- 學位論文
骨髓間質幹細胞併合富含血小板纖維蛋白促進骨組織再生-以兔子為實驗動物
Functionalized bone-graft substitutes with platelet-rich fibrin and bone marrow-derived mesenchymal stem cells for enhanced bone tissue regeneration-with rabbit animal model
摘要
多孔性的補骨材料，因為具有極佳的生物活性，生物相容性，還有骨質生成傳導活性，所以在骨組織的再生上扮演了極重要的角色。本研究探討是否骨粉成分(BG)，三鈣磷酸鹽/烴磷灰石(beta-TCP/HA)(40/60)的合成物，與富含血小板纖維蛋白(PRF)與骨髓間質幹細胞(BMSCs)以及/或明膠-軟骨素-透明質酸的三合聚合體支架Scaffold能促進骨頭的再生。在骨頭創傷後，這四組不同的處置治療之間，包括(Ⅰ)BG+PRF骨粉＋富含血小板纖維蛋白，(Ⅱ)BG+PRF+BMSCs骨粉＋富含血小板纖維蛋白＋骨髓間質幹細胞，(Ⅲ)BG+Scaffold，骨粉＋明膠-軟骨素-透明質酸的三合聚合體支架(Ⅳ)BG+Scaffold+BMSCs骨粉+明膠-軟骨素-透明質酸的三合聚合體支架＋骨髓間質幹細胞，是否能有更快速的新骨再生比較。 MSCs間質幹細胞是由兔子的股骨骨髓所取得，並加以純化製備而來。在兔子的兩側下顎骨角各做成一個直徑5公厘全厚度的骨缺損。不同組別的骨缺損處置如下：第一組（數目為三），BG骨粉1.7克和0.78立方公分的PRF富含血小板纖維蛋白。第二組（數目為四）BG骨粉1.7克加0.78立方公分的PRF富含血小板纖維蛋白以及2乘以10的6次方BMSCs骨髓間質幹細胞的混合物。第三組（數目為三）BG骨粉1.7克加大約0.78立方公分的Scaffold明膠-軟骨素-透明質酸的三合聚合體支架。第四組（數目為四）BG骨粉1.7克加0.78立方公分的Scaffold明膠-軟骨素-透明質酸的三合聚合體支架以及2乘以10的6次方BMSCs骨髓間質幹細胞的混合物。在術後的三個月犧牲兔子動物已取得檢體。在植入區分別以肉眼，組織切片以及Ｘ光片加以檢查評估。 組織切片的檢查評估顯示第二組及第四組有典型的骨組織修復。相反的，第一組及第三組僅有少許的骨形成且伴隨纖維性的連接。BG+PRF+BMSCS骨粉＋富含血小板纖維蛋白＋骨髓間質幹細胞移植有最好的骨聯合。第二為BG+Scaffold+BMSCs骨粉＋明膠-軟骨素-透明質酸的三合聚合體支架＋骨髓間質幹細胞。第三為BG+Scaffold，骨粉+明膠-軟骨素-透明質酸的三合聚合體支架。最後為BG+PRF骨粉+富含血小板纖維蛋白。在實驗組與控制組之間則存在著統計學上的差異。 實驗證明PRF+BMSCS富含血小板纖維蛋白＋骨髓間質幹細胞移植在骨質再生方面有好的潛力。在極端條件的下顎骨缺損骨粉移植治療上，是一個新的替代治療技術。
並列摘要
Functionalized bone-graft substitutes with platelet-rich fibrin and bone marrow-derived mesenchymal stem cells for enhanced bone tissue regeneration-with rabbit animal model. Porous bone grafting materials, which possess excellent bioactivity, biocompatibility and osteoconductivity, have played an important role in bone tissue regeneration. This study examined whether a combination of bone graft extender (BG), beta-tricalcium phosphate/hydroxyapatite (40/60) composite, with platelet-rich fibrin (PRF), bone marrow-derived mesenchymal stem cells (BMSCs) and/or gelatin-chondroitin-hyaluronan tri-copolymer scaffold could improve bone regeneration; and whether these treatments yield faster new bone formation following bone injury among the four treated groups that include (I) BG+PRF, (II) BG+PRF+BMSCs, (III) BG+scaffold, (IV) BG+scaffold+BMSCs. MSCs were harvested and isolated from the bone marrow of rabbits. Full-thickness bony defects (diameter 5.0 mm) were created in the bilateral mandible angles of the rabbit. Treatments for bone defect in each group were as follows: group I (n=3), BG 1.7 g and 0.75 cm3 of PRF; group II (n=4), BG 1.7 g and 0.75 cm3 of PRF/ 2x106 BMSCs admixtures; group III (n=3), BG 1.7 g and approximately 0.78 cm3 of scaffold; and group IV (n=4), BG 1.7 g and 0.78 cm3 of scaffold/ 2x106 BMSCs admixtures. The mandibles were harvested from all specimens at 3 months, and the grafted sites were evaluated by gross, histologic, and X-ray examination. Histologic examination revealed that the defect was repaired by typical bone tissue in groups II and IV, whereas only minimal bone formation with fibrous connection was observed in the groups I and III group. The best results were obtained with BG+PRF+BMSCs transplantation that had better bone union, formation and new bone position than BG+scaffold+BMSCs in the second, BG+Scaffold in the third, and BG+PRF in the last group. A statistically significant difference between the experimental and control groups was observed. Evidently, PRF plus BMSCs have good potential for bone regeneration and the technique describes a new alternative to bone grafting procedures for the treatment of critical sized bony lesions of the mandible.