Sciatic Neurectomy Research Articles

Distinct gene expression profiles in the femora of rats exposed to speceflight, tail-suspension and denervation

Unloading conditions occasionally cause muscle atrophy and osteopenia in humans and animals. In general, transcriptional alternations in bone and skeletal muscle cells mediate, respectively, the bone and muscle atrophy caused by unloading. In this study, we examined comprehensive gene expression in the femur of rats exposed to spaceflight (STS-58), tail-suspension and denervation (sciatic neurectomy), to characterize unloading-induced changes in bone gene expression. Interestingly, the transcriptional responses of bone cells to spaceflight were different from those to the other conditions: (i) the expression of bone formation markers was suppressed by spaceflight and denervation, but only spaceflight significantly inhibited the expression of bone resorption markers; (ii) the numbers of the genes that exhibited more than a five-fold change in expression level in response to spaceflight, tail-suspension and denervation were 122 (up, 14 genes; down, 108 genes), 74 (up, 72 genes; down, 2 genes) and 23 (up, 20 genes; down, 3 genes), respectively; (iii) spaceflight preferentially down-regulated the expression of signal transduction and gene-expression regulatory genes, especially GTP-binding protein-signaling molecules involved in bone formation; and (iv) only spaceflight up-regulated the expression of a predicted ubiquitin ligase, F-box protein 30. Our present study identifies novel key genes with which to elucidate further the mechanism of osteopenia caused by unloading, especially spaceflight.

Time response of the canal network in cortical bone to sciatic neurectomy in growing rats: a synchrotron radiation μCT study

Time response of the canal network in cortical bone to sciatic neurectomy in growing rats: a synchrotron radiation μCT study

210 若年ラットにおける坐骨神経切断の皮質骨微細構造への影響 : 単色放射光μCTを利用した脈管路構造の三次元解析(OS1-2 再生医工学(2),オーガナイズドセッション1:細胞・組織・器官のバイオメカニクス/再生医工学,学術講演)

210 若年ラットにおける坐骨神経切断の皮質骨微細構造への影響 : 単色放射光μCTを利用した脈管路構造の三次元解析(OS1-2 再生医工学(2),オーガナイズドセッション1:細胞・組織・器官のバイオメカニクス/再生医工学,学術講演)

Sympathetic Nervous System Does Not Mediate the Load-Induced Cortical New Bone Formation

The contribution of the SNS to bone's response to mechanical loading is unclear. Using a noninvasive model of axial loading of the murine tibia, we found that sciatic neurectomy enhances load-induced new cortical bone formation and that pharmacological blockade of the SNS does not affect such responses, indicating that the SNS does not mediate the osteogenic effects of loading in cortical bone. There is increasing evidence that the sympathetic nervous system (SNS) contributes to the regulation of bone mass and may influence remodeling by modulating bones' response to mechanical load-bearing. The aim of this study was to examine the effect of sciatic neurectomy (SN) on the changes in cortical bone formation induced in response to mechanical loading and to investigate whether the SNS is directly involved in such load-induced responses. Accordingly, load-induced responses were compared in tibias of growing and adult control C57Bl/J6 mice and in mice submitted to unilateral SN; noninvasive axial loading that induced 2,000 microstrain on the tibia lateral midshaft cortex was applied cyclically, 5 or 100 days after surgery, for 7 minutes, 3 days/week for 2 weeks, and mice received calcein on the third and last days of loading. Tibias were processed for histomorphometry, and transverse confocal images from diaphyseal sites were analyzed to quantify new cortical bone formation. Chemical SNS inactivation was achieved by prolonged daily treatment with guanethidine sulfate (GS) or by the introduction of propranolol in drinking water. Our results show that new cortical bone formation is enhanced by loading in all tibial sites examined and that load-induced periosteal and endosteal new bone formation was greater in the SN groups compared with sham-operated controls. This SN-related enhancement in load-induced cortical bone formation in tibias was more pronounced 100 days after neurectomy than after 5 days, suggesting that longer periods of immobilization promote a greater sensitivity to loading. In contrast, the increases in new bone formation induced in response to mechanical loading were similar in mice treated with either GS or propranolol compared with controls, indicating that inactivation of the SNS has no effect on load-induced cortical new bone formation. This study shows that SN, or the absence of loading function it entails, enhances loading-related new cortical bone formation in the tibia independently of the SNS.

Effects of risedronate on femoral bone mineral density and bone strength in sciatic neurectomized young rats

Immobilization induces a rapid loss of bone density and bone strength in rats. The purpose of the present study was to examine the effects of risedronate (Ris) on the femoral bone density and bone strength of sciatic neurectomized young rats. Forty male Sprague-Dawley rats, 6 weeks of age, were randomized by the stratified weight method into the following four treatment groups of 10 rats each: sham-operation, bilateral sciatic neurectomy (NX), NX + low-dose Ris (0.25 mg/kg/day, orally), and NX + high-dose Ris (0.5 mg/kg/day, orally). After 8 weeks of feeding, the volumetric bone mineral density (vBMD) and stress strain index (SSI) of the femoral distal metaphysis and middiaphysis of the rats were measured by peripheral quantitative computed tomography. The mechanical properties of the femoral distal metaphysis and middiaphysis were measured by the compression and three-point bending tests, respectively. The femoral length was also measured. As compared with the findings in the sham-operated controls, NX resulted in a loss of femoral length, cancellous vBMD, SSI, maximum load, stiffness, and breaking energy of the femoral distal metaphysis; there was also loss of cortical thickness, SSI, maximum load, and stiffness of the femoral middiaphysis, with no significant effects on the cortical vBMD or breaking energy of the femoral middiaphysis. High-dose Ris increased the vBMD to values higher than those in the sham-operated controls, and prevented the loss of SSI, maximum load, and stiffness of the femoral distal metaphysis, while low-dose Ris prevented the loss of cancellous vBMD of the femoral distal metaphysis. Neither high- nor low-dose Ris affected any of the cortical bone parameters of the femoral middiaphysis, except for cortical thickness, or the femoral length. These findings suggest that Ris may prevent immobilization-induced loss of cancellous bone density and bone strength in a dose-dependent manner without interfering with bone growth, but has no apparent effects on the cortical bone in sciatic neurectomized young rats. The results of the present preclinical study should be taken into consideration prior to the commencement of Ris treatment for disabled children.

Osteoactivin upregulates expression of MMP-3 and MMP-9 in fibroblasts infiltrated into denervated skeletal muscle in mice

In this study, we examined pathophysiological roles of osteoactivin, a functionally unknown type I membrane glycoprotein, in mouse skeletal muscle atrophied by denervation (sciatic neurectomy). Denervation increased the amounts of osteoactivin, vimentin, matrix metalloproteinase-3 (MMP-3), and MMP-9 in mouse gastrocnemius muscle. Interestingly, immunohistochemical analysis revealed that vimentin, MMP-3, and MMP-9 were mainly present in fibroblast-like cells infiltrated into denervated mouse gastrocnemius muscle, whereas osteoactivin was expressed in the sarcolemma of myofibers adjacent to the fibroblast-like cells. On the basis of these findings, we reasoned that osteoactivin in myocytes was involved in activation of the infiltrated fibroblasts. To address this issue, we examined effects of osteoactivin on expression of MMPs in fibroblasts in vitro and in vivo. Overexpression of osteoactivin in NIH-3T3 fibroblasts induced expression of MMP-3, but not in mouse C(2)C(12) myoblasts, indicating that osteoactivin might functionally target fibroblasts. Treatment with recombinant mouse osteoactivin increased the amounts of collagen type I, MMP-3, and MMP-9 in mouse NIH-3T3 fibroblasts. The upregulated expression of these fibroblast marker proteins was significantly inhibited by heparin, but not by an integrin inhibitor, indicating that a heparin-binding motif in the extracellular domain might be an active site of osteoactivin. In osteoactivin-transgenic mice, denervation further enhanced expression of MMP-3 and MMP-9 in fibroblasts infiltrated into gastrocnemius muscle, compared with wild-type mice. Our present results suggest that osteoactivin might function as an activator for fibroblasts infiltrated into denervated skeletal muscles and play an important role in regulating degeneration/regeneration of extracellular matrix.

Monochromatic synchrotron radiation μCT reveals disuse-mediated canal network rarefaction in cortical bone of growing rat tibiae

The purpose of this study was to demonstrate the ability of computed microtomography based on monochromatic synchrotron radiation (SRmuCT) in microstructural analysis of cortical bone. Tibial diaphyses of growing rats (14 wk, n = 8) undergoing unilateral sciatic neurectomy 8 wk before study were imaged with spatial volume resolution of 5.83 x 5.83 x 5.83 microm3 by SRmuCT (20 keV) at the synchrotron radiation facility (SPring-8). Reconstructed image data were translated into local mineral densities by using a calibrated linear relationship between linear absorption coefficients and concentrations of homogeneous K2HPO4 solution. Pure bone three-dimensional images, produced by simple thresholding at a bone mineral density of 0.82 g/cm3, were analyzed for macro- and microscopic structural properties. In neurectomized hindlimbs, cortical canal network rarefaction as well as bone atrophy were found. The former was characterized by 30% smaller porosity, 11% smaller canal density in transverse section, and 38% smaller canal connectivity density than those in contralateral bone. On the other hand, no difference was found in bone mineral density between neurectomized and intact hindlimbs (1.37 vs. 1.36 g/cm3). In conclusion, SRmuCT is a promising method for the three-dimensional analysis of cortical microstructure and the degree of mineralization in small animals.

The effects of low-intensity ultrasound on growing bone after sciatic neurectomy

The purpose of this study was to investigate the therapeutic effects of low-intensity ultrasound (US) on the sciatic-neurectomy-induced bone mass decrement in growing rats. A total of 20 male Sprague-Dawley rats (166.7 ± 11.3 g) underwent right leg sciatic neurectomy. They were randomly assigned into two groups, US treatment group (UST) and control group (CON). The rats receiving US treatment were treated with a 125 mW/cm 2 continuous low-intensity US stimulation for 15 min/day on the lateral site of the right leg. The control rats did not receive any US treatment. All the animals were euthanized after 4-week US treatment. Both the original data and bilateral difference ratio of femur or tibia weight, histomorphometry data and bone densitometry data showed that the sciatic neurectomy obviously reduced bone mass in the operated limbs of both groups. However, the continuous low-intensity US treatment did not ameliorate the neurectomy-induced loss of bone mass. Thus, the low-intensity US generated micromechanical strains might not induce enough bone formation activity to reverse the bone loss in this model of intact bones. (E-mail: winli@ntu.edu.tw)

Effect of Mornidae Radix extracts on experimental osteoporosis in sciatic neurectomized mice

The preventive and therapeutic effects of aqueous extracts of Mornidae Radix (MR) were observed in sciatic neurectomized mice, a disused osteoporotic model. The right hind limbs of 80 mice were neurectomized and 20 mice were sham-operated and served as a sham control. Then 50, 100 and 200 mg/kg of MR extracts were dosed 3 days after neurectomy for 6 weeks in the prevention study and were dosed 2 weeks after neurectomy for 12 weeks for the therapeutic study. After dosing with the MR extracts, the thickness of the hind limbs, tibia failure load, tibia bone mineral density (BMD), serum osteocalcin levels, tibia calcium (Ca) and phosphorus (P) contents were monitored with histomorphometrical changes of the tibia. In both the prevention and therapeutic studies, the MR extracts significantly and dose-dependently suppressed the decrease in hind limb thickness, tibia failure load, BMD, tibia Ca and P contents with an increase in serum osteoclacin levels. In addition, the MR extracts also significantly and dose-dependently suppressed the decrease in histomorphometrical parameters of the tibia such as volume, length and thickness of trabecular bone and thickness of cortical bone with an increase in osteoclast cells in both the prevention and therapeutic studies. Based on these results, the MR extracts may act as both a suppressor of bone resorption and an enhancer of bone formation in vivo and may have some favorable effects for preventing and treating the osteoporosis induced by sciatic neurectomy.

Osteoclast development in immobilized bone is suppressed by parathyroidectomy in mice

We tested the hypothesis that signaling of parathyroid hormone (PTH) facilitates osteoclastogenesis in bone marrow cells after immobilization, thereby reducing trabecular bone volume. We performed histomorphometric analyses in immobilized limbs after right sciatic neurectomy (IM) and in the contralateral limbs after sham surgery (M). Mice underwent thyroparathyroidectomy (TPTX) and then 0.2 microg/body of thyroxine was given three times a week, or the mice were subjected to sham surgery (sham). Six-week-old male ddY mice were assigned to four groups, as follows, after acclimatization for 1 week: M + sham, IM + sham; M + TPTX, and IM + TPTX. Bilateral tibial samples were used for analysis. Trabecular bone volume (BV/TV) in the secondary spongiosa of the proximal tibias in IM + sham was significantly reduced compared to that in M + sham. Osteoclast surface (Oc.S/BS) and number (Oc.N/BS) in IM + sham transiently increased at 3 and 4 weeks after IM. In contrast, TPTX partially prevented the IM-related reduction of BV/TV and completely suppressed the transient increases of Oc.S/BS and Oc.N/BS. In the bone marrow cells, the mRNA expression of RANKL was elevated in IM + sham, but not in IM + TPTX, compared to that in M + sham. The percentage of Mac-1-positive bone marrow cells, osteoclast precursors, was not altered after IM. There were no significant differences in the concentrations of interleukin (IL)-1alpha in the tibial bone marrow cell culture medium between M + sham and IM + sham. Our data demonstrated that significant increases in osteoclast surface and number after IM were suppressed in TPTX mice, closely associated with a reduction in the high expression of RANKL mRNA in the tibial bone marrow cells. We speculate that enhanced osteoclastogenesis due to limb immobilization may be related to the elevation of RANKL expression by the facilitation of parathyroid hormone signaling in bone marrow cells.

214 片足坐骨神経切断による脛骨皮質骨微細構造への影響(OS1-06 : 生体マイクロ・ナノ構造メカニクス(2),マイクロ・ナノバイオメカニクス)

214 片足坐骨神経切断による脛骨皮質骨微細構造への影響(OS1-06 : 生体マイクロ・ナノ構造メカニクス(2),マイクロ・ナノバイオメカニクス)

Menatetrenone rescues bone loss by improving osteoblast dysfunction in rats immobilized by sciatic neurectomy

Menatetrenone (MK-4) is a vitamin K 2 homologue that has been used as a therapeutic agent for osteoporosis in Japan. However, there is no far any reported evidence that MK-4 ameliorates a pre-existing condition of reduced bone mineral density (BMD) in vivo. In this study, we evaluated the effect of MK-4 in a rat model of established bone loss through immobilization caused by sciatic neurectomy. Unilateral sciatic neurectomy (SNx) was performed in rats, and 10 or 30 mg/kg of MK-4 or vehicle was administered to the rats three weeks after operation. Seven weeks after operation, the rats were sacrificed and BMD and bone histomorphometric parameters were measured to assess the effects of MK-4. While BMD of the distal femoral metaphysis was significantly decreased after SNx, MK-4 administration increased BMD in the neurectomized rats. Bone formation was decreased continuously and bone resorption was initially increased in SNx rats. Four weeks treatment of MK-4 increased bone formation and suppressed bone resorption. In addition, increased carboxylated osteocalcin and decreased undercarboxylated osteocalcin in serum were observed in MK-4-administered rats. These results indicated that MK-4 rescued bone volume by improving osteoblast dysfunction and accelerating gamma carboxylation of osteocalcin. MK-4 may be useful for treating disuse osteopenia.

Pinealectomy exaggerates and melatonin treatment suppresses neuroma formation of transected sciatic nerve in rats: gross morphological, histological and stereological analysis

At present, an intensive effort for prevention of neuroma formation following peripheral nerve section continues. It has been recently suggested that surgical pinealectomy (Px) induces elevation of the collagen content in the granulation tissue of a wound, while melatonin application after Px suppresses elevation of the collagen accumulation in the tissue. The aim of the present study was to assess whether melatonin had the ability to suppress collagen production and neuroma formation following peripheral nerve transection. A total of 40 male rats (four groups of 10) were left intact (intact controls) or sham operated (sham group), were Px, or were Px and given melatonin (Px + melatonin group). All animals underwent a surgical intervention consisting of right sciatic nerve neurectomy. After 4 wk, the animals were killed following intracardiac perfusion. Gross morphology of neuroma formation in the proximal nerve segment was examined and proximal neuroma evaluated. Macroscopic and microscopic findings revealed that Px caused a proliferation of connective tissue and large neuroma formation at the proximal ends of transected nerves. Stereological analysis showed that there was a statistically significant reduction in connective tissue content of the same region in Px animals treated with melatonin (P < 0.005). The results achieved in a rodent model of sciatic nerve neuroma formation showed that there was a positive correlation between macroscopic and microscopic observations, and that melatonin enhanced axonal regeneration presumably due to its inhibitory effect on neuroma formation.

Dramatic Decrease of Innervation Density in Bone after Ovariectomy

Recent studies have demonstrated that bone is highly innervated and contains neuromediators that have functional receptors on bone cells. However, no data exist concerning the quantitative changes of innervation during bone loss associated with estrogen withdrawal. To study the involvement of nerve fibers in the regulation of bone remodeling, we have evaluated the modifications of innervation in a classical in vivo model of osteopenia in rats, ovariectomy (OVX). Skeletal innervation was studied by immunocytochemistry using antibodies directed against specific neuronal markers, neurofilament 200 and synaptophysin, and the neuromediator glutamate. Sciatic neurectomy, another model of bone loss due to limb denervation and paralysis, was used to validate our quantitative image analysis technique of immunostaining for nerve markers. Female Wistar rats at 12 wk of age were sham-operated (SHAM) or ovariectomized (OVX). Bone mineral density measurement and bone histomorphometry analysis of tibiae 14 d after surgery demonstrated a significant bone loss in OVX compared with SHAM. We observed an important reduction of nerve profile density in tibiae of OVX animals compared with SHAM animals, whereas innervation density in skin and muscles was similar for OVX and control rats. Quantitative image analysis of immunostainings demonstrated a significant decrease of the percentage of immunolabeling per total bone volume of neurofilament 200, synaptophysin, and glutamate in both the primary and secondary spongiosa of OVX rats compared with SHAM. These data indicate for the first time that OVX-induced bone loss in rat tibiae is associated with a reduction in nerve profile density, suggesting a functional link between the nervous system and the bone loss after ovariectomy.

Effects of unilateral sciatic neurectomy on growing rat femur as assessed by peripheral quantitative computed tomography, Fourier transform infrared spectroscopy and bending test.

We studied the effects of unilateral sciatic neurectomy (USN) on the development of the femoral shaft in the 30 growing Wistar-derived rats aged 5 weeks. Rats were allocated to three groups. One of these was immediately used for measurements, and the remaining 2 groups underwent USN of internal control. Specimens obtained from each group were divided into 2 subgroups: left femurs of each group served as the control subgroup (CONT) and right femurs from each group as the USN-operated subgroup (USN-OP). The bone mineral density (BMD), bone mineral content (BMC), bone area, periosteal circumference and endosteal circumference were measured by peripheral quantitative computed tomography (pQCT) and the mineral/matrix ratio was evaluated by Fourier transform infrared spectroscopy (FTIR). A three-point bending test was performed to analyze the biomechanical effects of sciatic neurectomy. USN-OP showed a significant decrease in cortical BMC, bone area, and periosteal circumference compared with CONT. The mineral/matrix ratio of cortical bone did not differ significantly between USN-OP and CONT. Strength and stiffness were significantly decreased in USN-OP compared with CONT. The results showed that USN inhibited periosteal bone formation, but has no significant effects on the mineral/matrix ratio of cortical bone in femurs.

坐骨神経切除によるラット脛骨骨幹端の骨梁構造変化並びに骨格筋の萎縮

Disused rat hindlimb caused by sciatic neurectomy is characterized by skeletal muscle atrophy and bone loss accompanying alterations in trabecular bone architecture. We studied the time course responses of the two-dimensional architecture of trabecular bone and the relationships between bone loss and muscle atrophy using a unilateral sciatic nerve denervation model of disuse atrophy.Fischer-344 male rats aged 11 weeks were denervated. The right and left tibiae, soleus and extensor digitorum longus muscle of denervated and control rats were obtained at 1, 3, 7, 10 days and 2, 3, 4, 5, 6, 7, 8, 10 weeks after surgery. Histomorphometric analyses were performed on longitu-dinal sections of proximal tibial metaphyseal secondary spongiosa.Marked trabecular bone loss and muscle weight loss were observed mainly at 7-10 days and 1-3 days after denervation, respectively. Time courses of the percent decrease in trabecular width and length from 0 day (i, e., thinning rate; TR and fragmentation rate; FR) were not matched at 7-10 days and 5-6 weeks after denervation. The scatterplot for TR; FR showed that the FR changed stepwise with the first threshold between 12-25% of TR.

A substance P receptor (NK 1) antagonist enhances the widespread osteoporotic effects of sciatic nerve section

The long-term effects of sciatic nerve section on bone mineral density (BMD) were studied using dual-energy X-ray absorptiometry (DEXA) in skeletally mature rats. Unilateral sciatic neurectomy caused the rapid loss of cancellous bone in the proximal and distal femur and tibia in the ipsilateral hindlimb and, to a lesser extent, in the contralateral intact hindlimb. The reduction in BMD rapidly progressed for 4 weeks after sciatic section and then gradually stabilized with no evidence of recovery at 12 weeks. The development of osteoporosis in the contralateral intact hindlimb was a novel finding. There was no evidence of disuse in the normal contralateral hindlimb after unilateral sciatic section; grid-crossing activity over a 24-h interval was unchanged and there was no reduction in weight bearing on the contralateral normal hindpaw during the stance phase of ambulation. Unilateral peripheral nerve lesions have well-documented effects on substance P content and function in the corresponding contralateral intact nerve. We hypothesized that after sciatic section a reduction in substance P signaling might contribute to bone loss in the contralateral hindlimb. Daily administration of the substance P receptor (NK 1) antagonist LY303870 for 2 weeks caused significant loss of cancellous bone in the denervated and the contralateral hindlimb, evidence that substance P signaling sustained bone density after nerve section. After sciatic neurectomy there was a 33% reduction in sciatic nerve stimulation-evoked extravasation in the contralateral intact hindlimb, indicating transmedian inhibition of substance P signaling after nerve injury. Furthermore, there was a 50% reduction in the substance P content in both tibias after unilateral sciatic section. Collectively, these data support the hypothesis that a widespread reduction in substance P content in bone contributes to the osteoporotic effects of sciatic neurectomy and that residual substance P signaling maintains bone integrity after nerve section in both the denervated and contralateral intact hindlimb.

Effect of vitamin K2 on cortical and cancellous bones in orchidectomized and/or sciatic neurectomized rats.

We examined the effect of vitamin K2 on cortical and cancellous bones in orchidectomized and/or sciatic neurectomized rats. Ninety male Sprague-Dawley rats, 3 months of age, were randomized by stratified weight method into nine groups with 10 rats in each group: baseline control (BLC), age-matched intact control (IN), IN+vitamin K2 administration (K), orchidectomy (ORX), ORX+K, unilateral sciatic neurectomy (NX), NX+K, ORX+NX (ONX), and ONX+K. Vitamin K2 (menatetrenone) was administered orally twice a week at a dose of 30 mg/kg each. After 10 weeks of feeding, the tibial shaft and proximal tibia were processed for cortical and cancellous bone histomorphometric analyses, respectively. An ORX-induced reduction in maturation-related cortical bone gain and ORX-induced cancellous bone loss were attributable to increased endocortical and trabecular bone turnover, respectively. NX- and ONX-induced reductions in maturation-related cortical bone gain were attributable to decreased periosteal bone formation and increased endocortical bone turnover, while NX- and ONX-induced cancellous bone loss was attributable to increased bone resorption and decreased bone formation. ORX-induced cancellous bone loss was more pronounced when combined with immobilization. Vitamin K2 administration did not significantly alter any parameters in IN rats. Vitamin K2 administration in ORX rats suppressed endocortical bone resorption and trabecular bone turnover, retarding a reduction in maturation-related cortical bone gain and cancellous bone loss. This effect on cancellous bone loss was primarily because of prevention of a reduction of trabecular thickness. Vitamin K2 administration in NX and ONX rats suppressed bone resorption and stimulated bone formation (mineralization), with retardation of a reduction of trabecular thickness without any significant effect on cancellous bone mass, and suppressed endocortical bone resorption, retarding a reduction in maturation-related cortical bone gain. The present study provides evidence indicating that vitamin K2 has the potential to suppress bone resorption or bone turnover and/or stimulate bone formation in vivo in ORX and/or NX rats.

Menatetrenone ameliorates reduction in bone mineral density and bone strength in sciatic neurectomized rats.

Vitamin K2 (menaquinone) acts on the bone metabolism. Menatetrenon (MK-4) is a vitamin K2 homologue that has been used as a therapeutic agent for osteoporosis in Japan. Rat models of immobilization induced by sciatic neurectomy are characterized by transiently increased bone resorption and sustained reduction in bone formation. Using such a rat model, we investigated the efficacy of MK-4 on bone loss. Male Sprague-Dawley rats were subjected to unilateral sciatic neurectomy and administered MK-4 for 28 d beginning day 21 after operation. The effect of MK-4 on the immobilized bone was assessed by measuring the bone mineral density of the femur, breaking force of the femoral diaphysis, and bone histomorphometry in tibial diaphysis. The BMD on both the femoral distal metaphysis and diaphysis was reduced by sciatic neurectomy. The administration of MK-4 ameliorated this reduction in a dose-dependent manner. The administration of 30 mg/kg MK-4 ameliorated the reduction in bone strength. An improvement in bone formation was observed following the administration of MK-4. These results suggest that MK-4 has a therapeutic potential for immobilization-induced osteopenia.

Glutamatergic innervation in bone.

Bone is highly innervated, and evidence for a regulation of bone metabolism by nerve fibers has been suggested by many clinical and experimental studies. However, the nature of the neuromediators involved in these processes has not been well documented. Glutamate (Glu), a major neuromediator of the central nervous system (CNS), was recently identified in nerve fibers running in bone marrow in close contact with bone cells, suggesting that Glu may also act as a neuromediator in this tissue. During the last few years, all the machinery required for glutamate signalling in the CNS was demonstrated in bone. Osteoblasts and osteoclasts express ionotropic Glu receptors (iGluR) (NMDA, AMPA, and Kainate) and metabotropic Glu receptors (mGluR) as well as Glu transporters. Electrophysiological studies have demonstrated that NMDA receptors (NMDAR) and mGluR are functional on bone cells. NMDAR are involved in osteoclast formation and bone resorption and preliminary studies suggest that they may also participate in mechanisms underlying osteoblast proliferation or differentiation, providing evidence for a direct action of Glu on bone cells. The bone loss induced in a model of sciatic neurectomy in growing rats is associated with a decrease of glutamatergic innervation, suggesting that Glu released by nerve fibers may contribute to the regulation of bone remodeling. The manipulation of Glu action in bone may, therefore, represent a new therapeutic target for pathologies associated with modifications of bone remodeling.