• Maxillofacial Plastic and Reconstructive Surgery
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• 제32권3호
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• pp.199-206
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• 2010

This study investigated the effects of strontium on osteoblastic phenotypes of cultured human periostealderived cells. Periosteal tissues were harvested from mandible during surgical extraction of lower impacted third molar. Periosteal-derived cells were introduced into cell culture. After passage 3, the periostealderived cells were further cultured for 28 days in an osteogenic induction DMEM medium supplemented with fetal bovine serum, ascorbic acid 2-phosphate, dexamethasone and at a density of $3{\times}10^4$ cells/well in a 6-well plate. In this culture medium, strontium at different concentrations (1, 5, 10, and 100 ${\mu}g$/mL) was added. The medium was changed every 3 days during the incubation period. We examined the cellular proliferation, histochemical detection and biochemical measurements of alkaline phosphatase (ALP), the RT-PCR analysis for ALP and osteocalcin, and von Kossa staining and calcium contents in the periostealderived cells. Cell proliferation was not associated with the addition of strontium in periosteal-derived cells. The ALP activity in the periosteal-derived cells was higher in 5, 10, and 100 ${\mu}g$/ml strontium-treated cells than in untreated cells at day 14 of culture. Among the strontium-treated cells, the ALP activity was appreciably higher in 100 ${\mu}g$/ml strontium-treated cells than in 5 and 10 ${\mu}g$/ml strontium-treated cells. The levels of ALP and osteocalcin mRNA in the periosteal-derived cells was also higher in strontium-treated cells than in untreated cells at day 14 of culture. Their levels were increased in a dose-dependent manner. Von Kossa-positive mineralization nodules were strongly observed in the 1 ${\mu}g$/ml strontium-treated cells at day 21 and 28 of culture. The calcium content in the periosteal-derived cells was also higher in 1 ${\mu}g$/ml strontium-treated cells at day 28 of culture. These results suggest that low concentration of strontium stimulates the osteoblastic phenotypes of more differentiated periosteal-derived cells, whereas high concentration of strontium stimulates the osteoblastic phenotypes of less differentiated periosteal-derived cells. The effects of strontium on osteoblastic phenotypes of periosteal-derived cells appear to be associated with differentiation-extent.

• 대한소아치과학회지
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• 제35권2호
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• pp.205-215
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• 2008

외상이나 치주 질환, 유전적 결함 등에 의하여 치아가 상실되었을 경우, 보철 수복이나 임플란트로 치료하고 있다. 그러나 보철 수복이나 임플란트는 자연치가 아닌 인공치를 이용한 치료로서 기능성과 심미성 등에서 자연치에 비하여 한계가 있으므로 자연치를 이용한 이식 방법에 관심이 증가하고 있으며 치아 이식의 성공률을 높이기 위한 이식 치아의 발육과 성장에 대한 연구가 진행 중이다. 이 연구는 세 가지 단계의 치아싹을 발치와에 동종 이식하고 치아싹이 발육하여 석회화가 이루어지는지 평가하기 위하여 임신 15일과 17일, 출생 후 3일 된 흰쥐의 치아싹을 분리하여 11주 된 흰쥐의 발치와에 이식하고 4주와 8주 후 방사선학적 그리고 병리조직학적으로 석회화 과정을 관찰하여 다음과 같은 결과를 얻었다. 1. 발치와에 이식된 임신 15일과 17일, 출생 후 3일 된 치아싹에서 4주와 8주 후 석회화된 조직이 형성되었다. 2. 임신 15일과 17일, 출생 후 3일 된 치아싹을 발치와에 이식하고 4주 후 치아종 같은 비정상적인 석회화 조직과 인접 치조골과 유착된 골양상아질이 관찰되었다. 3. 임신 15일과 17일 된 치아싹을 발치와에 이식하고 8주 후 골양상아질이 보였으며 치주인대로 둘러싸인 것도 관찰되었다. 4. 출생 후 3일 된 치아싹을 발치와에 이식하고 8주 후 골양상아질과 백악질이 형성되었다. 이 연구는 치아싹을 흰쥐의 발치와에 이식한 후 치아싹의 발육을 관찰한 것으로 이식 조직에서 크기가 작고 무정형의 석회화 조직이 형성되었다. 정상적인 치아보다 형태가 불규칙하고 발육이 늦었으므로 이식된 치아싹이 정상적인 치아 형태로 석회화되도록 하기 위해서는 이식 조직의 발육 정도와 이식 조직을 절제할 때 외과적 손상의 감소, 치아싹으로의 풍부한 혈류 공급, 이식 기간, 이식 조직의 고정을 고려하여 더 많은 연구가 필요할 것으로 생각된다.

• Maxillofacial Plastic and Reconstructive Surgery
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• 제29권3호
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• pp.197-205
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• 2007

Angiogenesis is a essential part for bone formation and bone fracture healing. Vascular endothelial growth factor (VEGF), one of the most important molecules among many angiogenic factors, is a specific mitogen for vascular endothelial cells. VEGF-mediated angiogenesis is required for bone formation and repair. However, the effect of VEGF on osteoblastic cells during osteogenesis is still controversial. In recent days, substantial progress have been made toward developing tissue-engineered alternatives to autologous bone grafting for maxillofacial bony defects. Periosteum has received considerable interest as a better source of adult stem cells. Periosteum has the advantage of easy harvest and contains various cell types and progenitor cells that are able to differentiate into a several mesenchymal lineages, including bone. Several studies have reported the bone formation potential of periosteal cells, however, the correlation between VEGF signaling and cultured human periosteal cell-derived osteogenesis has not been fully investigated yet. The purpose of this study was to examine the correlation between VEGF signaling and cultured human periosteal-derived cells osteogenesis. Periosteal tissues of $5\;{\times}\;20\;mm$ were obtained from mandible during surgical extraction of lower impacted third molar from 3 patients. Periosteal-derived cells were introduced into the cell culture and were subcultured once they reached confluence. After passage 3, the periosteal-derived cells were further cultured for 42 days in an osteogenic inductive culture medium containing dexamethasone, ascorbic acid, and ${\beta}-glycerophosphate$. We evaluated the alkaline phosphatase (ALP) activity, the expression of Runx2 and VEGF, alizarin red S staining, and the quantification of osteocalcin and VEGF secretion in the periosteal-derived cells. The ALP activity increased rapidly up to day 14, followed by decrease in activity to day 35. Runx2 was expressed strongly at day 7, followed by decreased expression at day 14, and its expression was not observed thereafter. Both VEGF 165 and VEGF 121 were expressed strongly at day 35 and 42 of culture, particularly during the later stages of differentiation. Alizarin red S-positive nodules were first observed on day 14 and then increased in number during the entire culture period. Osteocalcin and VEGF were first detected in the culture medium on day 14, and their levels increased thereafter in a time-dependent manner. These results suggest that VEGF secretion from cultured human periosteal-derived cells increases along with mineralization process of the extracellular matrix. The level of VEGF secretion from periosteal-derived cells might depend on the extent of osteoblastic differentiation.