應用於硬骨再生之聚己內酯/氫氧基磷灰石奈米複合膜材經油酸調節之體外生物相容性評估

中山醫學大學機構典藏 CSMUIR > 口腔醫學院 > 口腔科學研究所 > 博碩士論文 > Item 310902500/6824

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题名:	應用於硬骨再生之聚己內酯/氫氧基磷灰石奈米複合膜材經油酸調節之體外生物相容性評估 In vitro biocompatibility evaluation of polycaprolactone / hydroxyapatite nanocomposite membrane by oleic acid mediation
作者:	陳柏翔 Po-Hsiang Chen
贡献者:	中山醫學大學口腔材料科學研究所;湯正明
关键词:	聚己內酯;氫氧基磷灰石;奈米複合膜材;油酸;模擬體液;人類臍帶間葉幹細胞;成骨細胞株 polycaprolactone;hydroxyapatite;nanocomposite;oleic acid;simulated body fluids;human umbilical cord mesenchymal stem cells;osteoblasts cell line
日期:	2013
上传时间:	2013-12-23T03:20:59Z (UTC)
摘要:	牙周病末期患者容易產生齒槽骨缺損，因軟組織生長較硬骨組織為快，臨床治療需要適合的膜材，提供阻隔性與骨引導性以加快硬骨修補速度。本研究選擇具生物可降解性之聚己內酯做為複合膜材的基材，以油酸調解氫氧基磷灰石在高分子基質中的分散性，製備出聚己內酯/油酸/氫氧基磷灰石的奈米陶瓷複合膜材，將膜材浸泡於模擬體液中7天以模擬生物礦化過程，結果發現低結晶度之聚己內酯/氫氧基磷灰石膜材較有高結晶度之聚己內酯/氫氧基磷灰石膜材有明顯且均勻的礦化層形成，而以油酸調解氫氧基磷灰石所製備之膜材產生團狀上帶針型的礦化層。以低掠角繞射量測表面礦化層具有氫氧基磷灰石特性繞射峰，而以反射式傅立葉轉換紅外線光譜分析礦化層具有磷酸根官能基吸收。材料生物相容性評估分別以人類臍帶間葉幹細胞(hUCMSC)與成骨細胞株 (MC3T3) 進行細胞存活率測定， hUCMSC培養在膜材上24小時後， PCL-HA膜材與PCL-C18-HA膜材之細胞存活率表現比TCPS高，而MC3T3培養PCL-C18-HA膜材表現最高。PCL-HA 膜材與PCL-C18-HA膜材上有觀察到細胞產生明顯的絲狀與扇狀偽足。將hUCMSC培養在複合膜材表面10天後，以茜素紅S染色觀察在膜材上的細胞分化，結果發現PCL-HA膜材與PCL-C18-HA膜材表面有明顯鈣離子沉積生成，證實hUCMSC細胞受到HA奈米粉體刺激而產生分化。另外，測定細胞在複合膜材上過氧化物質(ROS)的產生量，發現PCL-C18-HA膜材對hUCMSC細胞過氧化物質表現量顯著高於TCPS，但對MC3T3細胞卻沒有顯著差異，推測與油酸的釋放有關。本研究所製備之複合膜材證實能提供良好的隔絕性並引導硬骨再生，可做為組織導引再生膜以及硬骨組織工程等方面之應用。 A serious periodontal disease prone to alveolar bone defects due to growth rate of soft tissue faster than hard bone tissue. A suitable membrane providing guiding and barrier to accelerate bone repair bony speed in clinical therapy. In our study, polycaprolactone/oleic acid/hydroxyapatite nanocomposite membrane was prepared by a biodegradable polycaprolactone combined hydroxyapatite by oleic acid mediated. The membrane was immersed in SBF for 7 days to simulate the process of biomineralization. The results showed a low degree of crystallinity of polycaprolactone composite membranes compared with a high degree of crystallinity of polycaprolactone composite membranes significantly mineralized layer and uniform, needle-type mineralized layer was formation on polycaprolactone/oleic acid/hydroxyapatite nanocomposite surface. Mineralized layer showed presence of phosphate ion absorption and Hydroxyapatite diffraction peaks by reflection Fourier transform infrared spectroscopy and small-angle diffraction measure. In order to assess biocompatibility of composite, human umbilical cord mesenchymal stem cells (hUCMSC) and osteoblast cell line (MC3T3) for cell viability determination. The results show hUCMSC culture after 24 hours on the membrane, PCL-HA membrane and PCL-C18-HA membrane of the cell survival rate higher than TCPS, and MC3T3 cultured on PCL-C18-HA membrane performance up. PCL-HA membrane and PCL-C18-HA membranes have observed significant cell filopodia and lamellipodia. The hUCMSC cultureed on composite membrane after 10 days then cell staining by alizarin red S, and found that calcium deposition was generated on PCL-HA membrane and PCL-C18-HA surface membrane, confirmed hUCMSC cells are stimulated to produce HA nanopowders differentiation. Peroxidation species (ROS) generation of cells was determined, found ROS release of PCL-C18-HA membrane peroxidation on hUCMSC expression was significantly higher than TCPS, but no significant difference was MC3T3 cells. The composite provide good barrier properties and guide bony regeneration guided tissue regeneration membrane can be used as well as the hard bone tissue engineering and other aspects of the application.
URI:	https://ir.csmu.edu.tw:8080/ir/handle/310902500/6824
显示于类别:	[口腔科學研究所] 博碩士論文

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