Hashimoto K.,New York Medical College |
Hashimoto K.,Tohoku University |
Otero M.,New York Medical College |
Imagawa K.,University of Southampton |
And 7 more authors.
Journal of Biological Chemistry | Year: 2013
The role of DNA methylation in the regulation of catabolic genes such as MMP13 and IL1B, which have sparse CpG islands, is poorly understood in the context of musculoskeletal diseases. We report that demethylation of specific CpG sites at -110 bp and -299 bp of the proximal MMP13 and IL1B promoters, respectively, detected by in situ methylation analysis of chondrocytes obtained directly from human cartilage, strongly correlated with higher levels of gene expression. The methylation status of these sites had a significant impact on promoter activities in chondrocytes, as revealed in transfection experiments with site-directed CpG mutants in a CpG-free luciferase reporter. Methylation of the -110 and -299 CpG sites, which reside within a hypoxia-inducible factor (HIF) consensus motif in the respective MMP13 and IL1B promoters, produced the most marked suppression of their transcriptional activities. Methylation of the -110 bp CpG site in the MMP13 promoter inhibited its HIF-2-driven transactivation and decreased HIF-2α binding to theMMP13proximal promoter in chromatin immunoprecipitation assays. In contrast to HIF-2α, MMP13 transcriptional regulation by other positive (RUNX2, AP-1, ELF3) and negative (Sp1, GATA1, and USF1) factors was not affected by methylation status. However, unlike the MMP13 promoter, IL1B was not susceptible to HIF-2α transactivation, indicating that the -299 CpG site in the IL1B promoter must interact with other transcription factors to modulate IL1B transcriptional activity. Taken together, our data reveal that the methylation of different CpG sites in the proximal promoters of the human MMP13 and IL1B genes modulates their transcription by distinct mechanisms. © 2013 by The American Society for Biochemistry and Molecular Biology, Inc.
Mugnai R.,Modena Policlinic |
Digennaro V.,Modena Policlinic |
Ensini A.,Istituto Ortopedico Rizzoli |
Leardini A.,Centro Of Ricerca Codivilla Putti |
Catani F.,Modena Policlinic
Knee Surgery, Sports Traumatology, Arthroscopy | Year: 2014
Purpose: In a retrospective comparative analysis in patients undergoing primary guided-motion total knee arthroplasty (TKA), the authors have evaluated whether different TKA implant design would influence the clinical and functional outcomes. Methods: Between 2007 and 2009, 227 computer-assisted primary TKAs were performed in 219 consecutive patients. Patients received one of the two different fixed-bearing guided-motion TKA designs assisted by navigation surgery: the Scorpio Non-Restrictive Geometry (NRG) knee system and the Journey Bi-Cruciate Stabilized (BCS) knee systems. Results: Data were available for 180 patients (187 knees). No significant differences were observed between the two groups with respect to preoperative demographic characteristics, range of motion (ROM) and radiographic knee alignment. At a mean follow-up of 29 months, the Journey BCS group had higher mean Knee Injury and Osteoarthritis Outcome Score (KOOS) in all subscales and a greater ROM than the Scorpio NRG group. This difference was statistically significant for the KOOS subscales of pain (p = 0.007) and knee-related quality of life (p = 0.045), as well as for postoperative ROM (p = 0.018). Considering the overall complications, 1 patient of Scorpio NRG group (0.5 %) and 5 in Journey BCS (2.7 %) had stiffness. Anterior knee pain was reported in 4 cases of Scorpio NRG group (2.1 %). In the Journey BCS group were observed 2 cases (1.1 %) of frontal plane instability and 1 case (0.5 %) of synovitis pain. Conclusions: The bearing geometry and kinematic pattern of different guided-motion prosthetic designs can affect the clinical-functional outcome and complications type in primary TKA. Level of evidence: Clinical study, Level III. © 2013 Springer-Verlag Berlin Heidelberg.
Catani F.,Istituto Ortopedico Rizzoli |
Belvedere C.,Centro Of Ricerca Codivilla Putti |
Ensini A.,Istituto Ortopedico Rizzoli |
Feliciangeli A.,Istituto Ortopedico Rizzoli |
And 2 more authors.
Journal of Orthopaedic Research | Year: 2011
Total knee replacement designs claim characteristic kinematic performance that is rarely assessed in patients. In the present study, in vivo kinematics of a new prosthesis design was measured during activities of daily living. This design is posterior stabilized for which spine-cam interaction coordinates free axial rotation throughout the flexion-extension arc by means of a single radius of curvature for the femoral condyles in the sagittal and frontal planes. Fifteen knees were implanted with this prosthesis, and 3D video-fluoroscopic analysis was performed at 6-month follow-up for three motor tasks. The average range of flexion was 70.1° (range: 60.1-80.2°) during stair-climbing, 74.7° (64.6-84.8°) during chair-rising, and 64.1° (52.9-74.3°) during step-up. The corresponding average rotation on the tibial base-plate of the lines between the medial and lateral contact points was 9.4° (4.0-22.4°), 11.4° (4.6-22.7°), and 11.3° (5.1-18.0°), respectively. The pivot point for these lines was found mostly in the central area of the base-plate. Nearly physiological range of axial rotation can be achieved at the replaced knee during activities of daily living. © 2011 Orthopaedic Research Society. Published by Wiley Periodicals, Inc.
Imbert P.,Clinique Notre Dame De La Merci |
Belvedere C.,Centro Of Ricerca Codivilla Putti |
Leardini A.,Centro Of Ricerca Codivilla Putti
BioMedical Engineering Online | Year: 2014
Background: The comprehension of human knee laxity and of the failures of relevant surgical reconstructions of the anterior cruciate ligament (ACL) can be enhanced by the knowledge of the laximetric status of the contralateral healthy knee (CHK). Rarely this is available in patients, directly from the skeletal structures, and for a number of the standard clinical tests. The general aim of this study was to measure the extent to which laxity occurs immediately before surgery in the ACL deficient knee (ADK) with respect to CHK, in a number of standard clinical evaluation tests.Method: Thirty-two patients with ACL deficiency were analyzed at ADK and at CHK by a navigation system immediately before reconstructions. Knee laxity was assessed based on digitized anatomical references during the antero-posterior drawer, Lachman, internal-external rotation, varus-valgus, and pivot-shift tests. Antero-posterior laxity was normalized based on patient-specific length of the tibial plateau.Results: In the drawer test, statistical significance (p < 0.05) was found for the larger antero-posterior laxity in ADK than in CHK, on average, of 54' in the medial and 47' in the lateral compartments, when measured in normalized translations. In the Lachman test, these were about 106' and 68'. The pivot-shift test revealed a significant 70' larger antero-posterior central laxity and a 32' larger rotational laxity. No statistically relevant differences were observed in the other tests.Conclusion: The first conclusion is that it is important to measure also the antero-posterior and rotational laxity of the uninjured contralateral knee in assessing the laxity of the injured knee. A second is that the Lachman test shows knee laxity better than the AP drawer, and that the pivot-shift test was the only one able to reveal rotational instability. The present original measurements and analyses contribute to the knowledge of knee joint mechanics, with possible relevant applications in biomedical and clinical research. © 2014 Imbert et al.; licensee BioMed Central Ltd.
Teti G.,University of Bologna |
Cavallo C.,Centro Of Ricerca Codivilla Putti |
Grigolo B.,Centro Of Ricerca Codivilla Putti |
Giannini S.,University of Bologna |
And 4 more authors.
Microscopy Research and Technique | Year: 2012
The main purpose of this article was to describe the morphology of mesenchymal stem cells (MSCs) differentiated in vitro towards osteogenic and chondrogenic lineages and to focus on the ultrastructural features associated with these processes. Human mononuclear cells (hMNC) were isolated, expanded, and analyzed for the expression of specific cell surface markers to demonstrate their stem cell characteristics. Human mononuclear cells were differentiated in vitro in an osteogenic and in a chondrogenic sense for 7, 14, 21, and 28 days. Subsequently, they were processed using electron microscopic analysis (FEISEM). Alizarin red and alcian blue staining were carried out to demonstrate the deposition of mineral salts and proteoglycans in the extracellular matrix. Undifferentiated MSCs showed a cell surface covered by filopodia and ondulopodia. During differentiation, the MSCs changed their shape from a round to a fibroblastic-like shape. At the end of the differentiation, several filaments with a parallel orientation in the osteogenic samples as well as a network organization in the chondrogenic samples were detected in the extracellular spaces. This study demonstrated that there are morphological features associated with the undifferentiated and differentiated states of the MSCs, which could be utilized as new parameters for identifying and classifying these cells. © 2011 Wiley Periodicals, Inc.