骨缺損（Bone defect）所導致的創傷，在臨床上通常會以骨替代物的移植手術作為主要治療手段，常見的骨替代物選擇大致可分成三類；自體移植手術（Autografts）、異體異種移植手術（Allografts、Xenografts）、合成材料的替代物移植手術（Synthetic materials subsites），這些方案都各有部分缺點：自體移植手術受限於可使用的替代部位缺少，雖然具有良好生物活性，但並不能治療大範圍的骨缺損；異體及異種移植手術，可能來自於他人捐贈骨或是動物來源的豬骨、牛骨，經過高溫燒結加工處理後，再進行移植手術，但因此也缺少了生物活性；合成材料的優勢是可以被大量製造，大多來自於高分子或是陶瓷材料，能被使用的損傷部位有限。有鑑於目前臨床治療上有不足，科學家嘗試使用以組織工程（Tissue engineering）技術為核心，結合細胞治療（Cell therapy），期望打造出具有生物活性的類組織來滿足臨床需求；因此本研究建立體外三維（Three-dimensional）共同培養（Co-culture）系統，期望培養同時具有硬骨特徵且具有血管結構的類組織，選用具有能分化成成骨細胞（Osteoblasts）的人類間質幹細胞（Human mesenchymal stem cells）以及能夠形成血管（Blood vessels）的人臍靜脈內皮細胞（Human umbilical vein endothelial cells）；在二維（Two-dimensional）培養確認兩株細胞以1：1比例共培養具有最佳的生物性礦化表現，隨後進行了三維動態培養，有效促進細胞增生、存活率提升、尺寸團提升、分化之礦化物提升的效果。期許本技術能夠製作出符合個人化需求的生物活性硬骨移植功能性類器官，以改善目前骨替代物移植手術的不足之處。;For current clinical practice, bone substitute transplantation is the main therapeutic choice for bone defect treatments. Common options of bone substitute can be roughly divided into four categories： autografts, allografts, xenografts, and synthetic bone substitutes. Each of these options has some benefits and disadvantages. Autologous transplantation is limited by the lack of alternative tissue source, although it is still the gold standard in clinical. Allografts and xenografts may come from other people′s donated bones or even other animal-derived bone tissues, which are sintered at high temperature for removing immune responses but lacks biological activity. The advantage of synthetic materials (such as polymer-based or ceramic-based materials) is that they can be manufactured in large quantities, however, these synthetic materials can be only applied on specific injury sites. In view of the current lack of optimal clinical treatment, scientists are trying to use tissue engineering technique combined with cell therapy, hoping to create bioactive tissue-like substitutes to meet clinical needs. The aim of this study is to cultivate human mesenchymal stem cells (hMSCs) and human umbilical vein endothelial cells (HUVECs) forming biomineralized and vascular structure in-vitro via three-dimensional dynamic co-culture system. From two-dimensional culture condition, we have confirmed the best biomineralization of the two cells co-cultured at a 1:1 ratio. Afterwards, we have confirmed the cell proliferation, differentiation and biomineralization were improved in three-dimensional dynamic co-culture system. This study is expected to be able to create bioactive and functional bone-like tissues that meet individual needs and improve the shortcomings of current bone substitute transplants.
