The Control Release of Chondroitinase ABC in Chitosan Based Scaffold and PDLLA Microsphere

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Project title

Code/計畫編號

NSC97-2314-B019-001

Translated Name/計畫中文名

利用幾丁聚醣支架與聚乳酸微球體進行軟骨素分解酵素之控制釋放

Project Coordinator/計畫主持人

Yi-Cheng Huang

Funding Organization/主管機關

National Science and Technology Council

Department/Unit

Department of Food Science

Website

https://www.grb.gov.tw/search/planDetail?id=1720319

Year

2008

Start date/計畫起

01-11-2008

Expected Completion/計畫迄

01-07-2009

Bugetid/研究經費

600千元

ResearchField/研究領域

食品科技(工)

"脊髓受傷時，被活化的星狀細胞會移至受傷處，並分泌細胞外基質；這些細胞外基質被稱為疤痕組織，會阻礙新生的神經細胞與軸突的生長，阻礙神經修復。軟骨素分解酵素(Chondroitinase ABC, 簡稱 ChABC)已被證實可分解疤痕組織中醣蛋白結構的醣鏈，有助於神經細胞與軸突的向外生長，產生功能性的回覆，並幫助神經修復。然而，有鑒於ChABC的不穩定，且需要長時間使用ChABC才可有效分解醣鏈，因此，本實驗利用幾丁聚醣支架及聚乳酸微小球探討Ch ABC的控制釋放，以維持Ch ABC的穩定及活性，達到神經修復的目的。關於利用幾丁聚醣支架探討Ch ABC的控制釋放：我們利用幾丁聚醣及明膠製備的神經導管為固定化酵素的材料，將Ch ABC以離子鍵結或共價鍵結固定於導管上。過去我已利用幾丁聚醣製備神經導管，應用於神經再生的研究，然而，導管孔隙度的大小明顯影響神經修復的結果，因此，本實驗我將加入明膠，利用明膠與幾丁聚醣形成複合性材料，來控制導管的孔隙度。此外，這部分實驗亦將比較兩種不同的化學鍵結(離子鍵結與共價鍵結)對於酵素固定的成效。關於聚乳酸微小球探討Ch ABC的控制釋放：我們利用雙層乳化法(Double Emulsion Method)製備微小球，包覆水溶性的分解酵素Ch ABC。如何提高包覆酵素的活性以及延長釋放時間，是這方法必需克服的重要問題。過去，血清白蛋白是維持酵素活性常用的穩定劑，但在本實驗並不適用，因為血清白蛋白在脊髓會引起發炎反應。因此，本實驗將著重於尋找適當的方式，維持Ch ABC 的穩定性及活性，達到控制釋放的目的。" "In spinal cord injury, activated astrocytes migrate to the injury site and secrete the extracellular matrix (ECM), called scar tissue. The scar tissue will impede the re-growth of neurons and axons, and then inhibit nerve regeneration. It has been proved that Chondroitinase ABC (Ch ABC) can digest the polysaccharide of proteoglycan in scar tissue, and promote the re-growth of neuron, axon and behavior recovery. In order to digest the polysaccharide effectively, it is important to sustain release of Ch ABC. Furthermore, maintain the stability and activity of Ch ABC while sustained release is also a key factor. In this research, we will discuss the control release of Ch ABC in chitosan nerve conduit and PDLLA microspheres. As for the control release of Ch ABC in a chitosan nerve conduit, I will fabricate the nerve conduit by chitosan and gelatin. And then, I will immobilize Ch ABC on the surface of nerve conduits by ionic bonding or covalent bonding. I have used chitosan as the material to fabricate nerve conduit for nerve regeneration. However, I found that the pore size of conduits affect the result of nerve regeneration. Therefore, in this research, I will mix another material, gelatin, to chitosan to control the pore size of conduits. In addition, I will compare the results of immobilizing Ch ABC by the ways: ionic bonding and covalent bonding. As for the control release of Ch ABC in PDLLA microsphere, I will fabricate microspheres by double emulsion method to encapsulate the water dissolvable enzyme, Ch ABC. However, there are two key problems should be overcome, sustained release of Ch ABC and keep the activity of Ch ABC. Albumin is a frequently used stabilizer for protecting enzyme and maintaining its activity while releasing in past researches. Nonetheless, it is not suitable in my system because of inflammation reactions in spinal cord induced by albumin. This part of my research will focus on finding a way to stabilize Ch ABC for sustained release"

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The Control Release of Chondroitinase ABC in Chitosan Based Scaffold and PDLLA Microsphere

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