Mechanical Stimulation (mechanical + stimulation)

Distribution by Scientific Domains

Selected Abstracts

Dual Mechanism of Intercellular Communication in HOBIT Osteoblastic Cells: A Role for Gap-Junctional Hemichannels

Milena Romanello
Abstract Intercellular communication allows tissue coordination of cell metabolism and sensitivity to extracellular stimuli. Paracrine stimulation and cell-to-cell coupling through gap junctions induce the formation of complex cellular networks, which favors the intercellular exchange of nutrients and second messengers. Intercellular Ca2+ signaling was investigated in human osteoblast-like initial transfectant (HOBIT) cells, a human osteoblastic cell line in which cells retain most of the osteoblastic differentiation markers. HOBIT cells express connexin43 (Cx43) clustered at the cell-to-cell boundary and display functional intercellular coupling as assessed by the intercellular transfer of Lucifer yellow. Mechanical stimulation of a single cell induced a wave of increased Ca2+ that was radially propagated to surrounding cells. Treatment of cells with thapsigargin blocked mechanically induced signal propagation. Intercellular Ca2+ spreading and dye transfer were inhibited by 18,-glycyrrhetinic acid (18-GA), showing the involvement of gap junctions in signal propagation. Pretreatment of cells with suramin or with apyrase decreased the extent of wave propagation, suggesting that ATP-mediated paracrine stimulation contribute to cell-to-cell signaling. The functional expression of gap-junctional hemichannels was evidenced in experiments of Mn2+ quenching, extracellular dye uptake, and intracellular Ca2+ release, activated by uptake of inositol 1,4,5-trisphosphate (InsP3) from the external medium. Gap-junctional hemichannels were activated by low extracellular Ca2+ concentrations and inhibited by 18-GA. A role for Cx hemichannels in adenosine triphosphate (ATP) release and paracrine stimulation is suggested. [source]

Location of proliferating gingival cells following toothbrushing stimulation

ORAL DISEASES, Issue 1 2007
T Tomofuji
Objectives:, Mechanical stimulation by toothbrushing promotes healing of gingivitis through accelerating cell proliferation. Junctional epithelium proliferates at periodontal pocket formation. A question is arisen whether toothbrushing contributes to the repair of gingival inflammation or deterioration of pocket formation. The location of proliferating cells in gingiva stimulated mechanically by toothbrushing was investigated. Materials and methods:, A total of 24 teeth of dogs underwent daily plaque removal with a curette (plaque removal) or both plaque removal and toothbrushing (toothbrushing). Proliferative activity of gingival cells in six individual zones was evaluated by assaying expression of proliferating cell nuclear antigen (PCNA). Results:, Toothbrushing increased densities of PCNA-positive basal cells in the junctional epithelium, connective tissues adjacent to the junctional epithelium, the alveolar bone of the oral epithelial side and the oral epithelium. However, the densities of PCNA-positive cells at the apical portion of the junctional epithelium, connective tissues adjacent to the cementum and the alveolar bone of the periodontal ligament side did not increase following toothbrushing. Conclusions:, Toothbrushing promotes proliferation of gingival cells other than fibroblasts in periodontium and basal cells in the apical portion of the junctional epithelium. The repair of periodontal tissues might be promoted by toothbrushing within the limit of the direct mechanical stimulation. [source]

Real-time Monitoring of Force Response Measured in Mechanically Stimulated Tissue-Engineered Cartilage

Orahn Preiss-Bloom
Abstract:, Mechanical stimulation improves tissue-engineered cartilage development both in terms of biochemical composition and structural properties. However, the link between the compositional changes attributed to mechanical stimulation and the changing structural properties of the engineered cartilage is poorly understood. We hypothesize that transient events associated with construct stiffening can be documented and used to understand milestones in construct development. To do this, we designed and built a mechanical stimulation bioreactor that can continuously record the force response of the engineered construct in real time. This study documents the transient changes of the stiffness of tissue-engineered cartilage constructs over the first 14 days of their development under cyclic loading. Compressive strain stimulation (15%, 1 Hz) was applied to poly(ethylene glycol) (PEG) hydrogels seeded with primary articular chondrocytes. The average compressive modulus of strain-stimulated constructs was 12.7 ± 1.45 kPa after 2 weeks, significantly greater (P < 0.01) than the average compressive moduli of both unstimulated constructs (10.7 ± 0.94 kPa) and nonviable stimulated constructs (11.2 ± 0.91 kPa). The system was able to document that nearly all of the stiffness increase occurred over the last 2 days of the experiment, where live-cell constructs demonstrated a rapid 20% increase in force response. The system's ability to track significant increases in stiffness over time was also confirmed by Instron testing. These results present a novel view of the early mechanical development of tissue-engineering cartilage constructs and suggest that the real-time monitoring of force response may be used to noninvasively track the development of engineered tissue. [source]

Critical and sensitive periods for reversing the effects of mechanosensory deprivation on behavior, nervous system, and development in Caenorhabditis elegans

Susan Rai
Abstract In these studies the nematode Caenorhabditis elegans was used as a model to investigate ways to reverse the effects of mechanosensory deprivation on behavior and development. Rose et al. (J Neurosci 2005; 25:7159,7168) showed that worms reared in isolation responded significantly less to a mechanical tap stimulus, were significantly smaller, and expressed significantly lower levels of a postsynaptic glutamate receptor subunit on the command interneurons of the tap response circuit and a presynaptic vesicle marker in the tap sensory neurons compared with worms raised in groups. Here, brief mechanical stimulation at any time throughout development reversed the effects of isolation on the response to tap and on postsynaptic glutamate receptor expression on the command interneurons, suggesting there is no critical period for these measures. In contrast to the high level of plasticity in glutamate receptor subunit expression on the interneurons, low levels of stimulation only rescued vesicle expression in the tap sensory neurons early in development and progressively higher levels of stimulation were required as the worm developed, suggesting a sensitive period immediately after hatching, followed by a period of decreasing plasticity. Stimulation during the first three stages of larval development, but not later, rescued the effects of isolation on worm length, suggesting there is a critical period for this measure that ends in the third larval stage. These results indicate that different effects of early isolation required different amounts and/or timing of stimulation to be reversed. © 2007 Wiley Periodicals, Inc. Develop Neurobiol, 2007. [source]

Expression of multiple P2Y receptors by MDCK-D1 cells: P2Y1 receptor cloning and signaling

Richard J. Hughes
The Madin Darby canine kidney (MDCK) cell line, a well-differentiated renal epithelial cell line, is a useful model to examine P2Y receptor signaling and response. Our studies with MDCK-D1, a clonal isolate, demonstrate that these cells release ATP in response to mechanical stimulation and activation of certain G-protein-coupled receptors. Reverse transcriptase-polymerase chain reaction (RT-PCR) studies document that MDCK cells express multiple P2Y receptors, including P2Y1, P2Y2, P2Y6, and P2Y11 receptors. We isolated cDNAs for several of the P2Y receptor genes and expressed these in cells, such as the 1321N1 astrocytoma cell line, that lack native P2Y receptor expression. We report here the molecular cloning of the MDCK P2Y1 receptor, heterologous expression in 1321N1 cells, and the ability of the heterologously expressed receptors to increase intracellular calcium and phosphoinositide hydrolysis. ADP, methylthioATP, and ADP,S are agonists with the greatest potency, while ATP and ATP,S show lower potency and efficacy, and benzoylbenzoylATP, UTP, and UDP lack efficacy at the cloned P2Y1 receptor. Several antagonists, including MRS2179, A3P5PS, suramin, and PPADS blocked response at the cloned P2Y1 receptors. With their ability to respond to ADP and ATP, P2Y1 receptors, along with other P2Y receptors expressed in MDCK cells, contribute to the response of these cells to ATP (or its breakdown product, ADP) released from the cells and to exogenously added nucleotides. Drug Dev. Res. 59:1,7, 2003. © 2003 Wiley-Liss, Inc. [source]

Neuronal disinhibition in the trigeminal nucleus caudalis in a model of chronic neuropathic pain

Yasmina B. Martin
Abstract The mechanisms underlying neuropathic facial pain syndromes are incompletely understood. We used a unilateral chronic constriction injury of the rat infraorbital nerve (CCI-IoN) as a facial neuropathic model. Pain-related behavior of the CCI-IoN animals was tested at 8, 15 and 26 days after surgery (dps). The response threshold to mechanical stimulation with von Frey hairs on the injured side was reduced at 15 and 26 dps, indicating the presence of allodynia. We performed unitary recordings in the caudalis division of the spinal trigeminal nucleus (Sp5C) at 8 or 26 dps, and examined spontaneous activity and responses to mechanical and thermal stimulation of the vibrissal pad. Neurons were identified as wide dynamic range (WDR) or low-threshold mechanoreceptive (LTM) according to their response to tactile and/or noxious stimulation. Following CCI-IoN, WDR neurons, but not LTM neurons, increased their spontaneous activity at 8 and 26 dps, and both types of Sp5C neurons increased their responses to tactile stimuli. In addition, the on,off tactile response in neurons recorded after CCI-IoN was followed by afterdischarges that were not observed in control cases. Compared with controls, the response inhibition observed during paired-pulse stimulation was reduced after CCI-IoN. Immunohistochemical studies showed an overall decrease in GAD65 immunoreactivity in Sp5C at 26 dps, most marked in laminae I and II, suggesting that following CCI-IoN the inhibitory circuits in the sensory trigeminal nuclei are depressed. Consequently, our results strongly suggest that disinhibition of Sp5C neurons plays a relevant role in the appearance of allodynia after CCI-IoN. [source]

Sensitization of meningeal nociceptors: inhibition by naproxen

Dan Levy
Abstract Migraine attacks associated with throbbing (manifestation of peripheral sensitization) and cutaneous allodynia (manifestation of central sensitization) are readily terminated by intravenous administration of a non-selective cyclooxygenase (COX) inhibitor. Evidence that sensitization of rat central trigeminovascular neurons was also terminated in vivo by non-selective COX inhibition has led us to propose that COX inhibitors may act centrally in the dorsal horn. In the present study, we examined whether COX inhibition can also suppress peripheral sensitization in meningeal nociceptors. Using single-unit recording in the trigeminal ganglion in vivo, we found that intravenous infusion of naproxen, a non-selective COX inhibitor, reversed measures of sensitization induced in meningeal nociceptors by prior exposure of the dura to inflammatory soup (IS): ongoing activity of A,- and C-units and their response magnitude to mechanical stimulation of the dura, which were enhanced after IS, returned to baseline after naproxen infusion. Topical application of naproxen or the selective COX-2 inhibitor N -[2-(cyclohexyloxy)-4-nitrophenyl]-methanesulfonamide (NS-398) onto the dural receptive field of A,- and C-unit nociceptors also reversed the neuronal hyper-responsiveness to mechanical stimulation of the dura. The findings suggest that local COX activity in the dura could mediate the peripheral sensitization that underlies migraine headache. [source]

Unilateral Black Hairy Tongue in Trigeminal Neuralgia

HEADACHE, Issue 9 2004
William P. Cheshire Jr. MD
Unilateral hairlike discoloration of the tongue is described in a patient with ipsilateral mandibular division trigeminal neuralgia. This unusual physical sign coincided with the patient's painful trigger zone and was attributed to hypertrophy of keratinized filiform papillae, where guarded avoidance of mechanical stimulation over time prevented normal desquamation. [source]

Aged mice have enhanced endocortical response and normal periosteal response compared with young-adult mice following 1 week of axial tibial compression

Michael D Brodt
Abstract With aging, the skeleton may lose its ability to respond to positive mechanical stimuli. We hypothesized that aged mice are less responsive to loading than young-adult mice. We subjected aged (22 months) and young-adult (7 months) BALB/c male mice to daily bouts of axial tibial compression for 1 week and evaluated cortical and trabecular responses using micro,computed tomography (µCT) and dynamic histomorphometry. The right legs of 95 mice were loaded for 60 rest-inserted cycles per day to 8, 10, or 12,N peak force (generating mid-diaphyseal strains of 900 to 1900 µ, endocortically and 1400 to 3100 µ, periosteally). At the mid-diaphysis, mice from both age groups showed a strong anabolic response on the endocortex (Ec) and periosteum (Ps) [Ec.MS/BS and Ps. MS/BS: loaded (right) versus control (left), p,<,.05]. Generally, bone formation increased with increasing peak force. At the endocortical surface, contrary to our hypothesis, aged mice had a significantly greater response to loading than young-adult mice (Ec.MS/BS and Ec.BFR/BS: 22 months versus 7 months, p,<,.001). Responses at the periosteal surface did not differ between age groups (p,>,.05). The loading-induced increase in bone formation resulted in increased cortical area in both age groups (loaded versus control, p,<,.05). In contrast to the strong cortical response, loading only weakly stimulated trabecular bone formation. Serial (in vivo) µCT examinations at the proximal metaphysis revealed that loading caused a loss of trabecular bone in 7-month-old mice, whereas it appeared to prevent bone loss in 22-month-old mice. In summary, 1 week of daily tibial compression stimulated a robust endocortical and periosteal bone-formation response at the mid-diaphysis in both young-adult and aged male BALB/c mice. We conclude that aging does not limit the short-term anabolic response of cortical bone to mechanical stimulation in our animal model. © 2010 American Society for Bone and Mineral Research [source]

Tower Climbing Exercise Started 3 Months After Ovariectomy Recovers Bone Strength of the Femur and Lumbar Vertebrae in Aged Osteopenic Rats,

Takuya Notomi
Abstract To determine both the preventive and recovery effects of tower climbing exercise on mass, strength, and local turnover of bone in ovariectomized (OVX) rats, we carried out two experiments. In experiment I, 60 Sprague-Dawley rats, 12 months of age, were assigned to four groups: a Baseline Control, Sham-Operated Sedentary, OVX-Sedentary and OVX-Exercise rats. Rats voluntarily climbed a 200-cm tower to drink water from a bottle set at the top. At 3 months, OVX elevated both the femoral cortex and lumbar trabecular turnover, leading to a reduction in bone mass and strength. However, in OVX-Exercise rats, those values were maintained at the same level as in the Sham-Sedentary rats. Thus, the climbing exercise, started after 3 days of OVX, prevented OVX-induced cortical and trabecular bone loss by depressing turnover elevation. After confirming the preventive effect, we evaluated the recovery effect of exercise. In experiment II, 90 Sprague-Dawley rats, 12 months of age, were assigned to six groups: a Baseline control, two groups of Sham-Operated Sedentary and OVX-Sedentary, and OVX-Exercise rats. The exercise started 3 months after the OVX operation. At 3 months, OVX increased the trabecular bone formation rate and osteoclast surface, leading to a decrease in compressive strength. In the midfemur, the cross-sectional area, moment of inertia, and bending load values decreased. At 6 months, in the OVX-Exercise rats, the parameters of breaking load in both the lumbar and midfemur, lumbar bone mass, and the total cross-sectional area recovered to the same levels as those in the Sham-Sedentary rats. However, the cortical bone area did not recover. Periosteal bone formation increased, while endosteal bone formation decreased. These results showed that the climbing exercise had both a preventive and recovery effect on bone strength in OVX rats. In the mid-femur, effects on bone formation were site-specific, and the cross-sectional morphology was improved without an increase in cortical bone area, supporting cortical drift by mechanical stimulation. [source]

Human Bone Cell Hyperpolarization Response to Cyclical Mechanical Strain Is Mediated by an Interleukin-1, Autocrine/Paracrine Loop

D. M. Salter
Abstract Mechanical stimuli imparted by stretch, pressure, tension, fluid flow, and shear stress result in a variety of biochemical responses important in bone (re)modeling. The molecules involved in the recognition and transduction of mechanical stimuli that lead to modulation of bone cell function are not yet fully characterized. Cyclical pressure-induced strain (PIS) induces a rapid change in membrane potential of human bone cells (HBC) because of opening of membrane ion channels. This response is mediated via integrins and requires tyrosine kinase activity and an intact actin cytoskeleton. We have used this electrophysiological response to further study the signaling events occurring early after mechanical stimulation of HBC. Stimulation of HBC at 0.33Hz PIS, but not 0.104 Hz PIS, results in the production of a transferable factor that induces membrane hyperpolarization of unstimulated HBC. The production of this factor is inhibited by antibodies to ,1-integrin. Interleukin-1, (IL-1,) and prostaglandin E2 (PGE2) were identified as candidate molecules for the transferable factor as both were shown to induce HBC hyperpolarization by opening of small conductance calcium-activated potassium channels, the means by which 0.33 Hz PIS causes HBC hyperpolarization. Antibodies to IL-1,, but not other cytokines studied, inhibit the hyperpolarization response of HBC to 0.33 Hz PIS. Comparison of the signaling pathways required for 0.33 Hz PIS and IL-1,-induced membrane hyperpolarization shows that both involve the phospholipase C/inositol triphosphate pathway, protein kinase C (PKC), and prostaglandin synthesis. Unlike 0.33 Hz PIS-induced membrane hyperpolarization, IL-1,-induced hyperpolarization does not require tyrosine kinase activity or an intact actin cytoskeleton. These studies suggest that 0.33 Hz PIS of HBC induces a rapid, integrin-mediated, release of IL-1, with a subsequent autocrine/paracrine loop resulting in membrane hyperpolarization. IL-1, production in response to mechanical stimuli is potentially of importance in regulation of bone (re)modeling. [source]

Toothbrushing promotes gingival fibroblast proliferation more effectively than removal of dental plaque

Masazumi Horiuchi
Abstract Objectives: Removal of dental plaque is an essential element of periodontal treatment. However, there have also been studies of the effects of the mechanical stimulation provided by toothbrushing on gingival host-defense mechanisms. The aim of the study was to evaluate the effects of toothbrushing on gingival fibroblast proliferation in dogs over time, compared to effects of plaque removal without brushing. Methods: The mouths of six mongrel dogs were divided into four quadrants: two for daily toothbrushing, and two for daily plaque removal with a curette. After 1, 3 and 5 weeks of treatment, histometrical analyses were performed to assess inflammatory cell infiltration, proliferating cell nuclear antigen (PCNA)-positive fibroblasts, procollagen type I-positive fibroblasts in the subepithelial connective tissue of junctional epithelium. Results: Toothbrushing increased the number of PCNA-positive fibroblasts in the first week, increased the number of type I procollagen-positive fibroblasts at the fifth week, and reduced inflammatory cell infiltration at the third week. Conclusion: These findings suggest that mechanically stimulated fibroblasts begin proliferating within a week, and this cell division results in an increased number of fibroblasts at the third week. It takes 5 weeks before differences in collagen synthesis between brushing and plaque removal areas are detectable. Zusammenfassung Die Proliferation der gingivalen Fibroblasten wird durch Zähneputzen wirkungsvoller gefördert als durch Plaqueentfernung Ziele: Die Entfernung von Zahnplaque ist ein essenzieller Bestandteil der Parodontalbehandlung. Es gibt jedoch auch Studien über die Wirkung einer durch Zähneputzen bewirkten mechanischen Stimulation der gingivalen Abwehrmechanismen. Ziel dieser Studie war es, bei Hunden die Wirkung des Zähneputzen auf die Proliferation der gingivalen Fibroblasten über eine gewisse Zeit zu untersuchen und mit der Wirkung einer Plaqueentfernung ohne Zähneputzen zu vergleichen. Methoden: Das Maul von 6 Mischlingshunden wurde in vier Quadranten unterteilt: zwei mit täglichem Zähneputzen und zwei mit täglicher Plaqueentfernung mittels Kürette. 1, 3 und 5 Wochen nach der Behandlung wurden histometrische Analysen durchgeführt um das entzündliche Zellinfiltrat, die proliferierenden Cell-Nuclear-Antigen (PCNA)-positiven Fibroblasten und die Prokollagen-I-positiven Fibroblasten des subgingivalen Bindegewebes des Saumepithels zu bestimmen. Ergebnisse: Zähneputzen erhöhte in der ersten Woche die Anzahl der PCNA-positiven Fibroblasten, erhöhte bis zur fünften Woche die Anzahl der Type-I-Prokollagen-positiven Fibroblasten und reduzierte das entzündliche Zellinfiltrat bis zur dritten Woche. Schlussfolgerung: Diese Ergebnisse lassen annehmen, dass mechanisch stimulierte Fibroblasten während einer Woche zu proliferieren beginnen und diese Zellteilung eine erhöhte Anzahl von Fibroblasten in der dritten Woche zum Ergebnis hat. Es dauert fünf Wochen bevor zwischen den Bereichen mit Zähneputzen und Plaqueentfernung Unterschiede in der Kollagensynthese nachweisbar sind. Résumé Le brossage dentaire favorise la prolifération des fibroblastes gingivaux d'une manière plus efficace que l'enlèvement de la plaque dentaire L'enlèvement de la plaque dentaire est un élément essentiel dans le traitement parodontal. Cependant, des études ont été menées sur les effets de la stimulation mécanique produit par le brossage dentaire sur les mécanismes de défense de l'hôte au niveau gingival. Le but de l'étude présente a été d'évaluer les effets du brossage dentaire sur la prolifération des fibroblastes gingivaux chez les chiens dans le temps, comparés aux effets de l'enlèvement de la plaque dentaire sans brossage. Les bouches de six chiens bâtards ont été divisés en quatre quadrants : deux pour un brossage dentaire journalier et deux pour l'enlèvement journalier de la plaque à l'aide d'une curette. Après une, trois et cinq semaines de traitement, les analyses histométriques ont été effectuées pour évaluer l'infiltration cellulaire inflammatoire, les fibroblastes positifs à l'antigène du noyau cellulaire proliférant (PCNA), les fibroblastes positifs au procollagène-I dans le tissu conjonctif sous-épithélial de l'épithélium de jonction. Le brossage dentaire augmentait le nombre de fibroblastes positifs (PCNA) durant la première semaine, augmentait le nombre de fibroblastes positifs au collagène type-1 à la cinquième semaine et réduisait l'infiltration cellulaire inflammatoire à la troisième semaine. Ces découvertes suggèrent que les fibroblastes stimulés mécaniquement commencent à proliférer en une semaine, et cette division cellulaire abouti en un nombre plus important de fibroblastes à la troisième semaine. Il faut attendre cinq semaines avant que des différences dans la synthèse du collagène entre les zones de brossage et d'enlèvement de la plaque dentaire ne soient détectables. [source]

Oral mucosal versus cutaneous sensory testing: a review of the literature

R. Jacobs
summary, The innervation of skin and oral mucosa plays a major physiological role in exteroception. It also has a clinical interest as illustrated by sensory changes after neurosurgical procedures. These sensory changes often rely only on the patients' subjective reports, although objective assessments are possible. This review compares the neurophysiological features of the trigeminal sensory pathways with those of cutaneous sensory innervation. In this review, three receptor groups will be discussed: mechanoreceptors, thermoreceptors and nociceptors. Differences between receptors in the glabrous skin, the hairy skin and the oral mucosa will be highlighted. Sensory testing devices have been developed to quantify psychophysiological parameters such as the threshold level for receptor activation upon mechanical stimulation, but such devices have been merely developed to determine the threshold of skin receptors (tactile, thermal). Later on, some have been adapted to suit the particularities of the oral environment. This review attempts to compare the available literature on test devices for oral versus cutaneous tactile function. It summarizes what is common or rather particular to the devices used to study either cutaneous or oral receptors. [source]

Improved bioengineered cartilage tissue formation following cyclic compression is dependent on upregulation of MT1-MMP

J. N. Amrith De Croos
Abstract The generation of bioengineered cartilage tissue suitable for transplantation is a potential therapy to treat damaged cartilage. We have shown previously that the physical and biomechanical properties of bioengineered cartilage can be improved by the application of 30,min of cyclic compression by a mechanism involving sequential upregulation of gene and protein levels of membrane type-1 matrix metalloproteinase (MT1-MMP) and MMP-13. In the current study, we demonstrated that MT1-MMP is critical to this response, as blocking the upregulation of MT1-MMP prevented the improvement in tissue formation. MT1-MMP seems to act by inducing tissue remodeling as evidenced by the presence of aggrecan degradation products by Western blot analysis and increased release of matrix molecules into the media. Release of these molecules was diminished when MT1-MMP upregulation was prevented. This matrix degradation was likely due to MT1-MMP, as under conditions where MMP-13 expression is maintained (stimulation in the presence of MT1-MMP siRNA) the release of these matrix molecules into the media was still prevented. It also appears that MT1-MMP does not regulate MMP-13 gene expression, as MT1-MMP-siRNA pretreatment had no effect on MMP-13 expression following mechanical stimulation. Further analysis of the anabolic genes and proteins involved in mechanically stimulated cartilage will lead to better understanding of the mechanism(s) underlying tissue formation yielding improved bioengineered cartilage. © 2010 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 28:921,927, 2010 [source]

Pulsating fluid flow modulates gene expression of proteins involved in Wnt signaling pathways in osteocytes

Ana Santos
Abstract Strain-derived flow of interstitial fluid activates signal transduction pathways in osteocytes that regulate bone mechanical adaptation. Wnts are involved in this process, but whether mechanical loading modulates Wnt signaling in osteocytes is unclear. We assessed whether mechanical stimulation by pulsating fluid flow (PFF) leads to functional Wnt production, and whether nitric oxide (NO) is important for activation of the canonical Wnt signaling pathway in MLO-Y4 osteocytes. MC3T3-E1 osteoblasts were studied as a positive control for the MLO-Y4 osteocyte response to mechanical loading. MLO-Y4 osteocytes and MC3T3-E1 osteoblasts were submitted to 1-h PFF (0.7,±,0.3 Pa, 5 Hz), and postincubated (PI) without PFF for 0.5,3 h. Gene expression of proteins related to the Wnt canonical and noncanonical pathways were studied using real-time polymerase chain reaction (PCR). In MLO-Y4 osteocytes, PFF upregulated gene expression of Wnt3a, c-jun, connexin 43, and CD44 at 1,3-h PI. In MC3T3-E1 osteoblasts, PFF downregulated gene expression of Wnt5a and c-jun at 0.5,3-h PI. In MLO-Y4 osteocytes, gene expression of PFF-induced Wnt target genes was suppressed by the Wnt antagonist sFRP4, suggesting that loading activates the Wnt canonical pathway through functional Wnt production. The NO inhibitor L-NAME suppressed the effect of PFF on gene expression of Wnt target genes, suggesting that NO might play a role in PFF-induced Wnt production. The response to PFF differed in MC3T3-E1 osteoblasts. Because Wnt signaling is important for bone mass regulation, osteocytes might orchestrate loading-induced bone remodeling through, among others, Wnts. © 2009 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 27:1280,1287, 2009 [source]

Functional tissue engineering for tendon repair: A multidisciplinary strategy using mesenchymal stem cells, bioscaffolds, and mechanical stimulation,

David L. Butler
Abstract Over the past 8 years, our group has been continuously improving tendon repair using a functional tissue engineering (FTE) paradigm. This paradigm was motivated by inconsistent clinical results after tendon repair and reconstruction, and the modest biomechanical improvements we observed after repair of rabbit central patellar tendon defects using mesenchymal stem cell-gel-suture constructs. Although possessing a significantly higher stiffness and failure force than for natural healing, these first generation constructs were quite weak compared to normal tendon. Fundamental to the new FTE paradigm was the need to determine in vivo forces to which the repair tissue might be exposed. We first recorded these force patterns in two normal tendon models and then compared these peak forces to those for repairs of central defects in the rabbit patellar tendon model (PT). Replacing the suture with end-posts in culture and lowering the mesenchymal stem cell (MSC) concentration of these constructs resulted in failure forces greater than peak in vivo forces that were measured for all the studied activities. Augmenting the gel with a type I collagen sponge further increased repair stiffness and maximum force, and resulted in the repair tangent stiffness matching normal stiffness up to peak in vivo forces. Mechanically stimulating these constructs in bioreactors further enhanced repair biomechanics compared to normal. We are now optimizing components of the mechanical signal that is delivered in culture to further improve construct and repair outcome. Our contributions in the area of tendon functional tissue engineering have the potential to create functional load-bearing repairs that will revolutionize surgical reconstruction after tendon and ligament injury. © 2007 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 26:1,9, 2008 [source]


P. Von Dassow
Many marine planktonic dinoflagellates emit flashes of light in response to either laminar or turbulent flows as well as direct mechanical stimulation. The production of a flash of light is known to be mediated by a proton-mediated action potential across the vacuolar membrane; the mechanotransduction process initiating this action potential is unknown. Here we report on an investigation into the role of Ca+2 in the mechanotransduction process regulating bioluminescence in the red tide dinoflagellate Lingulodinium polyedrum. Calcium ionophores and low concentrations of the membrane-disrupting agent digitonin stimulated bioluminescence only when calcium was present in the media or added with the agent, indicating that the flash-triggering vacuolar action potential is specifically stimulated by a calcium influx. A variety of known calcium channel blockers or antagonists inhibited mechanically stimulated bioluminescence but did not affect cellular bioluminescent capacity. In many cases the inhibitory affect occurred after only a brief exposure. In addition, gadolinium (Gd+3), a blocker of many stretch-activated ion channels, caused potent inhibition of mechanically stimulated bioluminescence. The order of potency of the transition metals tested was La+3 > Gd+3 > Co+2 > Mn+2 > Ni+2, similar to their potency as blockers of known calcium channels. Experiments with a quantified shear flow demonstrated that flow-stimulated bioluminescence depended on the level of extracellular calcium. Future work will elucidate the signaling pathway involving calcium-mediated flow-stimulated mechanotransduction. Our goal is to use bioluminescence as a proxy for the initial cellular mechanotransduction events triggered by fluid flow. [source]

Current opportunities and challenges in skeletal muscle tissue engineering

Merel Koning
Abstract The purpose of this article is to give a concise review of the current state of the art in tissue engineering (TE) of skeletal muscle and the opportunities and challenges for future clinical applicability. The endogenous progenitor cells of skeletal muscle, i.e. satellite cells, show a high proneness to muscular differentiation, in particular exhibiting the same characteristics and function as its donor muscle. This suggests that it is important to use an appropriate progenitor cell, especially in TE facial muscles, which have a exceptional anatomical and fibre composition compared to other skeletal muscle. Muscle TE requires an instructive scaffold for structural support and to regulate the proliferation and differentiation of muscle progenitor cells. Current literature suggests that optimal scaffolding could comprise of a fibrin gel and cultured monolayers of muscle satellite cells obtained through the cell sheet technique. Tissue-engineered muscle constructs require an adequate connection to the vascular system for efficient transport of oxygen, carbon dioxide, nutrients and waste products. Finally, functional and clinically applicable muscle constructs depend on adequate neuromuscular junctions with neural cells. To reach this, it seems important to apply optimal electrical, chemotropic and mechanical stimulation during engineering and discover other factors that influence its formation. Thus, in addition to approaches for myogenesis, we discuss the current status of strategies for angiogenesis and neurogenesis of TE muscle constructs and the significance for future clinical use. Copyright © 2009 John Wiley & Sons, Ltd. [source]

Magnetic resonance elastography in the liver at 3 Tesla using a second harmonic approach

D.A. Herzka
Abstract Magnetic resonance elastography (MRE) using mechanical stimulation has demonstrated diagnostic value and clinical promise in breast, liver, and kidney at 1.5 Tesla (T). However, MRE at 1.5T suffers from long imaging times and would benefit from greater signal-to-noise for more robust postprocessing. We present an MRE sequence modified for liver imaging at 3.0T. To avoid artifacts in the phase images, the sequence maintains a short TE by using a second harmonic approach, including stronger motion encoding gradients, shorter radio frequency pulses and an echo-planar readout. Scan time was decreased by a factor of ,2 relative to 1.5T by using an EPI readout and a higher density sampling of the phase waveform was used to calculate shear stiffness and viscosity. Localized (small region of interest) and global (whole-liver region of interest) measurements in normal healthy subjects compared very favorably with previously published results at 1.5T. There was no significant difference between global and localized measures. Magn Reson Med, 2009. © 2009 Wiley-Liss, Inc. [source]

Peripheral synapses and giant neurons in whip spiders

Rainer Foelix
Among invertebrates the synapses between neurons are generally restricted to ganglia, i.e., to the central nervous system (CNS). As an exception, synapses occur in the sensory nerves of arachnid legs, indicating that some nervous integration is already taking place far out in the periphery. In the antenniform legs of whip spiders (Amblypygi), a very special synaptic circuit is present. These highly modified legs contain several large interneurons (giant neurons) that receive mechanosensory input from 700,1,500 tarsal bristles. Some of the sensory cell axons contact a giant neuron at its short, branched dendrite, a few at the soma, but most synapse onto the long giant axon. The fine structure of these synapses resembles that of typical chemical synapses in other arthropods. Although thousands of sensory fibers converge on a single giant neuron, there is no reduction in the actual number of sensory fibers, because these afferent fibers continue their course to the CNS after having made several en passant synapses onto the giant neuron. Touching a single tarsal bristle is sufficient to elicit action potentials in a giant neuron. Owing to the large diameter of the giant axon (10,20 ,m), the action potentials reach the CNS within 55 ms, at conduction velocities of up to 7 m/s. However, mechanical stimulation of the tarsal bristles does not elicit a fast escape response, in contrast to giant fiber systems in earthworms, certain insects, and crayfishes. A quick escape is observed in whip spiders, but only after stimulation of the filiform hairs (trichobothria) on the regular walking legs. Although the giant fiber system in the antenniform legs undoubtedly provides a fast sensory pathway, its biological significance is not clearly understood at the moment. Microsc. Res. Tech. 58:272,282, 2002. © 2002 Wiley-Liss, Inc. [source]

Polycystins: what polycystic kidney disease tells us about sperm

Abraham L. Kierszenbaum
Abstract Experimental evidence indicates that the membrane-associated proteins polycystin-1 and polycystin-2 operate as a receptor-calcium channel complex that regulates signaling pathways essential for modulation of renal tubulogenesis. Polycystic kidney disease is characterized by defective renal tubular structure and results from mutations in either PKD1 or PKD2 genes. Recent data suggest that polycystin-1 and polycystin-2 might localize to primary cilium in principal cells of renal collecting tubules and are thought to act as mechanosensors of fluid flow and contents. Ciliary bending by fluid flow or mechanical stimulation induce Ca2+ release from intracellular stores, presumably to modulate ion influx in response to tubular fluid flow. Polycystins are also emerging as playing a significant role in sperm development and function. Drosophila polycystin-2 is associated with the head and tail of mature sperm. Targeted disruption of the PKD2 homolog results in nearly complete male sterility without disrupting spermatogenesis. Mutant sperm are motile but are unable to reach the female storage organs (seminal receptacles and spermathecae). The sea urchin polycystin-1-equivalent suPC2 colocalizes with the polycystin-1 homolog REJ3 to the plasma membrane over the acrosomal vesicle. This localization site suggests that the suPC2-REJ3 complex may function as a cation channel mediating acrosome reaction when sperm contact the jelly layer surrounding the egg at fertilization. Future studies leading to the identification of specific ligands for polycystins, including the signaling pathways, might define the puzzling relationship between renal tubular morphogenesis and sperm development and function. Mol. Reprod. Dev. 67: 385,388, 2004. © 2004 Wiley-Liss, Inc. [source]

Recent advances in enteric neurobiology: mechanosensitive interneurons

T. K. Smith
Abstract, Until recently, it was generally assumed that the only intrinsic sensory neuron, or primary afferent neuron, in the gut was the after-hyperpolarizing AH/Type II neuron. AH neurons excited by local chemical and mechanical stimulation of the mucosa appear to be necessary for activating the peristaltic reflex (oral excitation and anal inhibition of the muscle layers) and anally propagating ring like contractions (peristaltic waves) that depend upon smooth muscle tone. However, our recent findings in the guinea-pig distal colon suggest that different neurochemical classes of interneuron in the colon are also mechanosensitive in that they respond directly to changes in muscle length, rather than muscle tone or tension. These interneurons have electrophysiological properties consistent with myenteric S-neurons. Ascending and descending interneurons respond directly to circumferential stretch by generating an ongoing polarized peristaltic reflex activity (oral excitatory and anal inhibitory junction potentials) in the muscle for as long as the stimulus is maintained. Some descending (nitric oxide synthase +ve) interneurons, on the other hand, appear to respond directly to longitudinal stretch and are involved in accommodation and slow transit of faecal pellets down the colon. This review will present recent evidence that suggests some myenteric S interneurons, in addition to AH neurons, behave as intrinsic sensory neurons. [source]

Location of proliferating gingival cells following toothbrushing stimulation

ORAL DISEASES, Issue 1 2007
T Tomofuji
Objectives:, Mechanical stimulation by toothbrushing promotes healing of gingivitis through accelerating cell proliferation. Junctional epithelium proliferates at periodontal pocket formation. A question is arisen whether toothbrushing contributes to the repair of gingival inflammation or deterioration of pocket formation. The location of proliferating cells in gingiva stimulated mechanically by toothbrushing was investigated. Materials and methods:, A total of 24 teeth of dogs underwent daily plaque removal with a curette (plaque removal) or both plaque removal and toothbrushing (toothbrushing). Proliferative activity of gingival cells in six individual zones was evaluated by assaying expression of proliferating cell nuclear antigen (PCNA). Results:, Toothbrushing increased densities of PCNA-positive basal cells in the junctional epithelium, connective tissues adjacent to the junctional epithelium, the alveolar bone of the oral epithelial side and the oral epithelium. However, the densities of PCNA-positive cells at the apical portion of the junctional epithelium, connective tissues adjacent to the cementum and the alveolar bone of the periodontal ligament side did not increase following toothbrushing. Conclusions:, Toothbrushing promotes proliferation of gingival cells other than fibroblasts in periodontium and basal cells in the apical portion of the junctional epithelium. The repair of periodontal tissues might be promoted by toothbrushing within the limit of the direct mechanical stimulation. [source]

Insulin Is Essential for the Recovery from Allodynia Induced by Complete Freund's Adjuvant

PAIN MEDICINE, Issue 9 2010
Gregory P. Casey PhD
Abstract Objective., To determine the effect of streptozotocin (STZ)-induced diabetes on the development and recovery of thermal and mechanical hyperalgesia associated with inflammation induced by subcutaneous injection of complete Freund's adjuvant (CFA). Background., The response to nociceptive injury in diabetes differs from that seen in normal individuals in that diabetic patients have increased susceptibility to infections and recover slowly or incompletely from infections and tissue injury due to an abnormal inflammatory response. We have chosen to examine the effect of STZ-induced hypoinsulinemia on the hyperalgesia associated with the enhanced inflammatory state that is induced by the subcutaneous injection of CFA to delineate the potential role of insulin in the development of chronic pain. Methods., STZ- and vehicle-treated Sprague-Dawley rats were tested using thermal and mechanical stimulation after subcutaneous injection of CFA. The behavioral response was compared with that similarly determined in non-diabetic controls and insulin-depleted rats that received insulin replacement. Results., Recovery of the thermal hyperalgesic response to baseline levels occurred over a period of 9,14 days, but the allodynic response to mechanical stimulation persisted for the duration of the study in STZ-treated rats. Insulin replacement prevented the delay in recovery of mechanical allodynia, but had no obvious effect on nociception in uninflamed tissue. Conclusions., Normal insulin function is essential for recovery from mechanical allodynia associated with inflammation induced by CFA. Altered insulin metabolism may selectively influence fiber-type specific mechanisms related to mechanical allodynia associated with inflammation and wound healing. [source]

Molecular control of ethylene production by cyanide in Arabidopsis thaliana

Jennifer McMahon Smith
Although cyanide has long been recognized as a co-product of ethylene synthesis, little attention has been given to its potential physiological and molecular roles. In the present work, the long-term effects of cyanide on growth and development were observed in Arabidopsis thaliana. Two days after a single 20-min application of cyanide, plants demonstrated visible signs of stress. Long-term detrimental effects on growth and photosynthetic capabilities were noted, including low chlorophyll accumulation and stunted growth. Because of the relationship between cyanide and ethylene production, we chose to evaluate the results of cyanide treatment on genes encoding proteins involved in ethylene synthesis. We have found that only the 1-aminocyclopropane-1-carboxylic acid (ACC) synthase gene, ACS6, is rapidly activated in response to cyanide treatment, while other ACS genes were unaffected. This same gene has previously been shown to be transcriptionally activated in response to touch and other environmental stimuli. Cyanide was capable of activating ACS6 transcription within 10 min of treatment, and the amount of transcript correlated positively with the cyanide dosage. Due to the toxic nature of cyanide, plant in vivo concentrations are generally maintained lower than 10 ,M, but can increase under certain stresses. In the present work, we observed that physiologically relevant concentrations as low as 1 ,M HCN, considered metabolically ,safe', were capable of initiating ACS6 transcription. ACS6 transcripts were not substantially reduced as a result of multiple cyanide treatments, which is in contrast with the effects of mechanical stimulation on transcription. Our results suggest a relationship between cyanide production during ethylene synthesis and the molecular control of ethylene synthesis. This work corresponds with earlier experiments that have demonstrated that ethylene and cyanide can elicit some similar physiological responses. It is possible that cyanide may play an active role in ethylene regulation under conditions where rapid cyanide accumulation occurs. Since cyanide can rapidly activate ethylene synthesis, it is possible that it is involved in the positive-feedback regulation of ethylene that occurs in some plant tissues. [source]

Pharmacological properties of the methanol extract from Mentha suaveolens Ehrh.

Lucrecia Moreno
Abstract The present study analyses the pharmacological activity in in vivo and in vitro models of the methanol extract obtained from the leaves and steams of Menta suaveolens Ehrh. This extract lacked toxicity, but exhibited a central nervous system depressant action; an analgesic effect in models of chemical and mechanical stimulation suggesting the induction of a peripheral analgesic response. The extract also exhibited an antiinflammatory action inhibiting the rat paw oedema induced by carrageenin. Moreover, the in vitro studies showed a significant diminution in the contractile effects induced by histamine, serotonin and acetylcholine. Copyright © 2002 John Wiley & Sons, Ltd. [source]

Effect of steady torque twisting on the orientation of cortical microtubules in the epidermis of the sunflower hypocotyl

PLANT BIOLOGY, Issue 4 2008
J. Elsner
Abstract Orientation of cortical microtubules (cMTs) is suggested to be affected by mechanical stress existing in cell walls. However, in mutants exhibiting helical (chiral) growth, there is a correlation between orientation of cMTs in outer tissues and helical growth direction. The aim of this research was to examine the effect of a chiral mechanical stimulation on cMTs. For this purpose, the orientation of cMTs was investigated in hypocotyls subjected to either a right- or a left-handed twist, resulting from a steady torque. cMTs were visualised in fixed material using the immunofluorescence method. The cMTs in untouched control hypocotyls were mostly transverse with respect to the cell long axis. In immobilised, but not twisted control hypocotyls, the transverse orientation was also most frequent, while applied twisting resulted in a change in cMT orientation from transverse to oblique. The data provide additional evidence that changes in tissue stress can be reorganized by cortical microtubules. [source]

Mimosa pudica: Electrical and mechanical stimulation of plant movements

ABSTRACT Thigmonastic movements in the sensitive plant Mimosa pudica L., associated with fast responses to environmental stimuli, appear to be regulated through electrical and chemical signal transductions. The thigmonastic responses of M. pudica can be considered in three stages: stimulus perception, electrical signal transmission and induction of mechanical, hydrodynamical and biochemical responses. We investigated the mechanical movements of the pinnae and petioles in M. pudica induced by the electrical stimulation of a pulvinus, petiole, secondary pulvinus or pinna by a low electrical voltage and charge. The threshold value was 1.3,1.5 V of applied voltage and 2 to 10 µC of charge for the closing of the pinnules. Both voltage and electrical charge are responsible for the electro-stimulated closing of a leaf. The mechanism behind closing the leaf in M. pudica is discussed. The hydroelastic curvature mechanism closely describes the kinetics of M. pudica leaf movements. [source]

Jr-ZFP2, encoding a Cys2/His2-type transcription factor, is involved in the early stages of the mechano-perception pathway and specifically expressed in mechanically stimulated tissues in woody plants

ABSTRACT Plants respond to environmental mechanical stimulation, such as wind, by modifying their growth and development. To study the molecular effects of stem bending on 3-week-old walnut trees, a cDNA-AFLP approach was developed. This study allowed the identification of a cDNA, known as Jr-ZFP2, encoding a Cys2/His2-type two-zinc-fingered transcription factor. Reverse transcriptase-polymerase chain reaction analysis confirmed that Jr-ZFP2 mRNA accumulation is rapidly and transiently induced after mechanical stimulation. After bending, Jr-ZFP2 transcript increase was restricted to the stem, the organ where the mechanical solicitation was applied. Furthermore, other abiotic factors, such as cold or salt, did not modify Jr-ZFP2 mRNA accumulation in walnut stems under our experimental conditions, whereas growth studies demonstrated that salt stress was actually perceived by the plants. These results suggest that the regulation of Jr-ZFP2 expression is more sensitive to mechanical stimulus. This gene will be a good marker for studying the early stages of mechanical perception in woody plants. [source]

Neonatal nociceptive somatic stimulation differentially modifies the activity of spinal neurons in rats and results in altered somatic and visceral sensation

Adrian Miranda
The role of intramuscular, low pH saline injections during the neonatal period in the development and maintenance of visceral hyperalgesia has not been systematically studied. We aimed to investigate alterations in visceral sensation and neural circuitry that result from noxious stimuli in early life. Neonatal male Sprague,Dawley rats received sterile saline injections of pH 4.0 or 7.4 in the gastrocnemius muscle starting at postnatal day 8. Injections were given unilaterally every other day for 12 days ending on postnatal day 20. A third group received needle prick only on the same shedule as the second group, while a fourth group was left naïve. At 2 months of age, rats underwent assessment of cutaneous and deep somatic sensitivity using von Frey filaments and gastrocnemius muscle pinch, respectively. A visceromotor response (VMR) to graded colorectal distension (CRD; 10,80 mmHg for 30 s with 180 s interstimulus intervals) was recorded. Extracellular single-unit recordings from the thoracolumbar spinal neurons (T13,L1) were performed in adult pH 4.0 injected and naïve controls. There was no difference in the threshold for response to mechanical stimulation of the paw in rats injected with pH 4.0 saline compared to all other groups. Conversely, rats treated with pH 4.0 saline showed a significant bilateral reduction in withdrawal threshold to muscle pinch as adults (P < 0.05). At colorectal distensions , 20 mmHg, an increase in the VMR was observed in the pH 4.0 injected group compared to all other groups (P < 0.05). Spinal neurons were classified as short latency abrupt (SL-A) or short latency sustained (SL-S). Spontaneous firing of SL-S (20.6 ± 2.2 impulses s,1), but not SL-A neurons (5.3 ± 0.9 impulses s,1) in the pH 4.0 treated rats was significantly higher than in control rats (SL-S, 2.6 ± 0.8 impulses s,1; SL-A, 3.1 ± 0.7 impulses s,1). The response of SL-S neurons to CRD in the pH 4.0 group was significantly higher at distension pressures , 20 mmHg. Nociceptive somatic stimulation in neonatal rats results in chronic deep somatic and visceral hyperalgesia in adulthood. Colorectal distension-sensitive SL-S neurons are primarily sensitized to neonatal somatic stimulation. [source]