Takeshi Kokubu

Low-intensity pulsed ultrasound initiates bone healing in rat nonunion fracture model

Low‐intensity pulsed ultrasound exposure has been shown clinically to shorten the fracture repair process and to induce healing of nonunions in humans, but its mechanism of action remains unclear. In this study we investigated the effect and mechanism of low‐intensity pulsed ultrasound on nonunion fracture healing in rat tibias. A consistently reproducible nonunion was produced in rat tibias by muscle interposition without osteotomy. This model was produced by creating a closed tibial fracture with only the distal end of the tibialis anterior muscle interposed into the fracture site. One limb was noninvasively exposed to low‐intensity pulsed ultrasound (a 200‐millisecond burst of sine waves of 1.5 MHz, repeating at 1.0 kHz) for 20 minutes daily. The incident intensity was approximately 30 mW/cm2. Rats were killed at intervals between 2 and 6 weeks. The events were assessed by radiographs, microfocus X‐ray computed tomograms, and histologic examination. After 6 weeks of exposure, 7 of 14 nonunion fractures showed healing on radiologic assessment. The results of three‐dimensional microfocus X‐ray computed tomographic reconstruction and histologic examination also supported this finding. On the other hand, all control tibias remained in a state of nonunion during the same period. These results indicate that low‐intensity pulsed ultrasound promotes healing in the rat nonunion fracture model.

Electrospun Synthetic Polymer Scaffold for Cartilage Repair Without Cultured Cells in an Animal Model

PURPOSE The purpose of our study was to explore the possibility that an electrospun bioabsorbable scaffold could be used in the treatment of a full-thickness articular defect without the addition of exogenous cells in a rabbit model. METHODS Two types of poly(D,L-lactide-co-glycolide) (PLG) scaffolds, a solid cylindrical type and a cannulated tubular type, were made with the electrospinning method. Osteochondral defects, 5 mm in diameter and 5 mm in depth, made on the femoral condyles of rabbits were filled with these scaffolds, and the repair process was investigated histologically. RESULTS In the groups in which the defect was filled with the scaffold, fibrous tissue at the articular surface of the scaffold was observed at postoperative week 2. Thereafter cartilage at the articular surface and bone at the subchondral zone were regenerated, and the repaired cartilage was maintained through postoperative week 24. By contrast, the untreated defect was filled with hematoma at postoperative week 2; thereafter regenerated cartilage and bone were observed. However, the surface of the articular cartilage was not regular, and regenerated cartilage was not well organized. The histologic scores of the groups in which the defect was filled with cannulated tubular electrospun PLG scaffolds were significantly higher than those of the untreated defect group at postoperative weeks 12 and 24 (P < .01). CONCLUSIONS The electrospun PLG scaffold could repair a 5-mm osteochondral defect created in the rabbit model without exogenous cultured cells. CLINICAL RELEVANCE The electrospun PLG scaffold could repair full-thickness osteochondral defects. The cannulated type of PLG scaffold has the possibility to lead not only to good regeneration of cartilage but also to easy transplantation by use of a guidewire through the cannulas in the scaffold.

Repair of osteochondral defects with a new porous synthetic polymer scaffold

We developed a new porous scaffold made from a synthetic polymer, poly(DL-lactide-co-glycolide) (PLG), and evaluated its use in the repair of cartilage. Osteochondral defects made on the femoral trochlear of rabbits were treated by transplantation of the PLG scaffold, examined histologically and compared with an untreated control group. Fibrous tissue was initially organised in an arcade array with poor cellularity at the articular surface of the scaffold. The tissue regenerated to cartilage at the articular surface. In the subchondral area, new bone formed and the scaffold was absorbed. The histological scores were significantly higher in the defects treated by the scaffold than in the control group (p<0.05). Our findings suggest that in an animal model the new porous PLG scaffold is effective for repairing full-thickness osteochondral defects without cultured cells and growth factors.

Platelet-rich plasma protects rotator cuff-derived cells from the deleterious effects of triamcinolone acetonide.

Triamcinolone acetonide (TA) injections are widely used to treat enthesopathy, but they may induce adverse effects such as tendon impairment and rupture. Platelet‐rich plasma (PRP) is a blood fraction containing high platelet concentrations and various growth factors that play a role in tissue repair processes. The purpose of this study is to investigate whether TA has deleterious effects on human rotator cuff‐derived cells, and if PRP can protect these cells from the effects of TA. Human rotator cuff‐derived cells were cultured with and without TA and PRP, and the culture without any additive served as the control. Cell morphology was assessed at days 7 and 21. Cell viability was evaluated at days 1, 7, 14, and 21 by a water‐soluble tetrazolium salt assay. Induction of apoptosis was measured by immunofluorescence staining and flow cytometry at day 7. Induction of cleaved caspase‐3 was measured by immunofluorescence staining at day 7. The cells cultured with TA had a flattened and polygonal shape at day 7. The cells cultured with both TA and PRP were similar in appearance to control cells. Exposure to TA also significantly decreased cell viability, but cell viability did not decrease when PRP was added along with TA. The number of apoptotic cells increased with TA exposure, while addition of PRP prevented cell apoptosis. In conclusion, the deleterious effect of TA was prevented by PRP, which can be used as a protective agent for patients receiving local TA injections.

Regeneration of rotator cuff tear using electrospun poly(d,l-Lactide-Co-Glycolide) scaffolds in a rabbit model.

PURPOSE The purpose of this study was to evaluate an application of poly(d,l-lactide-co-glycolide) (PLG) scaffold created by electrospinning in a rabbit rotator cuff defect model. METHODS Forty-two Japanese white rabbits were used in this study. Defects of the infraspinatus tendon were created, and the PLG scaffolds were implanted. Contralateral infraspinatus tendons were reattached without creating defects. Histologic analyses were performed 4, 8, and 16 weeks after the operation, and mechanical evaluations were performed 0, 4, 8, and 16 weeks after the operation. RESULTS Scaffold fibers remained without dissolution and spindle-shaped cells were observed inside of the scaffold at 4 weeks postoperatively. At 8 weeks, the PLG scaffold had dissolved and bone formation was observed at the scaffold-bone interface. At 16 weeks, the scaffold-bone interface matured and expression of type II collagen was observed. A statistical difference in ultimate failure load was not seen between the scaffold group and reattachment group or normal tendon after 8 weeks postoperatively. The stiffness in the scaffold group was not significantly different from that in the reattachment group at each time point. However, it was significantly weaker than normal tendon at each time point. CONCLUSIONS Transplantation of cell-free PLG scaffold showed cell migration and type II collagen and proteoglycan expression at the scaffold-bone junction by 16 weeks postoperatively with a sufficient ultimate failure load in a rabbit rotator cuff defect model. CLINICAL RELEVANCE The PLG scaffold could be applied to bridge rotator cuff defects. The results showed that bridging with scaffold can be equivalent to reattachment.

Temporary inductions of matrix metalloprotease‐3 (MMP‐3) expression and cell apoptosis are associated with tendon degeneration or rupture after corticosteroid injection

Corticosteroid injections are widely used to treat enthesopathy and tendinitis, but are also associated with possible side effects, such as tendon degeneration or rupture. However, the mechanism of tendon degeneration or rupture after corticosteroid injection remains controversial. The purpose of this study was to reveal the mechanism of tendon degeneration or rupture after injection of triamcinolone acetonide (TA) or prednisolone (PSL). Forty‐two rats were divided into 3 groups: A normal saline injection group (control group), a TA injection group, and a PSL injection group; the normal saline or corticosteroid was injected around the Achilles tendon. One or 3 weeks after injection, the tendons were subjected to biomechanical testing and histological analysis. At 1 week, the biomechanical strength was significantly lower in the corticosteroid groups. Histological analysis, at 1‐week post‐injection, showed collagen attenuation, increased expression of MMP‐3 and apoptotic cells in the corticosteroid groups. The histological changes and biomechanical weaknesses of the tendon were not seen at 3 weeks. These alterations appeared to be involved in tendon degeneration or rupture after corticosteroid injection.

Application of layered poly (L-lactic acid) cell free scaffold in a rabbit rotator cuff defect model

BackgroundThis study evaluated the application of a layered cell free poly (L-lactic acid) (PLLA) scaffold to regenerate an infraspinatus tendon defect in a rabbit model. We hypothesized that PLLA scaffold without cultivated cells would lead to regeneration of tissue with mechanical properties similar to reattached infraspinatus without tendon defects.MethodsLayered PLLA fabric with a smooth surface on one side and a pile-finished surface on the other side was used. Novel form of layered PLLA scaffold was created by superimposing 2 PLLA fabrics. Defects of the infraspinatus tendon were created in 32 rabbits and the PLLA scaffolds were transplanted, four rabbits were used as normal control. Contralateral infraspinatus tendons were reattached to humeral head without scaffold implantation. Histological and mechanical evaluations were performed at 4, 8, and 16 weeks after operation.ResultsAt 4 weeks postoperatively, cell migration was observed in the interstice of the PLLA fibers. Regenerated tissue was directly connected to the bone composed mainly of type III collagen, at 16 weeks postoperatively. The ultimate failure load increased in a time-dependent manner and no statistical difference was seen between normal infraspinatus tendon and scaffold group at 8 and 16 weeks postoperatively. There were no differences between scaffold group and reattach group at each time of point. The stiffness did not improve significantly in both groups.ConclusionsA novel form of layered PLLA scaffold has the potential to induce cell migration into the scaffold and to bridge the tendon defect with mechanical properties similar to reattached infraspinatus tendon model.

Localization of vascular endothelial growth factor during the early stages of osteochondral regeneration using a bioabsorbable synthetic polymer scaffold.

Vascular endothelial growth factor (VEGF) plays a critical role in chondrogenic differentiation in the growth plate of the epiphysis. This function is necessary for chondrocyte survival in cartilage development. We investigated the localization of VEGF in the osteochondral regeneration process using a bioabsorbable polymer scaffold. Osteochondral defects (5 mm in diameter and 5 mm in depth) were made on the femoral condyle of forty‐eight skeletally mature female Japanese white rabbits. In total, twenty‐four defects were filled with poly(DL‐lactide‐co‐glycolide) scaffolds and the others were left untreated. The regeneration process was investigated macroscopically, histologically, immunohistochemically, and by gene expression analysis. In the early stages of osteochondral regeneration, bone ingrowth was observed in the deep zone of the scaffold with continuous VEGF expression; cartilage regeneration was observed in the superficial zone of the scaffold with decreased VEGF expression. In contrast, when the defect was left untreated, VEGF localization was observed throughout the entire defect area, and cartilage regeneration at the articular surface was delayed. We conclude that decrease in localization of VEGF at the articular surface in the postoperative early stage is closely related to the progression of cartilage regeneration in osteochondral defects.

Usefulness of MRI for diagnosis of painful snapping elbow.

Painful snapping of the elbows is rare. We report on a 12-year-old boy with a painful snap in both elbows. High-resolution magnetic resonance imaging of the elbow using microscopy coils detected a synovial fold interposed in each humeroradial joint and was very helpful in establishing the cause of symptoms. Resection of the synovial folds was performed with subsequent relief of symptoms.

Electrophysiological Assessment of Carpal Tunnel Syndrome in Elderly Patients: One-Year Follow-Up Study

PURPOSE To objectively assess elderly patients with carpal tunnel syndrome to characterize their preoperative severity and prognosis after carpal tunnel release using a electrophysiological severity scale. METHODS Electrophysiologic assessment was performed preoperatively and 1 year postoperatively following carpal tunnel release in 112 hands in patients over 70 years of age prospectively by the use of the following electrophysiological severity scale: stage 1, normal distal motor latency (DML) and normal sensory conduction velocity (SCV); stage 2, DML ≥ 4.5 milliseconds and normal SCV; stage 3, DML ≥ 4.5 milliseconds and SCV < 40.0 m/s; stage 4, DML ≥ 4.5 milliseconds and non-measurable SCV; stage 5; non-measurable DML and non-measurable SCV. Additionally, the outcomes of clinical symptoms of pain, nocturnal symptoms, numbness, loss of 2-point discrimination in the median nerve territory, and thenar atrophy were assessed. RESULTS The mean age of patients was 77 years at the time of the operation. Preoperatively, the most common severity was stage 5 (70 of 112 hands, 63%), and clustering stage 4 and 5 together as severe resulted in 103 hands (92%). One year postoperatively, 97 hands (87%) demonstrated at least one stage improvement, and the numbers of mild (stage 1 or 2) increased from 3 (3%) to 45 hands (40%). Parallel with the electrophysiological improvement, pain and nocturnal symptoms resolved in 17 of 17 hands and 11 of 11 hands, respectively, in whom they were present preoperatively. Numbness, loss of 2-point discrimination, and thenar atrophy demonstrated the improvement in 96 of 112 (86%) hands, in 58 of 112 (52%) hands, and in 80 of 96 (83%) hands. CONCLUSIONS We observed electrophysiologic improvement in 86% of elderly patients following carpal tunnel release. Electrophysiologic outcomes correlated with improvement in clinical variables. TYPE OF STUDY/LEVEL OF EVIDENCE Therapeutic IV.