The biomechanical effects of limb lengthening and botulinum toxin type A on rabbit tendon

Numerous studies have examined the effects of distraction osteogenesis (DO) on bone, but relatively fewer have explored muscle adaptation, and even less have addressed the concomitant alterations that occur in the tendon. The purpose herein was to characterize the biomechanical properties of normal and elongated rabbit (N=20) tendons with and without prophylactic botulinum toxin type A (BTX-A) treatment. Elastic and viscoelastic properties of Achilles and Tibialis anterior (TA) tendons were evaluated through pull to failure and stress relaxation tests.All TA tendons displayed nonlinear viscoelastic responses that were strain dependent. A power law formulation was used to model tendon viscoelastic responses and tendon elastic responses were fit with a microstructural model. Distraction-elongated tendons displayed increases in compliance and stress relaxation rates over undistracted tendons; BTX-A administration offset this result. The elastic moduli of distraction-lengthened TA tendons were diminished (p=0.010) when distraction was combined with gastrocnemius (GA) BTX-A administration, elastic moduli were further decreased (p=0.004) and distraction following TA BTX-A administration resulted in TA tendons with moduli not different from contralateral control (p>0.05). Compared to contralateral control, distraction and GA BTX-A administration displayed shortened toe regions, (p=0.031 and 0.038, respectively), while tendons receiving BTX-A in the TA had no differences in the toe region (p>0.05). Ultimate tensile stress was unaltered by DO, but stress at the transition from the toe to the linear region of the stress–stretch curve was diminished in all distraction-elongated TA tendons (p<0.05). The data suggest that prophylactic BTX-A treatment to the TA protects some tendon biomechanical properties.     

Muscle lengthening surgery causes differential acute mechanical effects in both targeted and non-targeted synergistic muscles

Epimuscular myofascial force transmission (EMFT) is a major determinant of muscle force exerted, as well as length range of force exertion. Therefore, EMFT is of importance in remedial surgery performed, e.g., in spastic paresis. We aimed to test the following hypotheses: (1) muscle lengthening surgery (involving preparatory dissection (PD) and subsequent proximal aponeurotomy (AT)) affects the target muscle force exerted at its distal and proximal tendons differentially, (2) forces of non-operated synergistic muscles are affected as well, (3) PD causes some of these effects.In three conditions (control, post-PD, and post-AT exclusively on m. extensor digitorum longus (EDL)), forces exerted by rat anterior crural muscles were measured simultaneously. Our results confirm hypotheses (1–2), and hypothesis (3) in part: Reduction of EDL maximal force differed by location (i.e. 26.3% when tested distally and 44.5% when tested proximally). EDL length range of active force exertion increased only distally. Force reductions were shown also for non-operated tibialis anterior (by 11.9%), as well as for extensor hallucis longus (by 8.4%) muscles. In tibialis anterior only, part of the force reduction (4.9%) is attributable to PD. Due to EMFT, remedial surgery should be considered to have differential effects for targeted and non-targeted synergistic muscles.     

Distraction osteogenesis for obstructive apneas in patients with congenital craniofacial malformations

Infants with congenital craniofacial malformations often have associated severe mandibular hypoplasia causing obstruction of the hypopharynx by retroposition of the base of the tongue into the posterior pharyngeal airway. Management depends on the severity of the airway obstruction. Most cases can be managed by prone positioning until the infant outgrows the problem at 3 to 6 months of age. In more critical cases, monitoring of oxygen saturation, temporary placement of a nasopharyngeal tube, and placement of an endotracheal tube will be useful procedures. Tracheotomy is an effective method for more severe cases, but longstanding tracheotomies result in high morbidity and occasional mortality. Mandibular distraction was performed in seven patients, ranging in age from 1 to 18 months, with critical obstructive apnea secondary to mandibular hypoplasia characterized by an apnea/hypopnea index greater than 20 apneas per hour and oxygen saturation below 80 percent. Two patients were tracheotomized previously. Mandibular lengthening, from 16 to 25 mm on the left side and from 10 to 22 mm on the right, was achieved in 21 to 25 days. Improvement of airway obstruction parameters was measured on polysomnograms and lateral cephalograms. Mandibular lengthening by gradual distraction is a successful method for young patients with severe mandibular hypoplasia causing critical obstructive apneas. Avoidance of tracheotomy or early decannulation in previously tracheotomized patients is a great advantage for patients with congenital craniofacial malformation.     

Knee joint reaction force during tibial diaphyseal lengthening: a study on a rabbit model

The in vivo passive knee joint reaction force was measured in a rabbit model of tibial diaphyseal lengthening. This was based on the assumption that limb lengthening creates soft tissue tension that compresses the joint surface and generates the joint contact force. A measurement method was developed that involved the distraction of the joint and the determination of the distraction force that just separates the joint surfaces. Sixteen immature (mean+/-SD age=9+/-0.6 weeks) New Zealand White rabbits underwent 30% (left) tibial diaphyseal lengthening at a rate of two 0.4mm incremental lengthenings per day. The knee joint reaction force was measured at the end of lengthening (8 rabbits, mean+/-SD age=14+/-0.6 weeks) and five weeks after lengthening (8 rabbits, mean+/-SD age=19+/-0.7 weeks). An instrumented bilateral distractor and an extensometer were fixed cross the knee joint. The joint distraction force and distraction displacement were measured when the joint was distracted in steps and after the section of the Achilles tendon. The joint reaction force on the lengthened side was significantly higher than the control side at both time points (mean+/-SD 44.4+/-7.8 N v. 27.2+/-4.0 N at the end of lengthening, 44.3+/-S6.5 N v. 31.3+/-3.0 N at 5 weeks after lengthening). The contribution of the gastrocnemius to the joint reaction force on the lengthened side was also significantly higher than the control side at both time points (mean+/-SD 9.0+/-1.3N v. 2.8+/-0.8 N at the end of lengthening, 5.3+/-1.4N v. 2.7+/-0.5N at 5 weeks after lengthening). There were significant knee and ankle joint contractures at the end of lengthening, as evidenced by decreased range of motion (mean+/-SD 27+/-8 degrees and 36+/-13 degrees, respectively), which remained 5 weeks after lengthening (mean+/-SD 26+/-6 degrees and 35+/-8 degrees, respectively). The gastrocnemius contributed about 20% of the joint reaction force, indicating that changes in the other intra- and extra-articular structures due to joint contracture may be more important in generating the joint reaction force.     

Myofascial force transmission: muscle relative position and length determine agonist and synergist muscle force.

Equal proximal and distal lengthening of rat extensor digitorum longus (EDL) were studied. Tibialis anterior, extensor hallucis longus, and EDL were active maximally. The connective tissues around these muscle bellies were left intact. Proximal EDL forces differed from distal forces, indicating myofascial force transmission to structures other than the tendons. Higher EDL distal force was exerted (ratio approximately 118%) after distal than after equal proximal lengthening. For proximal force, the reverse occurred (ratio approximately 157%). Passive EDL force exerted at the lengthened end was 7-10 times the force exerted at the nonlengthened end. While kept at constant length, synergists (tibialis anterior + extensor hallucis longus: active muscle force difference approximately -10%) significantly decreased in force by distal EDL lengthening, but not by proximal EDL lengthening. We conclude that force exerted at the tendon at the lengthened end of a muscle is higher because of the extra load imposed by myofascial force transmission on parts of the muscle belly. This is mediated by changes of the relative position of most parts of the lengthened muscle with respect to neighboring muscles and to compartment connective tissues. As a consequence, muscle relative position is a major codeterminant of muscle force for muscle with connectivity of its belly close to in vivo conditions.     

Tensile properties of calcified and uncalcified avian tendons

The mechanical properties of turkey and heron leg tendons have been investigated in dynamic tensile tests. Heron tendons have properties similar to those found for various mammalian tendons. The Young's modulus and the density of turkey tendons increase with increasing calcification. Ultimate tensile stresses are similar to those found for uncalcified tendon, but Young's modulus may reach about 16 GPa, a value normally associated with bone. Calcification lowers the amount of strain energy that can be stored temporarily in the tendons of the legs. The contribution made by elastic strain energy storage to lowering the cost of running is reduced.     

Glycation-induced matrix stability in the rabbit achilles tendon

Connective tissue susceptibility to nonenzymatic glycation was examined following 0, 2, 4, 6, 8, and 10 weeks of incubating the rabbit Achilles tendon in phosphate-buffered saline containing ribose (glycated). The biomechanical integrity of the glycated tendons was then compared to control tendons incubated in phosphate-buffered saline (non-glycated) at each time interval, while the biochemical stability of both groups of tendons was determined by examining collagen extractability and the formation of pentosidine at 8 weeks. Whereas there were no significant biomechanical differences between control and glycated tendons at 0- and 2-week intervals (P > 0.05), moderately significant increases in maximum load, energy to yield, and toughness of glycated tendons were observed at 4 weeks. Beyond 4 weeks of incubation, the differences between glycated and non-glycated tendons became highly significant, as glycated tendons withstood more load and tensile stress (P < 0.01 for each variable), attained significantly higher modulus of elasticity (P < 0.01), absorbed more energy (P < 0.01), and became tougher (P < 0.01) than controls. These differences in the biomechanical indices of the effects of glycation were stable between the 6th and 10th week of glycation. The maximum increases in the biomechanical measurements as a result of glycation were 29% for maximum load, 125% for stress, 19% for strain, 106% for Young's modulus of elasticity, 14% for energy to yield, and 57% for toughness. Biochemical analysis showed a 61% reduction in the extractability of neutral salt-soluble collagen, a 48% decrease in acid-soluble collagen, and a 29% decline in pepsin-soluble collagen in glycated tendons (P < 0.01). In contrast, there was a 28% increase in the amount of insoluble collagen and significantly higher amounts of pentosidine (P < 0.01) in glycated tendons. Collectively, these biomechanical and biochemical results suggest that nonenzymatic glycation may explain the altered stability of connective tissue matrix induced by the processes of diabetes and aging.     

Assessment of biomechanical parameters of the shoulder joint at the operated side versus non-operated side in patients treated surgically for breast cancer

Aim

Isokinetic assessment of biomechanical parameters of the shoulder joint at the operated side versus non-operated side in patients treated surgically for breast cancer according to the type of surgery performed.

Influence of oocyte donor and embryo recipient conditions on cloning efficiency in dogs

To determine factors that affect the efficiency of dog cloning by somatic cell nuclear transfer, the present study was performed to investigate 1) the effects of surgical history (non-operated/operated) and parity (nullipara/multipara) on the recovery of in vivo canine oocytes; 2) the effects of surgical history and parity of recipients on the pregnancy and delivery; and 3) the effects of synchronization state (AA, advanced asynchrony; SY, synchrony; RA, retarded asynchrony) between oocytes donor and recipient on the pregnancy and delivery. Oocyte recovery rate was significantly higher in non-operated dogs compared to operated dogs (93.8 vs. 89.6%, P < 0.05) and not different between nulliparous dogs and multiparous dogs. Delivery rate was also significantly higher in non-operated dogs compared to operated dogs (2.8 vs. 1.0%, P < 0.05) and in nulliparous dogs than multiparous dogs (3.0 vs. 1.7%, P < 0.05). Even though SY showed increased pregnancy and delivery rate (20.0% and 3.0%) compared to AA (15.0% and 2.0%) and RA (0.0% and 0.0%), there was no significant difference. In conclusion, we recommend non-operated dogs as experimental dogs and nulliparous dogs as recipient dogs to increase delivery rate after transfer of somatic cell nuclear transferred embryos, but further study is needed to find out appropriate synchrony status at the transfer.     

Autologous rabbit adipose tissue-derived mesenchymal stromal cells for the treatment of bone injuries with distraction osteogenesis

Background aimsAdipose tissue-derived mesenchymal stromal cells (MSCs) have a higher capacity for proliferation and differentiation compared with other cell lineages. Although distraction osteogenesis is the most important therapy for treating bone defects, this treatment is restricted in many situations. The aim of this study was to examine the therapeutic potential of adipose tissue-derived MSCs and osteoblasts differentiated from adipose tissue-derived MSCs in the treatment of bone defects.MethodsBone defects were produced in the tibias of New Zealand rabbits that had previously undergone adipose tissue extraction. Tibial osteotomy was performed, and a distractor was placed on the right leg of the rabbits. The rabbits were placed in control (group I), stem cell (group II) and osteoblast-differentiated stem cell (group III) treatment groups. The rabbits were sacrificed, and the defect area was evaluated by radiologic, biomechanical and histopathologic tests to examine the therapeutic effects of adipose tissue-derived MSCs.ResultsRadiologic analyses revealed that callus density and the ossification rate increased in group III compared with group I and group II. In biomechanical tests, the highest ossification rate was observed in group III. Histopathologic studies showed that the quality of newly formed bone and the number of cells active in bone formation were significantly higher in group III rabbits compared with group I and group II rabbits.ConclusionsThese data reveal that osteoblasts differentiated from adipose tissue-derived MSCs shorten the consolidation period of distraction osteogenesis. Stem cells could be used as an effective treatment for bone defects.     

Stress deprivation simultaneously induces over-expression of interleukin-1beta, tumor necrosis factor-alpha, and transforming growth factor-beta in fibroblasts and mechanical deterioration of the tissue in the patellar tendon

To test the hypothesis that stress deprivation induces over-expression of cytokines in the patellar tendon, 40 rats were divided into the following two groups. In the stress-shielded group, we slackened the patellar tendon in the right knee by drawing the patella toward the tibial tubercle with flexible wires. In the control group, we performed a sham operation on the right knee. Animals were killed at 2 or 6 weeks for immunohistological evaluation and biomechanical examination. For IL-1beta, TNF-alpha and TGF-beta, the ratio of positively stained specimens to total specimens was significantly higher in the stress-shielded tendons than in the control tendons. The elastic modulus of the stress-shielded tendon was significantly lower than that of the control tendon, while the cross-sectional area of the stress-shielded tendon was significantly greater than that of the control tendon. Therefore, the present study indicated that stress shielding induced the over-expression of IL-1beta, TNF-alpha and TGF-beta in patellar tendon fibroblasts with mechanical deterioration of the tendon. Regarding clinical relevance, the present study suggests a possible application of an anti-IL-1beta or anti-TNF-alpha strategy for reducing the mechanical deterioration of tendons and ligaments in response to stress deprivation, although this study did not directly show that over-expression of IL-1beta or TNF-alpha in response to stress deprivation was the causation of mechanical deterioration of tendons.     

The diagnostic value of Western blot method in patients with cystic echinococcosis

Cystic echinococcosis (CE) is the larval cystic stage (called echinococcal cysts) of a small taeniid-type tapeworm (Echinococcus granulosus). Carnivores such as dogs are usually definitive hosts. Intermediate hosts are typically herbivores such as sheep and cattle. CE can be detected using various imaging techniques such as ultrasonography or radiology. Moreover the primary diagnosis has to be confirmed by serological tests since the clinical signs of the disease are non-specific. This study examined the antigenic band patterns useful for serologic diagnosis of hydatidosis. We also report on the post-operative evolution of patients treated for this disease and also determined the diagnostic performance of Western blot IgG kit. Twenty-five (16 females and 9 males) non-operated patients with hydatid cysts (NOP) and 33 (21 females and 12 males) operated patients with hydatid cysts (OP) were included as study group and 22 healthy individuals (14 females and 8 males) with no known chronic diseases were included as a control group. The ages of the patients and control group individuals were between 16-83 years. Patient and control groups were matched for age and sex. Cyst hydatid IgG antibodies were detected in the sera from all patient groups but no antibodies were found in the sera from the control group using ELISA IgG method. Twenty-three (92%) non-operated patients and 18 (54.5%) operated patients exhibited positive results when Western blot IgG kit was used. The P7 band pattern was detected in the sera from all operated and non-operated patients. Twenty-seven of these positive cases had p7 and (p7+p16/18), (p7+p24/26) or (p7+p16/18+p24/26). No antibodies against p7, p16/18 ve p24/26 band patterns were seen in sera from the control group A statistically significant difference was detected between operated and nonoperated patients for Western blot positivity.(p<0.01). p: 0.018- X2=5,604- OR: 0.176- 95% CI: 0.037- 0.841. The sensitivity, specificity, positive prediction and negative prediction values of Echinococcus granulosus Western blot kit for 25 cases with CE and 22 healthy controls were calculated as 92%, 100%, 100% and 91.7%, respectively. In conclusion, we suggest that monitoring p7 in all non-operated patients may be useful to determine the efficiacy of medical treatment and that monitoring p7 antibodies using serological and Western blot methods in operated patients may be useful for the screening of post-operative evolution in patients with hydatid cyst.     

Role of BMP, betaig-h3, and chitosan in early bony consolidation in distraction osteogenesis in a dog model

The purpose of this investigation was to study the effect of bone morphogenetic protein (BMP), transforming growth factor beta-induced gene h3 (betaig-h3), and chitosan on early bony consolidation in distraction osteogenesis in a dog model. Sixteen dogs were used for this study. The lateral surface of the mandibular body was exposed in the subperiosteal plane and the vertical osteotomy on the mandibular body was extended downward. An external distraction device was applied to the mandibular body, and the mandibular distraction was started 5 days after the operation at a rate of 2 mm/day up to a 10-mm distraction after 5 days. The experimental group was then divided into a control group, a BMP group, a betaig-h3 group, and a chitosan group, depending on the type of implantation material used in the distracted area. On the same day after completing the distraction, BMP, betaig-h3, or chitosan was implanted into the distracted area. No material was implanted into the distracted area in the control group. After implanting the materials, the distraction device was left in place for 7 weeks to allow for bony consolidation. Four dogs were allocated to each group. Two dogs in each group, a total of eight dogs, were killed 4 weeks after completing the distraction and the other eight dogs were killed after 7 weeks. Serial radiographs were obtained every week after completing the distraction. New bone was generated in the distracted zone in all groups. In the BMP group, the formation of active woven bone was observed throughout the distracted zone, and the new bone appeared to be nearly normal cortical bone 7 weeks after implantation. In the betaig-h3 and chitosan groups, the development of new bone was observed in the distracted zone after 7 weeks; however, the amount was less than that in the BMP group. In the control group, the new bone was observed at the edges of the distracted zone. These findings suggest that BMP seems to be very effective in early bony consolidation in distraction osteogenesis.     

The effects of lyophilization on flexural stiffness of extrasynovial and intrasynovial tendon

Tendon or ligament reconstructions often use autologous or allogenic tendons from either extrasynovial or intrasynovial sources. Allograft tendons must be lyophilized for preservation before transplantation, a process which can impact mechanical properties of the graft. Reconstituted graft properties that are similar to native tendon are desirable. Although tensile and compressive properties of tendons have been investigated, there is a paucity of information describing flexural properties of tendon, which can impact the gliding resistance. This study aims to design a testing method to quantify tendon flexural modulus, and investigate the effects of lyophilization/rehydration procedures on tendon flexibility. A total of 20 peroneus longus tendons (extrasynovial) and 20 flexor digitorum profundus tendons (intrasynovial) were collected. Ten of each tendon were processed with 5 freeze–thaw cycles followed by lyophilization and rehydration with saline solution (0.9%). Bend testing was conducted on tendons to quantify the flexural modulus with and without processing. As canine FDP tendons contain fibrous and fibrocartilaginous tissue regions, the flexural moduli were measured in both regions. Flexural modulus of rehydrated, lyophilized extrasynovial PL tendon was significantly lower than that of similarly processed intrasynovial FDP tendon (p < 0.001). Flexural moduli of both the fibrocartilaginous and non-fibrocartilaginous regions of intrasynovial tendon significantly increased after lyophilization (p < 0.001). The flexural modulus of the fibrocartilaginous region was significantly higher than that of the non-fibrocartilaginous region in intrasynovial tendon (p < 0.001). Lyophilization significantly increases the flexural modulus of extrasynovial and intrasynovial tendons, and flexural modulus differs significantly between these two tendon types. Increases in stiffness caused by lyophilization may impact the mechanical performance of the allograft in vivo.     

Fatigue damage of human tendons

The study was designed to examine the effects of partial fatigue on specific mechanical parameters which characterise human tendons in vitro. Specimens prepared from 12 intact Extensor digitorum longus tendons of the foot were subjected to partial fatigue, equivalent to 25% of the median fatigue life, by a cyclic square tension-tension stress waveform at the physiological frequency of 4 Hz. The maximum stress was set at a value corresponding to 20% of the calculated ultimate tensile strength (UTS) of 100 MPa. The minimum stress was set at 1% of the UTS. Dynamic characterisation was performed at stress levels of 10% and 20% of the UTS prior to and following partial mechanical fatigue. Subsequent quasi-static tests were performed on some of the specimens. Comparative analysis of the damage ratios (DRs) of quasi-static and dynamic mechanical parameters suggested the use of the DR for dynamic tensile modulus as a good indicator of damage inflicted by mechanical fatigue. Such an approach might be used for an in vivo assessment of tendon damage.     

Strain-related bone remodeling in distraction osteogenesis of the mandible

Distraction osteogenesis has become a mainstay in craniofacial surgery. However, there are several unresolved problems concerning the biology of bone regeneration. We investigated the biomechanical effects of mandibular lengthening in 32 rabbits on a cellular and histologic level. The mandible was subjected to a corticotomy, held in a neutral position for 4 days, and then lengthened at various strain rates and frequencies for 10 days. Radiographic, histologic, and electron microscopic examinations showed a strain-related bone regeneration. Application of physiologic strain rates (2000 microstrains or 0.2 percent) led to a bridging of the artificial fracture exhibiting woven ossification, whereas at 20,000 microstrains trabecular bone formation was demonstrated. In contrast, hyperphysiologic strain magnitudes (200,000 microstrains and 300,000 microstrains) showed a fibrous tissue formation. Multiple strain applications (10 cycles/day versus 1 cycle/day) increased the width of the distraction gap without changing the stage of bone regeneration. The gradual distraction of bone in physiologic magnitudes at higher frequencies seems to be desirable for a bony differentiation and may help to improve clinical applications.     

The characteristics of osteogenesis under conditions of stretch tension]

Structure of rough-fibrillar, netted and laminated bone tissues, formed in distraction regenerates on various stages of leg lengthening, was studied experimentally on dogs, using light, transmission and scanning electron microscopy and electron probe microassay. It was ascertained, that the first two types of tissues belong to the category of provisory, rapidly renewing ones, the volume increase of which is possible not only at the expense of appositional growth but also interstitially. Their formation in the area of the regenerate, more proximally and more distally than the medial layer, provided longitudinal growth of primary osteons forming here and thereby the growth of the whole bone regenerate.     

Tendon mineralization is progressive and associated with deterioration of tendon biomechanical properties,and requires BMP-Smad signaling in the mouse Achilles tendon injury model

Ectopic tendon mineralization can develop following tendon rupture or trauma surgery. The pathogenesis of ectopic tendon mineralization and its clinical impact have not been fully elucidated yet. In this study, we utilized a mouse Achilles tendon injury model to determine whether ectopic tendon mineralization alters the biomechanical properties of the tendon and whether BMP signaling is involved in this condition. A complete transverse incision was made at the midpoint of the right Achilles tendon in 8-week-old CD1 mice and the gap was left open. Ectopic cartilaginous mass formation was found in the injured tendon by 4 weeks post-surgery and ectopic mineralization was detected at 8 to 10 weeks post-surgery. Ectopic mineralization grew over time and volume of the mineralized materials of 25-weeks samples was about 2.5 fold bigger than that of 10-weeks samples, indicating that injury-induced ectopic tendon mineralization is progressive. In vitro mechanical testing showed that max force, max stress and mid-substance modulus in the 25-weeks samples were significantly lower than the 10-weeks samples. We observed substantial increases in expression of bone morphogenetic protein family genes in injured tendons 1 week post-surgery. Immunohistochemical analysis showed that phosphorylation of both Smad1 and Smad3 was highly increased in injured tendons as early as 1 week post-injury and remained high in ectopic chondrogenic lesions 4-weeks post-injury. Treatment with the BMP receptor kinase inhibitor (LDN193189) significantly inhibited injury-induced tendon mineralization. These findings indicate that injury-induced ectopic tendon mineralization is progressive, involves BMP signaling and associated with deterioration of tendon biomechanical properties.     

Characterization of midface maxillary membranous bone formation during distraction osteogenesis

The purpose of the study was to follow the early events in bone formation and neovascularization during maxillary distraction and after the consolidation period and to define the characterization of the new bone in the distracted area. Maxillary osteotomy was performed in seven sheep. In five animals, an external distraction device was used for maxillary lengthening of 20 mm at a rate of 1 mm/day for 20 days. Another two animals served as controls without distraction. Sequential biopsies were performed. The methods used for analysis were histologic, immunohistochemical, and ultrastructural by transmission electron microscopy. During the 5 days of latency, a fibrin clot was formed that after 5 days of distraction was replaced by granulation tissue, proliferating mesenchyme-like cells, and capillaries. After 10 days of distraction, the regenerated tissue could be divided into three main zones and two transitional areas: a central zone occupied by many polygonal mesenchyme-like cells and spindle-shaped cells that proliferated intensively; two paracentral zones on both sides of the central zone in which many cells showed morphologic signs of apoptosis leading to a decreased number of fibroblast-like cells embedded in wavy collagen fibers; a transitional area from the central to the paracentral zone in which concentric cellular colonies were believed to represent a novel form of vasculogenesis; distal-proximal zones, located on both sides of the paracentral zones and in continuation with the old bone, showed delicate new woven bone trabeculae that grew continuously in the direction of lengthening and gradually became mineralized; and a transitional area from the paracentral to the distal-proximal zones in which there was recruitment of preosteoblasts from the distracted tissue to the trabecular tips. These further differentiated into osteoblasts that contributed to the trabecular growth. The histologic feature pattern was similar after 15 and 20 days of continuous distraction. At the end of lengthening, after 20 days, delicate longitudinally oriented trabeculae continued to grow by recruiting preosteogenic cells from the central distracted tissue, became mineralized, and were rimmed by osteoblasts. After 6 weeks of retention, the trabeculae thickened and consisted of a mixture of lamellar and woven bone. In conclusion, the distraction force creates a pool of undifferentiated mesenchyme-like cells with osteogenic potential and triggers capillary formation, a clear zonation can be observed during active lengthening, and new bone trabeculae begin to form between 5 and 10 days after distraction, soon become aligned with osteoblasts, and continue to grow as long as distraction force is applied. This characterization may help in any exogenous involvement with growth factors to improve bone quality.     

A treadmill ergometer for three-dimensional ground reaction forces measurement during walking.

This report describes new treadmill ergometer designed to measure the vertical and horizontal ground reaction forces produced by the left and right legs during walking. It was validated by static and dynamic tests. Non-linearity was from 0.2% (left vertical force) to 1.4% (right antero-posterior force). The resonance frequency was from 219 (right vertical direction) to 58 Hz (left medio-lateral direction). A calibration "leg", an air jack in series with a strain gauge, was developed and used to produce force signals comparable to those obtained during human locomotion. The mean differences between the force measured by the calibration leg and treadmill ergometer at 5 km h(-1) were 3.7 N (0.7%) for the left side and 6.5 N (1.2%) for the right. Measurements obtained during human walking showed that the treadmill ergometer has considerable potential for analysing human gait.