- 學位論文
含鍶載體對乳牙幹細胞之骨分化研究
Osteo-differentiation of SHED cells in strontium containing scaffolds
摘要
近年來發現含鍶藥物在預防骨質疏鬆症產生的骨折達到效果，引起大家的重視；鍶可以促進骨細胞成熟，使新生骨量增多，同時可以抑制蝕骨細胞的活性。因此，本研究嘗試將鍶運用在骨修補材料中；將4%幾丁聚醣溶液混合0.7%磷酸鍶與0.3%氫氧基磷灰石，於-20℃冷凍一天，再以冷凍乾燥法製作成多孔立體載體，其孔洞直徑約50~80μm，孔隙度約90~95%，而壓縮強度與純幾丁聚醣相比從47kPa增強到96kPa。 乳牙幹細胞具有分化為成骨細胞的潛力。將乳牙幹細胞培養於本實驗室所製成的複合材料中，持續觀察21天。結果發現含鍶複合材料對於乳牙幹細胞的生長後貼附性良好，骨分化基因osteonectin和osteocalcin的表現量提升，在載體上產生較多的鈣化沉積，顯示含鍶複合材料能促進乳牙幹細胞分化為成骨細胞，並且與添加磷酸三鈣或是氫氧基磷灰石相比，添加磷酸鍶具有更大的分化效益。 最後，將載體放置於本研究設計之灌流式生物反應器，使培養基可以直接流經載體而改善質傳的速率。實驗結果顯示，灌流培養所伴隨的流體剪力提高osteocalcin表現，對於細胞的分化有一定的幫助。
並列摘要
There are much attention to the drug with strontium that has been shown to reduce the fracture risk of osteoporosis in recent years. Strontium could promote osteoblasts maturation, and new bone generation under the activity of osteoclasts inhibiting. In this study, we attempted to add strontium into bone repair materials. The 4% chitosan solution were agitated with 0.7% strontium phosphate and 0.3% hydroxyapatite. After mixing, the solution were placed in a -20℃ refrigerator overnight, and freeze-dried to form 3D porous scaffolds. The scaffolds were with 50~80μm pore diameter, and 90~95% in porosity. The compressive modulus increase to 96kPa compared to the pure chitosan scaffolds with 47kPa. Stem cells derived from pulp of human exfoliated deciduous teeth (SHEDs) have potential for osteogenic differentiation. The SHEDs were cultured in the scaffolds during 21 days. The results showed that the scaffold with strontium was suitable for SHED for attachment and growth, the gene expressions of osteonectin and osteocalcin were increased, and calcification was also increased. These results indicated that strontium could promote SHEDs in osteo-differentiation. The results shows that adding strontium has more benefits compare to tricalcium phosphate or hydroxyapatite,. Finally, we placed scaffolds in the perfusion bioreactor. It makes medium flowing through scaffolds directly to improve mass transfer rate. It was found that the flow shear stress of perfusion system increased osteocalcin expression, and help the SHEDs differentiation.
參考文獻
延伸閱讀
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