Seung Myung Moon

Neuroprotective effects of adipose-derived stem cells against ischemic neuronal damage in the rabbit spinal cord

Transplantation of adipose-derived stem cells (ASCs) is one of the possible therapeutic tools for ischemic damage. In this study, we observed the effects of ASCs against ischemic damage in the ventral horn of L(5-6) levels in the rabbit spinal cord. ASCs were isolated from rabbits, and cell type was confirmed by flow cytometry analysis, labeling with CM-DiI dye and differentiation into adipocytes in adipogenesis differentiation medium. ASCs were administered intrathecally into recipient rabbits (2 × 10⁵) immediately after reperfusion following a 15-min aortic artery occlusion in the subrenal region. Transplantation of ASCs significantly improved functions of the hindlimb and morphology of the ventral horn of spinal cord although CM-DiI-labeled ASCs were not observed in the spinal cord parenchyma. In addition, transplantation of ASCs significantly increased brain-derived neurotrophic factor (BDNF) levels at 72h after ischemia/reperfusion. These results suggest that transplantation of ASCs prevents motor neurons from spinal ischemic damage and reactive gliosis by increasing neurotrophic factors such as BDNF in the spinal cord.

Long-term changes in neuronal degeneration and microglial activation in the hippocampal CA1 region after experimental transient cerebral ischemic damage

Delayed neuronal death following transient cerebral ischemia is mixed with apoptosis and necrosis, and the activation of microglia are activated after the ischemic insult. In the present study, we examined the long-term changes in neuronal degeneration and microglial activation in the gerbil hippocampal CA1 region after 5min of transient cerebral ischemia using specific markers for neuronal damage and microliosis. Transient ischemia-induced neuronal death was shown in CA1 pyramidal cells 4days after ischemia/reperfusion (I/R). However, neuronal degeneration of the pyramidal cells were observed up to 45days in the CA1 region after I/R. Microglial activation was also observed in the CA1 region after I/R. Isolectin B4- (IB4) immunoreactive ((+)) microglia appeared in the CA1 region 4days after I/R. On the other hand, ionized calcium-binding adapter molecule 1 (Iba-1)(+) microglia was markedly increased after I/R, and peaked at 15days after I/R. Thereafter, Iba-1 immunoreactivity was decreased with time-dependant manner in the ischemic CA1 region. These results indicate that neuronal degeneration of CA1 pyramidal cells may last about 45days in the CA1 region after ischemic damage, and microglial activation may be diverse according to their function, such as phagocytosis, after I/R.

Chronological changes of N-methyl-D-aspartate receptors and excitatory amino acid carrier 1 immunoreactivities in CA1 area and subiculum after transient forebrain ischemia

We investigated changes of immunoreactivities of N-methyl-D-aspartate receptor (NR) and of excitatory amino acid carrier 1 (EAAC-1), the neuronal glutamate transporter, in the vulnerable CA1 area and the less vulnerable subiculum of the gerbil hippocampus at various times following transient forebrain ischemia. At 30 min after ischemia-reperfusion, the intensity of NR immunoreactivity increased markedly in neurons of CA1 and subiculum, particularly NR2A/B, while EAAC-1 immunoreactivity was reduced in CA1. At 3 hr after reperfusion, the density of NR1 immunoreactivity markedly decreased in CA1. In contrast EAAC-1 immunoreactivity increased in CA1 and in the subiculum. At 12 hr after reperfusion, the decrease of NR1 immunoreactivity was not detected whereas EAAC-1 immunoreactivities in the CA1 area were intensified. In the subiculum, both NR subunits immunoreactivities decreased significantly, in contrast to the maintenance of EAAC-1 immunoreactivity. At 24 hr after reperfusion, both NR2A/B and EAAC-1 immunoreactivities decreased markedly in CA1 and subiculum. We tentatively suggest that the increase of NR immunoreactivity in CA1 at early times after ischemia-reperfusion may increase the delayed neuronal death, and that the increase or maintenance of EAAC-1 immunoreactivity at early times after ischemia-reperfusion may be an important factor in survival of neurons.

Clinical outcome of posterior fixation of the C1 lateral mass and C2 pedicle by polyaxial screw and rod

OBJECTIVE Because of atlantoaxial complex has a unique and complicated anatomy and instability of this complex is very dangerous. We investigated the clinical results of posterior C1-C2 fixation with a polyaxial screw-rod system. METHODS Between July 2001 and December 2007, the authors treated 17 patients suffering from atlantoaxial deformity and instability. Atlantoaxial fusion was employed in 9 patients with upper cervical fracture and dislocation, in 6 patients with atlantoaxial subluxation, in 1 patient with pure transverse ligament injury, and in 1 patient with basilar invagination. The mean age at the time of surgery was 40.4 years (range, 15-68 years). RESULTS Operative times ranged from 165 to 420 min (average 306 min), and the postoperative mean VAS score was 2.4. The mean follow-up period was 26 months. Solid fusion was achieved in 15 patients at the last follow up; no injury of the vertebral artery or spinal cord and no operative mortality occurred in these cases. CONCLUSIONS We suggest that posterior atlantoaxial fixation using the polyaxial screw-rod system is an effective and relatively safe technique. The navigation guidance system employed during the surgical procedure was helpful methods. Future studies of the feasibility of navigation system-guided surgical procedures will be required.

Effects of treadmill exercise on cyclooxygenase-2 in the hippocampus in type 2 diabetic rats: Correlation with the neuroblasts

Cyclooxygenase (COX) is a rate-limiting enzyme in synthesis of prostaglandins from arachidonic acid. In this study, we observed the effects of a physical exercise on COX-2 immunoreactivity in the hippocampus using immunohistochemistry in rats. In addition, we examined effects of administration of a COX-2 inhibitor, celecoxib, on neuroblast differentiation. At 6weeks of age, Zucker lean control (ZLC) and Zucker diabetic fatty (ZDF) rats were put on a treadmill with or without running for 1h/session/day for 5weeks. The running speed was gradually increased from 16 to 22m/min with 2m/min per 2weeks. In the ZLC and ZDF rats, COX-2 immunoreaction was detected in the granule cell layer of the dentate gyrus and in the stratum pyramidale of the CA2/3 region; COX-2 immunoreaction in the CA1 region was hardly detected. In the exercised-ZLC and ZDF rats, COX-2 immunoreactivity was significantly increased compared to that in the ZLC and ZDF rats, showing that COX-2 immunoreactivity in the exercised-ZDF rats was slightly low than that in the exercised-ZDF rats. In addition, weak COX-2 immunoreactivity was shown in the CA1 region by exercise. On the other hand, the repeated oral administration of celecoxib to 4-week-old ZDF rats significantly decreased the neuroblasts in the subgranular zone of the dentate gyrus. These results suggest that COX-2 may be associated with the increase of synaptic plasticity or contacts in the hippocampus.

c-Myb Immunoreactivity, Protein and mRNA Levels Significantly Increase in the Aged Hippocampus Proper in Gerbils

Myb genes are a family of transcription factors and have been implicated in the control of the proliferation and differentiation of normal and transformed cells. c-Myb is the best characterized member of the myb family. In the present study, we investigated age-dependent changes of c-myb immunoreactivity, its protein and mRNA level in the hippocampus proper (CA1–3 regions) at various age stages in gerbils. In the postnatal month 1 (PM 1) group, c-myb immunoreactivity was detected in non-pyramidal neurons of the strata oriens and radiatum as well as in pyramidal neurons of the stratum pyramidale. At PM 3, c-myb immunoreactivity and its protein level were similar to those at PM 1. Thereafter, c-myb immunoreactivity and its protein level were increased with time. In the PM 24 group, c-myb immunoreactivity, its protein and mRNA levels were highest. These results suggest that the significant increase of c-myb immunoreactivity, protein and mRNA levels in the aged hippocampus may be associated with neuronal aging.

Expression and changes of calbindin D-28k immunoreactivity in the ventral horn after transient spinal cord ischemia in rabbits

We examined ischemia-related changes of calbindin D-28k (CB) immunoreactivity in L(7) of the spinal ventral horn after transient spinal cord ischemia in rabbits. In the sham-operated group, CB immunoreactivity was not present in the spinal ventral horn, but CB immunoreactivity was detectable in the dorsal horn. CB immunoreactivity was detectable in the ventral horn at 30 min after ischemia: the CB immunoreactivity was found in glial cells identified as astrocytes. At 1 h after ischemia, CB immunoreactivity was highest and present at a few somata located in the lamina VII as well as many glial cells. CB immunoreactivity was lower in the lamina VII at 3 h after ischemia compared to 1 h post-ischemic group. By 2 days after ischemia, CB immunoreactivity was decreased in this region. In addition, the result of Western blot result showed the pattern of CB expression similar to that of immunohistochemistry. In conclusion, the ischemia-related changes of CB immunoreactivity in neurons and glial cells in the ischemic spinal ventral horn in rabbits may be related to modulation of intracellular calcium following transient ischemia.

Differences in neuronal damage and gliosis in the hippocampus between young and adult gerbils induced by long duration of transient cerebral ischemia

Response to cerebral ischemia in young animals was very different from that in the adult. The aim of this study was to investigate differences in neuronal death and gliosis in the hippocampal CA1 region (CA1) between adult and young gerbils following 5 and 15 min of transient cerebral ischemia. Delayed neuronal death (DND) of pyramidal cells occurred in the CA1 was similar in all the adult gerbils after 5 and 15 min of ischemia: the DND occurred 4 days after ischemia. In the young groups, DND of pyramidal cells in the CA1 region occurred 7 and 3 days after 5 and 15 min of ischemia, respectively. On the other hand, the activation of GFAP-immunoreactive ((+)) astrocytes and Iba-1(+) microglia was different in the young groups from the adult groups after ischemia. The change pattern of GFAP immunoreactivity in the adult groups was similar in both the adult groups after ischemia; in the young groups, the activation of GFAP(+) astrocytes after 5 min of ischemia was much delayed than that after 15 min of ischemia. Activated Iba-1(+) microglia were aggregated in the stratum pyramidale 4 days after ischemia in all the adult ischemia-operated groups; in the young groups, activated Iba-1(+) microglia were aggregated in the stratum pyramidale 7 days after 5 min of ischemia and 3 days after 15 min of ischemia. These observations indicate that DND in young animals is very different from the adult according to different duration of transient cerebral ischemia and glial activation is very different in young animals after different duration of transient ischemia.

Neurons in the hippocampal CA1 region, but not the dentate gyrus, are susceptible to oxidative stress in rats with streptozotocin-induced type 1 diabetes

In this study, we investigated the effects of streptozotocin-induced type 1 diabetes on antioxidant-like protein-1 immunoreactivity, protein carbonyl levels, and malondialdehyde formation, a marker for lipid peroxidation, in the hippocampus. For this study, streptozotocin (75 mg/kg) was intraperitoneally injected into adult rats to induce type 1 diabetes. The three experimental parameters were determined at 2, 3, 4 weeks after streptozotocin treatment. Fasting blood glucose levels significantly increased by 20.7-21.9 mM after streptozotocin treatment. The number of antioxidant-like protein-1 immunoreactive neurons significantly decreased in the hippocampal CA1 region, but not the dentate gyrus, 3 weeks after streptozotocin treatment compared to the control group. Malondialdehyde and protein carbonyl levels, which are modified by oxidative stress, significantly increased with a peak at 3 weeks after malondialdehyde treatment, and then decreased 4 weeks after malondialdehyde treatment. These results suggest that neurons in the hippocampal CA1 region, but not the dentate gyrus, are susceptible to oxidative stress 3 weeks after malondialdehyde treatment.

Calbindin D-28k is expressed in the microvascular basal lamina in the ventral horn at early time after transient spinal cord ischemia in the rabbit.

Much evidence has been accumulated that the increased expression of calbindin D-28k (CB) is involved in the blockade of calcium-evoked excitotoxicity in cerebral ischemia. We investigated the expression of CB in the basal lamina of microvessels in the ventral horn of the rabbit spinal cord after transient spinal cord ischemia. Spinal cord sections at the level of L7 were immunostained using monoclonal antibody raised against CB at light and electron microscopic levels. CB immunoreactivity was detected in the basal lamina of microvessels at 30 min after ischemic insult. By 3 h after ischemia, CB immunoreactivity was increased in the basal lamina of the microvessels. CB immunoreactivity began to decrease at 6 h after ischemia and nearly disappeared at 48 h after ischemic insult. For calcium detection in the blood vessels of spinal cord, we conducted an alizarin red staining. Alizarin red reactivity was detected in some microvessels at 3 h after ischemic insult. Our results suggest that the ectopic expression of CB in the microvascular basal laminae may be associated with the buffering of calcium in the endothelial cells of microvessels after ischemic damage.