應用骨髓幹細胞與牙髓幹細胞併合富含血小板纖維釋放液對坐骨神經缺損的修復與再生：以大鼠為模式

Abstract

坐骨神經的損傷對於患者來說不論是治療上或是社交上都會是一個沉重的經濟負擔，傳統的直接縫合以及移植治療並非適用於所有的患者。 近幾年，利用幹細胞進行組織再生的細胞性療法在全世界都是一個新興的研究，並且在許多的器官與組織的再生上都取得了相當不錯的成果，比如在治療膝蓋軟骨的損傷、急性腎衰竭、骨質疏鬆症、第一型的糖尿病上，在動物實驗中都已經證實具有治療的效果，此外在牙齒的再生上也已經得到非常顯著且樂觀的成果。 於此同時，由血液中經分離所得到的富含血小板纖維素（PRF），因為血小板內分泌顆粒含有數種生長因子，經過實驗證實其含有的生長因子對於組織修復和再生過程中，可幫助調節細胞的新陳代謝，以促進組織的修復和癒合。 針對大鼠坐骨神經的修復與再生，我們試驗了大鼠骨髓幹細胞分別及牙髓幹細胞各別混合了富含血小板纖維釋放液（PRFr）來進行治療。首先我們進行手術在大鼠的左後肢的坐骨神經上製造斷裂並在神經的兩端產生一段10mm寬的間隙，之後我們將實驗分組為1.)不做任何治療的對照組；2.)僅注射PRFr的治療組；3.)僅注射骨髓或牙髓幹細胞的治療組；4.)注射骨髓或牙髓幹細胞混合PRFr的治療組。注射的幹細胞量為1 x 106/0.1ml。注射治療後的三個月間每隔一個月進行大鼠的行走步伐測試，以藉此間接推測各組大鼠的坐骨神經在損傷後的恢復情形；注射治療後的三個月後犧牲大鼠取出左後肢的坐骨神經做肉眼及組織觀察及比較。結果顯示，不論是步伐測試的結果還是肉眼及組織切片的結果，三種治療方法均對不做任何治療的對照組有著顯著（p ＜0.05）的治療效果；而三種治療之間則是以幹細胞混合PRFr治療組的效果最好，其次為單獨幹細胞注射治療而PRFr的治療效果遜於另外兩者，而兩種幹細胞（骨髓幹細胞及牙髓幹細胞）之間彼此對於神經的再生效果並沒有顯著性的優劣（p ＞0.05），均是優於PRFr的治療但遜於幹細胞混合PRFr的治療。

The injury of sciatic nerves was a serious problem to the patients, such as costing money for treatment and even worse to change their life. However, there were many limitations in the traditional therapy and as a result a better treatment should be found out. Recently, the cell-based therapy has been focused to repair the broken-down nerves. In the mean times, using Platelet-rich fibrin releasate (PRFr) to repair damaged organs was an insight in Regenerative Medicine. Bone marrow derived stem cells (BMSCs) and Dental pulp derived stem cells (DPSCs) and PRFr were combined to treat with the animals. The 8 week old- injured nerve rats were separated into 6 groups. (1.) Control group, without any treatment; (2.) PRFr group,0.1 ml PRFr was injected; (3.) BMSCs group, 1x106 / 0.1 ml BMSCs were injected; (4.) DPSCs group, 1x106 / 0.1 ml DPSCs were injected; (5.) BMSC and PRFr group; (6.) DPSC and PRFr group, the rats were treated with 0.1 ml BMSCs and PRFr. Twelve weeks later, rats were sacrificed and the sciatic nerves regeneration was evaluated by gross and microscope. In gross, we measured the diameters of the regeneration sciatic nerves of every rats, then average every group and compared each other; in microscope, we calculate the axon density in the regeneration part of nerve fiber and average every group and compared. The result demonstrated that the combined treatment has significant impact than other groups. Following that was stem cells (BMSCs, DPSCs) treatment. However, the PRFr group was not efficiency as our expect but it still reached significant than the control group. In conclusion, cell based therapy may provide a new way to the nerve regeneration.