Toshiyuki Tashiro

Neutralization of fibroblast growth factor-2 reduces intraarticular adhesions.

Adhesion is a serious complication after trauma or surgery. Because adhesion formation is essentially a fibrogenetic process, a series of growth factors are assumed to be involved in its development. If this is true, it may be possible that inhibition of the growth factor activity suppresses adhesion formation. The current study was conducted to verify this hypothesis on fibroblast growth factor-2 using an intraarticular adhesion model in the rabbit knee. Forty Japanese White rabbits were used. They were divided randomly into five groups of eight animals, and in three of them, activity of endogenous fibroblast growth factor-2 was suppressed locally by a neutralizing antibody. The remaining two groups served as controls, and formation of adhesions was evaluated 4 weeks after surgery. The results showed that the administration of the antibody reduced the extent of adhesions macroscopically, whereas histologic observation and collagen content measurement suggested the adhesion tissue was not affected significantly. Corresponding to the macroscopic findings, contraction of the knee was improved in the antibody groups. The findings showed that suppression of fibroblast growth factor-2 activity reduces adhesions. It is expected that control of the cytokine activity may become a novel method for reducing adhesions.

Influence of Medial Hamstring Tendon Harvest on Knee Flexor Strength after Anterior Cruciate Ligament Reconstruction

Background The advantages of hamstring tendon autografts for anterior cruciate ligament reconstruction are well known; however, concerns have arisen regarding the influence of hamstring tendon harvest on postoperative weakness in knee flexion. Purpose To evaluate the influence of hamstring tendon harvest on knee flexion strength in patients undergoing anterior cruciate ligament reconstruction. Study Design Prospective randomized study. Methods Ninety patients were randomly assigned at surgery to undergo anterior cruciate ligament reconstruction with either a semitendinosus tendon autograft or a semitendinosus and gracilis tendon autograft. Quadriceps and hamstring muscle strength was tested before surgery and at 6, 12, and 18 months after surgery. Results There was no significant difference in clinical results between the groups and neither group showed a significant decrease in isokinetic hamstring muscle strength. However, when the subjects’ knees were at positions of 70° or more of flexion, both isokinetic and isometric measurements revealed a significant decrease in hamstring muscle strength in both groups. The strength in the group with semitendinosus and gracilis tendons was considerably less than that in the group with semitendinosus tendon alone at 18 months. Conclusions Tendon harvest causes significant weakness of hamstring muscle strength at high knee flexion angles, but such weakness can be minimized if the gracilis tendon is preserved.

Genome-wide DNA methylation profile implicates potential cartilage regeneration at the late stage of knee osteoarthritis

OBJECTIVE The aim of this work was to characterize the genome-wide DNA methylation profile of cartilage from three regions of tibial plateau isolated from patients with primary knee osteoarthritis (OA), providing the first DNA methylation study that reflects OA progression. METHODS The unique model system was used to section three regions of tibial plateau: the outer lateral tibial plateau (oLT), the inner lateral tibial plateau (iLT) and the inner medial tibial plateau (iMT) regions which represented the early, intermediate and late stages of OA, respectively. Genome-wide DNA methylation profile was examined using Illumina Infinium HumanMethylation450 BeadChip array. Comparisons of the iLT/oLT and iMT/oLT groups were carried out to identify differentially methylated (DM) probes (DMPs) associated with OA progression. DM genes were analyzed to identify the gene ontologies (GO), pathways, upstream regulators and networks. RESULTS No significant DMPs were identified in iLT/oLT group, while 519 DMPs were identified in iMT/oLT group. Over half of them (68.2%) were hypo-methylated and enriched in enhancers and OpenSea. Upstream regulator analysis identified many microRNAs. DM genes were enriched in transcription factors, especially homeobox genes and in Wnt/β-catenin signaling pathway. These genes also showed changes in expression when analyzed with expression profiles generated from previous studies. CONCLUSION Our data suggested the changes in DNA methylation occurred at the late stage of OA. Pathways and networks enriched in identified DM genes highlighted potential etiologic mechanism and implicated the potential cartilage regeneration in the late stage of knee OA.

αvβ5 Integrin promotes dedifferentiation of monolayer-cultured articular chondrocytes

OBJECTIVE When cultured in monolayers, articular chondrocytes undergo an obvious phenotypic change. Although the involvement of integrins has been suggested, the exact mechanisms of the change have not been determined. This study was undertaken to clarify the mechanisms underlying the loss of chondrocyte phenotype early after plating. METHODS Primary cultured human articular chondrocytes were used for the experiments. Involvement of respective integrins in the phenotypic change was investigated in RNA interference (RNAi) experiments. A signaling pathway involved in the change was identified in experiments using specific inhibitors and adenoviruses encoding mutated genes involved in the pathway. Adenoviruses carrying mutated GTPases were used to determine the involvement of small GTPases in the process. RESULTS In monolayer-cultured chondrocytes, suppression of αv or β5 integrin expression by RNAi inhibited morphologic changes in the cells and increased (or prevented a reduction in) the expression of various cartilage matrix genes. Consistent results were obtained in experiments using a blocking antibody and a synthetic inhibitor of αvβ5 integrin. The decrease in cartilage matrix gene expression in chondrocytes after plating was mediated by ERK signaling, which was promoted primarily by αvβ5 integrin. In articular chondrocytes, the affinity of αvβ5 integrin for ligands was regulated by the small GTPase R-Ras. R-Ras was gradually activated in monolayer-cultured chondrocytes after plating, which caused a gradual decline in cartilage matrix gene expression through enhanced αvβ5 integrin activation and the subsequent increase in ERK signaling. CONCLUSION Our findings indicate that αvβ5 integrin may be involved in the change that occurs in monolayer-cultured chondrocytes after plating.

Identification of DNA methylation changes associated with disease progression in subchondral bone with site-matched cartilage in knee osteoarthritis

Subchondral bone plays a key role in the development of osteoarthritis, however, epigenetics of subchondral bone has not been extensively studied. In this study, we examined the genome-wide DNA methylation profiles of subchondral bone from three regions on tibial plateau representing disease progression using HumanMethylation450 BeadChip to identify progression associated DNA methylation alterations. Significant differential methylated probes (DMPs) and differential methylated genes (DMGs) were identified in the intermediate and late stages and during the transition from intermediate to late stage of OA in the subchondral bone. Over half of the DMPs were hyper-methylated. Genes associated with OA and bone remodeling were identified. DMGs were enriched in morphogenesis and development of skeletal system, and HOX transcription factors. Comparison of DMGs identified in subchondral bone and site-matched cartilage indicated that DNA methylation changes occurred earlier in subchondral bone and identified different methylation patterns at the late stage of OA. However, shared DMPs, DMGs and common pathways that implicated the tissue reparation were also identified. Methylation is one key mechanism to regulate the crosstalk between cartilage and subchondral bone.

Oral hyaluronan relieves knee pain: a review

Hyaluronan (HA) is a component that is particularly abundant in the synovial fluid. Randomized, double-blinded, placebo-controlled trials carried out between 2008 and 2015 have proven the effectiveness of HA for the treatment of symptoms associated with synovitis, and particularly, knee pain, relief of synovial effusion or inflammation, and improvement of muscular knee strength. The mechanism by which HA exerts its effects in the living body, specifically receptor binding in the intestinal epithelia, has gradually been clarified. This review examines the effects of HA upon knee pain as assessed in clinical trials, as well as the mechanism of these effects and the safety of HA.

Anatomical reconstruction of the patellar tendon : A new technique with hamstring tendons and iliotibial tract

A new technique of patellar tendon reconstruction was performed in a patient who lost tendon and tibial tuberosity during wide excision surgery for a malignancy. In this procedure, the tendon was anatomically replaced by a graft composed of ipsilateral hamstring tendons and iliotibial tract, with the biomechanical conditions considered. Both ends of the graft were secured in the size-matched bone tunnels in the patella and tibia by screw post fixation, which is a technique established in ligament reconstruction surgery in the knee joint. At the twenty-month follow-up, the result was deemed successful.