Inhibitors of endopeptidase and angiotensin-converting enzyme lead to an amplification of the morphological changes and an upregulation of the substance P system in a muscle overuse model.

BACKGROUND: We have previously observed, in studies on an experimental overuse model, that the tachykinin system may be involved in the processes of muscle inflammation (myositis) and other muscle tissue alterations. To further evaluate the significance of tachykinins in these processes, we have used inhibitors of neutral endopeptidase (NEP) and angiotensin-converting enzyme (ACE), substances which are known to terminate the activity of various endogenously produced substances, including tachykinins. METHODS: Injections of inhibitors of NEP and ACE, as well as the tachykinin substance P (SP), were given locally outside the tendon of the triceps surae muscle of rabbits subjected to marked overuse of this muscle. A control group was given NaCl injections. Evaluations were made at 1 week, a timepoint of overuse when only mild inflammation and limited changes in the muscle structure are noted in animals not treated with inhibitors. Both the soleus and gastrocnemius muscles were examined morphologically and with immunohistochemistry and enzyme immunoassay (EIA). RESULTS: A pronounced inflammation (myositis) and changes in the muscle fiber morphology, including muscle fiber necrosis, occurred in the overused muscles of animals given NEP and ACE inhibitors. The morphological changes were clearly more prominent than for animals subjected to overuse and NaCl injections (NaCl group). A marked SP-like expression, as well as a marked expression of the neurokinin-1 receptor (NK-1R) was found in the affected muscle tissue in response to injections of NEP and ACE inhibitors. The concentration of SP in the muscles was also higher than that for the NaCl group. CONCLUSIONS: The observations show that the local injections of NEP and ACE inhibitors led to marked SP-like and NK-1R immunoreactions, increased SP concentrations, and an amplification of the morphological changes in the tissue. The injections of the inhibitors thus led to a more marked myositis process and an upregulation of the SP system. Endogenously produced substances, out of which the tachykinins conform to one substance family, may play a role in mediating effects in the tissue in a muscle that is subjected to pronounced overuse.

Epidemiology of acute knee injuries seen at the Emergency Department at Umeå University Hospital, Sweden, during 15 years.

PURPOSE: To describe the incidence and injury distribution of knee injuries in the general population of a European setting. METHODS: Retrospective study of all knee injuries registered at the Emergency Department at Umeå University Hospital, Sweden, during 1995-2009 in relation to age, sex, diagnosis, location and activity at the time of injury, mechanism of injury, and treatment and/or follow-up plan. RESULTS: During 1995-2009, 12,663 knee injuries were registered, 8% of all injuries. The incidence of knee injuries resulting in a visit to the Emergency Department was six cases per 1,000 person years. One-third of all injuries occurred during sports. And 30% were 15-24 years. More men than women were injured during sporting activities and women were mostly injured during transportation. CONCLUSION: Knee injuries in a general population are common and the injury distribution varies with age and sex. Sports activities and young age were prominent features of the injured population. LEVEL OF EVIDENCE: IV.

Effects on contralateral muscles after unilateral electrical muscle stimulation and exercise.

It is well established that unilateral exercise can produce contralateral effects. However, it is unclear whether unilateral exercise that leads to muscle injury and inflammation also affects the homologous contralateral muscles. To test the hypothesis that unilateral muscle injury causes contralateral muscle changes, an experimental rabbit model with unilateral muscle overuse caused by a combination of electrical muscle stimulation and exercise (EMS/E) was used. The soleus and gastrocnemius muscles of both exercised and non-exercised legs were analyzed with enzyme- and immunohistochemical methods after 1, 3 and 6 weeks of repeated EMS/E. After 1 w of unilateral EMS/E there were structural muscle changes such as increased variability in fiber size, fiber splitting, internal myonuclei, necrotic fibers, expression of developmental MyHCs, fibrosis and inflammation in the exercised soleus muscle. Only limited changes were found in the exercised gastrocnemius muscle and in both non-exercised contralateral muscles. After 3 w of EMS/E, muscle fiber changes, presence of developmental MyHCs, inflammation, fibrosis and affections of nerve axons and AChE production were observed bilaterally in both the soleus and gastrocnemius muscles. At 6 w of EMS/E, the severity of these changes significantly increased in the soleus muscles and infiltration of fat was observed bilaterally in both the soleus and the gastrocnemius muscles. The affections of the muscles were in all three experimental groups restricted to focal regions of the muscle samples. We conclude that repetitive unilateral muscle overuse caused by EMS/E overtime leads to both degenerative and regenerative tissue changes and myositis not only in the exercised muscles, but also in the homologous non-exercised muscles of the contralateral leg. Although the mechanism behind the contralateral changes is unclear, we suggest that the nervous system is involved in the cross-transfer effects.

VGluT2 and NMDAR1 expression in cells in the inflammatory infiltrates in experimentally induced myositis: evidence of local glutamate signaling suggests autocrine/paracrine effects in an overuse injury model.

It is not known whether a glutamate signaling system is involved in muscle inflammation (myositis). In the present study, we examined this question in the soleus muscle in a laboratory model of myositis resulting from repetitive overuse induced by electrical stimulation and injection of pro-inflammatory substances. Sections of rabbit soleus muscle with an induced myositis, i.e., exhibiting infiltration of inflammatory cells, were examined immunohistochemically using antibodies against vesicular glutamate transporter VGluT2 and the glutamate receptor NMDAR1. In situ hybridization for demonstration of VGluT2 mRNA was also performed. Specific reactions for both VGluT2 and NMDAR1 could be observed immunohistochemically in the same cells. In situ hybridization demonstrated the occurrence of VGluT2 mRNA in the cells. Double staining showed that the VGluT2 reactions were detectable in cells marked with T cell/neutrophil marker and in cells expressing eosinophil peroxidase. These data suggest the occurrence of previously unknown glutamate-mediated autocrine/paracrine effects within the inflammatory infiltrates during the development of muscle inflammation.

Do both areal BMD and injurious falls explain the higher incidence of fractures in women than in men?

The higher incidence of fractures in women than in men is generally attributed to the lower areal bone mineral density (areal BMD, g/cm(2)) of the former. The purpose of the present study was to investigate both areal BMD and injurious falls as risk factors for fractures. In a first cohort, areal BMD was measured in 5,131 men and women (age range 40-95 years). In a second cohort, consisting of 26,565 men and women (age range 40-69 years), a health survey was conducted including questions about lifestyle and medication. Main outcome measures included validated prospective injurious falls and fractures in both cohorts. The higher areal BMD and femoral neck BMD in men compared to women (P < 0.001) were explained by a higher diameter of the femoral neck. Importantly, the diameter of the femoral neck was not associated with fractures in either sex (hazard ratio [HR] 0.94-1.04, P > 0.05 for all), suggesting that a higher areal BMD and lower incidence of osteoporosis in men do not explain their lower incidence of fractures. In contrast, women were more prone to sustain injurious falls than men in both cohorts investigated (HR for women = 1.61 and 1.84, P < 0.001 for both), resulting in a higher incidence of fractures (HR for women = 2.24 and 2.36, P < 0.001 for both). The number of injurious falls and fractures occurring each month during the study period showed a very strong correlation in both women (r = 0.95, P < 0.00001) and men (r = 0.97, P < 0.00001). In summary, low areal BMD, and thus osteoporosis, may not explain the higher fracture incidence in women than in men. Instead, a higher incidence of injurious falls in women was strongly associated with the higher fracture risk.

Substance P accelerates hypercellularity and angiogenesis in tendon tissue and enhances paratendinitis in response to Achilles tendon overuse in a tendinopathy model.

BACKGROUND: Tenocytes produce substance P (SP), and its receptor (neurokinin-1 receptor (NK-1R)) is expressed throughout the tendon tissue, especially in patients with tendinopathy and tissue changes (tendinosis) including hypercellularity and vascular proliferation. Considering the known effects of SP, one might ask whether SP contributes to these changes. OBJECTIVES: To test whether development of tendinosis-like changes (hypercellularity and angiogenesis) is accelerated during a 1-week course of exercise with local administration of SP in an established Achilles tendinopathy model. METHODS: Rabbits were subjected to a protocol of Achilles tendon overuse for 1 week, in conjunction with SP injections in the paratenon. Exercised control animals received NaCl injections or no injections, and unexercised, uninjected controls were also used. Tenocyte number and vascular density, as well as paratendinous inflammation, were evaluated. Immunohistochemistry and in situ hybridisation to detect NK-1R were conducted. Results There was a significant increase in tenocyte number in the SP-injected and NaCl-injected groups compared with both unexercised and exercised, uninjected controls. Tendon blood vessels increased in number in the SP-injected group compared with unexercised controls, a finding not seen in NaCl-injected controls or in uninjected, exercised animals. Paratendinous inflammation was more pronounced in the SP-injected group than in the NaCl controls. NK-1R was detected in blood vessel walls, nerves, inflammatory cells and tenocytes. CONCLUSIONS: SP accelerated the development of tendinosis-like changes in the rabbit Achilles tendon, which supports theories of a potential role of SP in tendinosis development; a fact of clinical interest since SP effects can be effectively blocked. The angiogenic response to SP injections seems related to paratendinitis.

Tenocyte hypercellularity and vascular proliferation in a rabbit model of tendinopathy: contralateral effects suggest the involvement of central neuronal mechanisms.

OBJECTIVE: To determine whether there are objective findings of tendinosis in a rabbit tendinopathy model on exercised and contralateral (non-exercised) Achilles tendons. DESIGN: Four groups of six New Zealand white rabbits per group were used. The animals of one (control) group were not subjected to exercise/stimulation. INTERVENTIONS: Animals were subjected to a protocol of electrical stimulation and passive flexion-extension of the right triceps surae muscle every second day for 1, 3 or 6 weeks. MAIN OUTCOME MEASURES: Tenocyte number and vascular density were calculated. Morphological evaluations were also performed as well as in-situ hybridisation for vascular endothelial growth factor (VEGF) messenger RNA. RESULTS: There was a significant increase in the tenocyte number after 3 and 6 weeks of exercise, but not after 1 week, in comparison with the control group. This was seen in the Achilles tendons of both legs in experimental animals, including the unexercised limb. The pattern of vascularity showed an increase in the number of tendon blood vessels in rabbits that had exercised for 3 weeks or more, compared with those who had exercised for 1 week or not at all. VEGF-mRNA was detected in the investigated tissue, with the reactions being more clearly detected in the tendon tissue with tendinosis-like changes (6-week rabbits) than in the normal tendon tissue (control rabbits). CONCLUSIONS: There were bilateral tendinosis-like changes in the Achilles tendons of rabbits in the current model after 3 weeks of training, suggesting that central neuronal mechanisms may be involved and that the contralateral side is not appropriate as a control.

Unexpected presence of the neurotrophins NGF and BDNF and the neurotrophin receptor p75 in the tendon cells of the human Achilles tendon.

Neurotrophins are substances that have been shown to be important in growth and remodelling phases in different types of tissue. There is no information concerning the possible occurrences of neurotrophins and their receptors in tendons. In this study, sections of both chronic painful (tendinosis) and pain-free (non-tendinosis) human Achilles tendons were immunohistochemically stained with antibodies against the neurotrophins NGF and BDNF, and their receptors TrkA, TrkB and p75. There were marked immunoreactions for NGF and BDNF in the tendon cells (tenocytes) of both tendinosis and non-tendinosis specimens. The tenocytes were also reactive for the receptor p75, but not for the receptors TrkA and TrkB. In addition, p75 immunoreactions were seen in nerve fascicles and in the walls of arterioles. This is the first study to identify neurotrophins in the tenocytes of human tendon. It is clear from this study that the local cells of tendons are sources of neurotrophins. The neurotrophins may play an important role in the tendon through their interaction with the receptor p75 in the tenocytes. These interactions may regulate tropic modulatory, and apoptotic effects. In conclusion, the observations show a new concept concerning production and function of neurotrophins, namely in the tenocytes of tendons.

Treatment of midportion Achilles tendinosis: similar clinical results with US and CD-guided surgery outside the tendon and sclerosing polidocanol injections.

Sclerosing polidocanol injections targeting the area with vasculo/neural ingrowth on the ventral side of the tendon have previously been demonstrated to give good clinical results in patients with chronic painful midportion Achilles tendinosis. In this study, 20 consecutive patients (9 men and 11 women, mean age 46 years) with chronic painful midportion Achilles tendinosis were treated with either sclerosing polidocanol injections (Group A) or open surgical revision of the area with vasculo/neural ingrowth on the ventral side of the Achilles tendon (Group B). Before treatment, all patients had structural tendon changes and increased blood flow-neovascularisation demonstrated with US and colour Doppler. Under US and colour Doppler-guidance, both the injections and the surgical revision targeted the area with neovessels just outside the ventral part of the tendon. A maximum of two sclerosing injection treatments, with 6-8 weeks in-between, were given. For evaluation, the patients recorded the severity of Achilles tendon pain during tendon loading activity (their recreational or sport activity), before and after treatment, on a VAS. Patient global satisfaction with treatment was also assessed. At the 3 months follow-up 6/9 (one patient was excluded) patients in Group A, and 8/10 patients in Group B, were satisfied with the treatment and had a significantly reduced level of tendon pain (Group A VAS from 76 to 24, P < 0.05, Group B VAS from 75 to 21, P < 0.05). There was one deep infection in Group B. At the 6 months follow-up, 6/9 patients in Group A, and 10/10 patients in Group B were satisfied. In summary, both treatment with sclerosing polidocanol injections and open surgical revision outside the ventral Achilles midportion show promising short-term clinical results.

Periosteal transplantation to the rabbit patella.

Autologous periosteal transplantation (without chondrocyte cell transplantation) for treating traumatic articular cartilage defects of the patella gives pain relief in uncontrolled clinical studies. To study the whole transplanted area macroscopically and microscopically, animal studies are motivated. In this pilot study, we reproduce the surgical technique for periosteum transplantation on human patella to a rabbit model. A full-thickness cartilage defect of the whole patella was created in eight adult female rabbits. The defect was treated with autologous periosteal transplantation. After surgery, the rabbits were allowed free activity. This is the difference compared to the treatment in humans, where our group uses CPM for 5 days and non-weight-bearing for 12 weeks. After 21 weeks, there was a diffuse synovitis in all transplanted knees, and in five of eight knees there were signs of osteoarthritis in the patello-femoral joint. Histologically, in three animals, small islands of hyaline cartilage surrounded by fibrocartilage were seen in the transplanted area. In the other five animals, fibrocartilage was the predominant tissue. In contrast to previous experimental studies using a rabbit model, we did not achieve hyaline cartilage resurfacing.

Chronic Achilles tendon pain treated with eccentric calf-muscle training.

Injuries involving the Achilles tendon and manifested as chronic tendon pain are common, especially among recreational athletes. In a pilot study on a small group of patients with chronic painful mid-portion Achilles tendinosis, eccentric calf-muscle training was shown to give good clinical results. The aim of this prospective study was to investigate if the previously achieved good clinical results could be reproduced in a larger group of patients, and also to investigate the effects of eccentric calf-muscle training in patients with chronic insertional Achilles tendon pain. Seventy-eight consecutive patients, having chronic painful Achilles tendinosis at the mid-portion (2-6 cm level) in a total of 101 tendons (55 unilateral and 23 bilateral), and thirty consecutive patients with chronic insertional Achilles tendon pain in 31 tendons (29 unilateral and one bilateral) were treated with eccentric calf-muscle training for 12 weeks. Most patients were recreational athletes. Evaluation of the amount of tendon pain during activity was recorded on a visual analogue scale (VAS), before and after treatment. In 90 of the 101 Achilles tendons (89%) with chronic painful mid-portion Achilles tendinosis, treatment was satisfactory and the patients were back on their pre-injury activity level after the 12-week training regimen. In these patients, the amount of pain during activity, registered on the VAS-scale (mean+/-SD), decreased significantly from 66.8+/-19.4 to 10.2+/-13.7. On the contrary, in only ten of the tendons (32%) with chronic insertional Achilles tendon pain was treatment satisfactory, with a significant decrease on the VAS-scale (mean+/-SD), from 68.3+/-7.0 to 13.3+/-13.2. Our conclusion is that treatment with eccentric calf-muscle training produced good clinical results in patients with chronic painful mid-portion Achilles tendinosis, but not in patients with chronic insertional Achilles tendon pain.

Intratendinous glutamate levels and eccentric training in chronic Achilles tendinosis: a prospective study using microdialysis technique.

Microdialysis has shown intratendinous glutamate levels to be significantly higher in Achilles tendons with painful tendinosis than in normal pain-free tendons, and treatment with eccentric training has shown good clinical results with diminished tendon pain during activity. In six patients with chronic painful Achilles tendinosis we performed microdialysis for 2 h, before and after the 12-week eccentric training program. The treatment was successful in all six patients, and the mean VAS score (amount of pain during Achilles tendon loading) decreased from 69 before treatment to 17 after treatment. There was no significant difference between the intratendinous glutamate levels before and after treatment. Our results offer no obvious neurophysiological explanation but showed that successful treatment with eccentric training was not associated with lowered intratendinous glutamate levels.

Vitamin D receptor start codon polymorphism ( FokI) is related to bone mineral density in healthy adolescent boys.

Peak bone mass is considered a major determinant in the emergence of osteoporosis and is mainly genetically regulated. Several genes have been investigated, among them the vitamin D receptor ( VDR) gene. A single-nucleotide polymorphism (defined by the endonuclease FokI) located in the start codon of the VDR creates the alleles F and f, resulting in different proteins. A number of previous studies have proved the F allele to be more advantageous as concerns bone mineral density (BMD). In this longitudinal study of 88 adolescent boys, we have investigated whether the different genotypes are associated with BMD, bone mineral content (BMC), or bone area. BMD, BMC, and bone area of the right femoral neck, lumbar spine, and total body were measured using dual-energy X-ray absorptiometry. Differences in phenotypes in relation to the FokI polymorphism were calculated by means of an analysis of variance (ANOVA), with Bonferroni's correction for multiple comparisons. At the first examination, the FokI genotypes were significantly related to lumbar spine BMC and total body bone area in boys aged 16.9 +/- 0.3 years (mean +/- SD). There was a strong tendency towards significance as regards pubertal stage, total body and femoral neck BMC, weight, lean body mass, lumbar spine bone area, and lumbar spine BMD. There were no significant differences in height, fat mass, birth height and weight, total body and femoral neck BMD, and femoral neck bone area. Regression analysis proved the FokI genotypes to be independently related to lumbar spine BMD ( FF > ff; P < 0.01), and possibly total body BMD ( P = 0.06), but not femoral neck BMD. At the second examination, approximately 2 years later, our ANOVA results showed significance as regards femoral neck BMC and weight. Using multiple regression, the FokI genotypes were independently related to lumbar spine BMD ( FF > ff; P = 0.03), and total body BMD ( P < 0.05), but not femoral neck BMD. This study proves the FokI polymorphism to be an independent predictor of lumbar spine BMD are probably total body BMD, but not femoral neck BMD.

High intratendinous lactate levels in painful chronic Achilles tendinosis. An investigation using microdialysis technique.

In this investigation the microdialysis technique was used to study the concentrations of lactate in Achilles tendons with painful chronic tendinosis and in normal pain-free tendons. In four patients (mean age 40.7 years) with a painful thickening localized at the 2-6 cm level in the Achilles tendon (chronic Achilles tendinosis) and in five controls (mean age 37.2 years) with normal Achilles tendons the local concentrations of lactate were registered under resting conditions. All tendons were examined using ultrasonography. In the tendons with tendinosis the painful thickening corresponded to a widened tendon and structural tendinosis changes. Normal tendons showed no widening and a normal structure. A standard microdialysis catheter was inserted into the Achilles tendon under local anesthesia. Samplings were done every 15 min during a 4 h period. The results showed significantly higher mean concentrations of lactate in tendons with tendinosis compared to normal tendons (2.15 mmol/l vs. 1.14 mmol/l). The lactate concentrations in the tendons with tendinosis were stable, and approximately twofold higher than in the normal tendons during the whole 4 h investigation period. In conclusion, the higher concentrations of lactate in Achilles tendons with painful tendinosis indicate that there are anaerobic conditions in the area with tendinosis. The importance of this finding for the pathogenesis and pain mechanisms in this chronic condition needs to be further investigated.

Chronic tendon pain: no signs of chemical inflammation but high concentrations of the neurotransmitter glutamate. Implications for treatment?

Chronic musculo-tendinous pain syndromes are relatively common and associated with very high socio-economic costs. Their aetiology and pathogenesis are still unknown. In the athletic population, chronic tendon pain is most often seen among recreational male and female athletes in the age group between 30-60 years, and is considered to be associated with overuse of the aged tendon. Treatment is known to be difficult. In general, these chronic painful conditions have been considered to include an inflammatory component, and the nomenclature used (tendinitis, tendonitis) most often implies an inflammatory involvement. Despite that tendon biopsies have shown an absence of inflammatory cell infiltration, anti-inflammatory agents (NSAID'S, corticosteroidal injections) are most often included in the treatment. Our research has been focused on chronic painful conditions in the Achilles-, patellar-, and extensor carpi radialis brevis (ECRB)-tendons. We have demonstrated, for the first time, that it is possible to use the microdialysis technique for in vivo investigations of human tendons, and that the excitatory neurotransmitter glutamate exists in human tendons. We have identified and measured the concentrations of prostaglandin E2 (PGE2) in tendons as well. The results showed that chronic painful tendinosis (Achilles-, patellar-, ECRB-) had significantly higher concentrations of glutamate, but not PGE2, as compared to the pain-free normal tendons. With the use of immunohistochemical analyses and enzyme histochemistry of human Achilles- and patellar tendon biopsies, we have also, for the first time demonstrated that glutamate NMDARI-immunoreaction was present in nerve structures. These findings altogether, indicate that glutamate might be involved in chronic tendon pain, and further emphasizes that there is no chemical inflammation (normal PGE2 levels) in the chronic stage of these relatively common so-called tendinopathies. The findings of glutamate and it's NMDARI-receptors might have implications for treatment and be a potential target for drugs.

Painful conditions in the Achilles tendon region: a common problem in middle-aged competitive badminton players.

Overuse injuries are the most frequent type in badminton, generally localized in the legs. An earlier study found 32% of young Swedish elite badminton players to have experienced disabling pain in the Achilles tendon region during the previous 5 years. The present investigation examined the prevalence and characteristics of painful conditions in the Achilles tendon region in 32 middle-aged competitive badminton players by means of questionnaire and physiotherapist's examination. Pain in the Achilles tendon region was reported by 44%, either presently or during the past 5 years, generally localized in the middle portion of the tendon. Symptoms had lasted 2 weeks-1 year (96 days). On the competition days 22% of the reported pain currently in the region. Age was found to be correlated to Achilles tendon pain, but there was no relationship between symptoms of pain and body mass index, gender, training quantity, or years of playing badminton. In conclusion, Achilles tendon pain seems to be relatively common among Swedish middle-aged competitive badminton players, particularly in the older ones.

Painful conditions in the Achilles tendon region in elite badminton players.

The purpose of this study was to investigate the prevalence and characteristics of painful conditions in the Achilles tendon region in elite badminton players. The study group consisted of 66 players in the Swedish elite division (highest level) in badminton, 41 men (mean age, 24.4 years) and 25 women (mean age, 21.9 years). Twenty-one players (32%) reported the occurrence of a disabling painful condition in the Achilles tendon region during the previous 5 years, and 11 players (17%) had an ongoing painful condition. A majority of the painful conditions (12 of 21, or 57%) were described as involving the midportion of the Achilles tendon. The players who had a painful condition reported a significantly higher weekly training load as measured by the number of hours spent in total training, badminton training, and endurance and strength training. There were no differences in age, sex, and body mass index between the players with and without painful conditions in the Achilles tendon region.