Muscle Microcirculatory Responses to Incremental Exercises Are Correlated with Peak Oxygen Uptake in Individuals With and Without Type 2 Diabetes Mellitus.

Background: The role of impaired oxygen extraction on peak oxygen uptake (V̇O2peak) has been extensively studied using noninvasive and indirect methods in both diabetic patients and healthy participants. Methods: A total of 22 participants with type 2 diabetes mellitus [T2DM; median (range) age: 60 (47-70) years] and 22 controls [58 (52-69) years] with no history of diabetes were recruited (reference no. 201812135RINB). Subjects performed an exhaustive incremental exercise and were evaluated using a gas analyzer and near-infrared spectroscopy (NIRS) to determine V̇O2peak and changes in muscle oxygenation (SmO2) in the vastus lateralis, respectively. Measurements were taken at rest, warm-up, a period during exercise when SmO2 reached a minimum saturation plateau, and recovery. The microcirculatory responses of the vastus lateralis muscle during incremental exercise in patients with T2DM were compared with those in control individuals, and the correlation between changes in SmO2 and V̇O2peak was estimated. Results: The diabetic group demonstrated lower V̇O2peak, peak workload, peak heart rate, peak minute ventilation (all P < 0.05), and lower SmO2 during the rest, warm-up, and recovery phases (all P < 0.05) compared with the control group. A correlation was observed between the change in SmO2 between the warm-up and plateau value and the V̇O2peak (r = 0.608, P = 0.006). Conclusions: The results obtained in this study using NIRS support the feasibility of directly measuring changes in muscle SmO2 magnitudes to estimate the contributions of peripheral active muscle to systemic O2 uptake (V̇O2) during incremental exercise.

Cyclic mechanical stretch regulates the AMPK/Egr1 pathway in tenocytes via Ca2+-mediated mechanosensing.

PURPOSE: Mechanical stimuli are essential for the maintenance of tendon tissue homeostasis. The study aims to elucidate the mechanobiological mechanisms underlying the maintenance of tenocyte homeostasis by cyclic mechanical stretch under high-glucose (HG) condition. MATERIALS AND METHODS: Primary tenocytes were isolated from rat Achilles tendon and 2D-cultured under HG condition. The in vitro effects of a single bout, 2-h cyclic biaxial stretch session (1 Hz, 8%) on primary rat tenocytes were explored through Flexcell system. Cell viability, tenogenic gene expression, intracellular calcium concentration, focal adhesion kinase (FAK) expression, and signaling pathway activation were analyzed in tenocytes with or without mechanical stretch. RESULTS: Mechanical stretch increased tenocyte proliferation and upregulated early growth response protein 1 (Egr1) expression. An increase in intracellular calcium was observed after 30 min of stretching. Mechanical stretch phosphorylated FAK, calmodulin-dependent protein kinase kinase 2 (CaMKK2), and 5' adenosine monophosphate-activated protein kinase (AMPK) in a time-dependent manner, and these effects were abrogated after blocking intracellular calcium. Inhibition of FAK, CaMKK2, and AMPK downregulated the expression of Egr1. In addition, mechanical stretch reinforced cytoskeletal organization via calcium (Ca2+)/FAK signaling. CONCLUSIONS: Our study demonstrated that mechanical stretch-induced calcium influx activated CaMKK2/AMPK signaling and FAK-cytoskeleton reorganization, thereby promoting the expression of Egr1, which may help maintain tendon cell characteristics and homeostasis in the context of diabetic tendinopathy.

Greater Cortical Activation and Motor Recovery Following Mirror Therapy Immediately after Peripheral Nerve Repair of the Forearm.

Cortical reorganization occurs immediately after peripheral nerve injury, and early sensorimotor training is suggested during nerve regeneration. The effect of mirror therapy and classical sensory relearning on cortical activation immediately after peripheral nerve repair of the forearm is unknown. Six participants were randomly assigned to the mirror-therapy group or the sensory-relearning group. Sensorimotor training was conducted in a mirror box for 12 weeks. The mirror-therapy group used mirror reflection of the unaffected hand in order to train the affected hand, and the sensory-relearning group trained without mirror reflection. Semmes-Weinstein Monofilaments (SWM) test, static 2-point discrimination test (S-2PD), grip strength, and the Disabilities of the Arm, Shoulder and Hand (DASH) scores were measured at baseline, the end of the intervention (T1), and 3 months after the intervention (T2). Finger and manual dexterity were measured at T1 and T2, and a functional MRI (fMRI) was conducted at T1. All participants showed improvement in the SWM, S-2PD tests, upper extremity function, and grip strength after the intervention at T1, except for the participant who injured both the median and ulnar nerves in the sensory-relearning group. In addition, the mirror-therapy group had better outcomes in finger dexterity and manual dexterity, and fMRIs showed greater activation in the multimodal association cortices and ipsilateral brain areas during motor tasks. This study provides evidence-based results confirming the benefits of early sensorimotor relearning for cortical activation in peripheral nerve injury of the forearm and different neuroplasticity patterns between mirror therapy and classical sensor relearning.

Effects of Conscious Control of Scapular Orientation in Oral Cancer Survivors With Scapular Dyskinesis: A Randomized Controlled Trial.

OBJECTIVES: Spinal accessory nerve dysfunction is one of the complications of neck dissection in patients with oral cancer. This study aimed to explore the effects of long-term scapular-focused exercises and conscious control of scapular orientation on scapular movement and quality of life (QoL). METHODS: This study was a randomized controlled trial with concealed allocation, assessor blinding, and intention-to-treat analysis. Thirty-six patients with oral cancer were randomly allocated to the motor-control group (scapular-focused exercise + conscious control of scapular orientation) or the regular-exercise group (scapular-focused exercises only). Both groups received conventional physical therapy after neck dissection for 3 months. Shoulder pain intensity, active range of motion (AROM) of shoulder abduction, scapular muscle strength and activity under maximal voluntary isometric contraction (MVIC), scapular muscle activity when performing scapular movements, and QoL were measured at baseline, 1 month after the start of the intervention, and the end of the intervention. RESULTS: Both groups showed significant improvement in all outcomes except shoulder pain intensity. After the 3-month intervention, the motor-control group had more significant improvement in AROM of shoulder abduction with a 19° difference (95% CI: 10-29, P < .001), muscle strength of upper trapezius with an 11 N difference (95% CI: 2-20; P = .021), and QoL than the regular-exercise group. When performing shoulder horizontal adduction and flexion, the relative value (%MVIC) of serratus anterior was smaller in the motor-control group with a 106%MVIC difference (95% CI: 7-205, P = .037). CONCLUSIONS: Scapular-focused exercises have promising effects on spinal accessory nerve dysfunction. Combining scapular-focused exercises with conscious control of scapular orientation has more remarkable benefits on AROM of shoulder abduction, UT muscle strength, and muscle activation pattern than the scapular-focused exercises alone. Conscious control of scapular orientation should be considered to integrate into scapular-focused exercises in patients with oral cancer and scapular dyskinesis.Trial registry name and URL, and registration number: ClinicalTrials.gov (URL: https://clinicaltrials.gov; Approval No: NCT03545100).

Microcirculatory Responses to Muscle and Tendon Exercises in Individuals With and Without Type 2 Diabetes Mellitus and the Association Between Microcirculatory and Exercise Performance.

Background: This study aimed to measure and compare (1) the microcirculation and microcirculatory responses of the muscles and tendons at rest and during isometric muscle contractions in participants with and without diabetes mellitus (DM) and (2) to determine correlations between microcirculation and muscle strength. Methods: Sixty-three participants with type 2 DM and 42 physically matched controls were recruited. Baseline measurements of the microcirculation of the rectus femoris (RF) and medial gastrocnemius (MG) muscles and patellar (PT) and Achilles tendons (AT), as well as their microcirculatory changes during maximal isometric exercises, were performed and recorded by using near-infrared spectroscopy and a red laser. Data on various laboratory tests (including glycated hemoglobin, triglyceride, high-density cholesterol), the monofilament test, and the ankle-brachial index were also obtained. Results: The baseline measurements indicated that, compared with the controls, the diabetic participants had lower oxygen saturation (SpO2) in their RF and MG muscles (both P < 0.001), and the total hemoglobin in the diabetic PT and AT was higher (P = 0.001 and P = 0.01). The minimal SpO2 levels in the aforementioned muscles during isometric contractions were lower in the diabetes group than in the control group (P ≤ 0.001). Furthermore, there were correlations between the microcirculatory change of the RF muscle and the knee extension force. Conclusions: This study demonstrated the effects of diabetes on the microcirculation of skeletal muscles and tendons during baseline measurements and responses to maximal isometric exercises. The results support the need for preventive strategies for diabetic muscles to prevent adverse complications when performing resistance training.

Biomimetic Synthesis of Nanocrystalline Hydroxyapatite Composites: Therapeutic Potential and Effects on Bone Regeneration.

The development of a novel alloplastic graft with both osteoinductive and osteoconductive properties is still necessary. In this study, we tried to synthesize a biomimetic hydroxyapatite microspheres (gelatin/nano-hydroxyapatite microsphere embedded with stromal cell-derived factor-1: GHM-S) from nanocrystalline hydroxyapatites and to investigate their therapeutic potential and effects on bone regeneration. In this study, hydroxyapatite was synthesized by co-precipitation of calcium hydroxide and orthophosphoric acid to gelatin solution. The microbial transglutaminase was used as the agent to crosslink the microspheres. The morphology, characterization, and thermal gravimetric analysis of microspheres were performed. SDF-1 release profile and in vitro biocompatibility and relative osteogenic gene expression were analyzed, followed by in vivo micro-computed tomography study and histological analysis. The synthesized hydroxyapatite was found to be similar to hydroxyapatite of natural bone tissue. The stromal cell-derived factor-1 was embedded into gelatin/hydroxyapatite microsphere to form the biomimetic hydroxyapatite microsphere. The stromal cell-derived factor-1 protein could be released in a controlled manner from the biomimetic hydroxyapatite microsphere and form a concentration gradient in the culture environment to attract the migration of stem cells. Gene expression and protein expression indicated that stem cells could differentiate or develop into pre-osteoblasts. The effect of bone formation by the biomimetic hydroxyapatite microsphere was assessed by an in vivo rats' alveolar bone defects model and confirmed by micro-CT imaging and histological examination. Our findings demonstrated that the biomimetic hydroxyapatite microsphere can enhance the alveolar bone regeneration. This design has potential be applied to other bone defects.

Acute and Long-Term Effects of Mechanotherapy on the Outcome After an Achilles Repair: A Prospective Cohort Study With Historical Controls.

OBJECTIVES: To evaluate the effects of vibration on Achilles' tendon microcirculation and characteristics following surgical repair of Achilles' tendon rupture. DESIGN: Cohort study with historical controls. SETTING: A university institute. PARTICIPANTS: Participants (N=32), including 19 (16 men, 3 women; median [range] age: 43.0 [25.0-57.0] years) and 13 (10 men, 3 women; 44.00 [29.0-60.0] years) in the vibration (application to the ball of the foot, 30Hz, 2mm amplitude, 4kg pressure, and self-administration) and control groups, respectively, who underwent unilateral Achilles' tendon repairs were recruited. INTERVENTION: A 4-week vibration intervention in the vibration group. MAIN OUTCOME MEASUREMENTS: The tendon microcirculation was measured after the first session of vibration. The participants were evaluated repeatedly with bilateral follow-up measurements of tendon stiffness, 3 functional outcome tests, and a questionnaire survey. RESULTS: Acute effects of the vibration were observed immediately after the 5-minute vibration (P≤.001). Lower total hemoglobin and oxygen saturation were respectively observed (P=.043) in the repaired legs 3 and 6 months postsurgery in the vibration group as compared with the control group. The vibration group also showed greater tendon stiffness, heel raising height and hopping distance 3 or 6 months postoperation in both the repaired and noninjured legs (all P<.05). The microcirculatory characteristics 2 months postoperation were correlated with the outcomes at 6 months postoperation. CONCLUSIONS: Differences in microcirculatory characteristics and better rehabilitation outcomes were observed in the legs with an Achilles repair that underwent the early vibration intervention.

Muscular Morphomechanical Characteristics After an Achilles Repair.

BACKGROUND: The purpose of the study was to compare the morphomechanical and functional characteristics during maximal isometric, concentric, and eccentric contractions in the legs of patients that underwent unilateral Achilles tendon repair with those in their noninjured control legs. METHODS: Twenty participants (median age = 38.2 years; range, 21.1-57.3 years) who underwent Achilles repair between 3 and 12 months ago were recruited with the following measures: (1) mechanical stiffness of the aponeurosis and (2) electromyography and medial gastrocnemius fascicle angle and length, standing muscle and tendon length, and height of heel rise with isometric contraction. RESULTS: Compared to the noninjured legs, the repaired legs showed less resting fascicle length, standing muscle length, isometric plantarflexion torque, and heel raise distance ( Ps ranged between .044 and <.001). During the concentric and eccentric phases of the raising and lowering test, the repaired legs demonstrated less fascicle length ( P ≤ .028) but greater tendinous tissue length ( Ps ranged between .084 and <.001) and fascicle angle ( Ps ranged between .247 and .008) and fewer change magnitudes of the fascicle length and tendinous tissue length ( P ≤ .003). The change magnitudes of the morphological characteristics showed correlations with the torque or distance. CONCLUSION: Selecting the appropriate surgical repair and rehabilitation for Achilles tendon ruptures is recommended for restoring the length and mechanical strength of the muscle-tendon unit of plantar-flexion muscles. LEVEL OF EVIDENCE: Level III, comparative study.

Functional outcomes and quality of life after a 6-month early intervention program for oral cancer survivors: a single-arm clinical trial.

BACKGROUND: Advanced treatment of oral cancer increases survival rates; however, it also increases the risk of developing shoulder dysfunction, dysphagia, oral dysfunction, donor site morbidity and psychological issues. This single-arm preliminary pilot study aims to explore the effects of a six-month early intervention program following reconstructive surgery in oral cancer survivors. METHODS: A total of 65 participants were analyzed following reconstructive surgery. Outcome measurements were taken during the first visit, and at one, three and six months after reconstructive surgery. RESULTS: Scapular muscle strength and shoulder range of motion progressively improved during the 6-month follow-up. The mean Disability of the Arms, Shoulder and Hand (DASH) score showed significant improvement at 1 month (p < .001). Health related QoL showed significant differences between baseline and 6-months post-surgery scores on global health and on most of the function and symptom scales. The predicted return-to-work rate was 80% at one year after the operation. Return-to-work rate differs in different vocational types, with a higher rate of return in the skilled or semi-skilled (87.5%) and self-employed (86.7%). CONCLUSIONS: We suggest that early integrated intervention program with a follow-up of at least six months following reconstructive surgery may help develop and identify intervention guidelines and goals in the initial six months of treatment following neck dissection in oral cancer survivors.

Hyperglycemia Augments the Adipogenic Transdifferentiation Potential of Tenocytes and Is Alleviated by Cyclic Mechanical Stretch.

Diabetes mellitus is associated with damage to tendons, which may result from cellular dysfunction in response to a hyperglycemic environment. Tenocytes express diminished levels of tendon-associated genes under hyperglycemic conditions. In contrast, mechanical stretch enhances tenogenic differentiation. However, whether hyperglycemia increases the non-tenogenic differentiation potential of tenocytes and whether this can be mitigated by mechanical stretch remains elusive. We explored the in vitro effects of high glucose and mechanical stretch on rat primary tenocytes. Specifically, non-tenogenic gene expression, adipogenic potential, cell migration rate, filamentous actin expression, and the activation of signaling pathways were analyzed in tenocytes treated with high glucose, followed by the presence or absence of mechanical stretch. We analyzed tenocyte phenotype in vivo by immunohistochemistry using an STZ (streptozotocin)-induced long-term diabetic mouse model. High glucose-treated tenocytes expressed higher levels of the adipogenic transcription factors PPARγ and C/EBPs. PPARγ was also highly expressed in diabetic tendons. In addition, increased adipogenic differentiation and decreased cell migration induced by high glucose implicated a fibroblast-to-adipocyte phenotypic change. By applying mechanical stretch to tenocytes in high-glucose conditions, adipogenic differentiation was repressed, while cell motility was enhanced, and fibroblastic morphology and gene expression profiles were strengthened. In part, these effects resulted from a stretch-induced activation of ERK (extracellular signal-regulated kinases) and a concomitant inactivation of Akt. Our results show that mechanical stretch alleviates the augmented adipogenic transdifferentiation potential of high glucose-treated tenocytes and helps maintain their fibroblastic characteristics. The alterations induced by high glucose highlight possible pathological mechanisms for diabetic tendinopathy. Furthermore, the beneficial effects of mechanical stretch on tenocytes suggest that an appropriate physical load possesses therapeutic potential for diabetic tendinopathy.

High glucose alters tendon homeostasis through downregulation of the AMPK/Egr1 pathway.

Diabetes mellitus (DM) is associated with higher risk of tendinopathy, which reduces tolerance to exercise and functional activities and affects lifestyle and glycemic control. Expression of tendon-related genes and matrix metabolism in tenocytes are essential for maintaining physiological functions of tendon. However, the molecular mechanisms involved in diabetic tendinopathy remain unclear. We hypothesized that high glucose (HG) alters the characteristics of tenocyte. Using in vitro 2-week culture of tenocytes, we found that expression of tendon-related genes, including Egr1, Mkx, TGF-ß1, Col1a2, and Bgn, was significantly decreased in HG culture and that higher glucose consumption occurred. Down-regulation of Egr1 by siRNA decreased Scx, Mkx, TGF-ß1, Col1a1, Col1a2, and Bgn expression. Blocking AMPK activation with Compound C reduced the expression of Egr1, Scx, TGF-ß1, Col1a1, Col1a2, and Bgn in the low glucose condition. In addition, histological examination of tendons from diabetic mice displayed larger interfibrillar space and uneven glycoprotein deposition. Thus, we concluded that high glucose alters tendon homeostasis through downregulation of the AMPK/Egr1 pathway and the expression of downstream tendon-related genes in tenocytes. The findings render a molecular basis of the mechanism of diabetic tendinopathy and may help develop preventive and therapeutic strategies for the pathology.

Morphomechanical alterations in the medial gastrocnemius muscle in patients with a repaired Achilles tendon: Associations with outcome measures.

BACKGROUND: Functional deficits are found in ankles that have sustained an Achilles rupture. This study sought to evaluate and compare the morphomechanical characteristics of the medial gastrocnemius muscle in the legs of participants within six months of a unilateral Achilles repair to determine any correlations between those characteristics and objective outcomes and self-reported functional levels. METHODS: Fifteen participants were assessed via measurements of muscle morphologies (fascicle length, pennation angle, and muscle thickness) in a resting state, the mechanical properties of the proximal aponeurosis of the medial gastrocnemius muscle, the pennation angle during ramping maximal voluntary isometric contractions (MVIC), the heel raise test, and the Taiwan Chinese version of the Lower Extremity Functional Scale (LEFS-TC) questionnaire. Findings Compared with the non-injured legs, the repaired legs showed a lower muscle fascicle length (mean 4.4 vs. 5.0cm) and thickness (1.7 vs. 1.9cm), lower stiffness of the GM tendon and aponeurosis (174.1 vs. 375.6N/mm), and a greater GM pennation angle (31.2 vs. 28.9°) during 90% MVIC (all p≤0.05). Correlations were found between the morphomechanical results and maximal heel raise heights or the LEFS-TC score, and between the symmetry ratios of the fascicle lengths and the LEFS-TC score. Interpretation There are decreases in fascicle length, muscle thickness and mechanical properties in the medial gastrocnemius muscles of the participants within the first six months after an Achilles repair. These morphomechanical alterations demonstrate associations with functional levels in the lower extremities and indicated the need for early mobilization of the calf muscles after the repair.

Tissue transglutaminase is involved in mechanical load-induced osteogenic differentiation of human ligamentum flavum cells.

Mechanical load-induced osteogenic differentiation might be the key cellular event in the calcification and ossification of ligamentum flavum. The aim of this study was to investigate the influence of tissue transglutaminase (TGM2) on mechanical load-induced osteogenesis of ligamentum flavum cells. Human ligamentum flavum cells were obtained from 12 patients undergoing lumbar spine surgery. Osteogenic phenotypes of ligamentum flavum cells, such as alkaline phosphatase (ALP), Alizarin red-S stain, and gene expression of osteogenic makers were evaluated following the administration of mechanical load and BMP-2 treatment. The expression of TGM2 was evaluated by real-time PCR, Western blotting, and enzyme-linked immunosorbent assay (ELISA) analysis. Our results showed that mechanical load in combination with BMP-2 enhanced calcium deposition and ALP activity. Mechanical load significantly increased ALP and OC gene expression on day 3, whereas BMP-2 significantly increased ALP, OPN, and Runx2 on day 7. Mechanical load significantly induced TGM2 gene expression and enzyme activity in human ligamentum flavum cells. Exogenous TGM2 increased ALP and OC gene expression; while, inhibited TG activity significantly attenuated mechanical load-induced and TGM2-induced ALP activity. In summary, mechanical load-induced TGM2 expression and enzyme activity is involved in the progression of the calcification of ligamentum flavum.

Entry into vocational rehabilitation program following work-related hand injury: Potential candidates.

OBJECTIVES: This case-control study aimed to investigate the predictors of return to work (RTW) following work-related major forearm, wrist or hand injury at the preparation stage of return to work. MATERIAL AND METHODS: A total of 80 clients were recruited and divided into 2 groups depending on their readiness of RTW. The groups were compared with each other with regard to their demographics, compensation status, hand injury severity, health perception, and time off work (TOW) using correlation coefficient. Predictors of RTW were measured by logistic regression analysis. RESULTS: There were no significant differences in demographics and the severity of hand injury between 2 groups. Self-perceived physical functioning (p = 0.04), vitality (p = 0.01), mental health (p = 0.03) and TOW (p = 0.001) were significantly different between Action group and Preparation group. With binary logistic regression analysis, self-perceived vitality (odds ratio (OR) = 1.041) and TOW (OR = 0.996) were shown to be strongly predictive of RTW at the preparation stage of return to work. CONCLUSIONS: This study has shown that shorter TOW and better self-perceived vitality could predict early readiness for RTW after major work-related forearm, wrist or hand injury.

Effects of combined exercise on gait variability in community-dwelling older adults.

Training that focuses on strength, balance, and endurance, the so-called combined exercise, can enhance physical function, including gait, according to a literature review. However, the effects of combined exercise on improving gait variability are limited. The objective of this study is to investigate the effects of 12 weeks of combined exercise comprised of resistance, endurance, and balance training on gait performance in older adults. Twenty-nine community-dwelling older adults were recruited and assigned to either the experimental group (n = 17) or the control group (n = 12). The 12-week intervention was a combined exercise program at 1 h per day and 3 days per week. The participants received an assessment for both a 6-min walk and gait during both habitual walking and fast walking conditions at pre-intervention and after 8 and 12 weeks of exercise. The 6-min walk was used to assess gait endurance. GAITRite was used to evaluate gait. An analysis of covariance with the pretest score as the covariate was used to determine the difference in each dependent variable between groups. The level of significance was set as p less than 0.05. Our results showed significant between-group effects in the 6-min walk and velocity, stride time, and stride length in both conditions after 8 weeks of exercise and significant between-group effects in the 6-min walk test and all selected gait parameters in both conditions after 12 weeks of exercise. Our findings demonstrate that a 12-week combined exercise program may positively affect gait endurance and gait performance including gait variability in habitual walking and fast walking conditions among older adults. The current study provides important evidence of short-term combined exercise effects on improvements in gait performance.

Second messengers mediating the proliferation and collagen synthesis of tenocytes induced by low-level laser irradiation.

For decades, low-level laser therapy (LLLT) has widespread applications in tendon-related injuries. Although the therapeutic effect of LLLT could be explained by photostimulation of target tissue and cells, how tenocytes sense photonic energy and convert them into cascades of cellular and molecular events is still not well understood. This study was designed to elucidate the effects of LLLT on cell proliferation and collagen synthesis by examining the associated second messengers including ATP, Ca(2+), and nitric oxide using rat Achilles tenocytes. Moreover, proliferating cell nuclear antigen (PCNA) and transforming growth factor-ß1 (TGF-ß1) related to cell proliferation and matrix metabolism were also studied. The results showed that 904 nm GaAs laser of 1 J/cm(2) could significantly increase the MTT activity and collagen synthesis of tenocytes. Second messengers including ATP and intracellular Ca2+ were increased after laser treatment. Quantitative PCR analysis of tenocytes treated with laser revealed up-regulated expression of PCNA, type I collagen, and TGF-ß1. Besides, laser-induced TGF-ß1 expression was significantly inhibited by extracellular signal-regulated kinase (ERK) specific inhibitor (PD98059). The findings suggested that LLLT stimulated ATP production and increased intracellular calcium concentration. Directly or indirectly via production of TGF-ß1, these second messengers mediated the proliferation of tenocytes and synthesis of collagen.

Centrifugal force induces human ligamentum flavum fibroblasts inflammation through activation of JNK and p38 pathways.

Inflammation has been proposed to be an important causative factor in ligamentum flavum hypertrophy. However, the mechanisms of mechanical load on inflammation of ligamentum flavum remain unclear. In this study, we used an in vitro model of human ligamentum flavum fibroblasts subjected to centrifugal force to elucidate the effects of mechanical load on cultured human ligamentum flavum fibroblasts; we further studied its molecular and biochemical mechanisms. Human ligamentum flavum fibroblasts were obtained from six patients undergoing lumbar spine surgery. Monolayer cultures of human ligamentum flavum fibroblasts were subjected to different magnitudes of centrifugal forces. Cell viability, cell death, biochemical response, and molecular response to centrifugal forces were analyzed. It was found that centrifugal stress significantly suppressed cell viability without inducing cell death. Centrifugal force at 67.1 g/cm(2) for 60 min significantly increases the production of prostaglandin E2 and nitric oxide as well as gene expression of proinflammatory cytokines, including interleukin (IL)-1α, IL-1ß and IL-6, showed that centrifugal force-dependent induction of cyclooxygense-2 and inducible NO synthase required JNK and p38 mitogen-activated protein kinase, but not ERK 1/2 activities. This study suggested that centrifugal force does induce inflammatory responses in human ligamentum flavum fibroblasts. The activation of both JNK and p38 mitogen-activated protein kinase mechanotransduction cascades is a crucial intracellular mechanism that mediates cyclooxygense-2/prostaglandin E2 and inducible NO synthase/nitric oxide production.

Elastin-derived peptides induce inflammatory responses through the activation of NF-κB in human ligamentum flavum cells.

The formation of fibrotic tissue in the ligamentum flavum (LF) is usually preceded by breakdown of elastic fibers. Elastin-derived peptides (EDPs) from breakdown of elastic fibers display a wide range of biological activities in a variety of cells, but there is minimal information regarding the involvement in the processes of LF hypertrophy. The aim of this study is to elucidate the effects of EDPs on cultured human LF cells and to investigate their molecular and biochemical mechanisms. Human LF cells were obtained from 18 patients who underwent lumbar spine surgery. After treatment with different concentrations of EDPs with or without specific inhibitors in culture medium, the viability and proliferation of LF cells, genes expression, and the signaling pathways were evaluated and analyzed. It was found that 50 µg/ml EDPs significantly increased cell proliferation and synthesis of prostaglandin E(2). The gene expression and protein production of proinflammatory cytokines, including interleukin-1α (IL-1α), IL-1ß, and IL-6, were also upregulated. The levels of p-ERK (extracellular signal-regulated kinase) and NF-κB increased immediately following EDP treatment and sustained up to 90 min. It was also found that NF-κB inhibitor, but not ERK1/2 inhibitor, attenuated EDP-dependent induction of IL-1α, IL-1ß, and IL-6 expression, indicating that NF-κB pathways are required for EDP-induced IL-1α, IL-1ß, and IL-6 gene expression in human LF cells. The results of this in vitro experiment suggest that EDPs do induce inflammatory responses in human LF cells and plays the key role in the development of LF hypertrophy.

The cross-talk between transforming growth factor-beta1 and ultrasound stimulation during mechanotransduction of rat tenocytes.

Ultrasound is an effective noninvasive treatment for various tendinopathies. However, how tenocytes convert ultrasound stimulation into cascades of cellular and molecular events is not well understood. The purpose of this study is to elucidate the signaling pathways of tenocytes during ultrasound stimulation. Primary cultures of tenocytes were harvested from Achilles tendons of Sprague-Dawley rats. The viability and proliferation of tenocytes, their genes expression, and the signaling pathways after ultrasound treatment with or without specific inhibitors were evaluated and analyzed. The results showed that ultrasound treatment (100 mW/cm(2) for 20 min) significantly enhanced matrix metalloproteinase 13 (MMP-13), c-Fos, and c-Jun gene expression, increased JNK and p38, but not extracellular signal-regulated kinase-1/2 (ERK1/2), phosphorylation at 5 min, and sustained up to 60 min. JNK inhibitor and p38 inhibitor, but not ERK1/2 inhibitor, attenuated ultrasound-dependent induction of MMP-13 expression, indicating that the JNK and p38 pathways are required for ultrasound-induced MMP-13 expression in tenocytes. We also found that SB431542 (transforming growth factor-beta (TGF-ß) receptor kinases inhibitor) suppressed ultrasound-induced MMP?13 and c-Fos gene expression, and p38 phosphorylation. This study revealed that ultrasound treatment stimulates tenocytes proliferation and regulates their matrix metabolism through the cross-talk between TGF-ß and ultrasound-induced mitogen-activated protein kinases (MAPKs) signaling pathways.

Effects of low intensity pulsed ultrasound on rat Schwann cells metabolism.

The effects of low intensity pulsed ultrasound to tenocytes and osteocytes are well understood and applied clinically. However, its effects on cultured Schwann cells are still not well elucidated. This study was designed to elucidate the effects of low intensity pulsed ultrasound on cultured Schwann cells and their possible molecular mechanism. Schwann cells were harvested from sciatic nerves of 3-day-old Sprague-Dawley rats. Low intensity pulsed ultrasound stimulator (frequency: 1 MHz, duration: 2 min, duty cycle: 20%, total treatment time: 3 min) was applied to three different culture conditions: regular culture medium containing 0, 5, or 10% fetal bovine serum. The viability, damage, and differentiation of Schwann cells were examined; gene expression was also analyzed. In the presence of 0.3 W/cm(2) pulsed ultrasound stimulation, increases in cell viability and decreases in cell apoptosis were observed in the serum deprivation group; in this culture condition, interleukin-1, tumor necrosis factor-alpha, and protein zero genes expression were downregulated and Desert Hedgehog transcripts gene expression was upregulated. We concluded that intervention with low intensity pulsed ultrasound could promote Schwann cell proliferation, prevent cell death, and keep adequate phenotype presentation for peripheral nerve recovery. The low intensity pulsed ultrasound stimulation to an injured nerve site could be applied as early as possible especially when the microenvironment is almost serum-free to obtain the most benefit.