term, the "myofascial unit", to describe the bilateral dependent relationship, both anatomical and functional, that occurs between fascia, muscle and accessory elements. The aim of this narrative review is to understand the scientific support for this new term, and whether it is actually correct to consider the myofascial unit the physiological basic element for peripheral motor control.
Asunto(s)
Fascia , Músculo Esquelético , Músculo Esquelético/fisiología , Fascia/fisiología , Tejido Conectivo/fisiología , Fibras Musculares Esqueléticas , Contracción Muscular/fisiologíaRESUMEN
Evisceration in dendrochirotid sea cucumbers leads to expulsion of the digestive tract, pharyrngeal complex and coelomic fluid through rupture of the anterior body wall. This process involves failure of three mutable collagenous tissue (MCT) structures, the introvert, the pharyngeal retractor muscle tendon, and the intestine-cloacal junction. These are complex structures composed of several tissue strata. The MCT in the three autotomy structures contains collagen fibrils, unstriated microfibrils, and interfibrillar molecules. Neurosecretory-like processes (juxtaligamental-type) with large dense vesicles (LDVs) are prominent in the autotomy structures. Biomechanical tests show that these structures are not inherently weak. Failure of the autotomy structures can be elicited by manipulating the ionic environment and the changes are blocked by anaesthetics. Autotomy and evisceration are under neural control, but local neural elements and neurosecretory-like processes do not appear to be a source of factors that cause MCT destabilisation. The LDVs remain intact while the tissue destabilises. The coelomic fluid contains an evisceration inducing factor indicating a neurosecretory-like mediation of autotomy. This factor elicits muscle contraction and MCT destabilisation. As the autotomy structures are completely or partially surrounded by coelomic fluid, the agent(s) of change may be located in the coelom (systemic origin) as well as originate from cells within the MCT. The biochemistry and mechanism(s) of action of the evisceration factor are not known. This factor is a promising candidate for biodiscovery investigation.
Asunto(s)
Tejido Conectivo , Pepinos de Mar , Animales , Tejido Conectivo/fisiología , Matriz Extracelular , Tracto Gastrointestinal , IntestinosRESUMEN
The subsynovial connective tissue is an integral component of flexor tendon gliding in the carpal tunnel, which is strained during longitudinal tendon displacement. We tested the effects of repetition frequency and finger load on flexor tendon function throughout active finger movement. Eleven participants performed metacarpophalangeal joint flexion/extension of the long finger cyclically at three repetition frequencies (0.75, 1.00, 1.25 Hz) and two finger loads (3.5, 7 N). Relative displacement between the flexor digitorum superficialis tendon and subsynovial connective tissue was assessed as the shear-strain index with color ultrasound throughout the entire time history of finger flexion and extension. In addition, long finger joint angles were measured with electrogoniometry while flexor digitorum superficialis and extensor digitorum muscle activities were measured with fine-wire electromyography to characterize the finger movements. The shear-strain index increased with greater finger flexion (p = 0.001), representing higher relative displacement between tendon and subsynovial connective tissue; however, no changes were observed throughout finger extension. The shear-strain index also increased with higher repetition frequencies (p = 0.013) and finger loads (p = 0.029), further modulating time-dependent effects during finger flexion versus extension. Using ultrasound, we characterized the time-dependent response of the shear-strain index, in vivo, providing valuable data on flexor tendon function during active finger movement. Our results infer greater subsynovial connective tissue strain and shear during repetitive and forceful finger movements. Future research characterizing time-dependent effects in carpal tunnel syndrome patients may further elucidate the relations between subsynovial connective tissue function, damage, and carpal tunnel syndrome.
Asunto(s)
Síndrome del Túnel Carpiano , Humanos , Síndrome del Túnel Carpiano/diagnóstico por imagen , Tejido Conectivo/diagnóstico por imagen , Tejido Conectivo/fisiología , Tendones/fisiología , Dedos , ManoRESUMEN
Aging is accompanied by morphological and mechanical changes to the intramuscular connective tissue (IMCT) of skeletal muscles, but whether physical exercise can influence these changes is debated. We investigated the effects of aging and exercise with high or low resistance on composition and mechanical properties of the IMCT, including direct measurements on isolated IMCT which has rarely been reported. Middle-aged (11 months, n = 24) and old (22 months, n = 18) C57BL/6 mice completed either high (HR) or low (LR) resistance voluntary wheel running or were sedentary (SED) for 10 weeks. Passive mechanical properties of the intact soleus and plantaris muscles and the isolated IMCT of the plantaris muscle were measured in vitro. IMCT thickness was measured on picrosirius red stained cross sections of the gastrocnemius and soleus muscle and for the gastrocnemius hydroxyproline content was quantified biochemically and advanced glycation end-products (AGEs) estimated by fluorometry. Mechanical stiffness, IMCT content and total AGEs were all elevated with aging in agreement with previous findings but were largely unaffected by training. Conclusion: IMCT accumulated with aging with a proportional increase in mechanical stiffness, but even the relatively high exercise volume achieved with voluntary wheel-running with or without resistance did not significantly influence these changes.
Asunto(s)
Colágeno , Actividad Motora , Envejecimiento/fisiología , Animales , Colágeno/fisiología , Tejido Conectivo/fisiología , Ratones , Ratones Endogámicos C57BL , Músculo Esquelético/fisiologíaRESUMEN
Bone fractures commonly repair by forming a bridging structure called callus, which begins as soft tissue and gradually ossifies to restore rigidity to the bone. Virtual mechanical testing is a promising technique for image-based assessment of structural bone healing in both preclinical and clinical settings, but its accuracy depends on the validity of the material model used to assign tissue mechanical properties. The goal of this study was to develop a constitutive model for callus that captures the heterogeneity and biomechanical duality of the callus, which contains both soft tissue and woven bone. To achieve this, a large-scale optimization analysis was performed on 2363 variations of 3D finite element models derived from computed tomography (CT) scans of 33 osteotomized sheep under normal and delayed healing conditions. A piecewise material model was identified that produced high absolute agreement between virtual and physical tests by differentiating between soft and hard callus based on radiodensity. The results showed that the structural integrity of a healing long bone is conferred by an internal architecture of mineralized hard callus that is supported by interstitial soft tissue. These findings suggest that with appropriate material modeling, virtual mechanical testing is a reliable surrogate for physical biomechanical testing.
Asunto(s)
Huesos/fisiología , Curación de Fractura/fisiología , Fracturas Óseas/fisiopatología , Pruebas Mecánicas/métodos , Osteogénesis/fisiología , Animales , Fenómenos Biomecánicos , Huesos/diagnóstico por imagen , Tejido Conectivo/diagnóstico por imagen , Tejido Conectivo/fisiología , Análisis de Elementos Finitos , Ovinos , Tomografía Computarizada por Rayos X/métodosRESUMEN
The intention of this special edition is to highlight the benefits of a holistic approach to computational and experimental approaches in the context of aiding the diagnosis and remediation of disease and injury, especially in neurological and connective tissues and organs [...].
Asunto(s)
Fenómenos Biomecánicos/fisiología , Tejido Conectivo/fisiología , Humanos , Estrés MecánicoRESUMEN
Strenuous and unaccustomed exercise frequently lead to what has been coined "delayed onset muscle soreness" (DOMS). As implied by this term, it has been proposed that the associated pain and stiffness stem from micro-lesions, inflammation, or metabolite accumulation within the skeletal muscle. However, recent research points towards a strong involvement of the connective tissue. First, according to anatomical studies, the deep fascia displays an intimate structural relationship with the underlying skeletal muscle and may therefore be damaged during excessive loading. Second, histological and experimental studies suggest a rich supply of algogenic nociceptors whose stimulation evokes stronger pain responses than muscle irritation. Taken together, the findings support the hypothesis that DOMS originates in the muscle-associated connective tissue rather than in the muscle itself. Sports and fitness professionals designing exercise programs should hence consider fascia-oriented methods and techniques (e.g., foam rolling, collagen supplementation) when aiming to treat or prevent DOMS.
Asunto(s)
Tejido Conectivo/fisiología , Fascia/fisiología , Mialgia/fisiopatología , Ejercicio Físico/fisiología , Humanos , Contracción Muscular/fisiología , Músculo Esquelético/fisiopatología , Músculos/fisiopatología , Mialgia/metabolismo , Dolor/metabolismo , Dolor/fisiopatología , Factores de TiempoRESUMEN
Different factors were shown to alter the vibration characteristics of soft-tissue compartments during running. Changing pre-heel strike muscle activation or changing footwear conditions represents two possibilities to influence the vibration response via frequency shift or altered damping. Associated with the study of muscle pre-tuning is the difficulty in quantifying clean experimental data for the acceleration of soft-tissue compartments and muscle activities in heterogeneous populations. The purpose of this study was to determine the vibration and pre-tuning response to footwear across a wide range of participants during running and establish and describe groups formed according to the damping coefficient. 32 subjects were used for further analysis. The subjects ran at a self-selected speed (5 min) on a treadmill in two different shoes (soft & hard), while soft-tissue accelerations and muscle activation at the gastrocnemius medialis were quantified. Damping coefficients, total muscle intensity and dominant vibration frequencies were determined. Anthropometrics and skinfold measurements of the lower limbs were obtained. According to the damping coefficient response to the footwear intervention, three groups were formed, with most runners (n = 20) showing less damping in the hard shoe. Total muscle intensity, anthropometrics, and dominant vibration frequency across footwear were not different for these three groups. Most runners (84.4%) used the strategy of adjusting the damping coefficients significantly when switching footwear. Despite damping being the preferred adjustment to changes in footwear, muscle pre-tuning might not be the only mechanism to influence damping as previously suggested. Future studies should focus on the subject-specific composition of soft-tissue compartments to elucidate their contribution to vibrations.
Asunto(s)
Tejido Conectivo/fisiología , Talón/fisiología , Músculo Esquelético/fisiología , Carrera/fisiología , Aceleración , Adolescente , Adulto , Antropometría , Fenómenos Biomecánicos , Electromiografía , Prueba de Esfuerzo , Femenino , Talón/anatomía & histología , Humanos , Masculino , Persona de Mediana Edad , Zapatos/clasificación , VibraciónRESUMEN
The morphological and possible functional interactions between the connective tissue and enamel organ cells were examined during the maturation phase of enamel formation, using immunohistochemical techniques. Decalcified mandibular sections (10 µm) including incisors were used from Wistar rats ages 10-12 weeks. Sections were incubated with one or two primary antibodies targeting cell cytoskeleton (vimentin, α-actin, α-tubulin), dendritic marker (OX6), gap junctions (cx-43), enzymes (nitric-oxide synthase (nos1) and cyclooxygenase (cox1)), and the ion transporters (Na+/H+ exchanger (NHE1) and Na+/Ca2+ exchanger (NCX)) for 24 h, before incubation with the appropriate conjugated fluorescent secondary antibodies. Sections were examined by fluorescence microscopy. Haematoxylin-eosin slides were also employed. Cellular heterogeneity and morphological modulations were identified within enamel organ cells and connective tissue covering suggesting complex cellular interactions and indicating a new functional concept and possible complementary role during enamel maturation. Also, some ion transportation activity, and nos1 and cox1 signalling pathways have been identified, indicating intercellular communication between these regions. A hypothesis is suggested, to explain the morphological modulation of ameloblasts and papillary cells during enamel maturation which functions to increase the transporting membrane surface area to accomplish faster and bulker ion transportation to achieve controlled pH and to direct Ca2+ towards enamel.
Asunto(s)
Tejido Conectivo/anatomía & histología , Tejido Conectivo/fisiología , Órgano del Esmalte/anatomía & histología , Órgano del Esmalte/crecimiento & desarrollo , Epitelio/anatomía & histología , Epitelio/fisiología , Animales , Ciclooxigenasa 1/metabolismo , Incisivo/citología , Masculino , Mandíbula/citología , Modelos Biológicos , Óxido Nítrico Sintasa/metabolismo , Ratas WistarRESUMEN
BACKGROUND: Soft tissue balancing is essential for the success of total knee arthroplasty (TKA) and is mainly dependent on surgeon-defined assessment (SDA) or a gap-balancer (GB). However, an electronic sensor has been developed to objectively measure the gap pressure. This study aimed to evaluate the accuracy of soft tissue balancing using SDA and GB compared with a sensor. METHODS: Forty-eight patients undergoing TKA (60 knees) were prospectively enrolled. Soft tissue balancing was sequentially performed using SDA, a GB, and an electronic sensor. We compared the SDA, GB, and sensor data to calculate the sensitivity, specificity, and accuracy at 0°, 45°, 90°, and 120° flexion. Cumulative summation (CUSUM) analysis was performed to assess the surgeon's performance during the sensor introductory phase. RESULTS: The sensitivity of SDA was 63.3%, 68.3%, 80.0%, and 80.0% at 0°, 45°, 90°, and 120°, respectively. The accuracy of the GB compared with sensor data was 76.7% and 71.7% at 0° and 90°, respectively. Cohen's kappa coefficient for the accuracy of the GB was 0.406 at 0° (moderate agreement) and 0.227 at 90° (fair agreement). The CUSUM 0° line achieved good prior performance at case 45, CUSUM 90° and 120° showed a trend toward good prior performance, while CUSUM 45° reached poor prior performance at case 8. CONCLUSION: SDA was a poor predictor of knee balance. GB improved the accuracy of soft tissue balancing, but was still less accurate than the sensor, particularly for unbalanced knees. SDA improved with ongoing use of the sensor, except at 45° flexion.
Asunto(s)
Artroplastia de Reemplazo de Rodilla/métodos , Tejido Conectivo/fisiología , Electrónica Médica/instrumentación , Prótesis de la Rodilla , Osteoartritis de la Rodilla/fisiopatología , Osteoartritis de la Rodilla/cirugía , Rango del Movimiento Articular , Cirujanos , Fenómenos Biomecánicos , Estudios de Cohortes , Humanos , Curva de Aprendizaje , Sensibilidad y Especificidad , Resultado del TratamientoRESUMEN
Skeletal muscle tissue has a highly complex and heterogeneous structure comprising several physical length scales. In the simplest model of muscle tissue, it can be represented as a one dimensional nonlinear spring in the direction of muscle fibres. However, at the finest level, muscle tissue includes a complex network of collagen fibres, actin and myosin proteins, and other cellular materials. This study shall derive an intermediate physical model which encapsulates the major contributions of the muscle components to the elastic response apart from activation-related along-fibre responses. The micro-mechanical factors in skeletal muscle tissue (eg. connective tissue, fluid, and fibres) can be homogenized into one material aggregate that will capture the behaviour of the combination of material components. In order to do this, the corresponding volume fractions for each type of material need to be determined by comparing the stress-strain relationship for a volume containing each material. This results in a model that accounts for the micro-mechanical features found in muscle and can therefore be used to analyze effects of neuro-muscular diseases such as cerebral palsy or muscular dystrophies. The purpose of this study is to construct a model of muscle tissue that, through choosing the correct material parameters based on experimental data, will accurately capture the mechanical behaviour of whole muscle. This model is then used to look at the impacts of the bulk modulus and material parameters on muscle deformation and strain energy-density distributions.
Asunto(s)
Tejido Conectivo/fisiología , Matriz Extracelular/fisiología , Modelos Biológicos , Fibras Musculares Esqueléticas/fisiología , Músculo Esquelético/fisiología , Estrés Mecánico , Fenómenos Biomecánicos , Tejido Conectivo/química , Matriz Extracelular/química , HumanosRESUMEN
Advances in vibrational spectroscopy have propelled new insights into the molecular composition and structure of biological tissues. In this review, we discuss common modalities and techniques of vibrational spectroscopy, and present key examples to illustrate how they have been applied to enrich the assessment of connective tissues. In particular, we focus on applications of Fourier transform infrared (FTIR), near infrared (NIR) and Raman spectroscopy to assess cartilage and bone properties. We present strengths and limitations of each approach and discuss how the combination of spectrometers with microscopes (hyperspectral imaging) and fiber optic probes have greatly advanced their biomedical applications. We show how these modalities may be used to evaluate virtually any type of sample (ex vivo, in situ or in vivo) and how "spectral fingerprints" can be interpreted to quantify outcomes related to tissue composition and quality. We highlight the unparalleled advantage of vibrational spectroscopy as a label-free and often nondestructive approach to assess properties of the extracellular matrix (ECM) associated with normal, developing, aging, pathological and treated tissues. We believe this review will assist readers not only in better understanding applications of FTIR, NIR and Raman spectroscopy, but also in implementing these approaches for their own research projects.
Asunto(s)
Huesos/citología , Cartílago/citología , Tejido Conectivo/fisiología , Espectrometría Raman/métodos , Animales , Huesos/química , Cartílago/química , Tecnología de Fibra Óptica , Humanos , Espectroscopía Infrarroja por Transformada de Fourier , Espectroscopía Infrarroja CortaRESUMEN
BACKGROUND: Under certain conditions, the physiological repair of connective tissues might fail to restore the original structure and function. Optimized engineered connective tissues (ECTs) with biophysical properties adapted to the target tissue could be used as a substitution therapy. This study aimed to investigate the effect of ECT enforcement by a complex of multiwall carbon nanotubes with chitosan (C-MWCNT) to meet in vivo demands. MATERIALS AND METHODS: ECTs were constructed from human foreskin fibroblasts (HFF-1) in collagen type I and enriched with the three different percentages 0.025, 0.05 and 0.1% of C-MWCNT. Characterization of the physical properties was performed by biomechanical studies using unidirectional strain. RESULTS: Supplementation with 0.025% C-MWCNT moderately increased the tissue stiffness, reflected by Young's modulus, compared to tissues without C-MWCNT. Supplementation of ECTs with 0.1% C-MWCNT reduced tissue contraction and increased the elasticity and the extensibility, reflected by the yield point and ultimate strain, respectively. Consequently, the ECTs with 0.1% C-MWCNT showed a higher resilience and toughness as control tissues. Fluorescence tissue imaging demonstrated the longitudinal alignment of all cells independent of the condition. CONCLUSION: Supplementation with C-MWCNT can enhance the biophysical properties of ECTs, which could be advantageous for applications in connective tissue repair.
Asunto(s)
Quitosano/farmacología , Tejido Conectivo/fisiología , Nanotubos de Carbono/química , Ingeniería de Tejidos , Animales , Fenómenos Biomecánicos , Bovinos , Línea Celular , Quitosano/química , Módulo de Elasticidad , Fibroblastos/efectos de los fármacos , HumanosRESUMEN
Near-infrared (NIR) spectroscopy is a powerful analytical method for rapid, non-destructive and label-free assessment of biological materials. Compared to mid-infrared spectroscopy, NIR spectroscopy excels in penetration depth, allowing intact biological tissue assessment, albeit at the cost of reduced molecular specificity. Furthermore, it is relatively safe compared to Raman spectroscopy, with no risk of laser-induced photothermal damage. A typical NIR spectroscopy workflow for biological tissue characterization involves sample preparation, spectral acquisition, pre-processing and analysis. The resulting spectrum embeds intrinsic information on the tissue's biomolecular, structural and functional properties. Here we demonstrate the analytical power of NIR spectroscopy for exploratory and diagnostic applications by providing instructions for acquiring NIR spectra, maps and images in biological tissues. By adapting and extending this protocol from the demonstrated application in connective tissues to other biological tissues, we expect that a typical NIR spectroscopic study can be performed by a non-specialist user to characterize biological tissues in basic research or clinical settings. We also describe how to use this protocol for exploratory study on connective tissues, including differentiating among ligament types, non-destructively monitoring changes in matrix formation during engineered cartilage development, mapping articular cartilage proteoglycan content across bovine patella and spectral imaging across the depth-wise zones of articular cartilage and subchondral bone. Depending on acquisition mode and experiment objectives, a typical exploratory study can be completed within 6 h, including sample preparation and data analysis.
Asunto(s)
Tejido Conectivo/metabolismo , Tejido Conectivo/fisiología , Espectroscopía Infrarroja Corta/métodos , Animales , Cartílago Articular/química , Células del Tejido Conectivo/citología , Humanos , Proteoglicanos/química , Manejo de Especímenes/métodosRESUMEN
Tissue regeneration often requires recruitment of different cell types and rebuilding of two or more tissue layers to restore function. Here, we describe the creation of a novel multilayered scaffold with distinct fiber organizations-aligned to unaligned and dense to porous-to template common architectures found in adjacent tissue layers. Electrospun scaffolds were fabricated using a biodegradable, tyrosine-derived terpolymer, yielding densely-packed, aligned fibers that transition into randomly-oriented fibers of increasing diameter and porosity. We demonstrate that differently-oriented scaffold fibers direct cell and extracellular matrix (ECM) organization, and that scaffold fibers and ECM protein networks are maintained after decellularization. Smooth muscle and connective tissue layers are frequently adjacent in vivo; we show that within a single scaffold, the architecture supports alignment of contractile smooth muscle cells and deposition by fibroblasts of a meshwork of ECM fibrils. We rolled a flat scaffold into a tubular construct and, after culture, showed cell viability, orientation, and tissue-specific protein expression in the tube were similar to the flat-sheet scaffold. This scaffold design not only has translational potential for reparation of flat and tubular tissue layers but can also be customized for alternative applications by introducing two or more cell types in different combinations.