The purpose of this study was to evaluate the correlation between nicotine and the activity of bone forming cell. MG63 osteoblast-like cells were used for this study. Several factors were examined including the proliferation of cell, alkaline phosphatase activity, the formation of osteocalcin and osteoprotegerin. and the synthesis of its mRNA. MG63 osteoblast-like cells were incubated for 1, 2, 3 and 6 days with nicotine added to the culture medium in 1.0 ${\mu}M$, 1.0mM, 2.5mM, 5.0mM, 7.5mM, and 10.0mM concentrations. The proliferation of MG63 osteoblast-like cells was temporarily activated at the low nicotine concentrations. At high concentrations (>5.0 mM), however. it was suppressed. Alkaline phosphatase activity was suppressed in a dose-dependent manner as the concentration of nicotine increased. Osteocalcin decreased in a dose-dependent manner at high nicotine concentrations of more than 7.5mM and the same result was show when the osteoblasts were treated with low concentrations for longer than 3 days. There was a difference in the influence of nicotine on the synthesis of osteocalcin mRNA and formation of osteocalcin itself at 1 and 3 days. Generally, osteoprotegrin notably declined in all experimental groups. However, the level of its mRNA increased at high nicotine concentrations of more than 7.5mM after 3 days and more than 5.0mM after 6days.
Journal of the Korean Association of Oral and Maxillofacial Surgeons
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v.37
no.5
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pp.415-420
/
2011
Purpose: Calcium phosphate cement (CPC) is one of many useful materials for restoring tooth defects, periodontium and maxillofacial area. Chitosan is a biodegradable material that has been shown to promote the growth and differentiation of osteoblasts in culture. This study examined the interaction between odontoblasts and bio-calcium phosphate cement reinforced with chitosan. Materials and Methods: $5{\times}10^3$ odontoblastic cells were seeded into each well. Various concentrations of bio-calcium phosphate cement reinforced with chitosan (10, 20, 50, 100, 200, 500 ${\mu}g$/ml, 1, 2, 4 mg/ml) were diluted and added to the wells. The well was incubated for 24 h, 48 h and 72 h. After incubation, the number of cells was assessed to determine the cell viability. A cytokinesis-block micronucleus assay and chromosomal aberration test were carried out to estimate the extent of chromosomal abnormalities. Microscopic photographs and RT-PCR were performed to examine the adhesion potential of bio-calcium phosphate cement reinforced with chitosan. Results: Bio-CPC-reinforced chitosan did not show significant cytotoxicity. The number of damaged chromosomes in the cells treated with Bio-CPC-reinforced chitosan was similar to that in the control cells. There was no significant increase in the number of chromosomal aberrations in the Bio-CPC reinforced chitosan exposed cells. Microscopic photographs and RT-PCR confirmed the adhesive potential of bio-CPC reinforced chitosan to odontoblasts. Conclusion: Bio-CPC-reinforced chitosan did not affect the odontoblastic cell viability, and had no significant cytotoxic effect. Bio-CPC-reinforced chitosan showed adhesive potential to odontoblasts. These results are expected form the basis of future studies on the effectiveness of dental restorative materials in Bio-CPC reinforced with chitosan.
Journal of the Korean Association of Oral and Maxillofacial Surgeons
/
v.34
no.2
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pp.220-229
/
2008
Purpose: The present study was aimed to examine the effect of acellular dermal matrix ($AlloDerm^{(R)}$) grafted to the experimental tissue defect on tissue regeneration. Materials and Methods: Male albino rabbits were used. Soft tissue defects were prepared in the external abdominal oblique muscle. The animals were then divided into 3 groups by the graft material used: no graft, autogenous dermis graft, and $AlloDerm^{(R)}$ graft. The healing sites were histologically examined at weeks 4 and 8 after the graft. In another series, critical sized defects with 8-mm diameter were prepared in the right and left iliac bones. The animals were then divided into 5 groups: no graft, grafted with autogenous iliac bone, $AlloDerm^{(R)}$ graft, $AlloDerm^{(R)}$ graft impregnated with rhBMP-2, and $AlloDerm^{(R)}$ graft with rhTGF-${\beta}1$. The healing sites of bone defect were investigated with radiologic densitometry and histological evaluation at weeks 4 and 8 after the graft. Results: In the soft tissue defect, normal healing was seen in the group of no graft. Inflammatory cells and foreign body reactions were observed in the group of autogenous dermis graft, and the migration of fibroblasts and the formation of vessels into the collagen fibers were observed in the group of $AlloDerm^{(R)}$ graft. In the bone defect, the site of bone defect was healed by fibrous tissues in the group of no graft. The marked radiopacity and good regeneration were seen in the group of autogenous bone graft. There remained the traces of $AlloDerm^{(R)}$ with no satisfactory results in the group of $AlloDerm^{(R)}$ graft. In the groups of the $AlloDerm^{(R)}$ graft with rhBMP-2 or rhTGF-${\beta}1$, there were numerous osteoblasts in the boundary of the adjacent bone which was closely approximated to the $AlloDerm^{(R)}$ with regeneration features. However, the fibrous capsule also remained as in the group of $AlloDerm^{(R)}$ graft, which separated the $AlloDerm^{(R)}$ and the adjacent bone. Conclusions: These results suggest that $AlloDerm^{(R)}$ can be useful to substitute the autogenous dermis in the soft tissue defect. However, it may not be useful as a bone graft material or a carrier, since the bone defect was not completely healed by the bony tissue, regardless of the presence of osteogenic factors like rhBMP-2 or rhTGF-${\beta}1$.
Kim Soon Hee;Yun Sun Jung;Jang Ji Wook;Kim Moon Suk;Khang Gilson;Lee Hai Bang
Polymer(Korea)
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v.30
no.1
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pp.14-21
/
2006
Tissue engineering techniques require the use of a porous biodegradable/bioresorbable scaffold, which server as a three-dimensional template for initial cell attachment and subsequent tissue formation in both in vitro and in vivo. Small intestinal submucosa (SIS) has been investigated as a source of collagenous tissue with the potential to be used as biomaterials because of its inherent strength and biocompatibility. SIS-loaded poly(L-lactide-co-glicolide)(PLGA) scaffolds were prepared by solvent casting/particle leaching. Characterizations of SIS/PLGA scaffold were carried out by SEM, mercury porosimeter, and so on. Muscle-derived stem cells can be differentiated in culture into osteoblasts, chondrocytes, and even myoblasts by the controlling the culture environment. Cellular viability and proliferation were assayed by 3-(4,5-dimethylthiazole-2-yl)-2,5-diphenyltetrazolium-bromide(MTT) test. Osteogenic differential cells were analyzed by alkaline phosphatase(ALP) activity. SIS/PLGA scaffolds were implanted into the back of athymic nude mouse to observe the effect of SIS on the osteoinduction compared with controlled PLGA scaffolds. Thin sections were cut from paraffin embedded tissues and histological sections were conducted hematoxylin and eosin (H&E), Trichrome, and von Kossa. We observed that bone formatioin of SIS/PLGA hybrid scaffold as natural/synthetic scaffold was better thean that of only PLGA scaffold. It canb be explained that SIS contains various kinds of bioactive molecules for osteoinduction.
Yoon, Kang Hugh;Baek, Jong Min;Kim, Ju Young;Kwak, Seong Cheoul;Cheon, Yoon Hee;Jeon, Byung Hoon;Lee, Chang Hoon;Choi, Min Kyu;Oh, Jaemin;Lee, Myeung Su;Kim, Jeong Joong
Journal of Physiology & Pathology in Korean Medicine
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v.28
no.1
/
pp.29-34
/
2014
Bone homeostasis is maintained by balance between bone resorbing-osteoclasts and bone forming-osteoblasts. Excessive osteoclastic bone resorption plays a critical role in bone destruction in pathological bone diseases such as osteoporosis, rheumatoid arthritis, and periodontal disease. Many compounds derived from natural products have pharmacological applications and have therapeutic value for treating or preventing several bone diseases characterized by excessive bone resorption. To discover new compounds that can act as anti-resorptive agents, we screened for natural compounds that regulate osteclast differentiation, and found that water extract of Ziziphus Jujuba Mill (WEZJ) has inhibitory effects on osteoclast differentiation. In this study, WEZJ clearly inhibits the osteoclast differentiation in the presence of receptor activator of nuclear factor kB (RANKL), macrophage colony-stimulating factor (M-CSF) without cytoxicity by blocking activation of nuclear factor of activated T cells (NFAT)c1, and c-Fos. In signaling pathway, the phosphorylation of Akt, p38, c-Jun N-terminal kinases (JNK), extracellular signal-regulated kinases (ERK) and the expression of osteoclast-associated receptor (OSCAR), tartrate-resistant acid phosphates (TRAP), Integrin av, Integrin b3, Cathepsin K are suppressed, too. These result suggest that WEZJ may have therapeutic value for treating or preventing several bone diseases characterized by excessive bone destruction.
The periosteum contains multipotent cells that can differentiate into osteoblasts and chondrocytes. Cultured periosteum-derived cells (PDCs) have an osteogenic capacity. The purpose of this study was to evaluate the interaction of PDCs with bone graft biomaterial. After cell isolation from the calvarial periosteum of Sprague-Dawley rats, cultured PDCs were placed in critical-sized calvarial defects with beta-tricalcium phosphate (${\beta}$-TCP). All rats were sacrificed 8 weeks after bone graft surgery, and the bone regenerative ability of bone grafting sides was evaluated by plain radiography, micro-computed tomography (CT), and histological examination. PDCs grafted with ${\beta}$-TCP displayed enhanced calcification in the defect site, density of regenerated bone and new bone formation within the defect and its boundaries. Furthermore, these PDCs more efficiently regenerated new bone as compared to grafted ${\beta}$-TCP only. The results suggest that cultured PDCs have the potential to promote osteogenesis in bone defects.
Osteoporosis is a major bone disorder defined as having bone mineral density (BMD) of 2.5 standard deviations or more below the peak bone mass. Osteoporosis will increasingly be a major disorder that faces the aging mankind. It is the result of an imbalance in the bone remodeling system, where bone constantly undergoes a cycle of resorption by osteoclasts and formation by osteoblasts. Estrogen deficiency in women following menopause is identified as the predominant reason that causes disparity in this system. Current medical treatments for osteoporosis include hormone replacement therapy (HRT), biphosphonates, and teriparatide, but have various side effects that raise questions concerning their medical safety and practicality. Alternative treatments involving natural product sources are under study to find a safer therapy. Many natural sources including lactoferrin and isoflavones and numerous traditional herbal medicines exhibit anti-resorptive or anabolic effects on bone and thus show promises to provide therapeutic agents in treating osteoporosis. Unfortunately, the majority of natural product treatments are still in its preliminary stages to prove their efficacy even though the development pace of treatment for osteoporosis is astounding in the past few decades. Further progress in pre-clinical studies and the subsequent clinical studies will someday lead to a breakthrough that takes us another step forward in science.
Human bone marrow mesenchymal stem cells (hBM-MSCs) can differentiate into various cell types including osteoblasts, adipocytes, chondrocytes, and myocytes. Previous studies, including our own, have shown that MSCs can also differentiate into neuron-like cells. However, their rate of neuronal differentiation is not sufficient for application to stem cell therapy, which requires well-defined cell types. For this purpose, we first examined the expression of neuronal lineage markers (GFAP, MAP-2, KCNH1, Nestin, NF-M, and Tuj-1) by real-time PCR, western blot, and immunocytochemical staining. The expressions of the astrocyte marker GFAP and neuronal markers NF-M and Tuj-1 increased in neuronal differentiated MSCs (dMSCs). To improve the neuronal differentiation efficiency, PDE4, an important signaling intermediator in the progression of neuronal differentiation, was modulated using well-known inhibitors such as rolipram or resveratrol and then differentiated into neuronal cells (Roli- or RSV-dMSCs). The expressions of NF-M, Tuj-1 were increased while that of GFAP decreased in Roli- and RSV-dMSCs, which were examined by real-time PCR, western blot, and immunocytochemical staining. From these experiments, we have found that the neuronal differentiation efficiency can be ameliorated by the modulation of PDE4 activity.
Bone morphogenetic proteins (BMPs) are multifunctional cytokines that play important roles in a variety of cellular functions. Among BMP family members, BMP2 efficiently promotes osteoblast differentiation through Smad-mediated runt-related transcription factor 2 (Runx2) expression. Several recent studies suggest that BMPs are associated with clock genes, in particular Bmal1. Bmal1 protein heterodimerizes with Clock protein and then induces period 1 (Per1) expression. However, the role of Per1 on osteoblast differentiation remains unclear. In this study, we investigated whether Per1 is involved in osteoblast differentiation. MC3T3-E1 cells were treated with BMP2 for induction of osteoblastic differentiation. Osteogenic maker gene and Per1 mRNA expression were measured using real-time PCR. Interestingly, BMP2 treatment induced Per1 mRNA expression in MC3T3-E1 cells. To further investigate the function of Per1 on osteoblast differentiation, MC3T3-E1 cells were transiently transfected with pCMV-Per1. Per1 overexpression increased Runx2 mRNA and protein levels. Also, mRNA expression and promoter activity of osteocalcin were upregulated by Per1 overexpression. To investigate the effect of interaction between Per1 and osteogenic condition, MC3T3-E1 cells were cultured in osteogenic medium containing ascorbic acid and ${\beta}$-glycerophosphate. Osteogenic medium-induced ALP staining level and mineralization were synergistically increased by overexpression of Per1. Taken together, these results demonstrate that Per1 is a positive regulator of osteoblast differentiation.
Kim, Byung-Min;Sung, Iel-Yong;Park, Bong-Soo;Shin, Sang-Hoon;Kim, Jong-Ryoul
Journal of the Korean Association of Oral and Maxillofacial Surgeons
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v.29
no.6
/
pp.397-406
/
2003
The purpose of this study was to observe the titanium implant osseointegration in the osteoporosis-induced animal model. Seventy rats, 11 weeks of age, were divided into two groups : an ovariectomized group and a control group. Titanium screw implants(diameter, 2.0mm; length, 3.5mm) were placed into left tibias of 70 rats, 35 in the control group and 35 in the experimental group. The rats were sacrificed at different time interval (1, 2, 3, 4, 6, 8, and 12 weeks after implantation) for histopathologic observation, histomorphometric analysis and immunohistochemistry with fibronectin and CD34 antibody. The results obtained from this study were as follows: 1. Histopathologically findings, newly formed bone was seen at 3 weeks and became lamellar bone at 8 weeks, and mature trabecullar bone was seen at 12 weeks control group. In experimental group, thickness of regenerated bone increased till 8 weeks gradually and mature trabecullar bone was seen at 12 weeks. 2. By histomorphometric analysis, marrow bone density and contact ratio of marrow bone to implant decreased significantly from 8 to 12 weeks in experimental group compared to control group and also total bone to implant contact ratio decreased significantly from 8 to 12 weeks in experimental group. 3. Fibronectin immunoreactivity was strong at 3 weeks control group and reduced after 8weeks gradually. But it was continuously strong from 3 to 8 weeks in experimental group. 4. CD34 immunoreactivity was very strong in the newly formed osteoblasts from 3 to 8 weeks control group. But it reacted minimally later. While in experimental group, it reacted continuously strong from 3 to 12 weeks. The results of this study suggest that osteoporosis is not an absolute contraindication to dental implantation if sufficient period suggested after fixture installation till second stage surgery.
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