• Maxillofacial Plastic and Reconstructive Surgery
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• v.28 no.6
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• pp.511-519
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• 2006

Autogenous bone grafts have been considered the gold standard for maxillofacial bony defects. However, this procedure could entail a complicated surgical procedure as well as potential donor site morbidity. Possibly the best solution for bone-defect regeneration is a tissue engineering approach, i.e. the use of a combination of a suitable scaffold with osteogenic cells. A major source of osteogenic cells is the bone marrow. Bone marrow-derived mesenchymal stem cells are multipotent and have the ability to differentiate into osteoblastic, chondrocytic, and adipocytic lineage cells. However, the isolation of cells from bone marrow has someproblems when used in clinical setting. Bone marrow aspiration is sometimes potentially more invasive and painful procedure and carries of a risk of morbidity and infection. A minimally invasive, easily accessible alternative would be cells derived from periosteum. The periosteum also contains multipotent cells that have the potential to differentiate into osteoblasts and chondrocytes. In the present study, we evaluated the osteogenic activity and mineralization of cultured human periosteal-derived cells. Periosteal explants were harvested from mandibule during surgical extraction of lower impacted third molar. The periosteal cells were cultured in the osteogenic inductive medium consisting of DMEM supplemented with 10% fetal calf serum, 50g/ml L-ascorbic acid 2-phosphate, 10 nmol dexamethasone and 10 mM -glycerophosphate for 42 days. Periosteal-derived cells showed positive alkaline phosphatase (ALP) staining during 42 days of culture period. The formation of ALP stain showed its maximal manifestation at day 14 of culture period, then decreased in intensity during the culture period. ALP mRNA expression increased up to day 14 with a decrease thereafter. Osteocalcin mRNA expression appeared at day 7 in culture, after that its expression continuously increased in a time-dependent manner up to the entire duration of culture. Von Kossa-positive mineralization nodules were first present at day 14 in culture followed by an increased number of positive nodules during the entire duration of the culture period. In conclusion, our study showed that cultured human periosteal-derived cells differentiated into active osteoblastic cells that were involved in synthesis of bone matrix and the subsequent mineralization of the matrix. As the periosteal-derived cells, easily harvested from intraoral procedure such as surgical extraction of impacted third molar, has the excellent potential of osteogenic capacity, tissue-engineered bone using periosteal-derived cells could be the best choice in reconstruction of maxillofacial bony defects.

• Korean Journal of Animal Reproduction
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• v.24 no.2
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• pp.189-197
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• 2000

In this study, we investigated the preventive and therapeutic effects of antler-extract for osteoporosis. Rats were ovariectomized bilaterally and were fed up with Ca- and P-free diet in order to induce osteoporosis. Body weight, organ weight, the weight of femur and bone ash quantity were examined for 5 weeks. We also performed histological and electronical microscopic examinations. 1. After adminstration of female and male antler extract to osteoporosis-induced rats at the doses of 625 and 1250 mg/kg, respectively, the body weights were significantly increased compared with those of normal control group's which was 230.2:t2.3-281.0:t2.5g (p＜0.05). 2. The weights of both right and left femur of osteoporosis-induced rats, administered with female or male antler-extract, little decreased compared with those of normal control group. 3. The bone ash quanties of femur of osteoporosis-induced rats, administered with female or male antler-extract, little decreased compared with those of normal control group. 4. The weights of liver, spleen, and kidney of osteoporosis-induced rats, administered with female or male antler-extract, decreased compared with those of normal control group. 5. Histological and electronic microscopical findings were (1) that in normal control rats the connectional of lacunae appeared well and were without loss of bone mineral, (2) that in ovariectomized rats the connections of lacunae were mostly broken and were with loss of bone mineral compared with those of normal control rats, (3) that in osteoporosis-induced rats, administrated with female or male antler-extract, the shape of lacunae and the connections of them were similar to those of normal control rats. These findings suggest a possible protective and therapeutic effects of female or male antler extract against bone loss in ovariectomized rats.

• Maxillofacial Plastic and Reconstructive Surgery
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• v.31 no.4
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• pp.281-286
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• 2009

Introduction: Enamel matrix derivative (EMD) is a protein which is secreted by Hertwig root sheath and plays a major role in the formation of cementum and attachment of peridontium. Several studies have shown that EMD promoted the proliferation and differentiation of preosteoblasts, osteoblasts and periodontal ligament cells in vitro: however, reports showing the inhibition of osteogenic differentiation by EMD also existed. This study was designed to simultaneously evaluate the effect of EMD on the two cell lines (human mesenchymal stem cells: hMSC, human periodontal ligament derived fibroblasts: hPDLCs) by means of quantitative analysis of some bone related matrices (Alkaline phosphatase : ALP, osteopontin ; OPN, osteocalcin ; OC). Materials and Methods: hMSCs and hPDLCs were expanded and cells in the 4${\sim}$6 passages were adopted to use. hMSc and hPDLCs were cultured during 1,2,7, and 14 days with 0, 50 and 100 ${\mu}g/ml$ of EMD, respectively. ALP activity was assessed by SensoLyte ALP kit and expressed as values of the relative optical density. Among the matrix proteins of the bony tissue, OC and OPN were assessed and quantification of these proteins was evaluated by means of human OC immunoassay kit and human OPN assay kit, respectively. Results: ALP activity maintained without EMD at $1,2^{nd}$ day. The activity increased at $7^{th}$ day but decreased at $14^{th}$ day. EMD increased the activity at $14^{th}$ day in the hPDLCs culture. In the hMSCs, rapid decrease was noted in $7^{th}$ and $14^{th}$ days without regard to EMD concentrations. Regarding the OPN synthesis in hPDLCs, marked decrease of OPN was noted after EMD application. Gradual decrease tendency of OPN was shown over time. In hMSCs, marked decrease of OPN was also noted after EMD application. Overall concentration of OPN was relatively consistent over time than that in hPDLCs. Regarding the OC synthesis, in both of hPDLCs and hMSCs, inhibition of OC formation was noted after EMD application in the early stages but EMD exerted minimal effect at the later stages. Conclusion: In this experimental condition, EMD seemed to play an inhibitory role during the differentiation of hMSCs and hPDLCs in the context of OC and OPN formation. In the periodontium, there are many kinds of cells contributing to the regeneration of oral tissue. EMD enhanced ALP activity in hPDLCs rather than in hMSCs and this may imply that EMD has a positive effect on the differentiation of cementoblasts compared with the effect on hMSCs. The result of our research was consistent with recent studies in which the authors showed the inhibitory effect of EMD in terms of the differentiation of mineral colony forming cells in vitro. This in vitro study may not stand for all the charateristics of EMD; thus, further studies involving many other bone matrices and cellular attachment will be necessary.

• Journal of the Korean Society of Food Science and Nutrition
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• v.42 no.2
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• pp.182-187
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• 2013

This study was conducted to examine the effect of calcium extracted from salted anchovy (Engraulis japonicus) on the calcium metabolism of rats. Sprague-Dawley male rats were fed low-calcium diets (0.15%) for 2 weeks after the adjustment period. Rats were divided into five groups and were fed experimental diet for four weeks. Experimental diets were low calcium (LC, 0.15% $CaCO_3$), 0.5% $CaCO_3$ (CC), seaweed calcium (SC), calcium lactate (LC), anchovy calcium (AC). The low-calcium diet group (LC) showed the lowest weight gain and had no differences among the groups with adequate calcium intake. Calcium retention was lowest in the LC group and higher in the CL, SC, AC groups than in SC groups. Serum alkaline phosphatase (ALP) level was highest in LC group, and significantly low in the CC and AC groups (p<0.05). Parathyroid hormone and osteocalcin levels showed no differences among experimental groups. The urine deoxypyridinoline (DPD) level was lower in AC and CC groups compared to the LC group (p<0.05). The dry weight of the femur showed no significant differences among normal calcium groups. The bone mineral density of the femur in AC and CC group were significantly higher than the LC group (p<0.05). From these results, calcium extracted from salted anchovy can be useful as a calcium supplement comparable with calcium carbonate.

• Journal of the Korean Society of Food Science and Nutrition
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• v.35 no.10
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• pp.1363-1370
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• 2006

This study was designed to evaluate the effect of Agaricus blazei $\beta-glucan$ and egg shell calcium complex on bone metabolism in ovariectomized (OVX) rats. Forty Sprague-Dewley female rats, 10 weeks of age $(248{\pm}1.7g)$, were divided into 4 groups and fed on the experimental diets for 6 weeks: sham operated control treated with normal diet containing 0.5% calcium (Sham-C), OVX-control treated with normal diet containing 0.5% calcium (OVX-C), $OVX-\beta-glucan$ group treated with $\beta-glucan$ diet containing 0.5% calcium (OVX-G), and $OVX-\beta-glucan$ egg shell calcium complex treated with $OVX-\beta-glucan$ egg shell calcium complex containing 0.5% calcium (OVX-GE). Bone weight of femur was higher in the OVX-GE group than in the other OVX groups. Bone mineral density of femur was significantly different (p<0.05) among the experimental groups and showed the highest level in the OVX-GE group. Calcium absorption rate and retention were higher in the $\beta-glucan$ supplement groups than in the other groups (p<0.05). Alkaline phosphatase activities and osteocalcin levels of serum showed lower in the $\beta-glucan$ supplement groups than in the OVX-C group. Deoxypyridinoline crosslink values of urine, indicator of bone absorption, showed the lowest in the OVX-GE group. The $\beta-glucan$ supplemented groups had a lower bone resorption ratio than in the OVX-C group. We concluded that bioavailability of calcium is higher in $\beta-glucan$ supplement groups compared to those in OVX rats. From the above results, these findings suggest the possibility of using $\beta-glucan$ egg shell calcium complex as a functional food material related to bone metabolism, even though there is no significant difference between the groups of $\beta-glucan$ and $\beta-glucan-egg$ shell calcium complex supplementation.

• Journal of Periodontal and Implant Science
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• v.35 no.2
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• pp.321-333
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• 2005

1. 목적 in vitro 상에서 법랑기질유도체가 치주인대섬유아세포, 불멸화 조골세포와 백악질 유래세포의 증식과 유전자 발현에 미치는 영향을 알아보고자 하였다. 2. 연구방법 및 재료 <세포증식 연구> 교정을 목적으로 발거한 치아에서 분리, 배양한 치주인대섬유아세포와 백악질유래세포, 그리고 $SaOs_2$ 세포를 이용하였다. 법랑기질유도체가 세포 증식에 미치는 영향을 알아보기 위해, 35 mm Petri dish에 dish 당 $5{\times}10^3$ 개의 세포를 접종하였다. 대조군은 1% 항생제와 10% FBS를 포함한 DMEM 배지를 이용했고, 5mM 초산을 첨가한 군과 첨가하지 않은 두 개의 대조군이 이용되었다. 실험군은 100 ${\mu}g/ml$의 법랑기질유도제를 첨가한 군과 100 ${\mu}g/ml$의 법랑기질유도체와 5 mM의 초산을 첨가한 2개의 실험군이 이용되었다. 각 군은 세 개의 배양접시에 행해졌고, 1, 3, 8일에 세포의 수를 각각 측정하였다. 결과는 repeated measures ANOVA로 통계 처리하였다. <유전자 발현 연구> 각 세포의 형질 특성을 알아보기 위해 RT-PCR을 실시하여 조골세포 분화 표식자와 연관된 Human collagen type I(COL I), human osteopontin(OP), human osteocalcin(OC), human alkaline phosphatase(ALP)와 human bone sialoprotein(BSP)의 mRNA 발현을 실험 1, 3, 8일에 걸쳐, 세 군의 차이를 비교 관찰하였다. 3. 결과 <세포증식 연구> 치주인대세포와 백악질유래세포, 그리고 $SaOs_2$ 세포의 증식은 법랑기질유도체에 의해 영향을 받지 않았다. 대조군과 초산이 포함된 대조군 그리고 법랑기질유도체와 초산이 포함된 실험군에서 유의할 만한 세포 수의 차이가 실험 기간 1, 3, 8일에 걸쳐 나타나지 않았다(p<0.05). <유전자 발현 연구> ALP와 COL I은 세 군의 세포에서 모두 발현되었고, 발현 정도는 EMD에 영향을 받지 않았다. OC은 세 군에서 모두 비교적 약하게 발현되었고, 특히 $SaOs_2$ cell과 백악질유래세포에서 약하게 발현되었다. EMD는 OC의 발현정도를 약하게 하였다. OP은 백악질유래세포에서 1, 3, 8일에 걸쳐 EMD 유무에 관련 없이 발현되지 않았다. 그러나 치주인대세포와 $SaOs_2$ cell에서는 강하게 발현되었다. BSP는 치주인대세포와 $SaOs_2$ cell에서 1, 3, 8일에 걸쳐 비교적 고르게 발현되었다. EMD 배지에서 배양된 백악질유래세포는 8일에는 BSP가 발현되지 않았다. 4 결론 이번 실험 결과에 의하면 법랑기질유도체는 치주인대세포, 불멸화 조골세포와 백악질 유래세포의 증식에 있어 유의성 있는 효과를 나타내지 않았다. 그러나, 유전자 발현에 있어서는, 치주인대세포와 백악질유래세포, 그리고 $SaOs_2$ 세포 모두에서 OC mRNA의 발현을 억제하는 효과를 나타내었다. EMD는 세포의 증식에는 영향을 미치지 않지만, 유전자 발현에 있어 일부 영향을 미치는 것으로 보인다. 법랑기질유도체가 세포의 증식과 유전자 발현에 미치는 영향은 배양된 세포의 형질특성, 배양환경, 배양일수 등에 따라 달라질 수 있다. 그러므로 법랑기질유도체가 in vitro 상에서 세포에 미치는 영향은 보다 정량화된 연구가 필요하다.

• Journal of the Korean Society of Food Science and Nutrition
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• v.35 no.7
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• pp.853-859
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• 2006

This study was carried out to investigate the effect of egg shell calcium salt types and egg shell membrane on calcium metabolism in rats. Sprague-Dawley male rats, 4 weeks of age, were fed on free-calcium diets for 2 weeks after adjustment period. Rats weighing approximately $247{\pm}2.3g$ were divided into 6 groups and were fed on the experimental diets containing 0.2% calcium for 4 weeks. Experimental groups were as follows; {ES(M+)} (egg shell powder diet with egg shell membrane), {ES(M-)} (egg shell powder diet without egg shell membrane), {AC(M+)} (egg shell calcium acetate diet with egg shell membrance), {AC(M-)} (egg shell calcium acetate diet without eg shell membrane), {GC(M+)} (egg shell calcium glucuronate diet with egg shell membrane) and {GC(M-)} (egg shell calcium glucuronate diet without egg shell membrane). Bone length of femur was significantly different by the types (p<0.05) of egg shell calcium salts. Bone mineral density of femur showed the highest level in AC(M-) group. Calcium content of femur and calcium absorption rate were higher in egg shell calcium salt groups than in eg shell powder groups. Calcium absorption rate and retention were significantly different (p<0.05) among the types of eg shell calcium salts and were higher in the AC(M-) group than in the other groups. Alkaline phosphatase activity, parathyroid hormone and osteocalcin levels of serum showed no significant difference among the experimental groups. From the above results, it is concluded that bioavailability of calcium is higher in groups of egg shell calcium salts compared to those in egg shell powder, even though egg shell membrane has no effect on calcium metabolism. Thus, these findings suggest the possibility of using egg shell calcium salts as a functional food material related to calcium metabolism.

• The Korea Journal of Herbology
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• v.24 no.1
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• pp.59-71
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• 2009

Objectives：The present study has been undertaken to investigate the effects of Mantidis $O\ddot{O}theca$ and Mori Fructus on treatment of osteoporosis in ovariectomized rats. Methods： In this experiment, the rats were ovariectomized. Rats were administered by Mantidis $O\ddot{O}theca$ and Mori Fructus. The levels of bone mineral density, osteocalcin.ALP.calcium.phosphorus in serum, calcium. phosphorus.deoxypyridinoline in urine and calcium.phosphorus.ash weight in bone were measured. Results： 1. The levels of femoral and fibula-tibial bone mineral density were significantly increased in comparison with OVX group at 4, 8 weeks in Mantidis $O\ddot{O}theca$ group. And the levels of femoral and fibula-tibial bone mineral density were significantly increased in comparison with OVX group at 8 weeks in Mori Fructus group. 2. The levels of serum osteoclacin and ALP showed significant decrease in comparison with OVX group at 4, 8 weeks in Mantidis $O\ddot{O}theca$ and Mori Fructus group. The levels of serum calcium showed significant decrease in comparison with OVX group at 4 weeks in Mantidis $O\ddot{O}theca$ and Mori Fructus group. The levels of serum phosphorus showed significant decrease in comparison with OVX group at 4, 8 weeks in Mantidis $O\ddot{O}theca$ and Mori Fructus group. 3. The levels of urine calcium, phosphorus and deoxypyridinoline showed significant decrease in comparison with OVX group in Mantidis $O\ddot{O}theca$ and Mori Fructus group. 4. The levels of fibula-tibial calcium and phosphorus showed significant increase in comparison with OVX group in Mantidis $O\ddot{O}theca$ group and Mori Fructus group. The levels of femoral calcium and phosphorus showed significant increase in comparison with OVX group in Mori Fructus group. The levels of femoral and fibula-tibial ash weight showed significant increase in comparison with OVX group in Mantidis $O\ddot{O}theca$ group and Mori Fructus group. Conclusions： Reviewing these experimetal results, it appeared that Mantidis $O\ddot{O}theca$ and Mori Fructus had efficacy on treatment of osteoporosis.