Peracetic acid sterilized tendon and ligament allografts for knee reconstruction : For anterior cruciate ligament (ACL), posterior cruciate ligament (PCL) and complex knee surgery.

BACKGROUND: The use of allografts and autografts has been met with mixed views on whether allografts are a suitable alternative to autografts. QUESTION: We aimed to investigate if chemically sterilized allografts show similar rerupture rates to those reported in the literature for allografts and autografts in anterior (ACL) and posterior cruciate ligaments (PCL) and complex knee surgery. MATERIALS AND METHODS: Retrospective data on knee reconstructions performed between 2011 and 2015 with tendon/ligamnet allografts sterilized with peracetic acid were collected in the form of a questionnaire. The inclusion criteria of 2 years for each patient were met by 38 patients, representing 22 ACL reconstructions, 5 PCL reconstructions, 3 OTHER surgeries, including the Larson technique and medial patellofemoral ligament (MPFL) reconstruction and 8 COMPLEX surgeries. The main endpoints were rerupture and complication rate. Secondary endpoints included stability of the knee (Lachman test, Pivot shift test) and the range of motion. RESULTS: The rerupture rate was 7.9% (3 grafts). Reruptures only occurred in the ACL group. No reruptures were observed in the PCL, OTHER and COMPLEX surgery groups. Stability improved significantly after surgery and the range of motion returned to values similar to that of healthy knees. CONCLUSIONS: Tendon allografts sterilized with peracetic acid show promising low rerupture rates and good clinical scores and the results are comparable to the literature on autografts and other allografts.

3D osteotomies-improved accuracy with patient-specific instruments (PSI).

PURPOSE: Three-dimensional (3D) printed patient-specific instruments (PSI) have been introduced to increase precision and simplify surgical procedures. Initial results in femoral and tibial osteotomies are promising, but validation studies on 3D planning, manufacturing of patient-specific cutting blocks and 3D evaluation of the attained results are lacking. METHODS: In this study, patient-specific cutting blocks and spacers were designed, fabricated, and used to perform a high tibial osteotomy (HTO). After segmentation of CT data sets from 13 human tibiae, 3D digital planning of the HTO was performed with a medial opening of 8 mm. These 3D models were used to fabricate patient-specific cutting blocks and spacers. After the surgical procedure, accuracy was evaluated measuring 3D joint angles and surface deviations. RESULTS: The lowest mean deviation was found to be 0.57° (SD ± 0.27) for the MPTA. Medial and lateral tibial slope deviated from the 3D planning by an average of 0.98° (SD ± 0.53) and 1.26° (SD ± 0.79), respectively, while tibial torsion deviated by an average of 5.74° (SD ± 3.24). Color analysis of surface deviations showed excellent and good agreement in 7 tibiae. CONCLUSION: With 3D cutting blocks and spacers, the 3D planning of the HTO can be translated into reality with small deviations of the resulting joint angles. Within this study, the results of the individual steps are examined for errors and thus a critical evaluation of this new and promising method for performing patient-specific HTOs is presented.

[The importance of ligament reconstruction with existing osteoarthritis]. / Stellenwert der Bandrekonstruktion bei bestehender Arthrose.

Regarding the importance of ligament replacement in existing osteoarthritis three topics are highlighted: the development of osteoarthritis after ACL-tear or -replacement, ACL-replacement in existing osteoarthritis and ACL-replacement together with medial unicompartmental knee replacement. Concomitant lesions at the meniscus and cartilage, especially the patella-femoral cartilage are risk factors for the development and progression of osteoarthritis in ACL insufficiency. The treatment of a symptomatic ACL-insufficiency in existing osteoarthritis in the elderly patient is directly dependent on pre-existing degenerative changes. Medial unicompartmental knee replacement and ACL-replacement can, however, be well combined and lead to very good long-term results even in the young patient.

The influence of radio frequency ablation on intra-articular fluid temperature in the ankle joint - a cadaver study.

BACKGROUND: Radio frequency ablation devices have found a widespread application in arthroscopic surgery. However, recent publications report about elevated temperatures, which may cause damage to the capsular tissue and especially to chondrocytes. The purpose of this study was the investigation of the maximum temperatures that occur in the ankle joint with the use of a commercially available radio frequency ablation device. METHODS: Six formalin-fixed cadaver ankle specimens were used for this study. The radio frequency device was applied for 120 s to remove tissue. Intra-articular temperatures were logged every second for 120 s at a distance of 3, 5 and 10 mm from the tip of the radio frequency device. The irrigation fluid flow was controlled by setting the inflow pressure to 10 mmHg, 25 mmHg, 50 mmHg and 100 mmHg, respectively. The controller unit voltage setting was set to 1, 5 and 9. RESULTS: Maximum temperatures exceeding 50 °C/122 °F were observed for all combinations of parameters, except for those with a pressure of 100 mmHg pressure. The main critical variable is the pressure setting, which is highly significant. The controller unit voltage setting showed no effect on the temperature measurements. The highest temperature was 102.7 °C/215.6 °F measured for an irrigation flow of 10 mmHg. The shortest time span to exceed 50 °C/122 °F was 3 s. CONCLUSION: In order to avoid temperatures exceeding 50 °C/122 °F in the use of radio frequency devices in arthroscopic surgeries of the ankle joint, it is recommended to use a high irrigation flow by setting the pressure difference across the ankle joint as high as feasible. Even short intervals of a low irrigation flow may lead to critical temperatures above 50 °C/122 °F. LEVEL OF EVIDENCE: Level II, diagnostic study.

Left Ventricular Assist Device Therapy for Destination Therapy: Is Less Invasive Surgery a Safe Alternative?

INTRODUCTION AND OBJECTIVES: The number of older patients with congestive heart failure has dramatically increased. Because of stagnating cardiac transplantation, there is a need for an alternative therapy, which would solve the problem of insufficient donor organ supply. Left ventricular assist devices (LVADs) have recently become more commonly used as destination therapy (DT). Assuming that older patients show a higher risk-profile for LVAD surgery, it is expected that the increasing use of less invasive surgery (LIS) LVAD implantation will improve postoperative outcomes. Thus, this study aimed to assess the outcomes of LIS-LVAD implantation in DT patients. METHODS: We performed a prospective analysis of 2-year outcomes in 46 consecutive end-stage heart failure patients older than 60 years, who underwent LVAD implantation (HVAD, HeartWare) for DT in our institution between 2011 and 2013. The patients were divided into 2 groups according to the surgical implantation technique: LIS (n = 20) vs conventional (n = 26). RESULTS: There was no statistically significant difference in 2-year survival rates between the 2 groups, but the LIS group showed a tendency to improved patient outcome in 85.0% vs 69.2% (P = .302). Moreover, the incidence of postoperative bleeding was minor in LIS patients (0% in the LIS group vs 26.9% in the conventional surgery group, P < .05), who also showed lower rates of postoperative extended inotropic support (15.0% in the LIS group vs 46.2% in the conventional surgery group, P < .05). CONCLUSIONS: Our data indicate that DT patients with LIS-LVAD implantation showed a lower incidence of postoperative bleeding, a reduced need for inotropic support, and a tendency to lower mortality compared with patients treated with the conventional surgical technique.

Return to sports after plate fixation of humeral head fractures 65 cases with minimum 24-month follow-up.

BACKGROUND: Humeral head fractures requiring surgical intervention are severe injuries, which might affect the return to sports and daily activities. We hypothesize that athletic patients will be constrained regarding their sporting activities after surgically treated humeral head fractures. Despite a long rehabilitation program physical activities will change and an avoidance of overhead activities will be noticed. METHODS: Case series with 65 Patients, with a minimum follow-up of 24 months participated in this study. All patients were treated using a locking plate fixation. Their sporting activity was investigated at the time of the injury and re-investigated after an average of 3.83 years. The questionnaire setup included the evaluation of shoulder function, sporting activities, intensity, sport level and frequency evaluation. Level of evidence IV. RESULTS: At the time of injury 61 Patients (94%) were engaged in recreational sporting activities. The number of sporting activities declined from 26 to 23 at the follow-up examination. There was also a decline in sports frequency and duration of sports activities. CONCLUSION: The majority of patients remains active in their recreational sporting activity at a comparable duration and frequency both pre- and postoperatively. Nevertheless, shoulder centered sport activities including golf, water skiing and martial arts declined or were given up.

Balloon-guided inflation osteoplasty in the treatment of Hill-Sachs lesions of the humeral head: case report of a new technique.

BACKGROUND: The use of the extra-vertebral balloon osteoplasty is increasing and in the meanwhile it has become a safe surgical technique in the treatment of tibial head, distal radius and calcaneus fractures. In addition, we already could show in a biomechanical setup that the balloon osteoplasty might be a safe tool for reduction in the treatment of Hill-Sachs lesions, but clinical application has not been performed so far. CASE PRESENTATION: We report the case of a 53 year- old male patient who was referred to our Trauma department (level I trauma center) after the first manifestation of a posterior shoulder dislocation due to an epileptic seizure, originated in a- up to this date unknown -glioblastoma. After closed reduction of the dislocated shoulder the X-ray showed a subcapital fracture of the proximal humerus with a large reversed Hill-Sachs lesion. We performed an open surgery via a deltoideopectoral approach and balloon osteoplasty was used to reduce the impression fracture (Hill-Sachs lesion) before fixing the fracture with a locking plate. In the post-operative CT scan we could show an anatomical reduction of the Hill-Sachs lesion. At the follow-up examination one year after surgery the patient reached full range of motion and stated no re-dislocation of the shoulder nor instability or pain. CONCLUSION: The reduction of an impressed humeral head fracture by use of balloon osteoplasty is a safe technique. This technique could be a new option in the treatment of Hill-Sachs lesions and might be an alternative to well known standard procedures like the remplissage or tendon transfers without affecting rotation.

Dynamic quantification of tibio-femoral rotation in postero-lateral bundle insufficiency of the anterior cruciate ligament: a cadaver study.

PURPOSE: Anterior cruciate ligament (ACL) injury represents one of the most common diagnoses in orthopaedic sports medicine. In the past, anatomic knowledge about the different bundles within the cruciate ligaments triggered new treatment concepts, such as double-bundle ACL reconstruction. Recently, besides complete tearing, partial ACL ruptures and bundle augmentation became a focus. However, only little is known regarding rotational stability of the knee with an isolated torn postero-lateral (PL) bundle. Therefore, the aim of the present study was the torsiometric analysis of tibio-femoral restraint patterns of the PL-insufficient knee joint. METHODS: Fresh human whole body cadavers were enrolled. After diagnostic arthroscopy to ensure the structural integrity of the cruciate ligaments, knee joints underwent torsiometry at 0°, 30°, and 90° degree flexion. Then stepwise the PL bundle and the anteromedial (AM) bundle were arthroscopically resected, while torsiometry of the PL- as well as of the ACL-deficient knee joints was repeated. An area under the curve (AUC) was calculated. All statistical analyses were conducted using a p-value of 0.05 as level of significance. RESULTS: The comparison of charged and equilibrated curves during internal rotation revealed significant results at low flexion (30° flexion) angles between the ACL intact versus PL absent conditions (p = 0.04). In addition, charged and equilibrated curves during external rotation at 90° flexion, thus high angles, resulted in a significant difference when comparing the ACL-intact with the PL-deficient condition (p = 0.01). CONCLUSIONS: In the present cadaver study using the Torsiometer tool we found a distinct destabilization of the rotational restraints in full knee extension only after total ACL resection. In contrast, no significant findings resulted after an isolated dissection of the PL bundle during internal deflection. Nevertheless, a significant loss of stability was found during unstressed external deflection after isolated PL bundle dissection. Therefore patients, undergoing PL augmentation might benefit regarding rotational instability patterns.

Influence of extremely low frequency, low energy electromagnetic fields and combined mechanical stimulation on chondrocytes in 3-D constructs for cartilage tissue engineering.

Articular cartilage, once damaged, has very low regenerative potential. Various experimental approaches have been conducted to enhance chondrogenesis and cartilage maturation. Among those, non-invasive electromagnetic fields have shown their beneficial influence for cartilage regeneration and are widely used for the treatment of non-unions, fractures, avascular necrosis and osteoarthritis. One very well accepted way to promote cartilage maturation is physical stimulation through bioreactors. The aim of this study was the investigation of combined mechanical and electromagnetic stress affecting cartilage cells in vitro. Primary articular chondrocytes from bovine fetlock joints were seeded into three-dimensional (3-D) polyurethane scaffolds and distributed into seven stimulated experimental groups. They either underwent mechanical or electromagnetic stimulation (sinusoidal electromagnetic field of 1 mT, 2 mT, or 3 mT; 60 Hz) or both within a joint-specific bioreactor and a coil system. The scaffold-cell constructs were analyzed for glycosaminoglycan (GAG) and DNA content, histology, and gene expression of collagen-1, collagen-2, aggrecan, cartilage oligomeric matrix protein (COMP), Sox9, proteoglycan-4 (PRG-4), and matrix metalloproteinases (MMP-3 and -13). There were statistically significant differences in GAG/DNA content between the stimulated versus the control group with highest levels in the combined stimulation group. Gene expression was significantly higher for combined stimulation groups versus static control for collagen 2/collagen 1 ratio and lower for MMP-13. Amongst other genes, a more chondrogenic phenotype was noticed in expression patterns for the stimulated groups. To conclude, there is an effect of electromagnetic and mechanical stimulation on chondrocytes seeded in a 3-D scaffold, resulting in improved extracellular matrix production.

The functional outcome of forty-nine single-incision suture anchor repairs for distal biceps tendon ruptures at the elbow.

PURPOSE: The objective of this retrospective study was to evaluate the functional results of distal biceps tendon repair using suture anchors via a single-incision approach. METHODS: Forty-nine patients were re-examined at a mean follow-up of 44.2 ± 32.1 months (range, 12-119 months). Subjective and objective criteria included patient's satisfaction, active range of motion (ROM), maximum isometric strength in flexion (at 45° and 90°), and supination of both arms. Functional scoring included the Morrey elbow score (MES) and the QuickDASH. Furthermore, follow-up radiographs were performed. RESULTS: Eighty-six percent of patients were highly satisfied or satisfied with their outcome. Compared to contralateral, the active ROM of elbow flexion, extension, and pronation was not affected; however, supination was decreased by 3° (P < 0.001). The isometric maximum strengths showed significant deficits in all tested scenarios (at 45°, P = 0.002; at 90°, P < 0.001; for supination, P < 0.001). The MES and the QuickDASH were 97.2 ± 4.9 and 7.9 ± 13.9, respectively. Heterotopic ossifications (HO) were found in 39% of patients; however, with respect to scores and strength, no significant differences were seen compared to patients without HO. Moreover, four anchor failures were detected. CONCLUSIONS: Single-incision suture anchor repair provides high patient's satisfaction and good results with respect to ROM and functional scoring. Nevertheless, based on presented data, the patient has to be informed of postoperative HO and especially for supination strength weakness after surgery. Distal biceps tendon repair should be reserved for experienced upper extremity surgeons to avoid procedure-related complications.

X-treme CT analysis of cancellous bone at the rotator cuff insertion in human individuals with osteoporosis: superficial versus deep quality.

PURPOSE: Rotator cuff (RC) repair-especially in the elderly population-is problematic since the patients suffer to a high extent from bone mineral density loss at the reattachment site. Therefore, the study was primarily driven by the question whether it is possible to reach more or qualitatively better cancellous bone and thus a more stable postoperative result if anchors with greater length are used for RC repair and/or the conventional anchors are screwed deeper into the bone. In anatomical terms, the question is raised whether cancellous bone is of better quality close to or far off the RC enthesis. METHODS: Axial HRqCT scans (X-tremeCT, Scanco Medical) of 36 human cadaveric humeral heads (75 ± 11 years) were performed to determine the ratio of bone volume to total volume (BV/TV), trabecular thickness (Trab Th), number of trabecles (Trab N), trabecular separation (Trab Sp) as well as non-metric indices such as connectivity density (Conn Dens) and structure model index (SMI). Within the greater tuberosity (GT), 6 volumes of interest (VOI) (A1, B1, C1, A2, B2, C2), in the lesser tuberosity (LT) 2 VOIs (D1, D2) and one control VOI in the subchondral bone were set. The analyzed bone cylinder of each VOI was divided into a superficial and a deep portion. RESULTS: The parameters BV/TV, Trab N, Trab Th and Conn Dens in all volumes of the GT and LT revealed higher values in the superficial portion reaching different levels of significance (p < 0.001/<0.05). The only parameter presenting a higher value in the deep portion was Trab Sp, but this was the case for all GT and LT regions. Interestingly, the difference between the superficial and deep portion reached significance for the non-metric parameter SMI in no volume of the GT/LT, although the higher values were found superficially. CONCLUSIONS: Our data show that cancellous bone presents with decreasing bone quality when analyzing increasingly deeper portions of the bone cylinders of the GT and LT starting at the articular surface. This information seems to be crucial for shoulder surgeons, especially when treating elderly patients. Our results clearly prove that screwing in anchors to a deeper extent will not improve stability, since the deeper bone stock is of worse quality.

Plating of proximal humeral fractures.

The optimal treatment for proximal humeral fractures is controversial. Few data exist concerning the influence of the surgical approach on the outcome. The purpose of this study was to evaluate the clinical and radiological outcomes of proximal humeral fractures treated with locking plate fixation through a deltopectoral vs an anterolateral deltoid-splitting approach. Of 86 patients who met the inclusion criteria, 70 were available for follow-up examination. Thirty-three patients were treated through a deltopectoral approach and 37 through an anterolateral deltoid-splitting approach. In all cases, open reduction and internal fixation with a PHILOS locking plate (Synthes, Umkirch, Germany) was performed. Clinical follow-up included evaluation of pain, shoulder mobility, and strength. Constant score and Disabilities of the Arm, Shoulder and Hand (DASH) score were assessed. A clinical neurological examination of the axillary nerve was also performed. Consolidation, reduction, and appearance of head necrosis were evaluated radiographically. After a mean follow-up of 33 months, Constant scores, DASH scores, and American Shoulder and Elbow Surgeons scores showed no significant differences between the groups. Clinical neurologic examination of the axillary nerve revealed no obvious damage to the nerve in either group. Deltopectoral and anterolateral detoid-splitting approaches for plate fixation of proximal humeral fractures are safe and provide similar clinical outcomes. The results of this study suggest that the approach can be chosen according to surgeon preference.

Return to sports activity after tibial plateau fractures: 89 cases with minimum 24-month follow-up.

BACKGROUND: Tibial plateau fractures requiring surgery are severe injuries. For professionals, amateurs, and recreational athletes, tibial plateau fractures might affect leisure and professional life. HYPOTHESIS: Athletic patients will be affected in their sporting activity after a tibial plateau fracture. Despite a long rehabilitation time and program, physical activity will change to low-impact sports. STUDY DESIGN: Case series; Level of evidence, 4. METHODS: A total of 89 consecutive patients (age range, 14-76 years) were included in the study and were surveyed by a questionnaire. Inclusion criteria were surgical treatment of tibial plateau fractures between 2003 and 2009 with a minimum follow-up of 24 months. The sporting activity was determined at the time of injury, 1 year postoperatively, and at the time of the survey at an average of 52.8 months postoperatively. The clinical evaluation included the Lysholm score, the Tegner activity scale, the activity rating scale (ARS), and a visual analog scale (VAS) for pain perception. Fractures were classified and analyzed using both the Arbeitsgemeinschaft für Osteosynthesefragen (AO) and the Schatzker classifications. RESULTS: At the time of injury, 88.8% of all patients were engaged in sports compared with 62.9% 1 year postoperatively and 73.0% at the time of the survey. Of the professional or competitive athletes (n = 11 at the time of injury), only 2 returned to competition at the time of the survey. The number of different sporting activities declined from 4.9 at the time of injury to 3.6 at the time of the survey (P < .001). The sports frequency and the activity duration per week, being 2.8 sessions and 4.5 hours at the time of injury, respectively, declined to 2.4 sessions and 3.8 hours (P < .001 and P = .007, respectively) at the time of the survey, respectively. The Lysholm score (98.7 points before accident) and the VAS for pain perception (0.2 before accident) illustrated significant declines to 76.6 points for the Lysholm score and 2.6 for the VAS (P < .001 and P < .001, respectively) at the time of the survey. The high-energy traumas, Schatzker IV to VI, had significant worse results in the clinical scores compared with the low-energy traumas (Lysholm, P < .001; Tegner, P = .027). CONCLUSION: The majority of patients could not return to their previous level of activity, and for patients playing competitive sports, this injury can be a career ender. Overall, we noticed a postinjury shift toward activities with less impact. However, at the time of the survey, 73% of all patients were engaged in sports.

Reconstruction of displaced acromio-clavicular joint dislocations using a triple suture-cerclage: description of a safe and efficient surgical technique.

PURPOSE: In this retrospective study we investigated the clinical and radiological outcome after operative treatment of acute Rockwood III-V injuries of the AC-joint using two acromioclavicular (AC) cerclages and one coracoclavicular (CC) cerclage with resorbable sutures. METHODS: Between 2007 and 2009 a total of 39 patients fit the inclusion criteria after operative treatment of acute AC joint dislocation. All patients underwent open reduction and anatomic reconstruction of the AC and CC-ligaments using PDS® sutures (Polydioxane, Ethicon, Norderstedt, Germany). Thirty-three patients could be investigated at a mean follow up of 32±9 months (range 24-56 months). RESULTS: The mean Constant score was 94.3±7.1 (range 73-100) with an age and gender correlated score of 104.2%±6.9 (88-123%). The DASH score (mean 3.46±6.6 points), the ASES score (94.6±9.7points) and the Visual Analogue Scale (mean 0.5±0,6) revealed a good to excellent clinical outcome. The difference in the coracoclavicular distance compared to the contralateral side was <5 mm for 28 patients, between 5-10 mm for 4 patients, and more than 10 mm for another patient. In the axial view, the anterior border of the clavicle was within 1 cm (ventral-dorsal direction) of the anterior rim of the acromion in 28 patients (85%). Re-dislocations occured in three patients (9%). CONCLUSION: Open AC joint reconstruction using AC and CC PDS cerclages provides good to excellent clinical results in the majority of cases. However, radiographically, the CC distance increased significantly at final follow up, but neither the amount of re-dislocation nor calcifications of the CC ligaments or osteoarthritis of the AC joint had significant influence on the outcome. LEVEL OF EVIDENCE: Case series, Level IV.

Balloon osteoplasty--a new technique for minimally invasive reduction and stabilisation of Hill-Sachs lesions of the humeral head: a cadaver study.

PURPOSE: Traumatic shoulder dislocation may be complicated by concomitant bony injury of the glenoid rim or the humeral head. In Hill-Sachs lesions, reconstruction techniques vary widely and range from open reduction to tendon transposition or humeral head derotation. These operations are extensive and have questionable outcomes. With the expertise from vertebral compression fracture reduction by kyphoplasty, we examined in a cadaver feasibility study whether reduction of the Hill-Sachs lesion via hydraulic lift might be an anatomical and minimally invasive treatment option. We postulated that the use a of a balloon- assisted kyphoplasty reduction could achieve almost anatomical correction of the defect. METHODS: We created Hill-Sachs lesions in six humeral specimens and performed a computed tomography (CT) scan before and after reduction with the kyphoplasty system. The entry point at the greater tuberosity and balloon positioning was visualised by fluoroscopy. The size of the Hill-Sachs lesion before and after reduction was measured using CT scans in the axial orientation. RESULTS: Using the balloon kyphoplasty system, we achieved a statistically significant reduction (80 % ) of the Hill-Sachs lesion. CONCLUSION: In a preliminary cadaver study we show that using a balloon kyphoplasty system might be an alternative treatment option for Hill-Sachs lesions, with reduced collateral damage that can occur with other minimally invasive techniques. Future work is needed to evaluate the technique under arthroscopic conditions.

Balloon osteoplasty--a new technique for reduction and stabilisation of impression fractures in the tibial plateau: a cadaver study and first clinical application.

PURPOSE: Fractures of the tibial plateau are among the most severe injuries of the knee joint and lead to advanced gonarthrosis if the reduction does not restore perfect joint congruency. Many different reduction techniques focusing on open surgical procedures have been described in the past. In this context we would like to introduce a novel technique which was first tested in a cadaver setup and has undergone its successful first clinical application. METHODS: Since kyphoplasty demonstrated effective ways of anatomical correction in spine fractures, we adapted the inflatable instruments and used the balloon technique to reduce depressed fragments of the tibial plateau. RESULTS: The technique enabled us to restore a congruent cartilage surface and bone reduction. CONCLUSIONS: In this technique we see a useful new method to reduce depressed fractures of the tibial plateau with the advantages of low collateral damage as it is known from minimally invasive procedures.

MicroRNA miR-335 is crucial for the BRCA1 regulatory cascade in breast cancer development.

The expression of microRNAs is altered in various cancer types, leading to their definition as onco- and tumor-suppressor microRNAs. In our study, we investigated the role of miR-335 in the formation of sporadic human breast cancer and its involvement in the regulatory network of the breast cancer susceptibility gene BRCA1. To validate single components of the BRCA1 cascade, microRNA overexpression was performed in a cell culture model with subsequent protein analysis and luciferase reporter assays. Here, we were able to identify miR-335 as simultaneously regulating the known BRCA1 activators ERα, IGF1R, SP1 and the repressor ID4, including a feedback regulation of miR-335 expression by estrogens. Overexpression of miR-335 resulted in an upregulation of BRCA1 mRNA expression, suggesting a functional dominance of ID4 signaling. The relevance of the miR-335 regulation for human breast cancer was confirmed in primary sporadic breast cancer specimens with significantly decreased miR-335 levels (p < 0.05) in comparison to normal controls. Interestingly, the microRNA expression level correlated positively to the BRCA1 transcript level, supporting the hypothesis of a miR-335-mediated regulation of the tumor suppressor gene. Functionally, overexpression of miR-335 led to decreased cell viability and an increase in apoptosis, supporting its tumor-suppressive function. In summary, our data indicate that miR-335 affects different targets in the upstream BRCA1-regulatory cascade with impact on key cellular functions such as proliferation and apoptosis. Deregulation of the microRNA during breast cancer development and progression may thereby lead to an increased tumorigenic potential by inactivating crucial tumor-suppressive signals.

MicroRNA profiles of healthy basal and luminal mammary epithelial cells are distinct and reflected in different breast cancer subtypes.

In order to determine the microRNA expression pattern in normal basal and luminal breast epithelium and to analyze the relationship of this expression pattern to different breast cancer subtypes, laser-microdissected luminal and basal cells isolated from plastic surgery tissue samples were used for comprehensive expression profiling, measuring 664 microRNAs by low-density TaqMan arrays. In a test (n = 5) and validation set (n = 9) 10 differentially expressed microRNAs were identified by TaqMan RT-qPCR. These microRNAs were studied in laser-microdissected cells of luminal A (n = 5), luminal B (n = 5), basal-like subtypes of breast cancer (n = 10), and malignant myoepithelioma of the breast (n = 10). From 116 microRNAs unequivocally expressed in normal breast epithelial cells, we identified 8 basal microRNAs (let7c, miR-125b, miR-126, miR-127-3p, miR-143, miR-145, miR-146b-5p, and miR-199a-3p), preferentially expressed in normal basal cells, exceeding luminal cells by a factor from 4 to 1000. All of these microRNAs were also found to be significantly elevated in malignant myoepithelioma but not in basal-type of breast cancer. Two members of the miR-200 family (miR-200c and miR-429) were predominantly luminal. Both microRNAs were expressed in the luminal and basal type of breast cancer in contrast to malignant myoepithelioma, which revealed significantly lower levels potentially contributing to its mesenchymal phenotype. In conclusion, normal luminal and basal mammary epithelial cells exhibit a different microRNA expression profile. Malignant myoepithelioma seems to exhibit a basal pattern of microRNA expression, whereas the so-called basal-like breast cancer is clearly different and reveals a luminal type pattern.