• Journal of Periodontal and Implant Science
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• 제41권5호
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• pp.227-233
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• 2011

Purpose: The aim of this study was to compare osteoblast behavior on zirconia and titanium under conditions cultured with bone morphogenetic protein-2. Methods: MC3T3-E1 cells were cultured on sandblasted zirconia and sandblasted/etched titanium discs. At 24 hours after seeding MC3T3-E1, the demineralized bone matrix (DBM) gel alone and the DBM gel with bone morphogenetic protein-2 (BMP-2) were added to the culture medium. The surface topography was examined by confocal laser scanning microscopy. Cellular proliferation was measured at 1, 4, and 7 days after gel loading. Alkaline phosphatase activity was measured at 7 days after gel loading. The mRNA expression of ALPase, bone sialoprotein, type I collagen, runt-related transcription factor 2 (Runx-2), osteocalcin, and osterix were evaluated by real-time polymerase chain reaction at 4 days and 7 days. Results: At 1, 4, and 7 days after loading the DBM gel alone and the DBM gel with BMP-2, cellular proliferation on the zirconia and titanium discs was similar and that of the groups cultured with the DBM gel alone and the DBM gel with BMP-2 was not significantly different, except for titanium with BMP-2 gel. ALPase activity was higher in the cells cultured with BMP-2 than in the other groups, but there was no difference between the zirconia and titanium. In ALPase, bone sialoprotein, osteocalcin, Runx-2 and osterix gene expression, that of cells on zirconia or titanium with BMP-2 gel was much more highly increased than titanium without gel at day 7. The gene expression level of cells cultured on zirconia with BMP-2 was higher than that on titanium with BMP-2 at day 7. Conclusions: The data in this study demonstrate that the osteoblastic cell attachment and proliferation of zirconia were comparable to those of titanium. With the stimulation of BMP-2, zirconia has a more pronounced effect on the proliferation and differentiation of the osteoblastic cells compared with titanium.

• Journal of Periodontal and Implant Science
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• 제35권2호
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• pp.511-524
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• 2005

The regeneration of lost periodontal tissue is a major goal of therapy. Periodontal ligament cell(PDL) is a specialized connective tissue that connects cementum and alveolar bone to maintain and support teeth in situ and preserve tissue homoeostasis. Bone morphogenetic proteins(BMPs) have shown much potential in the reconstruction of the periodontum by stimulate new bone and new cementum formation. Limitiations of BMP administration to periodontal lesions is high dose delivery, BMP transient biological activity, and low bioavailability of factors at the wound site. Gene delivery method can be alternative treatment strategy to deliver BMPs to periodontal tissue. The purpose of this study is to investigate efficiency of BMP-2 gene delivery with cell-based therapy using PDL cells. PDL cell were transduced with adenoviruses encoding either BMP-2 or Lac-Z gene. To evaluate osteogenic activity of expressed BMP-2 on PDL cells, we investigated secreted BMP-2, cellular activity, ALPase, produced mineralized nodules. To evaluate collagen scaffold as carrier for transduced cell delivery, we examined morphology and secreted BMP-2 of transducd PDL cells on it. BMP-2 transducd PDL cells produced higher levels of BMP-2, ALPase, mineralized nodules than non transduced cells. Cellular activity of transduced cells was showed similar activity to non transduced cells. Transduce cells attached on collagen scaffold secreted BMP-2 at 7day and was showed similar morphology to non transduced cells. These results demonstrated that transduced PDL cells produced biologically active BMP-2 and collagen scaffold could be carrier of transducd cells.

• Journal of Periodontal and Implant Science
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• 제27권4호
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• pp.867-882
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• 1997

Safflower(Carthamus tinctorius $Linn\acute{e}$ has been traditionally used for the treatment of blood stasis, and Dipsasi Radix has been used as a drug for fracture in Chinese medicine. The purpose of present study was to examine the biologic effects of safflower extract and Disasi radix extracts on the periodontal. ligament cells and osteoblastic cells and on the wound healing of rat calvarial defect. The ethanolic extract of safflower blossom, safflower seed and Dipsasi Radix(125, 250, and 500 ${\mu}g/ml$) were prepared as test group, and PDGF-BB(lOng/ml) and unsafonifiable fraction of Zea Mays L.(125, 250, and 500 ${\mu}g/ml$) were employed as positive control. The effects of each agents on the growth and survival, ALPase activity, expression of PDGF-BB receptor, chemotactic response of PDL cell and ATCC human osteosarcoma MG63 cells in vitro were examined. The tissue regenerative effect of each extracts was evaluated by histomorphometric measuring of newly formed bone on the 8mm defect in rat calvaria after oral administration of 3 different dosages groups : 0.02, 0.1 and 0.35g/kg, per day. It was also employed the same dosages of unsaponifiable fraction of Zea Mays L. as positive controls. Safflower blossom extract, safflower seed extract, and Dipsasi Radix extract stimulate the cellular activity of MG63 cells in concentration range of $125-500{\mu}g/ml$, and safflower bolssom extract and safflower seed extract stimulate also the cellular activity of periodontal ligament cells in concentration range of $250-500{\mu}g/ml$. In activity of ALPase, $250-500{\mu}g/ml$ of safflower blossom extracts showed significant stimulating effects on MG63 cells, and the same concentration range of safflower seed extracts showed significant effect on periodontal ligament cells. In the recovery on PDGF-BB receptor expression which was depressed by $IL-1{\beta}$, $125-250{\mu}g/ml$ of safflower blossom extracts and $250-500{\mu}g/ml$ of safflower seed extracts showed significant increasing effect on MG63 cells, and $500{\mu}g/ml$ of safflower blossom extract and $250-500{\mu}g/ml$ of safflower seed extracts showed significant effect on periodontal ligament cells. In chemotactic response, among all tested group, safflower seed extracts only were chemotactic to MG63 cells and periodontal ligament cells in concentration range of $125-500{\mu}g/ml$. Also in the view of bone regeneration in rat calvarial defect model, the only group that was orally administrated 0.35g/kg, day of safflower seed extract showed significant new bone formation. These results suggested that safflower extracts might have a potential possibilities as an useful drug for adjunct to treatment for regeneration of periodontal defect.

• Journal of Periodontal and Implant Science
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• 제29권1호
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• pp.173-192
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• 1999

The purpose of this study was to evaluate the effects of bioactive glass and natural coral on the human periodontal ligament fibroblast(HPLF) behaviors during the regeneration process of peridontium. To determine the cellular events occuring in the presence of the particles of bioactive glass and natural coral, HPLF were isolated from healthy premolar teeth extracted for orthodontic treatment. Cells were cultured in ${\alpha}$MEM at 37$^{\circ}C$, 5% $CO_2$, 95% humidity incubator. Bioactive glass and natural coral were powdered, and each particles(<40${\mu}$m) were placed on the cultured cells at the concentration of 0.3mg/ml, and 1,0mg/ml for experimental group. In control group no particles were added. And each group was evaluated by examining the cell morphology under phase-contrast micrograph at 4 day and transmission electron micrograph(TEM) and scanning electron micrograph(SEM) at 14 day, alkaline phosphatase activity at 5 and 9 day, protain synthesis at 4 day, DNA synthesis at 1, 2, 3 and 4 day, cell proliferation at 1, 3, 5,7 and 9 day and the formation of bone nodule at 30 day after culturing all groups in mineralizing supplemented mediun, No significant changes in cell morphology by adding these two matirials were found under phase contrast microscopy and TEM. HPLF phagocytocized each particles suggesting that HPLF is involved in the process of resorbing each particles and that bioactive glass were more biocompatible than natural coral. The ALPase activity of bioactive glass 0.3 mg/ml was similar with control groups and all the rests of control groups were significantly low(P<0.01) indicating a transient dedifferentiation of HPLF in the presence of bioactive glass and natural coral particles. There were no significant differences of protein synthesis between all groups. The DNA synthesis in experimental groups were significantly lower than control groups at 1, 2 and 3 day (P<0.01) but became similar to control groups at 4 day. Between control groups, the DNA synthesis in bioactive glass O.3mglml group was significantly higher than other groups(P<0.01). Cell proliferation in natural coral 1.0mg/ml and bioactive glass 1.0mglml groups were significantly lower than control group at 3 day(P<0.05) and there were no differences at 5, 7, 9 day. There were more bone nodule formation in experimental groups than in control groups. In conclusion, these results indicated that bioactive glass and natural coral have some effects of a transient dedifferentiation on HPLF and regeneration of periodontal tissues, however any significant cytotoxic effect on HPLF by these two particles were not found.

• Journal of Periodontal and Implant Science
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• 제40권3호
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• pp.105-110
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• 2010

Purpose: It has been reported that low-level semiconductor diode lasers could enhance the wound healing process. The periodontal ligament is crucial for maintaining the tooth and surrounding tissues in periodontal wound healing. While low-level semiconductor diode lasers have been used in low-level laser therapy, there have been few reports on their effects on periodontal ligament fibroblasts (PDLFs). We performed this study to investigate the biological effects of semiconductor diode lasers on human PDLFs. Methods: Human PDLFs were cultured and irradiated with a gallium-aluminum-arsenate (GaAlAs) semiconductor diode laser of which the wavelength was 810 nm. The power output was fixed at 500 mW in the continuous wave mode with various energy fluencies, which were 1.97, 3.94, and 5.91 $J/cm^2$. A culture of PDLFs without laser irradiation was regarded as a control. Then, cells were additionally incubated in 72 hours for MTS assay and an alkaline phosphatase (ALPase) activity test. At 48 hours post-laser irradiation, western blot analysis was performed to determine extracellular signal-regulated kinase (ERK) activity. ANOVA was used to assess the significance level of the differences among groups (P<0.05). Results: At all energy fluencies of laser irradiation, PDLFs proliferation gradually increased for 72 hours without any significant differences compared with the control over the entire period taken together. However, an increment of cell proliferation significantly greater than in the control occurred between 24 and 48 hours at laser irradiation settings of 1.97 and 3.94 $J/cm^2$ (P<0.05). The highest ALPase activity was found at 48 and 72 hours post-laser irradiation with 3.94 $J/cm^2$ energy fluency (P<0.05). The phosphorylated ERK level was more prominent at 3.94 $J/cm^2$ energy fluency than in the control. Conclusions: The present study demonstrated that the GaAlAs semiconductor diode laser promoted proliferation and differentiation of human PDLFs.

• 한국섬유공학회:학술대회논문집
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• 한국섬유공학회 2003년도 The Korea-Japan Joint Symposium
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• pp.41-42
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• 2003

Osteoblasts (MC3T3-E1) were cultured on polysaccharide type polyelectrolyte complex (PEC). The growth of the MC3T3-E1 on the PEC with carbxyl group (c-type) was slightly suppressed and exhibited aggregation morphology. On the other hand, cell growth on the PEC with sulfate group (s-type) was enhanced and the cell exhibited spreading form. Differentiation markers of osteoblast (ALPase activity, calcification, expression of osteocalsin) were enhanced and localized around cell aggregates on c-type PECs. These results suggest that PEC has the ability to control osteoblast proliferation and differentiation.

• 한국수산과학회지
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• 제44권1호
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• pp.25-30
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• 2011

We investigated the effect of a lactic acid extract of Sargassum horneri (ExSL) as a calcium supplement on bone formation in 48 female Sprague-Dawley rats for 4 weeks of their growth phase. The rats were divided into four groups based on diet: two calcium-sufficient and two calcium-deficient diets. The normal control group (NC) was fed AIN-93G; the NCS group was fed the same diet containing 1% extract; the calcium-deficient control (DC) diet was based on AIN-93G; and the DCS group received the same calcium-deficient diet plus 1% extract. Bone formation in the rats was evaluated using the wet weight, length, diameter, and bone mineral density (BMD) of the femur. Serum parameters were also examined. The food intake among the groups did not differ significantly (P<0.05). The NCS group gained the most body weight, while the DC group gained much less weight than the other groups. The feeding efficiencies of the groups that received the extract (NCS and DCS) were slightly higher than those of the control groups (NC and DC). The calcium intakes of all groups depended on the amount of calcium in the feed; the NCS and DCS diets contained 12.15 mg more calcium than the NC and DC diets. The calcium absorption was lower in NCS than in DC and DCS, but significantly higher than in NC (P<0.05). The BMDs in the calcium-sufficient groups were not significantly different (P<0.05), while in the calcium-deficient groups the BMD was significantly higher in DCS than in DC (P<0.05). The serum calcium and phosphorus levels in all groups were not associated with markers of bone growth related to the extract. The osteocalcin content and alkaline phosphatase (ALPase) activity were higher in the calcium-deficient groups than in the normal groups (P<0.05). Ultimately, the osteocalcin content and ALPase activity were lower in DCS compared to DC. These results suggest that the addition of ExSL promotes bone formation and calcium absorption in growing rats.

• Journal of Periodontal and Implant Science
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• 제32권1호
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• pp.143-160
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• 2002

타이태늄은 뛰어난 생체적합성과 적절한 물리적 성질을 바탕으로 치과 및 정형외과 영역의 매식재로 널리사용되어져 왔으며, 골과 매식재 사이의 골 융합 정도를 증가시킬 목적으로 물리, 화학적인 방법을 이용한 타이태늄의 표면처리에 관한 많은 연구들이 진행되어 왔다. 최근에는 부착단백질 또는 성장인자를 이용한 생체재료의 표면개질을 통하여 조직적합성 및 치유 능의 개선을 위한 시도들이 있어왔다. Fibronectin(FN)은 주요 세포외기질중의 하나로 생체 내 널리 분포하여 세포의 부착, 이동 및 증식에 관여하는 거대 당단백으로, RGD및 PHSRN 펩타이드 서열이 세포의 인테그린과 결합하여 세포의 활성을 조절하는 것으로 알려져 있다. 이 연구에서는 FN으로 처리된 타이태늄이 조골세포의 부착, 증식 및 분화에 미치는 영향과 이에 따른 석회화 정도에 미치는 영향을 관찰하여 부착분자를 이용한 타이태늄 표면개질의 효과를 규명하고자 하였다. 상업용 순수 타이태늄을 gold thiol법을 이용하여 표면처리 후, 혈장 FN(plasma FN, pFN)과 유전자재조합법을 이용하여 얻은 FN조각(FN type III 7-10, FNIII 7-10)을 피복한 시편을 실험군으로, 아무런 처리를 하지 않은것(smooth surface, SS)과 산 부식(Sandblasted and acid etched, SLA)처리된것을 대조군으로 이용하였다. 배양된 조골세포주(MC3T3-E1)를 사용하여 타이태늄 표면 처리에 따른 세포의 증식, 형태변화, 알칼리성 인산분해효소(ALPase) 생산 및 세포면역형광법을 이용한 분화정도를 시간 경과에 따라 관찰하였다. 조골세포증식의 경우 FNIII 7-10 처리군에서 pFN 처리군 및 대조군에 비해 시간경과에 따라 유의성있는 세포수의 증식이 관찰되었으며(p<0.05), ALPase 생성의 경우에도 FNIII 7-10 처리 군에서 아무 처리도 하지 않은 군에 비해 유의성 있게 높은 효소의 생성이 관찰되었다(p<0.05). 주사전자현미경을 이용한 세포의 형태관찰결과 아무 처리도 하지 않은 군에서는 마름모형태를 나타내었으며, 산 부식 처리된 군에서는 세포가 가시모양의 형태를 보인 반면 FN으로 처리된 두 군에서는 세포의 부착 및 펴짐이 매우 발달되어 있는 모습이 관찰되었다. 세포의 분화정도를 관찰하기 위하여 국소부착키나제(focal adhesion kinase, FAK), 및 actin stress fiber의 분포양상을 세포면역형광법을 이용하여 관찰한 결과 FN으로 표면처리된 두 군에서 아무런 처리도 하지않은 군 및 산 부식처리 한 군에 비해 프라크의 발현이 높게 나타났으며 잘 발달된 actin stress fiber의 소견을 나타내었다. 이 실험의 결과들은 gold thiol 법을 이용한 표면처리 후 FN부착을 통한 타이태늄의 표면개질이 조골세포의 부착, 증식 및 분화에 중요한 역할을 담당하여 석회화 정도를 촉진시키는 것을 보여주었으며, 이런 결과들은 더 짧은 FN조각을 이용한 다른 생체재료의 표면개질에 폭 넓게 응용할 수 있으리라 생각된다.

• 한국식생활문화학회지
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• 제23권6호
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• pp.801-811
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• 2008

The principal objective of this study was to assess the effects of soy isoflavone supplementation on bone mineral density in 36 female college students with osteopenia for 12 weeks. The subjects were divided into three groups on the basis of bone mineral density. The experimental groups were provided supplements of either 80 mg of isoflavone (Iso-80) or 40 mg of isoflavone (Iso-40). To the placebo group, 40 mg of powdered glutinous rice was administered. It was determined that many subjects with osteopenia evidenced lower levels of activity as compared to the control group. Isoflavone supplementation was more effective in controlling total cholesterol and LDL-cholesterol than was observed in the placebo group. We noted no significant differences in serum osteocalcin concentration between Iso-40 and the placebo group, but significant differences in osteocalcin concentration were detected between Iso-80 and the placebo group. Bone quality indices (BQI) were correlated positively with mineral content, lean body mass, muscular mass, and blood components including albumin, Ca, Mg, ALPase, and osteocalcin. Both Iso-40 and Iso-80 supplementation for 12 weeks significantly increased protein and mineral content in the body. As lower intakes of Ca and folate were noted in the subjects, emphasis should be given to adequate intakes of these nutrients in the subjects. In conclusion, 12 week isoflavone supplementation in young females with osteopenia exerted positive effects on bone mineral density and bone turnover markers.

• Journal of Periodontal and Implant Science
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• 제39권sup2호
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• pp.279-286
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• 2009

Purpose: Gene therapy (ex vivo) has recently been used as a means of delivering bone morphogenetic proteins (BMPs) to sites of tissue regeneration. In the present study, we investigated the effect of co-transduction of adenoviruses expressing BMP-2 and BMP-7 on osteogenesisof C2C12 cells in vitro. Methods: A replication-defective human adenovirus 5 (Ad5) containing a cDNA for BMPs in the E1 region of the virus (Ad5BMP-2 and Ad5BMP-7) was constructed by in vivo homologous recombination. Functional activity of Ad5BMP-2 and Ad5BMP-7 were evaluated in mouse stromal cells (W20-17cells). C2C12 cells are transduced with various MOI (multiplicity of infection) of Ad5BMP-2 and Ad5BMP-7 to assess most effective and stable titer. Based on this result, C2C12 cells were transduced with Ad5BMP-2 and Ad5BMP-7 alone or by combination. BMPs expression, alkaline phosphatase (ALPase) activity, cell proliferation, and mineralization were assessed. Results: Ad5BMP-2 and Ad5BMP-7 are successfully transduced to W20-17 cells, and secreted BMPs stimulated cell differentiation. Also, C2C12 cells transduced with Ad5BMPs showed expression of BMPs and increased ALPaseactivity. In all groups, cell proliferation was observed over times. At 7days, cells co-transduced with Ad5BMP-2 and Ad5BMP-7 showed lower proliferation than the others. C2C12 cells co-transduced with Ad5BMP-2 and Ad5BMP-7 had greater ALPaseactivity than that would be predicted if effect of individual Ad5BMPs were additive. Little mineralized nodule formation was detected in cells transduced with individual Ad5BMPs. In contrast, Ad5BMP-2 and Ad5BMP-7 combination stimulated mineralization after culturing for 10 days in mineralizing medium. Conclusions: Present study demonstrated that adenoviruses expressing BMPs gene successfully produced BMPs protein and these BMPs stimulated cells to be differentiated into osteoblastic cells. In addition, the osteogenic activity of Ad5BMPs can be synergistically increased by co-transduction of cells with Ad5BMP-2 and Ad5BMP-7.