Complexity of IL-1β induced gene expression pattern in human articular chondrocytes

SummaryThe mRNA fingerprinting technique, differential display reverse transcription polymerase chain (DDRT-PCR), was used to detect changes in the overall pattern of gene expression in human articular knee chondrocytes induced by interleukin-1β (IL-1β), the prototypical inducer of catabolic responses in degenerate joint diseases. One hundred different primer combinations generated approximately 10 000 different PCR fragments for IL-1β treated, as well as for untreated human chondrocytes, cultivated in alginate beads. This represented 53% of all expressed chondrocyte genes as based on statistical considerations. Side by side comparisons of differential display patterns originating from two different donor tissues yielded 44 reproducibly, differentially-displayed cDNA fragments, which were subcloned and sequenced. Sequence homology searches revealed sequence identities to the human necrosis factor α (TNF-α) and IL-1 regulated gene TSG-6, fibronectin, osteopontin, calnexin, and the DNA repair enzyme ERCC5. The differential expression was confirmed with Northern and quantitative PCR analyses. The known function of these genes and their known IL-1 responsiveness indicate that the employed model system reflects the pleiotropic effects of IL-1 on the overall gene expression in human articular chondrocytes and identifies genes involved in very different biochemical pathways. Twenty-seven cDNAs lacked sequence homologies to known genes and may represent novel genes

Effect of constrained posterior screw and rod systems for primary stability: Biomechanical in vitro comparison of various instrumentations in a single-level corpectomy model

Cervical corpectomy is a frequently used technique for a wide variety of spinal disorders. The most commonly used approach is anterior, either with or without plating. The results for single-level corpectomy are better than in multilevel procedures. Nevertheless, hardware- or graft-related complications are observed. In the past, constrained implant systems were developed and showed encouraging stability, especially for posterior screw and rod systems in the lumbar spine. In the cervical spine, few reports about the primary stability of constrained systems exist. Therefore, in the present study we evaluated the primary stability of posterior screw and rod systems, constrained and non-constrained, in comparison with anterior plating and circumferential instrumentations in a non-destructive set-up, by loading six human cadaver cervical spines with pure moments in a spine tester. Range of motion and neutral zone were measured for lateral bending, flexion/extension and axial rotation. The testing sequence consisted of: (1) stable testing; (2) testing after destabilization and cage insertion; (3a) additional non-constrained screw and rod system with lateral mass screws, (3b) with pedicle screws instead of lateral mass screws; (4a) constrained screw and rod system with lateral mass screws, (4b) with pedicle screws instead of lateral mass screws; (5) 360°set-up; (6) anterior plate. The stability of the anterior plate was comparable to that of the non-constrained system, except for lateral bending. The primary stability of the non-constrained system could be enhanced by the use of pedicle screws, in contrast to the constrained system, for which a higher primary stability was still found in axial rotation and flexion/extension. For the constrained system, the achievable higher stability could obviate the need to use pedicle screws in low instabilities. Another benefit could be fewer hardware-related complications, higher fusion rate, larger range of instabilities to be treated by one implant system, less restrictive postoperative treatment and possibly better clinical outcome. From a biomechanical standpoint, in regard to primary stability the constrained systems, therefore, seem to be beneficial. Whether this leads to differences in clinical outcome has to be evaluated in clinical trials

Gene Expression of Stromelysin and Aggrecan in Osteoarthritic Cartilage

Objective: To analyze cartilage gene expression of patients with osteoarthritis (OA) in correlation with radiographic and histological findings. Materials and Methods: Twenty-one patients with OA of the knee admitted for total knee replacement were analyzed clinically and radiographically by the Kellgren and Lawrence system. During surgery, cartilage samples from the medial and lateral condyles and tibial plateaus were harvested separately. Specimens were analyzed histologically (Mankin score) and total RNA was extracted directly from cartilage tissue. Steady state levels of stromelysin (MMP-3), aggrecan (AGG) and the house-keeping gene β-actin were measured using quantitative PCR. Results: Histology of medial and lateral knee compartments corresponded to radiographic changes (Spearman correlation coefficient: r = 0.7 (p < 0.01)). There was a positive correlation between MMP-3 and AGG gene expression (r = 0.4; p < 0.01). We found considerable variation of expression levels of MMP-3 and AGG and no correlation of gene expression with histological or radiographic scoring. Conclusion: The positive correlation between AGG and MMP-3 suggests a common regulation of anabolic and catabolic metabolism. There was no simple dependency between gene expression and histological and radiological findings in cartilage.Dieser Beitrag ist mit Zustimmung des Rechteinhabers aufgrund einer (DFG-geförderten) Allianz- bzw. Nationallizenz frei zugänglich