隨著高齡社會來臨，年長者因鈣質流失、骨質疏鬆等問題加劇，骨損傷復原的速度明顯較一般人慢且不佳。傳統上的骨移植治療方式大多是自體、異體骨移植，或是骨粉等生物性材料，但卻存在免疫排斥或骨材料硬度篩選的問題，而現行的細胞療法多採用單細胞（single cell）的方式來治療患者，不但價格昂貴且效果受限，因此想藉由細胞治療以及組織工程的方式來加速傷者骨修復速度。根據衛福部公告，細胞治療已經在多家醫院實行，藉由選取特定細胞加以培養再植入患部，能夠達成減少手術次數、降低感染風險等優勢。臨床醫學案例統計上，發現因車禍意外造成腦損傷患者，其骨頭修復速度較一般患者快速，這與腦神經分泌物突破血腦屏障（blood-brain barrier, BBB）有密切關聯，因此我們使用人類腦神經幹細胞株（human neural progenitor cells/ReNcell™ CX cell line）與初代人類骨髓間質幹細胞（primary human mesenchymal stem cell, hMSCs）在生物反應器中來模擬上述環境，除了添加骨分化試劑外，利用培養神經幹細胞時所分泌出之生長因子，透過系統洄流滋養人類骨髓間質幹細胞，以刺激成骨生長速度加速硬骨類組織成熟化。為了維持神經細胞的生長，本次實驗使用了明膠（gelatin）及人類腦血管內皮細胞株 （human brain endothelial capillary cells, hCMEC/D3 cell line）來做為神經細胞的生物支架，配合海藻膠（alginate）生物支架來促進人類骨髓間質幹細胞形成類硬骨團塊（bone-like tissues）。透過實驗室自組裝的生物反應器，藉由生物反應器的回流裝置輸送生長因子與滋養物質，能夠更貼近真實的人類體循環，最後達到類硬骨團塊成熟化（maturation）之目的。希望透過建立此系統，來突破目前骨科細胞治療市場的困境，造福更多的患者。;With the growing share of aging population, the elderly has crucial problems such as calcium loss and osteoporosis. Thereupon, these aggravated problems lead to bone reconstruction more slowly than the average person. The traditional treatments of bone defects are mostly autologous/allogeneic bone transplantation, but there are problems of immune rejection. Or the clinical doctor would like to use biomaterial substitution, however, how to choose the appropriate material source is very confusing. Moreover, the current cell therapy is expensive and mostly restricted to single cells. According to the announcement of the Ministry of Health and Welfare in Taiwan, the cell therapy has been implemented in many hospitals. The statistics of clinical medical cases showed, it is found that the speed of bone repair in patients with traumatic brain injury (TBI) caused by car accidents is faster than that of ordinary patients. From previous reference we found that is closely related to the breakthrough of blood-brain barrier (BBB) by brain-derived neural secretions. In this study, human neural progenitor cell line (ReNcell™ CX) and primary human mesenchymal stem cells (hMSCs) were applied in the designed bioreactor system to biomimic the micro-environment. The neural-related growth factors secreted by ReNcell™ CX cell line and stimulated hMSCs toward osteogenesis and accelerated bone-like tissue maturation during the dynamic cultivation. In order to maintain the growth of ReNcell™ CX cells, gelatin scaffolds were used as biological scaffolds and human brain endothelial capillary cells (hCMEC/D3) were co-cultured with ReNcell™ CX cells in bioreactor system. At the same time, alginate scaffolds were applied hMSCs to form bone-like tissues. Through the self-designed bioreactor system, growth factors and nourishing substances are delivered through the refluxing device. It is closer to the real human physiological circulation, and finally achieve the purpose of in-vitro bone-like tissue maturation. We sincerely hope the strategy can break through the current predicament of the orthopedic cell therapy market and benefit more patients.
