Posterolateral spinal fusion (PSLSF) in rabbits is a challenging model for bone substitutes because the transverse processes are extremely thin and the space to be filled with bone is greater than critical and meiopragic in terms of vascularity. Several investigators have shown beneficial effects of PRP in bone and soft-tissue healing processes. However, controversial results have been reported in clinical setting analysing the effectiveness of PRP. Aim of the present study was to test the effectiveness of PRP in experimental model of PLSF in rabbits. MATERIAL AND METHODS. 20 White females New Zeland Rabbits were used. Seven rabbits (Group 1) had PRP plus carrier on the right side (Group 1A) and plus carrier and fresh bone marrow on the left side (Group 1B). Seven rabbits (Group 2) had carrier alone on the right side (Group 2A) and carrier plus fresh bone marrow on the left side (Group 2B). Six rabbits (Group 3) had sham operation on both right and left sides. Animals were sacrificed 6 months after surgery and the lumbar spine submitted to radiolographic and histologic analysis. Vascular density (VD) was also assessed in the different zone of the grafted material. RESULTS. Radiographs showed a complete fusion in 83% of group 1A and in 83% of group 1B, and in 86% of group 2A and 2B. Pseudarthrosis or non union, was observed in 1 specimen of group 1B and 2A and in all specimens of group 3 (sham). In contrast to radiographic results, no specimen showed a complete bony bridge between the transverse processes on histologic analysis. VD was significantly greater in the periapophyseal compared to the interapophyseal region of the graft material. However, no significant difference was found in the VD between groups. CONCLUSIONS. In this study PRP alone, or augmented with fresh bone marrow, failed to induce a histologically proved bony fusion in the PLSF model. Factors which may influence the effectiveness of PRP should be further addressed before applying PRP in the clinical setting

Purpose of this study is to create an experimental model of electrophysologic evaluation of the supraspinatus muscle on rats, after traumatic rupture of its tendon. The population of this study consisted of 10 male Sprague Dawley rats weighting 300–400g. Under general anaesthesia we proceeded with traumatic rupture of the supraspinatus tendon and exposure of the muscle. The scapula was immobilized, and the supraspinatus tendon was attached to a force transducer using a 3–0 silk thread. A dissection was performed in order to identify the suprascapular nerve, which was then stimulated with a silver electrode. Stimulations were produced by a stimulator (Digitimer Stimulator DS9A) and were controlled by a programmer (Digitimer D4030). Fiber length was adjusted until a single stimulus pulse elicited maximum force during a twitch under isometric conditions. Rectangular pulses of 0.5 ms duration were applied to elicit twitch contractions. During the recordings, muscles were rinsed with Krebs solution of approximately 37 8C (pH 7.2–7.4) and aerated with a mixture of 95% O2 and 5% CO2. The output from the transducer was amplified and recorded on a digital interface (CED). The following parameters were measured at room temperature (20–21 8C): single twitch tension; time to peak; half relaxation time; tetanic tensions at 10, 20, 40, 80 and 100 Hz; and fatigue index, which was evaluated using a protocol of low frequency (40 Hz) tetanic contraction, during 250 ms in a cycle of 1 s, for a total time of 180 s. The fatigue index value was then calculated by the formula [fatigue index=(initial tetanic tension − end tetanic tension) ∗ 100/(initial tetanic tension)]. In the end, the transducer was calibrated with standard weights and tensions were converted to grams. The mean single twitch was 8.2, the time to peak 0.034 msec and the half relaxation time 0.028 msec. The strength of titanic muscle contractures was 5.7 msec at 10Hz and 17.7 at 100Hz. Finally, the fatigue index was calculated at 48.4. We believe that electrophysiologic evaluation of the supraspinatus muscle in rats will help us understanding the pathology of muscle atrophy after rotator cuff tears and possibly the functional restoration after cuff repair

Introduction. The same cup orientation is classically applied to all cases of hip replacement (45° abduction, 20° anteversion). We hypothesize that this orientation must be adapted to the patient's hip range of motion. We tested this hypothesis by means of an experimental study with respect to hip range of motion, comparing the classical orientation (45° and 20°), and the orientation obtained with computer-assisted navigation. Material and Methods. The experimental model included a hemipelvis equipped with a femur whose mobility was controlled for three configurations: stiff (60°/0°, 15°/10°, 10°/10°), average (80°/10°, 35°/30°,35°/25°), mobile (130°/30°, 50°/50°, 45°/35°). The hemipelvis and the cup holder were equipped with an electromagnetic system (Fastrack ™) to measure cup orientation. The Pleos™ navigation system (equipping the hemipelvis, the femur, and the cup holder) guided the cup orientation by detecting the positions risking impingement through a kinematic study of the hip. Nine operators each performed 18 navigation-guided implantations (162 hip abduction, anteversion, and range of movement measurements) in two series scheduled 2 months apart. Results. The model used herein showed intra and interobserver reliability. Compared to the navigation-assisted surgery, the arbitrary orientation gave a mean anteversion error of only 1° ± 6° (−12 to +19°) but 5° ± 8° (−26° to +13°) for abduction. However, 16% of the errors were more than 10° in anteversion (1/2 in the mobile configuration) and 11% of the errors were more than 15° in abduction (for the most part in the mobile configuration). With arbitrary orientation, the errors consisted in excess anteversion and insufficient abduction. Discussion and Conclusion. The experimental model developed was reliable and can be used to evaluate different prosthetic configurations. This study emphasizes that the ideal arbitrary cup orientation cannot be applied to all hips. All the surgeons are very reproducible but the only way to integrate the range of motion in there ‘own way to do’ in vitro, is to use a navigation system witch can guide the surgeon so as to reduce the risk of impingement and instability

Compartment syndrome, a devastating consequence of limb trauma, is characterised by severe tissue injury and microvascular perfusion deficits. We hypothesised that leucopenia might provide significant protection against microvascular dysfunction and preserve tissue viability. Using our clinically relevant rat model of compartment syndrome, microvascular perfusion and tissue injury were directly visualised by intravital video microscopy in leucopenic animals. We found that while the tissue perfusion was similar in both groups (38.8% (standard error of the mean (sem) 7.1), 36.4% (sem 5.7), 32.0% (sem 1.7), and 30.5% (sem 5.35) continuously-perfused capillaries at 45, 90, 120 and 180 minutes compartment syndrome, respectively versus 39.2% (sem 8.6), 43.5% (sem 8.5), 36.6% (sem 1.4) and 50.8% (sem 4.8) at 45, 90, 120 and 180 minutes compartment syndrome, respectively in leucopenia), compartment syndrome-associated muscle injury was significantly decreased in leucopenic animals (7.0% (sem 2.0), 7.0%, (sem 1.0), 9.0% (sem 1.0) and 5.0% (sem 2.0) at 45, 90, 120 and 180 minutes of compartment syndrome, respectively in leucopenia group versus 18.0% (sem 4.0), 23.0% (sem 4.0), 32.0% (sem 7.0), and 20.0% (sem 5.0) at 45, 90, 120 and 180 minutes of compartment syndrome in control, p = 0.0005). This study demonstrates that the inflammatory process should be considered central to the understanding of the pathogenesis of cellular injury in compartment syndrome.

Cite this article: Bone Joint J 2015;97-B:539–43

There are a number of periprosthetic femoral fracture (PFF) fixation failures. In several cases the effect of fracture configuration on the performance of the chosen fixation method has been underestimated. As a result, fracture movement within the window that seems to promote callus formation has not been achieved and fixations ultimately failed. This study tested the hypothesis that: PFF configuration and the choice of plate fixation method can be detrimental to healing. A series of computational models were developed, corroborated against measurements from a series of instrumented laboratory models and in vivo case studies. The models were used to investigate the fixation of different fracture configurations and plate fixation parameters. Surface strain and fracture movement were compared between the constructs. A strong correlation between the computational and experimental models was found. Computational models showed that unstable fracture configurations increase the stress on the plate fixation. It was found that bridging length plays a pivotal role in the fracture movement. Rigid fixations, where there is clinical evidence of failure, showed low fracture movement in the models (<0.05mm); this could be increased with different screw and plate configurations to promote healing. In summary our results highlighted the role of fracture configuration in PFF fixations and showed that rigid fixations that suppress fracture movement could be detrimental to healing

Despite its clinical significance, metaphyseal fracture healing has received little attention in research and experimental models have been limited. In particular it is not known to what extent the mechanical environment plays a role in metaphyseal fracture healing. Recently, a new murine internal fixation plate has been developed to stabilise fractures in the distal femur under highly standardised conditions. Goal of the current study was to modify this design, in order to be able to evaluate the influence of the fixator bending stiffness on metaphyseal fracture healing in mice. Adapting the existing single body design, resulting in low flexibility fixation, two new plates were developed with a decreased bending stiffness of approximately 65% and 45% of the original implant (100%). Pilot experiments were performed on 54 animals, whereas the mice were sacrificed and fracture healing assessed radiologically and biomechanically after 14 and 28 days. MicroCT evaluation confirmed that the osteotomy was created in the trabecular, metaphyseal bone of the distal mouse femora. All bones showed progressive fracture healing over time, with decreased implant stiffness leading to increased periosteal callus formation. These implants represent an important new research tool to study molecular and genetic aspects of metaphyseal fracture healing in mice under standardized mechanical conditions, in order to improve clinical treatment in challenging situations, such as in osteoporotic bone

Introduction. Our previous study using microarray analysis showed that Rad (Ras associated with diabetes) was highly expressed in nonunion. The purpose of this study is to investigate the gene expression and immunolocalization of Rad, and other Ras-related G proteins: Rem1 and Rem2 in fracture/nonunion site using rat experimental models. Hypothesis. We hypothesized that Rad had a significant role in nonunion formation. Materials & Methods. For standard healing model, K-wire was inserted into the femur and a closed fracture was created. Nonunion model was produced by periosteal cauterization at the fracture site. At post-fracture days 3, 7, 10, 14, 21, and 28, RNA was extracted from callus or fibrous tissue for real-time PCR. At day 14, specimens were harvested for immunohistochemistry. Results. Significant difference of Rad gene expression was not observed between standard healing fracture and nonunion at the earlier time points. In contrast, significantly higher expression in nonunion was observed at the later time points. There were no significant differences between standard healing fracture and nonunion in gene expression of Rem1 and Rem2. In immunohistochemical analysis, Rad and Rem1 were detected in the fracture site, and Rem2 was not detected. On the other hand, Rad was only detected in fibrous tissue in nonunion. Discussion & Conclusion. Our results suggest a significant role of Rad in fracture healing and nonunion formation. Rad may become a target agent for treatment of nonunion

Objective. To assess the beneficial use of polypropylene mesh impregnated with vancomycin in an experimental model open fractures Gustilo IIIa in rabbits. Material and Method. We worked with 15 New Zeland White rabbits. All of them were carried out under general anaesthetic, a 5-cm incision longitudinal was made at the back of the right thigh. The femur was aproached and a fracture was performed with a shear, giving rise to a multifragment fracture. The wound remained open for 6 hours with the bone exposed, in a non-surgical ambient. Subsequently underwent surgical cleaning of the open fractures in two stages. The fracture was stabilized with an intramedular pin. The animals were sorted in 3 different therapeutic groups:. Group 1: (5 rabbits) without other treatment. Group 2: (5 rabbits) a polypropylene mesh was placed around the fracture. Group 3: (5 rabbits) a polypropylene mesh with vancomycin was placed around the fracture. The wound was closed with nylon stiches. Three weeks postoperative, the animals were intervened surgically under general anesthesia, after aseptic cure and placement of surgical fields, femoral bone biopsies, soft tissue and mesh were taken. The rabbits were sacrified. The samples were sent to pathology and bacteriology labs. Results. The bacteria isolated were as follows: Escherichia coli, Pasteurella multocida, Staphylococcus spp., Clostridium spp. Mamheinia spp. The Clostridium spp. is a common contaminant in the exposed fractures present in the environment. The Pasteurella mustocida is a microorganism present in the oral cavity of rabbits, as well as Escherichia coli is a germ present in the animal's digestive tract. Mannheimia spp. It is a beta-hemolytic organism, found in the nasal flora of these animals and their pathological role is not elucidated. Staphylococcus spp. is a germ that is found in the normal flora of the animals skin. Group 1 showed a relative risk for an infection. For Group 2 the relative risk was substantially greater than 1.4, while in Group 3, the relative risk was 0.6, significantly lower than the previous two groups. The results have shown a beneficial effect of the use of impregnated polypropylene mesh with vancomycin in this animal group. Conclusion. The use of polypropylene meshes with vancomycin could be useful in the treatment of muscle and ligamentary deficits in patients with open fractures Gustillo IIIa

We aimed to investigate the effect of Seprafilm®, a synthetic biomembran, on the intra-articular adhesion formation in an experimental arthrofibrosis model. Twenty male white rabbits were randomly allocated into two groups of 10 animals in each. A standard surgical procedure aiming at the development of arthrofibrosis and including medial parapatellar arthrotomy, lateral eversion of the patella, partial synovectomy and debridement of anterior of supracondylar area and patella joint surface by scalpel was performed on all rabbits' right knees. Group 1 rabbits served as controls, and in Group 2 rabbits a Seprafilm®, barrier placed into the described area. In both groups, after surgery, knee joint was immobilized by a no.5 wire suture passing from the ankle and groin and keeping the joint in 140° of flexion. At 6th week, all animals were sacrificed and adhesion formation was evaluated both macroscopically and histo-pathologically. All data were semi-quantified and analyzed statistically by Fisher's exact test. While all rabbits in control group displayed different rates of adhesion macroscopically (62.5% severe, 25% moderate, 12.5% mild), none in the study group had it. The average macroscopic adhesion score was 2.5 ± 0.75 in control group, and 0 in Seprafilm® group. Histopathologic evaluation also revealed microscopic adhesion in all rabbits in control group, but none in Seprafilm® group. Fibroblast proliferation in Seprafilm® group (100% mild) was significantly lower than in control group (62.5% severe, 37.5% moderate) (p<0.05). In conclusion, use of Seprafilm® as a mechanical barrier may be of value against the formation of arthrofibrosis in risky knees such as septic and traumatic ones

This article presents a unified clinical theory that links established facts about the physiology of bone and homeostasis, with those involved in the healing of fractures and the development of nonunion. The key to this theory is the concept that the tissue that forms in and around a fracture should be considered a specific functional entity. This ‘bone-healing unit’ produces a physiological response to its biological and mechanical environment, which leads to the normal healing of bone. This tissue responds to mechanical forces and functions according to Wolff’s law, Perren’s strain theory and Frost’s concept of the “mechanostat”. In response to the local mechanical environment, the bone-healing unit normally changes with time, producing different tissues that can tolerate various levels of strain. The normal result is the formation of bone that bridges the fracture – healing by callus. Nonunion occurs when the bone-healing unit fails either due to mechanical or biological problems or a combination of both. In clinical practice, the majority of nonunions are due to mechanical problems with instability, resulting in too much strain at the fracture site. In most nonunions, there is an intact bone-healing unit. We suggest that this maintains its biological potential to heal, but fails to function due to the mechanical conditions. The theory predicts the healing pattern of multifragmentary fractures and the observed morphological characteristics of different nonunions. It suggests that the majority of nonunions will heal if the correct mechanical environment is produced by surgery, without the need for biological adjuncts such as autologous bone graft.

Cite this article: Bone Joint J 2016;98-B:884–91.

MicroRNAs (miRNAs ) are small non-coding RNAs that regulate gene expression. We hypothesised that the functions of certain miRNAs and changes to their patterns of expression may be crucial in the pathogenesis of nonunion. Healing fractures and atrophic nonunions produced by periosteal cauterisation were created in the femora of 94 rats, with 1:1 group allocation. At post-fracture days three, seven, ten, 14, 21 and 28, miRNAs were extracted from the newly generated tissue at the fracture site. Microarray and real-time polymerase chain reaction (PCR) analyses of day 14 samples revealed that five miRNAs, miR-31a-3p, miR-31a-5p, miR-146a-5p, miR-146b-5p and miR-223-3p, were highly upregulated in nonunion. Real-time PCR analysis further revealed that, in nonunion, the expression levels of all five of these miRNAs peaked on day 14 and declined thereafter.

Our results suggest that miR-31a-3p, miR-31a-5p, miR-146a-5p, miR-146b-5p and miR-223-3p may play an important role in the development of nonunion. These findings add to the understanding of the molecular mechanism for nonunion formation and may lead to the development of novel therapeutic strategies for its treatment.

Cite this article: Bone Joint J 2015; 97-B:1144–51.

It is becoming increasingly common for a patient to have ipsilateral hip and knee replacements. The inter-prosthetic (IP) distance, the distance between the tips of hip and knee prostheses, has been thought to be associated with an increased risk of IP fracture. Small gap distances are generally assumed to act as stress risers, although there is no real biomechanical evidence to support this.

The purpose of this study was to evaluate the influence of IP distance, cortical thickness and bone mineral density on the likelihood of an IP femoral fracture.

A total of 18 human femur specimens were randomised into three groups by bone density and cortical thickness. For each group, a defined IP distance of 35 mm, 80 mm or 160 mm was created by choosing the appropriate lengths of component. The maximum fracture strength was determined using a four-point bending test.

The fracture force of all three groups was similar (p = 0.498). There was a highly significant correlation between the cortical area and the fracture strength (r = 0.804, p <  0.001), whereas bone density showed no influence.

This study suggests that the IP distance has little influence on fracture strength in IP femoral fractures: the thickness of the cortex seems to be the decisive factor.

Cite this article: Bone Joint J 2014;96-B:1378–84.