• Journal of Periodontal and Implant Science
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• v.38 no.sup2
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• pp.273-298
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• 2008

Purpose: The aim of this review is to introduce a novel bone-graft material for hard-tissue regeneration based on the calcium phosphate glass(CPG). Materials and Methods: CPG was synthesized by melting and subsequent quenching process in the system of CaO-$CaF_2-P_2O_5$-MgO-ZnO having a much lower Ca/P ratio than that of conventional calcium phosphates such as HA or TCP. The biodegradability and bioactivity were performed. Effects on the proliferation, calcification and mineralization of osteoblast-like cells were examined in vitro. Influence in new bone and cementum formations was investigated in vivo using calvarial defects of Sprague-Dawley rats as well as 1-wall intrabony defect of beagle dogs. The application to the tissue-engineered macroporous scaffold and in vitro and in vivo tests was explored. Results: The extent of dissolution decreased with increasing Ca/P ratio. Exposure to either simulated body fluid or fetal bovine serum caused precipitation on the surface. The calcification and mineralization of osteoblast-like cells were enhanced by CPG. CPG promoted new bone and cementum formation in the calvarial defect of Sprague-Dawley rats after 8 weeks. The macroporous scaffolds can be fabricated with $500{\sim}800{\mu}m$ of pore size and a three-dimensionally interconnected open pore system. The stem cells were seeded continuously proliferated in CPG scaffold. Extracellular matrix and the osteocalcin were observed at the $2^{nd}$ days and $4^{th}$ week. A significant difference in new bone and cementum formations was observed in vivo (p<0.05). Conclusion: The novel calcium phosphate glass may play an integral role as potential biomaterial for regeneration of new bone and cementum.

• Journal of Periodontal and Implant Science
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• v.35 no.2
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• pp.277-288
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• 2005

There is a potential role of collagenase-3 in alveolar bone loss and periodontal disease progression, we need to develope or find chemotherapeutic drugs or herbal agents which may regulate the expression of MMP-13. Ginseng saponin, one of the major components of Korea ginseng(panax ginseng) root, has many various biologic effects, such as cytotoxic effect, tumoricidal effects, cytokine regulations, and protein biosynthesis effect. The purpose of this study was to determine the effects of Korea red ginseng saponin on MMP-13 gene expression in osteoblasts. The experimental groups were cultured with ginseng saponin in concentration of 1.0, 10, 25, 50, 100, 250 and $500{\mu}g/ml$ for MTT assay. Primary rat calvarial cells were pre-treated for 1 hour with ginseng saponin(100 ${\mu}g/ml$) and then stimulated with $IL-1{\beta}(1.0ng/ml)$ and PTH(10 nM). MMP-13 gene expression was evaluated by RT-PCR. The results were as follows: Ginseng saponin was cytotoxic to osteoblast at concentration exceeding $250{\mu}g/ml$ for longer than 24 hours in tissue culture(p<0.01). In RT-PCR analysis, steady state MMP-13 mRNA levels were increased approximately 350% by $IL-1{\beta}$, and 400% by PTH when normalized to untreated control. $IL-1{\beta}-indued$ MMP-13 mRNA expression was reduced 50% by pretreatment with ginseng saponin. But ginseng saponin didn't inhibit MMP-13 expression from PTH stimulated cells. This results suggest that ginseng saponin Inhibit $IL-1{\beta}-indued$ MMP-13 mRNA expression.

• Journal of Periodontal and Implant Science
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• v.28 no.2
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• pp.281-291
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• 1998

Chitosan is a biodegradable and non-toxic material with a molecular weight of 800-1,500Kd which can be obtained in various forms with extraordinary chemical structures and biological characteristics of which enables it to be used in many fields as a biomaterial. Ion irradiation is a useful tool to modify chemical structures and physical properties of high molecular weight polymers. The basic hypothesis of this study is that when surface properties of chitosan in a sponge form are modified with ion beam-irradiation and cell adhesion properties of chitosan would improve and thereby increase the regenerative ability of the damaged bone. The purpose of this study was to illuminate the changes in the cytocompatibility of chitosan sponges after ion beam-irradiation as a preliminary research. Argon($Ar^+$) ions were irradiated at doses of $5{\times}10^{13}$, $5{\times}10^{15}$ at 35 keV on surfaces of each sponges. Cell adhesion and activity of alkaline phosphatases were studied using rat fetal osteoblasts. The results of this study show hat ion beam-irradiation at optimal doses($5{\times}10^^{13}\;Ar^+\;ion/cm^2$) is a useful method to improve cytocompatibility without sacrificing cell viability and any changing cell phenotypes. These results show that ion beam-irradiated chitosan sponges can be further applied as carriers in tissue engineering and as bone filling materials.

• Journal of Periodontal and Implant Science
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• v.47 no.5
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• pp.273-291
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• 2017

Purpose: Although static magnetic fields (SMFs) have been used in dental prostheses and osseointegrated implants, their biological effects on osteoblastic and cementoblastic differentiation in cells involved in periodontal regeneration remain unknown. This study was undertaken to investigate the effects of SMFs (15 mT) on the osteoblastic and cementoblastic differentiation of human osteoblasts, periodontal ligament cells (PDLCs), and cementoblasts, and to explore the possible mechanisms underlying these effects. Methods: Differentiation was evaluated by measuring alkaline phosphatase (ALP) activity, mineralized nodule formation based on Alizarin red staining, calcium content, and the expression of marker mRNAs assessed by reverse transcription polymerase chain reaction (RT-PCR). Signaling pathways were analyzed by western blotting and immunocytochemistry. Results: The activities of the early marker ALP and the late markers matrix mineralization and calcium content, as well as osteoblast- and cementoblast-specific gene expression in osteoblasts, PDLCs, and cementoblasts were enhanced. SMFs upregulated the expression of Wnt proteins, and increased the phosphorylation of glycogen synthase $kinase-3{\beta}$ ($GSK-3{\beta}$) and total ${\beta}-catenin$ protein expression. Furthermore, p38 and c-Jun N-terminal kinase (JNK) mitogen-activated protein kinase (MAPK), and nuclear $factor-{\kappa}B$ ($NF-{\kappa}B$) pathways were activated. Conclusions: SMF treatment enhanced osteoblastic and/or cementoblastic differentiation in osteoblasts, cementoblasts, and PDLCs. These findings provide a molecular basis for the beneficial osteogenic and/or cementogenic effect of SMFs, which could have potential in stimulating bone or cementum formation during bone regeneration and in patients with periodontal disease.

• Journal of Periodontal and Implant Science
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• v.36 no.2
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• pp.335-344
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• 2006

Osteoblast or bone marrow stromal cell-derived RANKL is the major effector molecule essential for osteoclastogenesis. Previous studies have shown that tetracyclines have beneficial therapeutic effects in the prevention and treatment of inflammatory bone disease including periodontal disease. Periodontal ligament cells are thought not only to play an important role in the progression of periodontal disease, but to play an important role in alveolar bone remodeling. Previous studies indicated that receptor activation of nuclear factor $\kappa\;B$ ligand (RANKL) and osteoprotegerin (OPG) are expressed in periodontal ligament cells by pro-inflammatory cytokine, such as $IL-1{\beta}$ and $TNF-{\alpha}$. This study was designed to investigate the inhibitory effect of doxycycline on RANKL and OPG mRNA in rat periodontal ligament cells induced by $IL-1{\beta}$ (1 ng/ml). The results are as follows; 1. MTT assay showed that doxycycline at the concentration of $1-50\;{\mu}g/m{\ell}$ didn't result in statistically significant cell death at day 1 and 3. 2. RANKL mRNA expression was increased to 2.6 folds by $IL-1{\beta}$. When cells were treated with doxycycline ($50{\mu}g/m{\ell}$), $IL-1{\beta}$ -induced mRANKL expression was reduced by 33%. In contrast to RANKL, OPG mRNA expression was not inhibited by pre-treatment with doxycycline. These results suggest that doxycycline decrease the expression of mRANKL resulting in regulation of osteoclastogenesisp in rat periodontal ligament cells.

• Journal of Periodontal and Implant Science
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• v.35 no.3
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• pp.747-760
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• 2005

Periodontal therapy has dealt primarily with attempts at arresting progression of disease. however, more recent techniques have focused on regenerating the periodontal ligament having the capacity to regenerate the periodontium. Recombinant human bone morphogenetic protein-7(rhBMP-7) can differentiate the osteoprogenitor cells and induce bone formation. The purpose of this study was to evaluate the effect of BMP-7 on rat periodontal ligament cells differentiation, in vitro. In the control group, cells was cultured with DMEM media. In the experimental groups, cells were cultured with rhBMP-7 in concentration of 10, 25, 50 and 100 ng/ml. Each group was characterized by examining alkaline phosphatase activity at 3 and 5 days of culture and the ability to produce mineralized nodules of rat calvarial cells at 14 days of culture. Synthesis of type I collagen(COL-I), osteocalcin(OCN), and bone sialoprotein(BSP) was evaluated by RT-PCR at 7 days of culture. Activation of Smad proteins and p38 MAP kinase was determined by western blot analysis of the cell lysates. Alkaline phosphatase activity was significantly increased in the concentration of BMP-7 50 ng/ml and 100 ng/ml compared to the control(p<0.05). The mineralized bone nodule formation was greater with addition of 50 ng/ml and 100 ng/ml BMP-7 than the control(p<0.01). In 7 days' culture, the expressions of COL-I, BSP, and OCN was increased by BMP-7 in concentration of 10 $ng/ml{\sim}100$ ng/ml. In western blot analysis, BMP-7 treated culture cells expressed Smad 1,5,8 in dose-dependent manner, whereas BMP-7 did not activate phosphorylated form of p38 MAP kinase. These result suggested that BMP-7 stimulate rat periodontal ligament cells to differentiate toward osteoblast phenotype and increase bone matrix production by activation of BMP-Smad pathway.

• Journal of Periodontal and Implant Science
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• v.31 no.1
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• pp.1-23
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• 2001

Chitosan is biodegradable natural polymer that has been demonstrated its ability to improve wound healing, and calcium metaphosphate(CMP) is a unique class of phosphate minerals having a polymeric structure. In this study, chitosan/CMP and platelet derived growth factor(PDGF-BB) loaded chitosan/CMP sponges were developed, and the effect of the sponges on bone regeneration and their possibility as scaffolds for bone formation by three-dimensional osteoblast culture were examined. PDGF-BB loaded chitosan/CMP sponges were prepared by freeze-drying of a mixture of chitosan solution and CMP powder, and soaking in a PDGF-BB solution. Fabricated sponge retained its 3-dimensional porous structure with $100-200\;{\mu}m$ pores. The release kinetics of PDGF-BB loaded onto the sponge were measured in vitro with $^{125}I-labeled$ PDGF-BB. In order to examine their possibility as scaffolds for bone formation, fetal rat calvarial osteoblastic cells were isolated, cultured, and seeded into the sponges. The cell-sponge constructs were cultured for 28 days. Cell proliferation, alkaline phosphatase activity were measured at 1, 7, 14 and 28 days, and histologic examination was performed. In order to examine the effect on the healing of bone defect, the sponges were implanted into rat calvarial defects. Rats were sacrificed 2 and 4 weeks after implantation and histologic and histomorphometrical examination were performed. An effective therapeutic concentration of PDGF-BB following a high initial burst release was maintained throughout the examination period. PDGF-BB loaded chitosan/CMP sponges supported the proliferation of seeded osteoblastic cells as well as their differentiation as indicated by high alkaline phosphatase activities. Histologic findings indicated that seeded osteoblastic cells well attached to sponge matrices and proliferated in a multi-layer fashion. In the experiments of implantation in rat calvarial defects, histologic and histomorphometric examination revealed that chitosan/CMP sponge promoted osseous healing as compared to controls. PDGF-BB loaded chitosan/CMP sponge further echanced bone regeneration. These results suggested that PDGF-BB loaded chitosan/CMP sponge was a feasable scaffolding material to grow osteoblast in a three-dimentional structure for transplantation into a site for bone regeneration.

• Journal of Periodontal and Implant Science
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• v.33 no.3
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• pp.457-474
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• 2003

The ultimate objective of periodontal treatment is to get rid of an on-going periodontal disease and further regenerate the supporting tissue, which is already destroyed, functionally. Currently, the bone grafting operation using various kinds of bone grafting materials and the operation for induced regeneration of periodontal tissue using the blocking membrane are performed for regeneration of the destroyed periodontal tissue. However, there are respective limitations Galenical preparations, which are used for regeneration of periodontal of tissue, has less risk of rejective reaction or toxicity that may be incidental to degradation and their effect is sustainable. Thus, in case they are applicable to a clinic, they can he used economically. Chitosan has such compatibility, biological actions including antibacterial activity, acceleration of wound treatment, etc., and excellent mechanical characteristics, which has recently aroused more interest in it. Also, it has been reported that it promotes osteogenesis directly or indirectly by functioning as a matrix to promote migration and differentiation of a specific precussor cell (for example, osteoblast) and further inhibiting the function of such a cell as fibroblast to prevent osteogenesis. In this study, the pure chitosan solution, which was obtained by purifying chitosan, was used. However, since this chitosan is of a liquiform, it is difficult to sustain it in a defective region. It is, therefore, essential to use a carrier for delivering chitosan to, and sustaining it gradually in the defective region. In the calvarial defect model of the Sprague-Dawley rat, it is relatively easy to maintain a space. Therefore, in this study, the chitosan solution with which ACS was wetted was grafted onto the defective region, For an experimental model, a calvarial defect of rat m s selected, and a critical size of the defective region was a circular defect with a diameter of 8 mm. A group in which no treatment was conducted for the calvarial defect was set as a negative control group. Another group in which treatment was conducted with ACS only was set as a positive control group (ACS group). And another group in which treatment was conducted was conducted with by grafting the pure chitosan solution onto the defective region through ACS which was wetted with the chitosan solution was set an experimental group (Chitosan/ACS group). Chitosan was applied to the Sprague-Dawley rat's calvarial bone by applying ACS which was wetted with the chitosan solution, and each Sprague-Dawley rat was sacrificed respectively 2 weeks and 8 weeks after the operation for such application. Then, the treatment results were compared and observed histologically and his tometrically. Thereby, the following conclusions were obtained. 1. In the experimental group, a pattern was shown that from 2 weeks after the operation, vascular proliferation proceeded and osteogenesis proceeded through osteoblast infiltration, and at 8 week after the operation, ACS was almost absorbed, the amount of osteogensis was increased and many osteoid tissue layers were observed. 2. At 2 weeks after the operation, each amount of osteogenesis appeared to be 8.70.8 %, 13.62.3 % and 4.80.7 % respectively in the experimental group, the positive control group and the negative control group. Accordingly, it appeared to be higher in the Experimental group and the positive control group than in the negative control group, but there was no significant difference statistically (p<0.01). 3. At 8 weeks after the operation, each amount of osteogenesis appeared to be 62.26.1%, 17.42.5 % and 8.21.4 % respectively in the experimental group, the positive control group and the negative control group. Accordingly, it appeared to be substantially higher in the experimental group than in the positive control group and the negative control group, and there was a significant difference statistically (p<0.01). As a result of conducting the experiment, when ACS was used as a carrier for chitosan, chitosan showed effective osteogenesis in the perforated defective region of the Sprague-Dawley rat's calvarial bone.

• The Journal of Korean Academy of Prosthodontics
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• v.46 no.6
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• pp.620-627
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• 2008

STATEMENT OF PROBLEM: A biochemical approach for surface modification has offered an alternative for physicochemical and morphological methods to obtain desirable bone-implant interfaces. PURPOSE: The purpose of the present study was to investigate cell responses to poly (D,L-lactide-co-glycolide) (PLGA)/$1{\alpha}$,25-(OH)$_2D_3$ coating with reference to cellular proliferation and differentiation in vitro. MATERIAL AND METHODS: 96 titanium discs were fabricated and divided into four groups. Group 1 was anodized under 300 V as control. Group 2, 3 and 4 were anodized then coated with 3 ml PLGA/$1{\alpha}$,25-(OH)$_2D_3$ solutions. Amount of the solutions were 2 ul, 20 ul and 200ul respectively. The osteoblast-like Human Osteogenic Sarcoma (HOS) cells were seeded and cultured for 1, 3 and 7 days. MTSbased cell proliferation assay and ALPase activity test were carried out. RESULTS: PLGA nanoparticles were observed as fine, smooth and round and HOS cells attached to the anodized surfaces through strand-like and sheet-like filopodia. After 3 days of culture, the dendritic filopodia were exaggerated and sheet-like cytoplasmic projections covered the coated titanium surfaces. After 3 days of culture, all of the groups showed increased cellular proliferation and the lowest proliferation rate was measured on group 2. Higher amount of incorporated $1{\alpha}$,25-(OH)$_2D_3$ (Group 3 and 4) improved cellular proliferation but the differences were not significant statistically (P > .05). But they increased the rate of ALP activities than the control group at day 3 (P < .05). CONCLUSION: Biodegradable PLGA nanoparticles incorporated with vitamin D metabolite positively affected proliferation and differentiation of cells on the anodized titanium surface.

• Journal of Periodontal and Implant Science
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• v.34 no.3
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• pp.475-487
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• 2004

Purpose: This study was aimed to evaluate the effect of the deproteinated bovine bone powder (DBBP) coated with calcium phosphate (Ca-P) on osseous regeneration in the calvarial bone defect of rat. Materials and Methods : The DBBP (Control group, n=6) and the Ca-P coated DBBP (Experimental group, n=6) were grafted in the critical sized calvarial bone defect (8 mm) of rat weighing 250 g. The animals were sacrificed at 1, 4 week. The biopsy specimens were decalcified with 5%formaldehyde and embedded in paraffin. The rats were sacrificed at 8 week received tetracycline (1 week), calcein blue (4 week), and alizarin red (7 week), and the biopsy specimens were taken. The specimens were embedded in methylmethacrylate and ground to 10 ${\mu}m$ thin sections were made. All of the specimens were stained with H & E and Masson's trichrome and examined under light microscope. The specimens at 8 week were examined under fluorescent microscope. Results : In the Control group, the grafted DBBP was surrounded with connective tissue, and osteoblasts were observed partially around the grafted particles at 1 week. At 4 week, some osteoid was observed and, new bone formation was observed at the periphery of grafted materials at 8 week, In the Experimental group, some osteoid was seen at the periphery of the grafted Ca-P coated DBBP at 1 week, and osteoblast and newly formed bone were observed around the grafted materials. At 8 week, newly formed bone was observed at the periphery of the grafted materials. Conclusion: These results suggest that Ca-P coated DBBP group was more and faster than DBBP group in new bone formation and Ca-P could contribute to enhance bone formation in the critical sized calvarial bone defect of rat.