Expression of alarmins in a murine rotator cuff tendinopathy model.

The aim of this study was to investigate the presence of alarmins in a novel murine rotator cuff tendinopathy model. Alarmins have been described as essential early activators of an immune response to tissue damage. Subacromial impingement was induced in both shoulders of 37 male C57Bl/6 mice by placement of a small metal clip in the subacromial space. Animals were allocated to different time points up to 6 weeks. The morphology and cellularity of the supraspinatus tendon were evaluated by hematoxylin-eosin staining, alcian blue, and picrosirius red. The expression and localization of alarmins interleukin-33 (IL-33), c (HMGB1), hypoxia-inducible factor-1 subunit α (HIF1α), and S100A9 were evaluated by immunohistochemical staining and quantitative polymerase chain reaction. The percentage of positively stained cells with HMGB1 and IL-33 was significantly increased in the impingement group at 1w, 4w, and 6w. HIF1α staining was higher in the impingement group at 1w and 6w compared with the control group. HMGB1 gene expression was higher in the 5d impingement group and 6w impingement group. The gene expression of HIF1α was upregulated at all-time points in the impingement group (5d, 2w, 4w, and 6w). The expression of the S100A9 gene was also upregulated in the 5d impingement group. This is the first study to demonstrate the involvement of alarmins in the early phase of tendinopathy using a reproducible animal model. Alarmins may play an important role in the early phases of the development of tendinopathy They may represent potential therapeutic targets for treatment of tendinopathy.

Identification of Inflammatory Mediators in Tendinopathy Using a Murine Subacromial Impingement Model.

Subacromial impingement is associated with a spectrum of disorders-including rotator cuff disease-but their relationship is complex. We have established a novel murine model of subacromial impingement to study supraspinatus tendinopathy. The purpose of this study was to evaluate changes in gene expression in this murine shoulder impingement model to further elucidate the mechanisms underlying the development of tendinopathy. Twenty-eight C57BL/6 mice were used in this study. All mice underwent bilateral surgery with insertion of a small metal clip in the subacromial space or a sham procedure. The supraspinatus tendons underwent histological analyses, biomechanical testing, and RNA extraction for multiplex gene expression analysis (NanoString, Seattle, WA). Histology demonstrated increased cellularity and disorganized collagen fibers of the supraspinatus tendon in the clip impingement group. Mean load to failure (5.20 vs. 1.50 N, p < 0.001) and mean stiffness (4.95 vs. 1.47 N/mm, p < 0.001) were lower in the impingement group than the sham group. NanoString analyses revealed 111 differentially expressed genes (DEGs) between the impingement and sham groups. DEGs of interest included Mmp3 (expression ratio [ER]: 2.68, p = 0.002), Tgfb1 (ER: 1.76, p = 0.01), Col3a1 (ER: 1.66, p = 0.03), and Tgfbr2 (ER: 1.53, p = 0.01). Statement of clinical significance: We identified 111 DEGs that may contribute to the development of tendinopathy in this model. Further studies of these specific genes will allow identification of their roles in the initiation and regulation of tendon damage, and their potential to serve as novel therapeutic targets in the treatment of rotator cuff disease. © 2019 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 37:2575-2582, 2019.

Metabolic profiling of serum and tissue from the rotator interval and anterior capsule in shoulder stiffness: a preliminary study.

BACKGROUND: Using mass spectrometry, we evaluated the metabolic profiles of patients who had rotator cuff tears with shoulder stiffness, or shoulder stiffness only, and compared these with samples from a control group. METHODS: This study enrolled 28 patients, including 10 patients with shoulder stiffness only (group I), nine patients with rotator cuff tear and stiffness (group II), and nine controls selected from patients diagnosed with impingement syndrome or long head of the biceps lesions without evident limitation of joint motion or rotator cuff tears. Serum and tissue from the rotator interval and anterior capsule were collected. In all, 82 samples were analyzed for metabolite profiling using the AbsoluteIDQ™p180 Kit. RESULTS: Comparison of 186 metabolites revealed that groups I and II had significantly higher concentrations of sphingolipids in serum (SM C24:1; group I = 65.16 µm, group II = 68.07 µm) than controls (55.37 µm, p = 0.005 & 0.015, respectively). Higher concentrations of sphingolipids were also present in the rotator interval tissue (SM C22:3) of groups 1 (0.0197 µm) and 2 (0.0144 µm) than controls (0.0081 µm, p = 0.012 & 0.014, respectively). The concentration of glycerophospholipid (PC aa C30:0) was higher in the anterior capsule tissue of groups I (0.850 µm) and II (1.164 µm) than controls (0.572 µm; p = 0.007) Total cholesterol was positively correlated with sphingolipid concentration in serum (SM C24:1, rho = 0.782, p = 0.008) and rotator interval tissue (SM C22:3, rho = 0.750, p = 0.017). There was no significant difference in the metabolites evaluated in groups I and II. CONCLUSION: Metabolic profiling showed that levels of lipid-related metabolites were increased in the anterior capsule tissue and rotator interval tissue of patients with shoulder stiffness. Sphingomyelin (SM C22:3) in the tissue of the rotator interval was positively correlated with the serum level of total cholesterol in patients with shoulder stiffness only. The level of glycerophospholipid (PC30:0) in the anterior capsule was positively correlated with the serum level of total cholesterol in patients who had rotator cuff tear with shoulder stiffness. The results indicate that serum total cholesterol may be related to shoulder stiffness. Future studies are needed to evaluate the role of serum cholesterol in the pathogenesis of shoulder stiffness. TRIAL REGISTRATION: KC12OISI0532. Registered Nov 15, 2012. approval by the Institutional Review Board of Seoul St. Mary's Hospital, the Catholic University of Korea.

Evaluating the role of subacromial impingement in rotator cuff tendinopathy: Development and analysis of a novel murine model.

Subacromial impingement of the rotator cuff is understood as a contributing factor in the development of rotator cuff tendinopathy. However, changes that occur in the impinged tendon are poorly understood and warrant further study. To enable further study of rotator cuff tendinopathy, we performed a controlled laboratory study to determine feasibility and baseline characteristics of a new murine model for subacromial impingement. This model involves surgically inserting a microvascular clip into the subacromial space in adult C57Bl/6 mice. Along with a sham surgery arm, 90 study animals were distributed among time point groups for sacrifice up to 6 weeks. All animals underwent bilateral surgery (total N = 180). Biomechanical, histologic, and molecular analyses were performed to identify and quantify the progression of changes in the supraspinatus tendon. Decreases in failure force and stiffness were found in impinged tendon specimens compared to sham and no-surgery controls at all study time points. Semi-quantitative scoring of histologic specimens demonstrated significant, persistent tendinopathic changes over 6 weeks. Quantitative real-time polymerase chain reaction analysis of impinged tendon specimens demonstrated persistently increased expression of genes related to matrix remodeling, inflammation, and tendon development. Overall, this novel murine subacromial impingement model creates changes consistent with acute tendonitis, which may mimic the early stages of rotator cuff tendinopathy. A robust, simple, and reproducible animal model of rotator cuff tendinopathy is a valuable research tool to allow further studies of cellular and molecular mechanisms and evaluation of therapeutic interventions in rotator cuff tendinopathy. © 2018 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 36:2780-2788, 2018.

Prevalencia del dolor de hombro en nadadores de competición: estudio piloto / Shoulder pain prevalence in competitive swimmers: a pilot study

Objetivos: conocer la prevalencia de dolor de hombro en nadadores de competición, sus características y su relación con factores antropométricos y deportivos. Métodos: estudio de prevalencia. Ciento cuarenta nadadores/as entre 12 y 24 años cumplimentaron un cuestionario durante una fase de entrenamiento específico. Resultados: el 25,7% revelaron padecer dolor de hombro. Se hallaron relaciones estadísticamente significativas entre el dolor de hombro y episodios previos de dolor (p<0,001), experiencia superior a tres años (p=0,014), Índice de Masa Corporal (p=0,015) y la especialidad estilo (p=0,008) y distancia (p=0,011). El dolor fue significativamente más intenso durante la actividad que en reposo (p<0,001).Conclusiones: el dolor de hombro en nadadores de competición entre 12 y 24 años parece ser un problema frecuente y repetitivo, que aumenta con la experiencia y que se asocia a la actividad, a un mayor Índice de Masa Corporal y a la especialidad del nadador (AU)

Objectives: to investigate the prevalence of shoulder pain in competitive swimmers and find out the characteristics of pain as well as its relation to anthropometric and sports factors.Methods: prevalence study. A hundred and forty competitive swimmers between 12 and 24 years old completed a questionnaire in a high intensity training phase. Results: 25.7% swimmers reported shoulder pain. There were significant statistical correlations between shoulder pain and previous episodes of pain (p<0.001), more than three years of experience (p=0.014), Body Mass Index (p=0.015) and stroke (p=0.008) and distance (p=0.011) specialty. Pain was statistically correlated with activity (p<0.001). Conclusions: shoulder pain seems to be a frequent and repetitive problem in competitive swimmers between 12 and 24 years old, which increases with years of practice. Furthermore, it seems to be associated with the activity, a higher Body Mass Index and the swimmers’ specialty (AU)

Proteomics perspectives in rotator cuff research: a systematic review of gene expression and protein composition in human tendinopathy.

BACKGROUND: Rotator cuff tendinopathy including tears is a cause of significant morbidity. The molecular pathogenesis of the disorder is largely unknown. This review aimed to present an overview of the literature on gene expression and protein composition in human rotator cuff tendinopathy and other tendinopathies, and to evaluate perspectives of proteomics--the comprehensive study of protein composition--in tendon research. MATERIALS AND METHODS: We conducted a systematic search of the literature published between 1 January 1990 and 18 December 2012 in PubMed, Embase, and Web of Science. We included studies on objectively quantified differential gene expression and/or protein composition in human rotator cuff tendinopathy and other tendinopathies as compared to control tissue. RESULTS: We identified 2199 studies, of which 54 were included; 25 studies focussed on rotator cuff or biceps tendinopathy. Most of the included studies quantified prespecified mRNA molecules and proteins using polymerase chain reactions and immunoassays, respectively. There was a tendency towards an increase of collagen I (11 of 15 studies) and III (13 of 14), metalloproteinase (MMP)-1 (6 of 12), -9 (7 of 7), -13 (4 of 7), tissue inhibitor of metalloproteinase (TIMP)-1 (4 of 7), and vascular endothelial growth factor (4 of 7), and a decrease in MMP-3 (10 of 12). Fourteen proteomics studies of tendon tissues/cells failed inclusion, mostly because they were conducted in animals or in vitro. CONCLUSIONS: Based on methods, which only allowed simultaneous quantification of a limited number of prespecified mRNA molecules or proteins, several proteins appeared to be differentially expressed/represented in rotator cuff tendinopathy and other tendinopathies. No proteomics studies fulfilled our inclusion criteria, although proteomics technologies may be a way to identify protein profiles (including non-prespecified proteins) that characterise specific tendon disorders or stages of tendinopathy. Thus, our results suggested an untapped potential for proteomics in tendon research.

The association between retraction of the torn rotator cuff and increasing expression of hypoxia inducible factor 1α and vascular endothelial growth factor expression: an immunohistological study.

BACKGROUND: Differing levels of tendon retraction are found in full-thickness rotator cuff tears. The pathophysiology of tendon degeneration and retraction is unclear. Neoangiogenesis in tendon parenchyma indicates degeneration. Hypoxia inducible factor 1α (HIF) and vascular endothelial growth factor (VEGF) are important inducers of neoangiogenesis. Rotator cuff tendons rupture leads to fatty muscle infiltration (FI) and muscle atrophy (MA). The aim of this study is to clarify the relationship between HIF and VEGF expression, neoangiogenesis, FI, and MA in tendon retraction found in full-thickness rotator cuff tears. METHODS: Rotator cuff tendon samples of 33 patients with full-thickness medium-sized rotator cuff tears were harvested during reconstructive surgery. The samples were dehydrated and paraffin embedded. For immunohistological determination of VEGF and HIF expression, sample slices were strained with VEGF and HIF antibody dilution. Vessel density and vessel size were determined after Masson-Goldner staining of sample slices. The extent of tendon retraction was determined intraoperatively according to Patte's classification. Patients were assigned to 4 categories based upon Patte tendon retraction grade, including one control group. FI and MA were measured on standardized preoperative shoulder MRI. RESULTS: HIF and VEGF expression, FI, and MA were significantly higher in torn cuff samples compared with healthy tissue (p < 0.05). HIF and VEGF expression, and vessel density significantly increased with extent of tendon retraction (p < 0.05). A correlation between HIF/VEGF expression and FI and MA could be found (p < 0.05). There was no significant correlation between HIF/VEGF expression and neovascularity (p > 0.05) CONCLUSION: Tendon retraction in full-thickness medium-sized rotator cuff tears is characterized by neovascularity, increased VEGF/HIF expression, FI, and MA. VEGF expression and neovascularity may be effective monitoring tools to assess tendon degeneration.

Tendinopathy and tears of the rotator cuff are associated with hypoxia and apoptosis.

The aim of this study was to investigate the occurrence of tissue hypoxia and apoptosis at different stages of tendinopathy and tears of the rotator cuff. We studied tissue from 24 patients with eight graded stages of either impingement (mild, moderate and severe) or tears of the rotator cuff (partial, small, medium, large and massive) and three controls. Biopsies were analysed using three immunohistochemical techniques, namely antibodies against HIF-1alpha (a transcription factor produced in a hypoxic environment), BNip3 (a HIF-1alpha regulated pro-apoptotic protein) and TUNEL (detecting DNA fragmentation in apoptosis). The HIF-1alpha expression was greatest in mild impingement and in partial, small, medium and large tears. BNip3 expression increased significantly in partial, small, medium and large tears but was reduced in massive tears. Apoptosis was increased in small, medium, large and massive tears but not in partial tears. These findings reveal evidence of hypoxic damage throughout the spectrum of pathology of the rotator cuff which may contribute to loss of cells by apoptosis. This provides a novel insight into the causes of degeneration of the rotator cuff and highlights possible options for treatment.

Rotura del supraespinoso en paciente con pinzamiento subacromial crónico / Supraspinous rupture in patients with chronic subacromial impingement

En el espacio subacromial se desliza el manguito de los rotadores que cubren la cabeza humeral. La lesión de estos tendones, especialmente del supraespinoso, es origen de molestias que obligan a tratamientos prolongados, incluyendo el quirúrgico. El pinzamiento subacromial es un compromiso de la inserción del supraespinoso bajo el borde anterior del acromion y ligamento acromiocoracoideo. Hay tres estadios evolutivos: tendinopatía, rotura parcial y rotura transfixiante. Lesiones, que en un primer momento pueden ser pequeñas, con el esfuerzo diario aumentan y se agravan. Clínicamente el dolor suele exacerbarse con la elevación del brazo, de predominio nocturno, asociándose a debilidad y limitación del movimiento. El diagnóstico de rotura del supraespinoso es pasado por alto en Atención Primaria, y su retraso en el tratamiento tiene mal pronóstico. La radiología es normal inicialmente y el diagnóstico se confirma con ecografía, tomografía y resonancia. En pacientes mayores de 65 años suele optarse por un tratamiento conservador (AU)

In the subacromial space, the rotator cuff that covers the humeral head slides. Injury to these tendons, especially of the supraspinous one, is the origin of discomforts that require prolonged treatments, including surgery. Subacromial impingement is a compromise of the supraspinous insertion under the anterior border of the acromion and coracoacromial ligament. There are three evolutive stages: tendonitis, partial rupture and full-thickness rupture. Injuries which may be small in the beginning may increase and become worse with exercise. Clinically, the pain may worsen with elevation of the arm, with nighttime predominance, this being associated to weakness and movement limitation. Diagnosis of supraspinous rupture is overlooked in Primary Care and delay in its treatment has a poor prognosis. The x-ray is normal initially and diagnosis is confirmed with ultrasonography, tomography and resonance. Conservative treatment should be chosen in patients over 65 years (AU)

Inducement of mitogen-activated protein kinases in frozen shoulders.

BACKGROUND: Mitogen-activated protein (MAP) kinases are well-known molecules that play key roles in mechanical stress signals during skeletal development. To test our hypothesis that the synovium in frozen shoulders is induced by MAP kinases, immunohistochemical analyses for detecting expression and signal transduction of MAP kinases were performed in synovial tissue obtained from the rotator interval (RI) in frozen shoulders. METHODS: Synovial tissues were examined from 10 frozen shoulder patients with a mean age of 55.4 years (46-62 years). Synovial tissues between the long head of the biceps tendon (LHB) and the RI in frozen shoulders were stained with hematoxylin and eosin (H&E) and then examined with immunohistochemical staining. Extracellular signal-regulated (ERK), the Jun N-terminal (JNK), and p38 mitogen-activated protein (MAP) kinases, nuclear factor kappaB (NF-kappaB), p50, CD29 (beta(1)-integrin), matrix metalloproteinase (MMP)-3, interleukin-6 (IL-6), CD56, CD68, S-100, and vascular endothelial growth factor (VEGF) were analyzed to detect expression patterns. RESULTS: H&E showed vascular proliferation with fibrin and fibrous tissue in the synovium of frozen shoulders. ERK was expressed in the epithelial cells of vascular tissue, and JNK was expressed strongly in the interstitial cells around vascular tissue; p38 MAPK was not expressed. NF-kappaB was expressed in vascular tissue, and IL-6 was expressed around vascular tissue. CD29 (beta1-integrin) was expressed in vascular tissue and in superficial cells of synovial tissue. MMP-3 and VEGF were expressed on the surface layer of synovial tissue and vascular tissue, and CD68 was expressed on the surface layer. Nerve-related proteins, CD56 and S-100, were expressed weakly. CONCLUSIONS: Mechanical stress on the LHB and RI in the shoulder may induce ERK and JNK to express NF-kappaB by CD29 to develop capsule contracture, producing MMP-3, IL-6, and VEGF.

Does the nature of deposited basic calcium phosphate crystals determine clinical course in calcific periarthritis of the shoulder?

OBJECTIVE: Two typical clinical courses of calcific periarthritis in the shoulder are known: acute, with severe inflammation, and chronic, in the form of impingement syndrome with secondary subacromial bursitis. It remains unclear what factors determine these clinical courses. Our objectives were to clarify whether the calcified deposits that occur in both acute and chronic cases are composed of carbonate apatite; and to compare the Ca:P molar ratio in the 2 forms and to determine if there was any correlation in this respect with the intensity of inflammation induced by basic calcium phosphate crystals. METHODS: Ten samples were aspirated from 10 women (ages 42-65 yrs) with acute inflammation. The average time from first attack to aspiration was 2.3 days. A further 10 samples were operatively removed from 10 women (ages 35-58 yrs) with refractory chronic subacromial bursitis, among whom an average of 7.8 months had passed since the onset of symptoms. All samples were analyzed by x-ray diffraction, Fourier transform infrared spectroscopy, and Raman spectroscopy, and Ca:P molar ratios were measured by x-ray fluorescence spectrometry. RESULTS: Calcified deposits from both acute and chronic cases were identified as carbonate apatite, and not hydroxyapatite, octacalcium phosphate, tricalcium phosphate, or dicalcium phosphate dihydrate. The average Ca:P molar ratio of calcified deposits was calculated as 1.71 +/- 0.16 in acute cases and 1.71 +/- 0.16 in chronic cases (statistically nonsignificant). CONCLUSION: Deposits around the shoulder in both acute and chronic calcific periarthritis are composed of carbonate apatite, Ca:P molar ratios being almost identical in the 2 forms. The results suggest that some factor other than the composition of the crystalline deposits may determine clinical course in calcific periarthritis of the shoulder.

[In vitro effects of diclofenac and selective cyclooxygenase-2 inhibitors on prostaglandin release from inflamed bursa subacromialis tissue in patients with subacromial syndrome]. / Effekte von Diclofenac und selektiven Cyclooxygenase-(COX-)2-Inhibitoren auf die Prostaglandinfreisetzung aus entzündetem Bursa-subacromialis-Gewebe von Patienten mit Subakromialsyndrom in vitro.

BACKGROUND: To compare the in vitro effects of selective COX-2 inhibitors (L-745,337, NS-398 and DFU) and of COX-unspecific diclofenac on release of PGE(2 )and 6-keto-PGF(1alpha) from inflamed bursa subacromialis tissue (IBST) obtained from a total of 35 patients with shoulder impingement syndrome (SIS). PATIENTS AND METHODS: Bursal specimens were incubated in the presence of drugs (0.01-1000 microM) for 20 min and 16 h. RESULTS: After 20 min 10 microM diclofenac significantly inhibited formation of PGE(2) and 6-keto-PGF(1alpha), whereas L-745,337 and NS-398 (10-1000 microM) induced significant inhibition only at concentrations > or =100 microM. In contrast to equimolar diclofenac, DFU (0.01-10 microM) induced no inhibition of bursal PGE(2) release but a dose-dependent, although statistically not significant inhibition after 16 h. The inhibitory potency of diclofenac (0.01-10 microM) was even more increased during long-term incubation showing greater inhibition than DFU at all concentrations studied. CONCLUSION: The data suggest that in IBST in SIS in vitro the majority of PG is generated via the COX-1 pathway.

Vascular endothelial growth factor 121 and 165 in the subacromial bursa are involved in shoulder joint contracture in type II diabetics with rotator cuff disease.

Vascular endothelial growth factor (VEGF) is a glycoprotein that plays an important role in neovascularization and increases vascular permeability. We reported that VEGF is involved in motion pain of patients with rotator cuff disease by causing synovial proliferation in the subacromial bursa (SAB). The present study investigates whether VEGF is also involved in the development of shoulder contracture in diabetics with rotator cuff disease. We examined 67 patients with rotator cuff disease, including 36 with complete cuff tears, 20 with incomplete tears, and 11 without apparent tears (subacromial bursitis). The patients were into groups according to the presence or absence of diabetes (14 type II diabetics and 53 non-diabetics). Specimens of the synovium of the SAB were obtained from all patients during surgery. Expression of the VEGF gene in the synovium of the subacromial bursa was evaluated by using the reverse transcriptase polymerase chain reaction. The VEGF protein was localized by immunohistochemistry, and the number of vessels was evaluated based on CD34 immunoreactivity. The results showed that VEGF mRNA was expressed in significantly more diabetics (100%, 14/14) than in non-diabetics (70%, 37/53) (P=0.0159, Fisher's test). Investigation of VEGF isoform expression revealed VEGF121 in all 14 diabetics and in 37 of the 53 non-diabetics, VEGF165 in 12 of the 14 diabetics and in 21 of the 53 non-diabetics, and VEGF189 in 1 of the 14 diabetics and in 2 of the 53 non-diabetics. No VEGF206 was expressed in either group. VEGF protein was localized in both vascular endothelial cells and synovial lining cells. The mean number of VEGF-positive vessels and the vessel area were also significantly greater in the diabetics (p<0.015, Mann-Whitney U test). Synovial proliferation and shoulder joint contracture were more common in the diabetics (P=0.0329 and P=0.073, respectively; Fisher's test). The mean preoperative range of shoulder motion significantly differed in terms of elevation between two groups: 103.8 degrees in diabetics and 124.9 degrees in no diabetics (p=0.0039 Mann-Whitney U test). In contrast, external rotation did not significantly differ: 44 degrees in diabetics and 49 degrees in non-diabetics (p=0.4957, Mann-Whitney U test). These results suggest that VEGF121 and VEGF165 expression in the SAB is responsible for the development of shoulder joint contracture, especially in elevation, among type II diabetic patients with rotator cuff disease.

Involvement of the subacromial bursa in impingement syndrome of the shoulder as judged by expression of tenascin-C and histopathology.

Our aim was to evaluate bursal involvement at different stages of the impingement syndrome as judged by conventional histopathological examination and expression of tenascin-C, which is known to reflect active reparative processes in different tissues and disorders. Samples of subacromial bursa were taken from 33 patients with tendinitis, 11 with a partial tear and 18 with a complete tear of the rotator cuff, and from 24 control shoulders. We assessed the expression of tenascin-C, the thickness of the bursa, and the occurrence and degree of fibrosis, vascularity, haemorrhage and inflammatory cells. The expression of tenascin-C was significantly more pronounced in the complete tear group (p < 0.001) than in the partial tear, tendinitis or control groups. It was more pronounced in the tendinitis group than in the control group (p = 0.06), and there was more fibrosis in all the study groups than in the control group. The changes in the other parameters were not equally distinctive. Expression of tenascin-C did not correlate with the conventional histopathological parameters, suggesting that these markers reflect different phases of the bursal reaction. Tenascin-C seems to be a general indicator of bursal reaction, being especially pronounced at the more advanced stages of impingement and this reaction seems to be an essential part of the pathology of impingement at all its stages.

Interleukin-1-induced subacromial synovitis and shoulder pain in rotator cuff diseases.

OBJECTIVE: To determine the relationship between the expression of interleukin-1beta (IL-1beta) and IL-1 receptor antagonists (IL-1ra) in the subacromial bursa and shoulder pain in rotator cuff diseases. METHODS: Synovial specimens were analysed using various methods including reverse transcriptase-polymerase chain reaction (RT-PCR), immunohistochemistry and in situ RT-PCR. Thirty-nine patients with rotator cuff diseases were candidates. The degree of their shoulder pain was evaluated using a visual analogue scale. RESULTS: The mRNA expression levels of the cytokines were significantly correlated with the degree of pain [IL-1beta: r=0.782; secreted IL-1ra (sIL-1ra): r=0.756; intracellular IL-1ra (icIL-1ra): r=0.806, P<0.001, respectively]. The combined results of immunohistochemistry and in situ RT-PCR analysis indicated that both synovial lining and sublining cells produce IL-1beta, while synovial lining cells predominantly produce icIL-1ra and sublining cells secrete sIL-1ra. CONCLUSIONS: The differential regulation of the two forms of IL-1ra mRNAs may play an important role in shoulder pain in rotator cuff diseases, regulating IL-1-induced subacromial synovitis.

Immunolocalization of cytokines and growth factors in subacromial bursa of rotator cuff tear patients.

Inflammation in the subacromial bursa causes pain in patients suffering from rotator cuff tear, with this long-lasting inflammation leading to fibrosis and thickening of the subacromial bursa. Both inflammatory cytokines and mechanical stress, and impingement in the subacromial space, might induce and worsen this inflammation. However, little is known of the mechanism of this inflammation. In this study, we used immunohistological staining to demonstrate the expression of Interleukin-1 beta (IL-1 beta), Tumor necrosis factor alpha (TNF-alpha), transforming growth factor beta (TGF-beta), and basic fibroblast growth factor (bFGF) in subacromial bursa derived from the patients suffering from rotator cuff tear. On the other hand the expression of these inflammatory cytokines and growth factors were little detected only to a small degree in patients with anterior shoulder instability who did not have severe shoulder pain and impingement in the subacromial space. Our findings suggest that those inflammatory cytokines and growth factors may play an important role in inflammation of the subacromial bursa. Controlling the expression of these cytokines and growth factors might be important for treating patients suffering from shoulder pain due to rotator cuff tear.

Vascular endothelial growth factor (VEGF) expression in the subacromial bursa is increased in patients with impingement syndrome.

Vascular endothelial growth factor (VEGF), which is known to be an angiogenetic factor, plays an important role in the inflammation of synovial tissue. To investigate the relationships between VEGF and clinical symptoms in rotator cuff disease, VEGF expression was examined using RT-PCR and immunohistochemical analysis in 50 patients with this disease (26 with full-thickness cuff tear, 12 with partial-thickness tear, and 12 with subacromial bursitis). VEGF mRNA expression was detected in 40 out of 50 patients by RT-PCR. VEGF mRNA expression was found more frequently in the patients with motion pain (39 out of 41) than in those without motion pain (1 out of 9) with statistical significance (Fisher's test, P < 0.001). Thirty-one out of 33 patients with synovial proliferation showed VEGF mRNA expression, whereas the expression of this transcript was found in 9 out of 17 patients without synovial proliferation. This association with synovial proliferation was also significant (Fisher's test, P = 0.0013). Thirty out of 41 patients with motion pain had synovial proliferation but 3 out of 9 patients without motion pain had synovial proliferation. In all these 30 patients with both motion pain and synovial proliferation, VEGF mRNA expression was detected. This association between motion pain and synovial proliferation was also significant (Fisher's test, P < 0.05). The mean vessel count and area in subacromial bursa expressing VEGF was significantly higher than in those without VEGF (Mann Whitney's U test, P < 0.01). These results suggested that VEGF expression is associated with vascularity, synovial proliferation and shoulder motion pain in the rotator cuff disease.

Localised deposition of amyloid in tears of the rotator cuff.

Age-related localised deposition of amyloid in connective tissue has been found in degenerative articular and periarticular tissue. Biopsies of the supraspinatus tendon of 28 patients undergoing repair of the rotator cuff were analysed histologically for the presence of localised deposition of amyloid. There was a long history of impingement in 20 patients, and eight patients had suffered an acute traumatic tear with no preceding symptoms. Localised deposition of amyloid identified by Congo Red staining was detected in 16 samples (57%). Amyloid was present in 14 (70%) of the degenerative tears, but in only two (25%) of the acute tears. Immunohistochemical staining showed that the amyloid deposits were positive for P component, but negative for kappa and lambda light chains, prealbumin, and beta2 microglobulin. Critical electrolyte staining revealed highly-sulphated glycosaminoglycans at sites of deposition of amyloid. The presence of localised deposition of amyloid in tears of the rotator cuff is likely to represent irreversible structural changes. These findings support the theory that impingement and tears are due to intrinsic degenerative changes within the tendons of the rotator cuff.

Characterisation of human deltoid muscle in patients with impingement syndrome.

Muscle biopsies from the anterior and medial parts of the deltoid of 11 male patients and six male controls were analysed with morphological and immunohistochemical methods. The distribution, area, and capillarization of the muscle fibres were determined, and the amount of connective tissue was measured with staining for type-III collagen. Compared with the controls, the patients with impingement syndrome had more type-I than type-II fibres, but the areas of the different types were almost the same. There was no difference in capillarization per fibre type between patients and controls, but the patients had more connective tissue. The results indicate that patients with impingement syndrome have morphological changes in the deltoid muscle, probably due to immobilisation and pain. They support the hypothesis that the deltoid muscle, the medial part in particular, is affected in patients with impingement syndrome.