利用電氣紡絲製備降解性纖維高分子成型物作為喉部組織工程之評估

Preparation of electrospun biodegradable fiber polymer scaffolds for larynx tissue engineering

周致弘(Chih-Hung Chou)

指導教授 ： 曾厚

臺北醫學大學/醫學院/醫學科學研究所/碩士(2013年)

https://doi.org/10.6831/TMU.2013.00067

摘要

喉癌在頭頸部的癌症當中佔第三位，造成原因有吸菸、飲酒或是外在環境致癌因子，喉癌治療分別為手術治療、放射線治療、化學治療和合併療法，喉癌初期時，只要需進行部份切除或是放射治療，部分切除仍可保留聲帶，保持發聲及吞嚥功能，而放射線治療的最大好處是保留喉部完整構造和功能。喉癌末期，由於聲門上癌或聲門下癌因淋巴豐富，癌細胞很容易經由淋巴轉移，為了防止癌細胞經由淋巴轉移及擴散到周圍組織，術前術及後進行放射治療，如果腫瘤侵犯淋巴結及軟骨組織時，需要施行全喉切除來根除腫瘤，呼吸改由前頸部的永久性氣管造口處進出，導致失去正常發聲、吞嚥、呼吸；為了解決目前全喉切除手術所造成生活品質影響，本研究利用電氣紡絲(electrospinning)，製備除喉嚨組織的細胞支架，並且利用生物玻璃(bioglass)製備出喉嚨軟骨，生物玻璃在過去研究中類似氫氧機磷灰石成分，藉由電氣紡絲技術製作出完整喉嚨組織細胞支架。
本實驗已經成功製作出喉嚨組織工程細胞支架，從喉嚨內部模擬內皮細胞排列到喉嚨結構及喉嚨軟骨成分及結構。製備配向電氣紡絲圓管纖維膜讓內皮細胞生長方向一致利用poly-L-lactic acid (PLLA， 10wt %)溶於DCM:DMF8:2混合；之後圓管支架結合PLLA coil來模擬氣管固定支架結構，為了可以讓細胞貼附移動及增生；我們利用同軸儀器(coaxial)將PLLA纖維披覆一層HA天然性高分子，除了具有親水性外細胞之架具完整性，製作三維細胞支架並依細胞特性製作出多層電器紡絲纖維，像是添加paraffin sphere可增加纖維的孔洞大小，並依照轉速調整及篩網大小控制在電紡絲的孔洞(350μm~500μm) ，將電氣紡絲混和paraffin sphere利用hexane將paraffin溶解，之後利用孔洞測定儀測定薄膜孔洞，喉嚨組織工程細胞支架具有多能性及應用性，具有臨床移植手術的潛能。

關鍵字

聚左乳酸 ； 同軸儀器 ； 電氣紡絲 ； 配向 ； 生物玻璃

並列摘要

Laryngeal cancer is one of the most common diseases for head and neck cancer. Risk factors, such as smoking, drinking, and external environmental carcinogen, may induce its tumor growth. Treatments of laryngeal cancer include surgery, radiotherapy, chemotherapy, and a combination of therapies. In the early stage, tumors can be removed partially by surgery or treated by chemotherapy. After surgery, the remaining vocal cords can be retained and swallowing and voice functions, maintained. The advantage of chemotherapy is the retention of the full structure and function of the throat. For malignant laryngeal cancer, such as subglottic and supraglottic cancers, metastasis of tumor cells is likely to happen via lymphatic tissue due to the abundant amount of lymph content. Pre-operative and post-operative radiotherapies are usually performed to prevent the spread of cancer cells into the surrounding tissue. For metastatic tumor cells already in the lymph nodes and cartilage soft tissue, total laryngectomy surgery is necessary to remove the whole tumor. The application of total laryngectomy surgery removes the entire larynx and will create a hole for respiration. This permanent hole goes through the front of the neck to the trachea, and will cause the loss of voice, swallowing, and breathing.

The purpose of this study is to investigate ways to improve the quality of life after total laryngectomy surgery. We chose hydroxyapatite-rich bioactive glass (bioglass) as our material to prepare a tissue-engineered throat cartilage scaffold by the electrospinning technique. The structure and the composition of the throat scaffold, and mimic of arrangement of endothelial cells in the inner layer of the scaffold are successfully prepared and well characterized. In this study, biodegradable poly-L-lactic acid (PLLA, 10wt %) was dissolved in a solvent mixture of dichloromethane (DCM) and N,N-dimethylformamide (DMF) at a 8:2 (v/v) ratio. The mimic of Trachea of PLLA coil was combined with electrospun tubular scaffold. In order to promote cell adhesion and proliferation, a layer of hyaluronic acid polymer was coated onto the PLLA fiber surface by using a coaxial technique to improve the integrity and hydrophilicity of the scaffold. Electrospun membranes were created to form a 3-D multi-layer shape , and further modified in accordance with different requirements, ie. increasing the pore size (350μm-500μm) by adding paraffin sphere based on mesh size and the control of rotation rate. Paraffin spheres were then dissolved in hexane solution. Pore sizes and distributions were characterized by using a capillary flow porometer; larynx tissue engineering scaffold has the versatility and applicability of clinical transplant surgery potential.

並列關鍵字

PLLA ； coaxial ； Electrospinning ； Alignment ； bioglass

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