論文名稱:

骨髓幹細胞及冷凍擠壓層積成型聚乳酸-甘醇酸強化骨形態發生蛋

 

4/血管內皮生長因子165膠原蛋白活化基因基質之協同骨生成作

 

 

The Synergistic Osteogenic Effect of BMSCs and FCDM-PLGA

 

Reinforced BMP4/VEGF165 Collagen GAM

研究生:

陳乃瑜  Nai-Yu Chen

指導教授:

陳恆理  Hen-Li Chen

學位類別(Degree):

碩士

學校名稱(School):

國立陽明大學

系所名稱(Department):

口腔生物研究所

學號(Student Number):

39517008

學年度(Academic Year):

96

語文別(Language):

中文

出版年(Thesis Year):

97

關鍵字:

組織工程  Tissue engineering

 

骨髓幹細胞  bone marrow stem cell

 

活化基因基質  gene-activated matrix

 

骨形態發生蛋白4  bone morphogenetic protein 4

 

血管內皮生長因子  vascular endothelial growth factor

 

聚乳酸甘醇酸  poly(lactic-co-glycolic acid)

全文說明(Fulltext description):

(本論文 20100827 對校內公開)

  論文頁數(Page):

73

摘要:

傳統治療骨缺損採用自體或同種異體移植,受到不少限制,在修補骨缺損時再生效果仍未理想,近來骨組織工程已被認為是骨移植具潛力之替代治療。組織工程主要是利用細胞、支架、訊號要素來促進組織再生,應用於大範圍缺損可能須採取用的策略以求最佳再生效果。骨髓幹細胞具有分化成多種細胞的能力,為骨組織工程常用的細胞來源。活化基因基質gene-activated matrixGAM)技術是一種被用以持續給予訊號的基因療法,活化基因基質是由基質與內含訊號基因之質體所組成。第一型膠原蛋白(collagen)利於細胞貼,為最常用之GAM基質材料,惜collagen GAM機械性質較差。以冷凍擠壓層積成型 (FCDM) 製成的聚乳酸甘醇酸(PLGA)支架機械性質佳,且能控制支架孔徑大小,可被用於強化collagen GAM。先前研究指出骨形態發生蛋白4 (BMP4)及血管內皮生長因子 (VEGF165)可協同促進骨生成。我們假設合併使用骨髓幹細胞與FCDM-PLGA強化的BMP4/VEGF165 collagen GAM可加成促進骨再生。本研究以嚴重免疫不全小鼠皮異位骨生成模型測試合併植入體內的療效。癒合3815後的標本以雙能量骨質密度儀及X光照射分析骨含量,結果證實骨髓幹細胞與FCDM-PLGA強化的BMP4/VEGF165 collagen GAM之骨生成作用有協同作用。本研究結果顯示所發展的組合治療將來有潛力可被使用於修補大型骨缺損。

 

 

 

Autogenic and allogenic bone grafts used traditionally for repairing bony defects have many limitations. Moreover, their regenerative effects in treating large bony defects are still less than ideal. Recently, bone tissue engineering has been regarded as a potential alternative for bone grafting. Cells, scaffolds and signaling molecules are employed in tissue engineering approach to enhance regeneration. In order to maximize the regenerative potential in treating large defects, the combined use of three elements in tissue engineering may be needed. Bone marrow stem cells (BMSCs) are multipotential cells widely used in bone tissue engineering. Gene-activated matrix (GAM) technology is a gene therapy strategy used for sustained delivery of signals. GAM is composed of matrix and plasmid DNA encoding gene of interest. Type I collagen, with excellent cell attachment property, is the most used matrix materials for GAMs. However, collagen GAMs suffer from poor mechanical strength. PLGA scaffolds fabricated by frozen compressed deposit manufacturing (FCDM), with superior mechanical properties and controllable pore size, can be used to reinforce the collagen GAM. Previous studies have indicated that both bone morphogenetic protein 4 (BMP4) and vascular endothelial growth factor 165 (VEGF165) enhance bone formation synergistically. We hypothesized that the combined use of human BMSCs and FCDM-PLGA reinforced collagen GAM encoding BMP4 and VEGF165 may act cooperatively on bone regeneration. The combined effect was determined in vivo using SCID mice subcutaneous ectopic bone formation model. The bone mineral content of samples after 3, 8, and 15 weeks of implantation, measured by dual-energy X-ray absorptiometry (DEXA) and radiographic analysis, demonstrated that BMSCs and FCDM-PLGA reinforced BMP4/VEGF165 collagen GAM exerted a synergistic effect in bone formation. The results suggested that the developed combinational approach may be useful in repairing large bony defects in the future.

 

 

 

 

論文目次
Table of Content
:

中文摘要..................................................1

 

英文摘要..................................................2

 

第一章 緒論...............................................4

 

第二章 實驗材料與方法....................................14

 

第三章 實驗結果..........................................38

 

第四章 討論..............................................47

 

第五章 結論..............................................50

 

附圖.....................................................51

 

參考文獻.................................................68