2012-08-012024-05-14https://scholars.lib.ntu.edu.tw/handle/123456789/657011摘要：關節軟骨缺損一直是臨床治療上一個棘手的問題，由於軟骨的自我修復能力極低，在臨床上多半採用自體移植或是人工關節來治療或取代，然而這些均不是一個長期有效的治療方式。近幾年由於組織工程學的興起，結合材料、生物與醫學等領域為關節軟骨缺損的治療提供了一個新的方向。如何幫助軟骨細胞成功地在體外生物醫材中立體培養，一直被熱烈地討論與研究，目前大多數提出的自體軟骨治療缺損模式均採用兩階段模式:第一階段為第一次手術取出健康可用之軟骨後結合組織工程於體外大量培養，第二階段為第二次手術植入體外培養之軟骨細胞與醫材來修補缺損。此治療模式之缺點為體外培養軟骨細胞有造成軟骨細胞變性之疑慮，且需進行二次手術，手術間有數週之軟骨缺損之等待期。故本研究團隊提出一次手術治療之觀念－自體即為最好的生物反應器，我們在一次手術中，先取出健康可用之軟骨，利用組織切片機將軟骨切碎，並將軟骨碎片放進本團隊先前研發之兩相骨材中，直接植入欲修補區域，利用自體之體內環境當作生物反應器以幫助植入軟骨片生長以進行修補。基於此，軟骨細胞或碎片如何在體內進行有效地增生與基質修補便是一個極為重要的課題。由於軟骨塊中的軟骨細胞被大量的細胞外基質包覆，不利細胞接收訊息及營養，經適當之酵素處理溶釋細胞外基質後可促使細胞增生，本計畫欲建立一臨床動物模式來探討酵素溶釋軟骨塊之處理對於軟骨在體內進行自我修補之影響。試驗條件分為:(A) 單純兩相載體、(B) 兩相載體加切碎之軟骨碎片與(C) 兩相載體加經由酵素處理之切碎軟骨碎片。我們將使用12 隻成年猪進行實驗，每隻猪隻之左右後腳股骨髁各進行一個試驗條件，試驗條件將隨機分配至24 個股骨髁，術後六個月與十二個月進行MRI 與採樣評估。此實驗結果不僅可協助改進本團隊提出之兩相骨材同時修補軟硬骨之一次手術流程，亦可為臨床軟骨治療提供重要參考資訊。<br> Abstract: Repair of articular cartilage defect is a significant issue in clinical field due to its poorrepair capacity. At present, the main treatments are autograft and allograft transplantationusing engineered cartilage fragments or chondrocytes. However, the results are notsatisfactory and long-standing. Due to the advancement in tissue engineering field, repair ofthe articular cartilage defect shifts to a new direction by combining expertise in materialscience, biology and medicine. Development of an effective method for 3-D chondrocyteculture in a biomaterial scaffold has drawn a lot of attention. Autogenous/allogenicchondrocyte implantation (ACI) is the current strategy for cartilage regeneration thatrequires two surgeries. The first surgery serves to isolate healthy chondrocytes forregeneration in 3-D scaffold in vitro. Such treatments include a biodegradable biomaterialscaffold to hold the implanted chondrocytes in place therefore are beyond cell therapy andpertain to tissue engineering. The second surgery is to fill the defect using scaffoldcontaining cultured chondrocytes. The disadvantage of this protocol is that the regeneratedchondrocyte in 3-D scaffold in vitro may induce chondrocyte transformation. In addition, Itrequire two sugeries and several-week in-between waiting period with the defect unrepaired.Based on the hypothesis that the body is the best bioreactor, our research team purposes asingle surgery protocol to repair the articular cartilage defect. At the surgery, the cartilage isisolated and chopped uniformly by a tissue cutter, treated with enzymes, placed into abi-phase scaffold, and then inserted into the defect site for autologous chondrocyteregeneration. The effectiveness of chondrocyte proliferation and extracellular matrix (ECM)secretion in vivo becomes very important in this treatment. Since the chondrocytes aresurrounded by abundant ECM that interfere the cells receiving growth signals and nutrition,appropriate enzyme treatment to loose chondrocytes should be able to promote regeneration.In this study, we will seed the cartilage fragments with or without enzyme treatment intobi-phase scaffold and implant into articular cartilage defect in pig animal model. Theproposed treatment groups in this project will include (A) bi-phase scaffold only; (B)bi-phase scaffold containing cartilage fragments; (C) bi-phase scaffold containing theenzyme-treated cartilage fragments. Twelve adult pigs will be used in the proposedexperiment. The rear femoral condyle (24 in total) will be randomly assigned to one of thethree treatment groups. All the procedure will be completed in one single surgery and theMRI and the biopsy evaluation will be retrieved six and twelve months after the singlesurgical treatment. The results will not only amend the protocol of bi-CRI (BiphasicCartilage Repair Implant) ongoing clinical treatment, but also provide important referenceinformation for clinical cartilage repair.軟骨細胞酵素溶釋軟骨修復兩相骨材