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標題: | 幾丁聚醣單體培養人類牙根尖乳突細胞之研究 Study of Human dental papilla cells cultured with Chitosan monomer. |
作者: | Chia-Tzu Liou 劉佳姿 |
指導教授: | 楊台鴻(Tai-Horng Young) |
關鍵字: | 幾丁聚醣,牙乳突細胞,礦化誘導,成骨分化, Chitosan,dental papilla cells,mineralization,osteogenic differentiation, |
出版年 : | 2008 |
學位: | 碩士 |
摘要: | 牙周組織為牙齒的支持組織—牙齒依靠特殊之支持結構連接在顎骨上,包括齒槽骨(alveolar bone)、牙周韌帶(periodontal ligament, PDL)、牙骨質(cementum)及其保護組織—牙齦(gingiva)。在牙周組織重建與再生的過程中,來自牙根尖乳突的前趨細胞扮演重要的角色。對於牙周和植入手術中,「骨再生」一直以來都是個具爭議性的課題,很多的程序是為促進再生的目的而發展出來,包括指標性的組織再生以及骨移植。幾丁聚醣是生物可分解性天然高分子,無毒且不會致免疫性。於很多研究中,幾丁聚醣被認為是可以促進止血(hemostasis)且可增進傷口癒合。從動物模式中也顯示能增進骨頭治療,亦即幾丁聚醣可能具有促進成骨的功能。已有研究顯示牙周組織細胞會受到幾丁聚醣刺激而促進成骨表型特徵。
本研究的目的在探討一般培養液含有不同濃度之幾丁聚醣單體時,牙根尖乳突細胞在不同培養時間(3天、7天、14天),藉由鹼性磷酸酶(Alkaline Phosphatase)染色與觀察、鹼性磷酸酶(Alkaline Phosphatase)活性、細胞基質礦化小體的染色(Alizarin Red Staining)與觀察及定量,和其細胞分化造骨細胞表型分化指標基因以及以動物實驗來檢驗幾丁聚醣單體對於骨生成的幫助。 本研究結果發現:(1) 無論在一般培養液或是促進鈣化的培養液培養,培養時,幾丁聚醣濃度若低於0.1mg/ml時,均不會對牙根尖乳突細胞造成傷害。(2)由鹼性磷酸酶染色、鹼性磷酸酶活性、Alizarin Red S(ARS)染色法,以及成骨指標蛋白的基因表現,得知:牙根尖乳突細胞在二種培養液加入幾丁聚醣單體濃度為0.1mg/ml時,其表現均較其它條件顯著增強許多。 (3)動物實驗也顯示,以骨髓調和幾丁聚醣單體粉末所形成的膏狀物來充填顱骨缺損,的確有助於骨生成,而若能加入支架(聚已內酯),則一方面讓缺陷內的幾丁聚醣可以被保留住而發揮其作用,另一方面可能促成骨膜等軟組織能有更完整的附著,進而達到較有效率的骨缺陷修復。綜合本研究結果可知:牙根尖乳突細胞具有造骨細胞表型特徵,受幾丁聚醣單體的刺激可以促進其成骨特性的表現,至於幾丁聚醣單體是否可有效促進牙周組織再生修復,仍有待未來研究作進一步探討。 Periodontal tissues which is included alveolar bone, periodontal ligament, cementum and gingival, is the supporting tissues for teeth. Teeth depend on specialized supporting structure (periodontal tissues) connect teeth to jaw bone. In the process of periodontal tissues regeneration, the progenitor cells which from dental papilla tissues plays an important role. For periodontal and implant surgery, bone regeneration is always critical issue that many processes are developing for promoting regeneration, including tissue regeneration and bone transplantation. Chitosan is biodegradable, non-toxic, non-immunogenic and natural polymer. Chitosan is thought to help hemostasis and promote wound healing in many studies. In animal models, it can also enhance bone therapy which means chitosan may have the functions of promoting osteogenic process. There is already study that cells from periodontal tissues will stimulate by chitosan to promote expressing more osteogenic phenotype. The purpose of this study is discussing dental papilla cells treat with different concentrations chitosan monomer in culture medium and different culture duration, by Alkaline Phosphatase staining, alkaline phosphatase activity, cell matrix mineralization foci staining(Alizarin Red staining) , cell differentiate osteogenic phynotype markers expression and animal experiment for examination the effect of chitosan on bone regeneration. Our result demonstrated that: (1) When the concentrations of chitosan monomer lower than 0.1mg/ml it won’t injure dental papilla cells. (2) By alkaline Phosphatase staining, alkaline phosphatase activity, ARS staining and gene expressions, chitosan monomer can improve osteogenic phenotype when condition is 0.1mg/ml. (3)Animal experiment also shows that filling with chiosan monomer into bony defects can promote bone formation; if bone defect covered with Polycaprolactone(PCL) scaffold, it can keep chiosan from losing and promote soft tissues, such as periosteum, to attach more completely, then reach to more effective regeneration in bone defect. These results suggest that dental papilla cells with osteogenic cells phenotype and stimulated by chitosan can promoting osteogenic expression. Whether or not chitosan monomer can effectively promote the regeneration of periodontal tissues needs more detailed study. |
URI: | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/42080 |
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顯示於系所單位: | 醫學工程學研究所 |
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