• 폴리머
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• 제30권1호
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• pp.14-21
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• 2006

조직공학 기술은 in vitro와 in vivo에서 초기 세포 부착과 차후의 조직형성을 위해 3차원적인 지지체로서 다공성의 생분해성 담체의 사용이 필수적이다. 소장점막하조직(small intestinal submucosa, SIS)은 고유의 인장력과 생체적합성 때문에 생체물질로서 사용될 잠재력을 가지고 있는 콜라겐 조직이다. 근육유래 줄기세포는 배양조건에 따라 골세포, 연골세포, 및 근육세포 등으로 분화가 가능하다고 알려져 있다. 본 연구에서는 SIS를 함유한 락타이드-글리콜라이드 공중합체(PLGA) 다공성 지지체를 용매캐스팅/염추출법으로 제조하였고, 전자주사현미경 및 수은다공측정계를 이용하여 특성을 결정하였다 세포의 생존율과 성장률은 MTT(3-(4,5-dimethylthiazole-2-yl)-2,5-diphenyltetrazolium-bromide) 분석 방법을 이용하였고 골로 분화된 세포를 알칼라인 포스파테이즈(ALP) 활성을 측정하여 확인하였다. SIS가 함유된 지지체와 SIS가 함유되지 않은 지지체를 면역결핍 쥐의 피하에 삽입하여 이들의 골형성 정도를 비교하여 보았다. 조직을 파라핀으로 고정시켜 슬라이드를 제조한 후 hematoxylin과 eosin, 트라이크롬 및 본쿠사 염색을 실시하였다. 천연/합성 하이브리드 담체로서의 SIS/PLGA 담체가 PLGA 단독으로 사용하였을 때와 비교하여 볼 때 골형성이 우수하였는데 이는 SIS 내에 함유하고 있는 여러 생체활성분자에 기인한 것으로 추측되었다.

• Archives of Plastic Surgery
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• 제34권3호
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• pp.298-304
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• 2007

Purpose: This study was undertaken to investigate the osteogenic induction potential of PGA & FBS mixture on a calvarial defect in the rabbit. Methods: Twenty New zealand white rabbit, weighing from 3.5-4kg were allocated into each of the three groups. Four 8 mm sized bone defects were made on the parietal bone by drilling. In group I, the bony defects were implanted with $50{\mu}m$ thickness film containing mixture of PGA and FBS. In group II, with PGA only film, & in group III, the bony defects were left with no implants. Results were evaluated by using morphologic change, radiographic study, biochemical study and histologic examination at 1 week (group I n=7, group II n=7, group III n=14), 2 weeks (group I n=6, group II n=6, group III n=12) and 3 weeks (group I n=7, group II n=7, group III n=14) following implantation. Results: In the morphologic & radiographic study, the formation and corticalization of callus were observed earlier in group I than in groups II and III (p < 0.05). In histological examination, group I showed more abundant and faster new bone formation than in group II and III. In biochemical analysis, group I displayed more activity than in group II and III. Group I also showed more abundant osteopontin, osteocalcin than groups II and III. Conclusion: In conclusion, the results demonstrate that the mixture of PGA and FBS has an effect on osteoblastic formation in the rabbit model. It is considered that further evaluation of long term results on resorption, immunologic tissue reaction and response of applied mixture in the human model will be needed.

• 한국방사선학회논문지
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• 제5권1호
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• pp.11-18
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• 2011

다능성 세포를 포함하는 골막은 골모세포와 연골세포로 분화될 수 있다. 그리고 배양된 골막유래세포는 골형성 능력을 가지고 있다. 이 연구의 목적은 골막유래 세포들과 골이식재 간의 상호작용을 평가하는 것이다. Sprague-Dawley 랫드의 두개골 골막에서 세포를 분리한 다음, 배양된 골막유래세포를 beta-tricalcium phosphate (${\beta}$-TCP)와 함께 임계결손부 크기의 두개결손부에 이식하였다. 모든 랫드는 골이식 수술 후 8주째에 희생되었으며, 골이식부의 골형성 능력은 일반방사선, micro CT 및 조직검사를 통해 평가되었다. ${\beta}$-TCP와 함께 이식된 골막유래세포는 골결손부에서 더욱 증가된 석회화작용을 나타내었으며, 골결손부 안쪽 및 가장자리에 골밀도 증가와 신생골이 형성되었다. 특히 골막유래세포는 ${\beta}$-TCP만 단독으로 이식하였을때보다 함께 이식 시 효과적으로 신생골을 형성하였다. 이러한 결과는 배양된 골막유래세포가 골결손부에서 골형성을 증진시킬 수 있는 가능성을 보였다.

• 대한두경부종양학회지
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• 제10권2호
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• pp.218-224
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• 1994

Soft tissue sarcoma of the head and neck is not frequent neoplasm, accounting for less than 1% of all malignant neoplasm in the region. The histological varieties include osteogenic sarcoma, malignant fibrous histiocytoma, rhabdomyosarcoma, fibrosarcoma, tenosynovial sarcoma, angiosarcoma and chondrosarcoma. Rhabdomyosarcomas of the head and neck usually occur in children under the age of 10 years (over 70%) and rarely develop in adults over the age of 20 years. The prevalent sites of involvement include the orbit, nasal cavity, external ear, paranasal sinus and soft tissue of mouth and the primary location of tumor is considered to be one of the important prognostic factors. Before the 1960s, when surgical resection was the only method of treatment, the 5-year survival rate was less than 20%, but recently it has been greatly improved by the multimodality treatment, combining surgery with chemotherapy and radiation therapy. Here we treated a rhabdomyosarcoma woman with three cycles of high dose chemotherapy followed by radiation therapy. After the, completion of preoperative treatments, successful result of more than partial response was achieved. Three months later total maxillectomy and radical neck dissection was performed. There was no evidence of tumor infiltration in the resected tumor and regional lymphnodes but metastasized tumor cells in cervical lymphnodes were detected. Tumor cell infiltration was also found on the bone marrow biopsy to evaluate the pancytopenia which occurred during postoperative recovery. Two months later she died of secondary bone marrow failure. We think that this multimodality treatment combining pre-operative chemotherapy, radiotherapy and surgery might play an important role in curative resection and eyeball preservation in patients with rhabdomyosarcoma involving the eyeball.

• Journal of Periodontal and Implant Science
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• 제28권1호
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• pp.145-160
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• 1998

The ultimate goal of periodontal therapy is the regeneration of periodontal tissue which has been lost due to destructive periodontal disease, and numerous kinds of materials and techniques have been developed to achieve this goal. Bone grafts include autografts, allografts, xenografts and synthetic grafts. Among the synthetic grafts, bioactive glass has been used in dentistry for more than ten years and Fetner reported improved new bone formation and more amount of new attachment after grafting PerioGlas, a kind of bioactive glass, in 2-wall defects of monkeys in 1994. It Is well known that 1-wall defects have less osteogenic potential and more epithelial migration, so we need to study the erect of bioactive glass in 1-wall dejects in dogs. The present study evaluates the effect of bioactive glass on the epithelial migration, alveolar bone regeneration, cementum formation and gingival connective tissue attachment in intrabony detects of dogs. Four millimeter deep and four millimeter wide 1-wall defects were surgically cheated in the mesial aspects of premolars. The test group received bioactive glass with a flap procedure and the control underwent flap procedure only. Histologic analysis after 8 weeks of healing revealed the following results: 1. The height of gingival margin was 1.30{\pm}0.73mm$ above CEJ in the control and $1.40{\pm}0.78mm$ in the test group. There was no statistically significant difference between the two group. 2. The length of epithelial growth (the distance from CEJ to the apical end of JE) was $1.74{\pm}0.47mm$ in the control and $1.12{\pm}0.36mm$ in the test group. These was a statistically significant difference between the two groups (P<0.01). 3. The length of new cementum was $2.06{\pm}0.73mm$ in the control and $2.62{\pm}0.37mm$ in the test group. There was no statistically significant difference between the two groups. 4. The length of new bone was $1.83{\pm}0.74mm$ in the control and $2.39{\pm}0.59mm$ in the test group. There was no statistically significant difference between the two groups. These results suggest that the use of bioactive glass 1-wall intrabony defects has significant effect on the prevention of junctional epithelium migration, but doesn't have any significant effect on new bone and new cementum formation.

• Journal of Periodontal and Implant Science
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• 제39권2호
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• pp.119-128
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• 2009

Purpose: The aim of this study was to evaluate the stemness of cells from canine dental tissues and bone marrow. Methods: Canine periodontal ligament stem cells (PDLSC), alveolar bone stem cells (ABSC) and bone marrow stem cells(BMSC) were isolated and cultured. Cell differentiations (osteogenic, adipogenic and chondrogenic) and surface antigens (CD146, STRO-1, CD44, CD90, CD45, CD34) were evaluated in vitro. The cells were transplanted into the subcutaneous space of nude mice to assess capacity for ectopic bone formation at 8 weeks after implantation. Results: PDLSC, ABSC and BMSC differentiated into osteoblasts, adipocytes and chondrocytes under defined condition. The cells expressed the mesenchymal stem cell markers differently. When transplanted into athymic nude mice, these three kinds of cells with hydroxyapatite /${\beta}$- tricalcium phosphate (HA/TCP) carrier showed ectopic bone formation. Conclusions: This study demonstrated that canine dental stem cells have stemness like bone marrow stem cells. Transplantation of these cells might be used as a therapeutic approach for dental stem cell-mediated periodontal tissue regeneration.

• Journal of Periodontal and Implant Science
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• 제38권sup2호
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• pp.325-334
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• 2008

Purpose: The carrier used as delivery agent for bone morphogenetic proteins(BMPs) should also act as a scaffold for new bone formation. Moreover, bone formation should be predictable in terms of the volume and shape. This study examined the osteogenic effect of macroporous biphasic calcium phosphate (MBCP) block combined with ePTFE membrane as a carrier for recombinant human bone morphogenetic proteins (rhBMP-2). In addition, the additive effect of ePTFE membrane on bone formation was evaluated. Materials and Methods: Eight-millimeter critical sized calvarial defects were created surgically in 28 male Sprague-Dawley rats. The animals were divided into 2 groups containing 14 animals each. The defects were treated with either rhBMP-2/MBCP block (rhBMP-2/MBCP group) or rhBMP-2/MBCP block/ePTFE membrane (rhBMP-2/MBCP/ePTFE group). A disc-shaped MBCP block (3 mm height and 8 mm diameter) was used as the carrier for the rhBMP-2 and ePTFE membrane was used to cover the rhBMP-2/MBCP block. The histologic and histometric parameters were used to evaluate the defects after 2- or 8-week healing period (7 animals/group/healing interval). Results: The level of bone formation in the defects of both groups was significantly higher at 8 weeks than that at 2 weeks (P < 0.05). The ePTFE membrane has no additional effect compared with the rhBMP-2/MBCP block only. However, at 8 weeks, rhBMP-2/MBCP/ePTFE group showed more even bone formation on the top of the MBCP block than the rhBMP-2/MBCP group. Conclusion: These results suggest that the ePTFE membrane has no additive effect on bone formation when a MBCP block is used as a carrier for rhBMP-2.

• Journal of Oral Medicine and Pain
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• 제26권1호
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• pp.39-50
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• 2001

In order to investigate the effect of Transforming growth factor ${\beta}1$(below TGF-${\beta}1$) and osteogenic protein-1(below Op-1) onto the myogenic differentiation, C2C12 satellite myoblastic cell line was cultured and treated with both growth factors. At first morphological changes with microscopical examination were examined, and isolated total RNA to analyse mRNA expression of bone marker proteins, muscle regulatory proteins, TGF-${\beta}$ receptor and their ligands by Northern blot analysis. And cellular proliferative inducibility of both growth factors was also tested to C2C12 cells. Incubating the cell with $5ng/m{\ell}$ of TGF-${\beta}1$ until 4 days almost inhibited multinucleated myotube formation expressing muscular regulatory proteins, and induced decreasing Id proteins. However, no osteoblastic phenotypes was induced by TGF-${\beta}1$ in C2C12 cells. The mRNA expression of TGF-${\beta}$ receptors with TGF-${\beta}1$ was conversed after 48 hours cultured. Type I TGF-${\beta}$ receptor was seemed to play a role in negative signalling for inhibition of myogenic differentiation. OP-1 dose dependently induced ALP activity, osteopontine production and bone sialoprotein production at concentrations above $100ng/m{\ell}$ and osteocalcin production at concentrations above $300ng/m{\ell}$. The concentration of OP-1 required to induce these osteoblastic phenotypes was the same as that required to almost completely inhibit myotube formation. Incubation with above $100ng/m{\ell}$ OP-1 suppressed the expression of mRNA for muscular egulatory proteins from 2 days after incubation. Expression of Id-1, 2, 3 mRNA were stimulated by OP-1 at concentration above $300ng/m{\ell}$. When C2C12 cells were treated with both growth factors, TGF-${\beta}1$ potentiated the inhibitory effect of OP-1 on myotube formation and expression of mRNA for myogenin at 12 days. And TGF-${\beta}1$ reduced osteocalcin and bone sialoprotein production induced by OP-1 at 12 days in C2C12 cells. Both growth factor had no mitogenic effect. These results indicate that OP-1 converts the differentiation pathway of C2C12 myoblasts into that of osteoblastic lineage cells and it's not heritable, but TGF-${\beta}1$ does not and has reversible inhibitory activity on the myogenic differentiation. TGF-${\beta}1$ and OP-1 play a role in myogenic differentiation via different mechanism between them.

• Restorative Dentistry and Endodontics
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• 제26권4호
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• pp.296-306
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• 2001

The purpose of this study was to observe the effect of dexamethasone and osteogenic protein-1(BMP-7) on bone, cementum and periodontal tissue regeneration. A total of 60 Sprague-Dawley white female mice were selected and beta-APN was used for five days to extract the maxillary first molar a traumatically. After the extraction of the teeth, the mesiobuccal root canal was filled with Caviton$^{\circledR}$. The teeth were etched with citric acid for 1 min and coated with one of four different experimental solutions : DEX(500nM/ml), DEX(1000nM/ml), OP-1(100$\mu\textrm{g}$/ml) and OP-1(500$\mu\textrm{g}$/ml) for three minutes depending on the group. All teeth were then replanted under microscope. All replantation procedures were done within 30 minutes. Teeth that were replanted after 30 minutes of bench dry only was used as positive control. All animals were sacrificed at 3 weeks following replantation and histologic observtion was done. The results were as follows ; 1. Active root resorption rate was decreased by the order of OP-1(500$\mu\textrm{g}$/ml), DEX(1000nM/ml), OP-1(100$\mu\textrm{g}$/ml), and DEX(500nM/ml). There was statistically less root resorption in OP-1 (500$\mu\textrm{g}$/ml) and DEX(1000nM/ml) group(P<0.05). 2. The group with higher concentration of dexamethasone(1000nM/ml) had statistically more bone union compared to positive control group(P<0.05),but there were no significant differences among four experimental groups. 3. OP-1(500$\mu\textrm{g}$/ml) and DEX(1000nM/ml) groups showed less degree of inflammation compared to the OP-1(100$\mu\textrm{g}$/ml). DEX(500nM/ml), and positive control group (P<0.05). In conclusion, the group with higher concentration of OP-1 had the best results on root resorption, bone ankylosis and anti-inflammatory effects compared to the other experimental groups, but a long-term study is also necessary to evaluate the exact pharmacological effects of the drugs in the future.

• Maxillofacial Plastic and Reconstructive Surgery
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• 제29권6호
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• pp.474-484
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• 2007

Purpose: The effect of platelet-rich plasma(PRP) on osteogenesis of marrow-derived osteoblasts on histomorphometric analysis in the mandible of rabbit was assessed. Materials and Method: Bone marrow cells were obtained from iliac bone of rabbits and were cultured in a Dulbecco's Modified Eagle's Medium(DMEM) with Dexamethasone, L-Ascortic acid, ${\beta}$-Glycerophosphate to proliferate and differentiate into osteoblasts for $4{\sim}5$ weeks. The expression of osteogenic mar-kers was detected by reverse transcription-polymerase chain reaction(RT-PCR) and silver nitrate stain. Then we prepared bony defects in the mandible of rabbit, 10.0mm in diameter and 4.0mm deep, by trephine bur. In the control group, the defects were filled with autogenous bone and cultured osteoblasts. In the experimental group, the defects were filled with autogenous bone, cultured osteoblasts and PRP. 2 weeks, 4 weeks, 8 weeks later, each group was evaluated with histological and histomorphometric analyses. Results: In vitro, osteoblasts were identified on RT-PCR and silver nitrate stain. According to histological observation, at 2 weeks well-developed anasto-mosing newly-formed woven bone was observed, at 4 weeks anastomosing newly-formed woven bone having osteoblastic activation was observed, and at 8 weeks thick newly-formed woven bone was observed in both control and experimental groups. According to histomorphometric analysis, there were 1.5% more newly-formed bone volume in experimental group than control group at 2 weeks, 28.4% more at 4 weeks, 4.3% more at 8 weeks. Particularly there were significant differences in bone volume at 4 weeks and 8 weeks new bone. Conclusion: Our results demonstrated PRP may enhance osteogenesis of marrow-derived osteoblasts at 4 weeks, 8 weeks.