Objectives: We aimed to evaluate the genotoxicity of a recently developed no-pain pharmacopuncture (NPP) targeting muscle relaxation and analgesia using the micronucleus test. Methods: To evaluate the potential of NPP extracts to induce micronuclei in rat bone marrow cells, a micronucleus test was performed using male Sprague-Dawley rats. The test substance NPP was administered intramuscularly at concentrations of 0.25, 0.5, and 1 mL/animal. Saline was used as the negative control and cyclophosphamide as the positive control. Results: No NPP treatment-related deaths or abnormal changes in general appearance were observed at any dose level during the experimental period. No statistically significant differences in body weight were observed in any of the NPP dose groups compared to the saline negative control group. NPP did not cause a significant increase in the incidence of micronucleated polychromatic erythrocytes (PCEs) and PCEs or in the ratio of PCE-to-total erythrocytes. Conclusion: The NPP extract did not exhibit genotoxic in Sprague-Dawley rat bone marrow cells under the conditions of this study. Further toxicity studies of the NPP extract are required.
Objective : This study was designed to validate the cell trafficking efficiency of the in vivo bioluminescence image (BLI) study in the setting of transplantation of the luciferase expressing bone marrow-derived mesenchymal stem cells (BMSC), which were delivered at each different time after transient middle cerebral artery occlusion (MCAO) in a mouse model. Methods : Transplanting donor BMSC were prepared by primary cell culture from transgenic mouse expressing luciferase (LUC). Transient focal infarcts were induced in 4-6-week-old male nude mice. The experiment mice were divided into five groups by the time of MSC transplantation : 1) sham-operation group, 2) 2-h group, 3) 1-day group, 4) 3-day group, and 5) 1-week group. BLI for detection of spatial distribution of transplanted MSC was performed by detecting emitted photons. Migration of the transplanted cells to the infarcted area was confirmed by histological examinations. Differences between groups were evaluated by paired t-test. Results : A focal spot of bioluminescence was observed at the injection site on the next day after transplantation by Signal intensity of bioluminescence. After 4 weeks, the mean signal intensities of 2-h, 1-day, 3-day, and 1-week group were $2.6{\times}10^7{\pm}7.4{\times}10^6$. $6.1{\times}10^6{\pm}1.2{\times}10^6$, $1.7{\times}10^6{\pm}4.4{\times}10^5$, and $8.9{\times}10^6{\pm}9.5{\times}10^5$, respectively. The 2-h group showed significantly higher signal intensity (p<0.01). The engrafted BMSC showed around the infarct border zones on immunohistochemical examination. The counts of LUC-positive cells revealed the highest number in the 2-h group, in agreement with the results of BLI experiments (p<0.01). Conclusion : In this study, the results suggested that the transplanted BMSC migrated to the infarct border zone in BLI study and the higher signal intensity of LUC-positive cells seen in 2 hrs after MSC transplantation in MCAO mouse model. In addition, noninvasive imaging in real time is an ideal method for tracking stem cell transplantation. This method can be widely applied to various research fields of cell transplantation therapy.
Journal of Korean Academy of Oral and Maxillofacial Radiology
/
v.23
no.1
/
pp.27-44
/
1993
This study was performed to investigate the changes of mandibular condyle by low calcium diet and the effects of irradiation on the bone in osteoporotic state. In order to carry out this experiment, 80 seven-week old Sprague-Dawley strain rats weighing about 150 gm were selected and equally divided into one experimental group of 40 rats and one control group with the remainder. The experimental group and the control group were then subdivided into two group and exposed to irradiation. The two irradiation groups received a single dose of 20 Gy on the jaw area only and irradiated with a cobalt-60 teletherapy unit. The rats in the control and experimental groups were serially terminated by fours on the 3rd, the 7th, the 14th, and the 21st day after irradiation. After termination, both sides of the dead rats mandibular condyle were removed and fixed with 10% neutral formalin. The bone mineral density of mandibular condyle was measured by use of dual energy X-ray bone densitometer(model DDX-alpha, Lunn Corp., U. S. A.). The mandibular condyle was radiographed with Hitex HA-80(Hitex Co., Japan). Thereafter, the obtained radiographs were observed, and the mandibular condyle was further decalcified and embedded in paraffin as the general method. The specimen sectioned and stained with hematoxylin-eosin, PAS and Rabbit Anti-Human Tumor Necrosis Factor-a observed by a light microscope. The obtained results were as follows: 1. In the non-irradiated group with the low calcium diet, the bone mineral density of the condyle was markedly decreased after 14 days, and decrease the number of trabeculae of the condyle and resorption of the calcified cartilaginous zone were observed after 3 days. On microscopic observations, the number nd size of trabeculae were decreased after 7 days of experiment. 2. In the irradiated group with the low calcium diet, the bone mineral density of the condyle was markedly decreased after 14 days and resorption of the calcified cartilaginous zone and decrease the number and coarse of the trabeculae of the condyle were observed. These findings were extended rather than in non-irradiated group with low calcium diet. On microscopic observations, many osteoclasts were detected and the number and size of trabeculae were somewhat decreased after 7 days. Also there was degenerative changes of tissues of bone marrow on the 14th day but that condition was restored on the 21st day of experiment. 3. In the irradiated group with normal diet, the bone mineral density of the condyle was somewhat decreased with times and degree of decrease of the number of trabeculae was somewhat larger than in the non-irradiated group with normal diet. On microscopic observations, the tissues of bone marrow were atrophic and degenerative changes but that condition was restored on the 21st day of experiment. 4. In immunocytochemical findings, in the irradiated and non-irradiated groups with low calcium diet, negative or partial positive response to TNF was observed, but positive response in the normal diet groups.
Objectives : In this study, erythrocyte micronucleus test of pomace Schisandra chinensis extracts was conducted in order to up-cycling to a high value-added industry using by-products discarded in the production process of Schisandra chinensis products and active ingredients such as dibenzocyclooctadiene lignans in Schisandra chinensis. Methods : The micronucleus test was performed according to the 'OECD Guidelines'. Including the negative control group(0 mg/kg) and the positive control group(CPA 70 mg/kg), pomace Schisandra chinensis extracts were orally administered to ICR mouse at doses of 500, 1,000, and 2,000 mg/kg. After sacrificing the experimental animals bone marrow cells were collected and micronucleated polychromatic erythrocyte were counted. And genetic toxicity was confirmed according to the frequency of micronucleus. Results : As a result of the micronucleus test, there were no changes in body weight, clinical signs, or death in any group. But, a significant increase was observed in the frequency of micronucleated polychromatic erythrocyte among polychromatic erythrocytes in the positive control group administered with CPA compared to the negative control group(p<0.05). Whereas, no significant increase was observed in the group administered with pomace Schisandra chinensis extracts compared to the negative control group. Conclusions : Pomace Schisandra chinensis extracts did not induce micronucleus in bone marrow cells of ICR mouse up to a concentration of 2,000 mg/kg, and it was judged that no genetic toxicity was observed.
Journal of the Korean Association of Oral and Maxillofacial Surgeons
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v.30
no.5
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pp.380-390
/
2004
Bone healing plays an important role in orthognathic and craniofacial surgery. Bone tissue repair and regeneration are regulated by an array of growth and morphogenetic factors. Bone formation and remodeling require continuous generation of osteoprogenitor cells from bone marrow stromal cells, which generate and respond to a variety of growth factors with putative roles in hematopoiesis and mesenchymal differentiation. In this study, the efficacy of a single application of hepatocyte growth factor to promote bone regeneration in 5-mm experimental calvarial defects of adult male rats was assessed histologically and immunohistochemically. The result of the experimental site were compared with those of the contralateral contral side. None of the control and experimental bone defects demonstrated complete bone closure. Bone regeneration was found close th the margine and central part of the defects. At 1, 2 weeks, there were found much significant cellural mitotic activity and many inflammatory cells and osteoblasts on the experimental site than control site. At 4, 6 weeks, new bone apposition was founded in both site but, more apposition was seen at experimental site. At 8, 12 weeks, also, some differences was found that more apposition of new bone and collagen fiber was seen on experimental site. Our results have some possibility that HGF do a early positive role to repair the bone defect. More study will be needed.
Tuan Anh Vuong;Yan Zhang;June Kim;Young-Eun Leem;Jong-Sun Kang
BMB Reports
/
v.57
no.7
/
pp.330-335
/
2024
Arginine methylation, which is catalyzed by protein arginine methyltransferases (Prmts), is known to play a key role in various biological processes. However, the function of Prmts in osteogenic differentiation of mesenchymal stem cells (MSCs) has not been clearly understood. In the current study, we attempted to elucidate a positive role of Prmt7 in osteogenic differentiation. Prmt7-depleted C3H/10T1/2 cells or bone marrow mesenchymal stem cells (BMSCs) showed the attenuated expression of osteogenic specific genes and Alizarin red staining compared to the wild-type cells. Furthermore, we found that Prmt7 deficiency reduced the activation of bone morphogenetic protein (BMP) signaling cascade, which is essential for the regulation of cell fate commitment and osteogenesis. Taken together, our data indicate that Prmt7 plays important regulatory roles in osteogenic differentiation.
Kadry, Dalia Y;Khorshed, Amira M;Rashed, Reham A;Mokhtar, Nadia M
Asian Pacific Journal of Cancer Prevention
/
v.17
no.4
/
pp.1705-1712
/
2016
Background: The aim of this study was to determine and evaluate the association of different viral infections, with hepatitis B and C viruses, Epstein-Barr virus, cytomegalovirus and human herpes virus-8 (HBV, HCV, EBV, CMV, HHV-8) with the risk of lymphomas (Hodgkin and non-Hodgkin) among Egyptian patients, and correlate with the histopathological staging and typing as well as the prevalence of combined infections. Materials and Methods: A total of 100 newly diagnosed lymphoma patients with 100 healthy age and sex matched normal controls were assayed for viral infection using enzyme linked immunosorbant assay (ELISA) followed by real time polymerase chain reaction (RT-PCR). Results: Our results showed a high statistical significant difference between cases and controls as regards clinical and laboratory findings (P<0.001 and=0.003). A high statistical difference was seen for the association of most viruses and lymphoma cases (p<0.001) except for positive HBs Ag, positive CMV IgG and HHV-8 (p=0.37, 0.70 and 1.0 respectively). No statistical significant difference was found between Hodgkin (HL) and non-Hodgkin (NHL) as regards viral prevalence except HCV antigen, 57.1% for HL and 26.5% for NHL (p = 0.03). Only, HBV DNA showed a high significant value among infiltrated bone marrow cases (p=0.003) and finally, a high significant association of 2 combined viral infections with infiltrated bone marrow lymphoma cases (p=0.04). Conclusions: Our results showed that infection with HBV, HCV, CMV and EBV were associated with increased risk of lymphoma among the Egyptian population. Detection of new associations between infectious agents and risk of cancer development will facilitate progress in elaboration of prophylactic measures, early diagnostic methods and, hopefully, novel therapy of malignant tumours.
Due to the great deal of effort that has gone into the study of osteoclastic differentiation and activation over the last few decades, the mechanisms of these two events have been discovered gradually. Nitric oxide($NO^-$), which is produced from arginine by a nitric oxide synthase, opened up a new area of biological research. Recently, it has been reported that $NO^-$ is produced by osteoblasts stimulated by lipopolysaccharide and several other cytokines. In this study, the effect of sodium nitroprusside(SNP), a donor of nitric oxide($NO^-$), on osteoclast-like cell formation and on mature osteoclast function was examined. To determine the mechanism of the inhibitory effects of SNP decreased not only the basal $^{45}Ca$ release but also thee bone resorption induced by PTH and 1,25-dihydroxyvitamin $D_3\;(1,25[OH]_{2}D_3)$. The inhibitory effect of SNP on bone resorption induced by PTH appeared 2 dyas after treatment, whereas SNP effect on inhibiting bone resorption induced by $1,25[OH]_{2}D_3$ appeared at the thhird days. When chicken and rat osteeoclasts were cultured on dentin slices, treatment of $300{\mu}M$ SNP resulted in a significant decrease in dentin resorption by osteoclasts in terms of total resolution area and average individual area. We also examind the effect of SNP on formation of osteoclast-like cells that is TRAP-positive multinucleated cells from chicken and rat bone marrow cells in the presence or absence of $10^{-8}\;M\;1,25[OH]_{2}D_3$. The addition of $300{\mu}M$ SNP inhibiteed the formation of TRAP-positive multinucleated cells. The present data suggest that SNP, possibly as a $NO^-$ donor, inhibits the osteoclastic differentiation and osteoclastic activity.
The endocrinology of type 2 diabetes (T2D) and its predisposing factors have been studied extensively while its skeletal effects have received negligible research despite this being a global disease. The cellular and molecular association between proximal humeral fractures and T2D has not been fully elucidated. We aimed to study bone cell quantities and immunolabel osteogenic and antiosteogenic cytokines. The study used 12-week-old rats (23 males) consisting of 8 Sprague Dawley (SD) and 15 Zucker Diabetic Sprague Dawley (ZDSD). Weekly mass measurements were taken while fasting blood glucose levels were recorded every 2 weeks with oral glucose tolerance tests conducted once every 4 weeks. Upon termination at the age of 28 weeks, humeri were fixed in 10% buffered formalin, prior to decalcification in ethylenediaminetetraacetic acid. The bone samples were then processed in ascending grades of alcohol using an automatic processor before embedding in paraffin wax. Sections were cut at 5 ㎛ thickness in a series for Haematoxylin and Eosin stain, and immunohistochemistry was performed with the anti-tartrate-resistant acid phosphatase (TRAP), anti-alkaline phosphatase (ALP), anti-bone morphogenetic protein 3 (BMP3), anti-transforming growth factor beta 1 (TGFβ1), anti-aged glycation end product (AGE) antibodies in the sequence. ZDSD rats had more adipocytes, BMP3 and AGEs expression with higher numbers of TRAP positive osteocytes and fewer ALP cells although no differences were found in TGFβ1 immunopositivity. We also found that T2D increases the number of AGEs immuno-positive cells, as well as its extracellular expression, thus providing a conducive environment for the interaction of the osteogenic cytokine and its antagonist to suppress osteoblastogenesis. ZDSD groups had higher adipocyte numbers therefore increased marrow adiposity in T2D.
Journal of the Korean Society of Food Science and Nutrition
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v.39
no.3
/
pp.363-368
/
2010
Artemisia princeps has been utilized as a traditional medicine for a variety of diseases in Korea. In this study, we investigated the effects of Artemisia princeps extract (APE) on bone metabolism both in vitro using primary mouse bone marrow-derived macrophage and in vivo using ovariectomized rats. APE decreased the number of tartrate-resistant acid phosphatase (TRAP)-positive multinucleated cells and TRAP activity. Also, APE inhibited bone resorptive activity of differentiated osteoclasts. In ovariectomized rats, APE alleviated the decrease in the trabecular bone mineral density. These results showed that APE might be useful for the prevention of postmenopausal bone loss.
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