Effect of dynamic compression on in vitro chondrocyte metabolism.

BACKGROUND: Chondrocytes can detect and respond to the mechanical environment by altering their metabolism. This study was designed to explore the effects of dynamic compression on chondrocyte metabolism. METHODS: Chondrocytes were harvested from newborn Wistar rats. After 7 days of expansion, chondrocytes embedded in agarose discs underwent uniaxial unconfined dynamic compression loads at different amplitudes (5%, 10%, and 15%) and frequencies (0.5 Hz, 1.0 Hz, 2.0 Hz, and 3.0 Hz) with a duration of 24 hours. The delayed effects on the chondrocytes were studied at 1, 3, and 7 days after the experiment. RESULTS: The results showed that at 10% strain, higher-frequency compression pressure can enhance the proliferation of chondrocytes. The synthesis of glycosaminoglycan (GAG) increased at 10%-15% strain and a 1-Hz load. The synthesis of nitric oxide (NO) increased at the 0.5-Hz load; while decreasing at the 15% strain. With 10% strain, 1 Hz dynamic compression, the proliferation of chondrocytes and GAG synthesis increased and persisted for 7 days, and NO synthesis decreased at the third and seventh days of culture. CONCLUSIONS: This study showed that chondrocytes respond metabolically to compressive loading, which is expected to modulate the growth and the resultant biomechanical properties of these tissue-engineered constructs during culture.

Scavenging effect of benzophenones on the oxidative stress of skeletal muscle cells.

Benzophenone is an ultraviolet (UV)-absorbing agent that has been used in industry and medicine for more than 30 years. Consumers of cosmetics and sunscreens containing UV-absorbers are exposed to benzophenones on a daily basis, owing to the widespread use of these compounds. However, the efficacy of these compounds as scavengers of oxidative stress is still not well established. In the present study, we investigate the antioxidative capacity of six sunscreen benzophenone compounds. A primary myoblast culture was mixed in vitro with 100 microM menadione. The cytotoxic effect by menadione-induced oxidative stress was monitored by the lucigenin- or luminol-amplified chemiluminescence, methylthiotetrazole (MTT) assay, and the antioxidative effects of various benzophenone compounds were evaluated. The results showed that the addition of menadione can induce oxidative stress on myoblasts by superoxide and hydrogen peroxide production, which can be eradicated by superoxide dismutase (SOD) and catalase, respectively, in a dose-dependent mode. The catalase has a protective effect on the cytotoxicity induced by menadione as measured by the MTT assay, while the SOD does not. The selected benzophenones also have a significant scavenging effect on the menadione-induced cell death on the myoblasts. The ortho-dihydroxyl structure and other hydroxy groups in the same ring have a stronger scavenging effect on the superoxide anion on myoblasts; thus, a stable penoxy radical may be formed. The mechanism of this effect remains to be clarified.

Effect of hydroxyapatite particle size on myoblasts and fibroblasts.

After surgery, the bone and soft tissues around integrated biomaterials can be adversely affected by implant-related factors acting over a period of years. However, few studies have directly addressed the effects upon the adjacent soft tissue. The present study was designed to test the biological effects of various sized hydroxyapatite (HA) particles on myoblasts and fibroblasts. Both the myoblasts and fibroblasts were mixed in in vitro culture with 0.1% (1 mg ml(-1)) of various sized HA particles (0.5-3.0, 37-63, 177-250, 420-841 microm) for 1 h, 3 h, 1 day, 3 days and 7 days to test their effects on the cell culture. The results show that adding HA particles into a cell culture can decrease the cell count significantly. The transforming growth factor-beta1 (TGF-beta1) concentrations in the culture medium decreased significantly on addition of HA particles. When calculated as a ratio to the cell number, the TGF-beta1 titre increased most significantly in the groups of medium-sized particles. The prostaglandin E2 (PGE2) concentrations in the medium increased significantly. The changes in TGF-beta1 and PGE2 concentrations with the smallest particles were most significant and persisted longer. The inhibitory effects of the HA particles on the cell culture were mediated by the increased synthesis of PGE2. Caution should be exercised before considering the use of an HA product which could easily break down into a fine powder.

Biochemical and histopathological changes in the mortality caused by acute ischemic limb injury: a rabbits' model.

Restoration of blood flow to an acute ischemic extremity may deteriorate the ischemic injury, lead to multiple organ dysfunction or even death. This paradox of continuing injury during reperfusion is not completely understood. The role of multi-organ damage in the mortality caused by ischemic limb injury is also still not clarified. The purpose of this study is to determine the biochemical and histopathological changes in the mortality caused by ischemic limb injury. After anesthesia, the hindlimbs of 14 New Zealand white rabbits were made ischemic and set into 8 hours or 12 hours of ischemia. Blood samples were obtained then the creatine kinase (CK) levels were determined and CK isoenzymes analyzed. All rabbits with 8 hours' ischemia survived well, and 5 of the 7 rabbits with 12 hours' ischemia expired within 8 hours after reperfusion. CK elevation was correlated most strongly with the time of the ischemic insults. The percentage of CK-MB isoenzyme remained unchanged after 8 hours' ischemia-reperfusion insult, while increased significantly after 12 hours' ischemia-reperfusion insult. Histologic examinations showed that the major systemic manifestation was massive destruction of the liver and kidney. The injuries are more obvious in areas with the greatest blood flow during reperfusion. We concluded that the ratio of CK-MB isoenzyme is most useful for distinguishing the risk of mortality caused by acute ischemic limb injury, and the cause of systemic complications are attributed to the multi-organ failure.

Biocompatibility and osteoconductivity of the pyrost bone substitutes.

The purpose of this study was to re-evaluate the bone regeneration power and the in vitro biocompatibility of the Pyrost bone substitute. Twenty-four adult New Zealand White rabbits were used. Bony defect over both iliac crest and mid-diaphyseal portion of the ulna bone were created. Appropriate sized-block of Pyrost bone substitute were implanted. Four of the animals were killed at each postoperative month to evaluate its bone regeneration power by histologic study. The Pyrost bones were co-cultured with osteoblasts to evaluate its biocompatibility. The results showed that Pyrost bone substitute was quite stable and incorporated well with active bone regeneration. The Pyrost heal better at the iliac crest than at the ulnar defect. The Pyrost was compatible to the osteoblasts. Osteoblasts had successfully seeded and mitotically expanded on the porous surface of the Pyrost bone graft. The result showed that Pyrost bone obviously exerts an intense stimulus on osteo-regeneration in the presence of osteoblasts. We consider Pyrost to be an alternate to the conventional preserved allografts that is occasionally necessary.

Antioxidant status following acute ischemic limb injury: a rabbit model.

Although ischemic injury to skeletal muscle is a matter of great clinical importance, relatively little is known about the mechanisms which determine systemic responses. One purpose of this study is to elucidate the systemic antioxidant status following an episode of acute ischemic limb injury and subsequent reperfusion. Twelve New Zealand white rabbits were used in this study. After the animals were anesthetized, an ischemic insult was created in the right hind limb for twelve hours, followed by four hours of reperfusion. Several series of blood samples were obtained. At the end of the experiment, the animals were killed and necropsies undertaken in order to evaluate the antioxidant status of various visceral organs. The results link ischemia and reperfusion injury to a significant decline in antioxidative activity in various tissues. The weakening in antioxidant status after ischemic limb injury was most pronounced in the heart tissue, followed in descending order by the spleen, skeletal muscle, lung, liver, and kidney tissue. The levels of specific antioxidants and reactive oxygen species in various organs changed significantly, and the changes were tissue specific. Endogenous radical scavenging systems were not entirely overwhelmed in most of the tissues studied. But higher levels of malondialdehyde (MDA) found in cardiac tissue suggest that the production of oxygen free radicals is accelerated by an ischemic injury. Based on the study, we believe that the cardiac tissue is particularly susceptible to the effects of ischemia and reperfusion injury. Damage to cardiac tissue is probably the major cause of mortality following acute ischemic injury in a limb.

Trunk muscle cocontraction: the effects of moment direction and moment magnitude.

This study investigated the cocontraction of eight trunk muscles during the application of asymmetric loads to the torso. External moments of 10, 20, 30, 40, and 50 Nm were applied to the torso via a harness system. The direction of the applied moment was varied by 30 degrees increments to the subjects' right side between the sagittally symmetric orientations front and rear. Electromyographic (EMG) data from the left and right latissimus dorsi, erector spinae, external oblique, and rectus abdominus were collected from 10 subjects. The normalized EMG data were tested using multivariate and univariate analyses of variance procedures. These analyses showed significant interactions between the moment magnitude and the moment direction for seven of the eight muscles. Most of the interactions could be characterized as due to changes in muscle recruitment with changes in the direction of the external moment. Analysis of the relative activation levels, which were computed for each combination of moment magnitude and direction, indicated large changes in muscle recruitment due to asymmetry, but only small adjustments in the relative activation levels due to increased moment magnitude.

Trunk muscle geometry and centroid location when twisting.

The trunk muscles of the lumbar region were studied using magnetic resonance imaging (MRI) in five male volunteers placed in neutral and in two twisted postures. Using a CAD digitizing system, the positions of trunk muscle centroids and the cross-sectional areas of these muscles were measured from the transverse scans at 1 cm intervals from L2 to S1. Muscle lines of action were created by connecting the muscle centroids from the successive sections. Changes in cross-sectional areas and displacements of centroids in a three-dimensional coordinate system and the local two-dimensional coordinate system were estimated at each disc level. In the three-dimensional coordinate system, all muscle centroids were displaced when twisting. Thus, the locations and orientations of all muscle lines of action changed. In the local two-dimensional coordinate system, only the muscle centroids of four abdominal muscles were displaced. Most of these displacements occurred in the first 25 degrees of twisting. Higher up in the lumbar spine, the displacement was greater. The changes in moment arms were sometimes as great as twofold. Only the abdominal oblique muscles (AOM) changed their cross-sectional area significantly; the area of the right AOM ipsilateral to the side of twisting increased, while that of the left AOM was decreased.

An ultra-weak chemiluminescence study on oxidative stress in rabbits following acute thermal injury.

It is not easy to detect oxygen free radicals directly because of their very short half-life. In the present study, a sensitive ultra-weak chemiluminescence detector was used to detect the generation of oxygen free radicals following thermal injury. Twelve New Zealand white rabbits were used in this study. After anesthesia, the bilateral hind-limbs were exposed to 100 degrees C water for 30 s. Six control animals were exposed to 22 degrees C water to act as a control. The chemiluminescence of whole blood and visceral organs were measured with both luminol-amplified t-butyl hydroperoxide-initiated and lucigenin-initiated methods. The results showed that chemiluminescence of blood was affected significantly by acute thermal injury. The chemiluminescence of blood increased significantly at 1 h following acute thermal injury, reached a peak at 2 h, then decreased but still remained above the control level at 4 h following thermal injury. The results for TBHP-initiated chemiluminescence from visceral organs following acute thermal injury were much higher than that of the control rabbits. The effects of lucigenin-initiated tissue chemiluminescence following acute thermal injury were not statistically significant. It is suggested that the decreased vascular antioxidant activity following local thermal injury is partially contributed by the superoxide pathway; while, the remote pathophysiologic events are mediated by the defective scavenging defenses.

Cushioning effect of heel cups.

Three types of heel cups, two rubber and one plastic, were evaluated in this study. The vertical forces under the heel were monitored using the Computer Dyno Graphy system in 16 normal subjects. Peak force reduction in walking and running after heel cup use was found for all three types of heel cups. The shock absorbency (peak force reduction as a ratio) of heel cups was better in walking (3.5 km h(-1)) than in running (10 km h(-1)). Pressure-sensitive film under the heel revealed that the pressure concentration at the location of calcaneal tuberosity could be smoothed out by the use of heel cups. When plastic heel cups were used, pedobarography showed that the contact area of the heel while standing decreased to 61% of that when barefoot. Roentgenographic study in six patients with heel pain syndrome showed that the thickness of the heel pad increased from 14.4 mm (SD 1.4 mm) to 17.0 mm (SD 1.2 mm) when plastic heel cups were used.

Failure sites and peak tensile forces of the composite triceps surae muscle by passive extension in the rabbit.

The triceps surae muscle-tendon unit of rabbit was tested by a passive extension technique. The effects of different stretch rates were evaluated. Functioning as a composite muscle-tendon unit, the triceps surae responded viscoelastically to the tensile loads. The sites of failure were consistent at the muscular portion near the musculotendinous junction. Failure occurred within 0.1-1.0 mm of the distal musculotendinous junction for the soleus muscle and within 5-10 mm proximal to the distal musculotendinous junction in the lateral head of gastrocnemius muscle. In the medial head of gastrocnemius muscle failure occurred within 15-30 mm proximal to the distal musculotendinous junction. There was a greater peak tensile force requirement and a greater energy absorption when tested with faster stretch rates. These results furnish evidence that the biomechanical property of composite musculotendinous unit is strain-rate dependent and behaves viscoelastically.

Effect of nerve function on the failure mechanism of the triceps surae muscle by passive extension in the rabbit.

The triceps surae muscle-tendon units of rabbits were tested in vivo by a passive extension technique using an MTS Bionix 858 machine. The effect of nerve function on the failure mechanism was evaluated. The sites of failure were always at the muscular portion near the musculotendinous (MT) junction. It always occurred within 0.1 to 1.0 mm of the distal MT junction of the soleus muscle and within 5 to 10 mm of the distal MT junction in the lateral head of the gastrocnemius muscle. In the medial head of the gastrocnemius muscle, it always occurred within 5 to 10 mm of the proximal MT junction. The loss of nerve function significantly reduced the peak force and the absorption of energy before the point of peak force, but it produced no significant changes in the failure length and the total energy absorption. The functioning nerve might have a protective mechanism which works on the muscle-tendon unit by increasing the peak force and the energy requirement before the muscle is injured.

Experimental structural scoliosis in rabbits.

Structural scoliosis was produced by purely posterolateral mechanical tethering of the spine with ligation of the scapula to the opposite pelvic bone in rabbits during the growing period. It is a simple and easy method without direct trauma to the spine. When the tethering force occurred discontinuously or only in the coronal plane by drawing of the scapula to the ipsilateral pelvic bone in 17 rabbits, spinal deformity could not be produced. The model provided evidence of biplanar deformity as one of the initial factors for the development of structural scoliosis.

Magnetic resonance imaging of vertebral compression fractures.

In this study, magnetic resonance imaging (MRI) was used to analyze the signal intensity and vascularity of compression fractures of vertebrae in 74 patients. The possibility of nonunion was assessed according to the specific image findings and clinical presentation. All patients had chronic back pain for more than 3 months and compression fractures of the vertebrae initially demonstrated by plain radiography. Pre-enhanced T1 and T2*-weighted images (*multiplaner gradient recall sequence) and postenhanced MRI were obtained. Images were divided into three categories according to the signal intensity of the fractured vertebrae such as hyperintensity (n = 35), hypointensity (n = 24) on T1-weighted image and necrotic type compression fractures of the vertebrae (n = 15). Of the 15 necrotic-type cases, 13 disclosed "fluid"-containing space at the collapsed vertebrae and two showed "air"-containing space at the vertebral body. We believe that these findings are pathognomonic signs of nonunion of the collapsed vertebrae. Surgical specimens were obtained from the four patients whose vertebrae showed necrosis and granulation tissue. After posterior spinal instrumentation, the collapsed vertebral body regained the height and presence of the open end-plate of the vertebra on postoperative lateral radiography. The superior capabilities of MRI offers useful criteria that make the diagnosis of nonunion in compression fractures of the spine possible. Thus, a space with "fluid" or "air" collection at the anterior aspect of a collapsed vertebra as well as strong enhancement with Gd-DTPA at the posterior aspect of the collapsed vertebra may be considered to be pathognomonic signs of nonunion of the fractured vertebra.

Superoxide anion scavenge effect of Quercus glauca Thunb. in whole blood of patients with ankylosing spondylitis.

Nine phenolic compounds, catechin (1), epicatechin (2), gallocatechin (3), epigallocatechin (4), procyanidin B-4 (5), catechin-3-O-rhamnoside (6), rutin (7), querglanin (8) and isoquerglanin (9) were isolated from oak leaves (Quercus glauca Thunb. Fagaceae), and the latter two (8, 9) were identified as new compounds. Several Quercus species have been used in folk medicine as an astringent for hemorrhoids and for treatment of inflammation, jaundice, and tumor. In this study, these compounds were tested for scavenging effects of the superoxide anion in the whole blood of patients with ankylosing spondylitis by means of an ultra-sensitive chemoluminescence (CL) analyzer and lucigenin amplification. The results showed that at a concentration of 2.3 x 10(-5) M, isoquerglanin (9) displayed the strongest inhibition activity (73.55%), followed by querglanin (8) (68.81%) and then gallocatechin (3) and epigallocatechin (4) (66.97 and 60.17% inhibition, respectively). In addition, the blood chemoluminescence (CL) level of patients with ankylosing spondylitis was inhibited by superoxide dismutase (SOD) but not by catalase, suggesting that superoxide anion is the major component of reactive oxygen species (ROS) involved in this assay system.

Entrapment of index flexor digitorum profundus and median nerve: an unusual complication of distal radial fracture--a case report.

A case of index flexor digitorum profundus and median nerve entrapment by bony fracture after reduction as a complication of distal radial fracture in an 11-year-old male is described. After release of tendon and nerve the patient had a good functional result.

Age differences in remote pointing performance.

The purpose of this study was to investigate age differences in remote pointing movements. The subjects were recruited from three age groups (ages 18-22 yr., 40-50 yr., and 60-70 yr., with 9 men and 9 women in each group). They were required to perform cursor-positioning tasks using a remote pointing device, in which the dependent measures were the time taken to reposition the cursor and the accuracy of subjects' movement trajectories. The movement time was further separated into two components, First Submovement duration and Adjustment Submovement duration. Analysis indicated that age groups showed reduced performance on remote pointing. Moreover, remote positioning movement for the young-adult group was mostly completed in their First Submovement phase, while the elderly subjects spent most of their movement time on the Adjustment Submovement phase. These results support the proposition that different age groups exhibit different kinds of movement patterns.

The in vitro effects of dehydroepiandrosterone on chondrocyte metabolism.

OBJECTIVE: To investigate the in vitro effects of dehydroepiandrosterone (DHEA) on neonatal rat chondrocytes. DESIGN: Chondrocytes isolated from neonatal rat cartilage were cultured in three-dimensionally agarose beads and were treated with DHEA. METHODS: Primary culture of chondrocytes was harvested from newborn Wistar rats. The DHEA effects on chondrocyte activities were evaluated by analyzing chondrocyte proliferation, matrix protein synthesis, gene expressions of collagen, matrix metalloproteinase-1, -3 and -13 (MMP-1, -3 and -13), and cyclooxygenase-2 (COX-II), and protein synthesis of interleukin-6 (IL-6), prostaglandin E2 (PGE2) and tissue inhibitor of metalloproteinase-1 (TIMP-1). RESULTS: The DHEA treatment did affect chondrocyte proliferation and glycosaminoglycan (GAG) synthesis. DHEA suppressed the expression of MMP-1, -3 and -13 genes and PGE2 protein synthesis enhanced by lipopolysaccharide (LPS) while the COX-II and inducible nitric oxide synthase (iNOS) gene expressions were down-regulated by DHEA. CONCLUSIONS: Our study demonstrates that DHEA has an ability to modulate the imbalance between MMPs and PGE2 in the neonatal chondrocytes which suggest that it has a potential protective role against articular cartilage damage.

Trunk muscle activation and cocontraction while resisting applied moments in a twisted posture.

Previous studies of twisting have revealed substantial cocontraction of agonist and antagonist muscles within the torso when torsional moments are generated. The objective of the current study was to quantify the activations and cocontraction of eight trunk muscles as subjects maintained an axially rotated trunk posture and resisted external applied bending moments. Ten subjects twisted their torsos 25 degrees to the right (clockwise) and resisted 20 and 40 Nm bending moments from 12 directions. The moment directions were in a transverse plane and labelled clockwise as viewed from above, ranging from 0 degrees (mid-saggital, anterior) to 330 degrees, in 30 degrees increments. RMS EMG amplitude data were collected using surface electrodes and normalized to maximal voluntary contractions. Significant changes were observed in the muscle responses due to the interaction of the moment direction and moment magnitude for six of the eight muscles tested. Comparison of the present data with that collected previously in neutral postures indicated: (1) a large increase in the activation levels of the right erector spinae and the left external oblique muscles; and (2) a counter-clockwise shift in the moment direction at which the peak activation of these same muscles occurs. Analysis of the relative activation levels (RALs), constructed from the NEMG data to quantify the cocontraction, indicated that the changes in cocontraction were more robust in response to changes in the bending moment's direction as opposed to changes in bending moment's magnitude.

Biomechanical analysis in ladder climbing: the effect of slant angle and climbing speed.

This paper presents the findings of a study conducted to determine the peak forces generated in the articulated joints and the foot/floor contact point of a ladder while an individual was engaged in ladder climbing. A kinematic description of joint movements and the calculation of forces acting on each body segment as well as the foot of the ladder were obtained on the basis of videographic data. Task variations were (1) slant angle (70 degrees and 75 degrees with the horizontal) and (2) climbing speed (86 steps/min and 106 steps/min). It was identified in this study that the ladder's friction forces had a time variant nature as a result of biodynamic movements. There were two peak friction coefficients, in opposite directions, occurring at 7% and 38% of each half of the strike time. The primary differences between climbing the ladders at 70 and 75 degrees were a greater posterior displacement of the body's center gravity and smaller peak center gravity shearing forces in climbing the 75 degree ladder. There were greater posterior displacement of the center gravity of body and a larger biomechanical load while climbing at 106 steps/min than at 86 steps/min.