本文選題：細(xì)胞相容性 + 改性聚乳酸��； 參考：《重慶大學(xué)》2012年碩士論文

【摘要】：改性聚乳酸材料與單純的PLA材料相比在理化性能及生物應(yīng)用等方面具有明顯的優(yōu)勢，因此成為組織工程材料研究的熱點(diǎn)。本研究是將具有良好生物性能的聚乙二醇(PEG)單體引入到聚乳酸中，結(jié)合均苯四甲酸酐共聚形成了PAP單體，進(jìn)一步縮合反應(yīng)生成了PAP-PLA共聚物。首先，采用靜態(tài)水接觸角及吸水率對材料的親/疏水性進(jìn)行了研究，其次對降解產(chǎn)物PAP片段及PMA的細(xì)胞相容性進(jìn)行了考察，同時(shí)對PAP-PLA材料膜的細(xì)胞相容性進(jìn)行了評價(jià)，主要檢測了細(xì)胞的粘附及鋪展形態(tài)、細(xì)胞的增殖、細(xì)胞的分化及礦化等，其中細(xì)胞的粘附及鋪展?fàn)顟B(tài)以顯微鏡及激光共聚焦觀察，細(xì)胞增殖通過CCK-8試劑盒檢測，細(xì)胞分化通過對堿性磷酸酶(ALP)的檢測來考察，而細(xì)胞礦化通過茜素紅法及膠原分泌量來檢測。實(shí)驗(yàn)得出以下主要結(jié)論： ①材料的靜態(tài)水接觸角及吸水率的實(shí)驗(yàn)結(jié)果表明，PAP-PLA共聚物材料的親水性相較于PLA材料有明顯的改善，親/疏水性能合適，而且其親疏水性能可以通過調(diào)節(jié)嵌段的不同比例進(jìn)行調(diào)解，這為材料在組織工程領(lǐng)域的應(yīng)用奠定了基礎(chǔ)。同時(shí)，由于PAP-PLA共聚物材料中所具有的多數(shù)結(jié)構(gòu)及基團(tuán)較為常見，具有良好的細(xì)胞相容性，，而聚酰亞胺由于研究較少，包括其降解產(chǎn)物PMA在內(nèi)的細(xì)胞相容性問題都還有待于檢測。 ②原代成骨細(xì)胞的培養(yǎng)與鑒定實(shí)驗(yàn)表明，獲取的原代成骨細(xì)胞形態(tài)良好，細(xì)胞增殖分化活性高，且具備了礦化能力，適合進(jìn)行相關(guān)細(xì)胞實(shí)驗(yàn)。 ③采用浸提液法直接考察了合成材料中新型嵌段PAP的細(xì)胞相容性。結(jié)果表明，使用PAP浸提不同天數(shù)的浸提液培養(yǎng)的細(xì)胞與對照組相比，細(xì)胞的形態(tài)、增殖、分化礦化等均表現(xiàn)正常，該材料表現(xiàn)出良好的細(xì)胞相容性。 ④同時(shí)采用浸提液法考察了合成材料的降解產(chǎn)物PMA的細(xì)胞相容性。即將PAP和兩組不同PAP-PLA樣品完全降解后可能產(chǎn)生的最大量降解產(chǎn)物PMA浸提于培養(yǎng)液中用于培養(yǎng)成骨細(xì)胞，結(jié)果表明，PAP降解后產(chǎn)生的PMA與對照組相比其分化活力略有下降，但增長趨勢相同，而PAP-PLA樣品組完全降解產(chǎn)生的PMA與對照組不論是形態(tài)、增殖和分化都沒有明顯差異。這些結(jié)果表明PAP-PLA材料降解產(chǎn)物中的PAP片段及PMA沒有明顯的細(xì)胞毒性，具有良好的細(xì)胞相容性。 ⑤對PAP-PLA材料膜上的細(xì)胞粘附及鋪展形態(tài)的檢測表明，過于親水或疏水的表面都不利于細(xì)胞的粘附，其中PAPD4/20與PAPD4/25兩種材料的親疏水性適中，細(xì)胞的粘附及鋪展?fàn)顟B(tài)較好，而PAPD4/15與PLA材料則相對較差。 ⑥對PAP-PLA材料膜上的細(xì)胞的增殖檢測表明，細(xì)胞在PLA上的增殖速率低于PAPD4/20和PAPD4/25，細(xì)胞在PAP-PLA材料的增殖狀況良好，其在各組材料表面上的增殖速率均呈上升趨勢。 ⑦對PAP-PLA材料膜上的細(xì)胞分化和礦化實(shí)驗(yàn)檢測表明，細(xì)胞在PAP-PLA共聚物材料上的分化、礦化及膠原分泌等都較PLA材料要好。 通過以上的檢測表明，PAP-PLA共聚物材料的細(xì)胞相容性良好，這為本改性聚乳酸材料及其他潛在的PAP改性材料的生物相容性評價(jià)提供了良好的借鑒。
[Abstract]:The modified poly (lactic acid) materials have obvious advantages in physical and chemical properties and biological applications compared with simple PLA materials. Therefore, it has become a hot spot in the research of tissue engineering materials. This study is to introduce PEG monomers with good biological properties into polylactic acid, and to form PAP monomers by copolymerization of benzylic anhydride. PAP-PLA copolymer was formed by step condensation reaction. First, the parent / hydrophobicity of the material was studied by the static water contact angle and water absorption. Secondly, the cell compatibility of the degradation product PAP fragment and PMA was investigated, and the cell compatibility of the PAP-PLA membrane was evaluated, and the cell adhesion and spreading morphology were mainly detected. Cell proliferation, cell differentiation and mineralization, in which cell adhesion and spreading state are observed by microscope and laser confocal, cell proliferation is detected by CCK-8 kit, cell differentiation is detected by the detection of alkaline phosphatase (ALP), and cell mineralization is detected by alizarin red method and collagen secretion. To conclude:
The experimental results of the static water contact angle and water absorption of the material show that the hydrophilic property of the PAP-PLA copolymer is obviously improved compared with the PLA material, and the hydrophobicity and hydrophobicity are suitable, and the hydrophobicity can be mediated by adjusting the different proportion of the block. This lays the foundation for the application of the material in the field of tissue engineering. Because most of the structures and groups in PAP-PLA copolymer are more common and have good cellular compatibility, the biocompatibility problems of polyimides, including their degradation product PMA, are still to be detected because of less research.
2. The culture and identification of primary osteoblasts showed that the acquired osteoblasts had good morphology, high proliferation and differentiation activity, and had the ability to mineralogate, which was suitable for the related cell experiments.
(3) the cell compatibility of the new block PAP in the synthetic material was investigated by the extraction method. The results showed that the cells cultured with PAP in different days were compared with the control group, and the cell morphology, proliferation and differentiation mineralization were all normal, and the material showed good cytocompatibility.
At the same time, the biocompatibility of the degradation product PMA of the synthetic material was examined by the leaching solution. The maximum degradation product of the PAP and two groups of different PAP-PLA samples, which may be produced after the complete degradation of the PAP-PLA samples, was extracted in the culture medium for the culture of osteoblasts. The results showed that the PMA produced after the degradation of PAP had a slight differentiation activity compared with the control group. The growth trend was the same, but the total degradation of PMA in the PAP-PLA group was not significantly different from the control group, no matter its morphology, proliferation and differentiation. These results showed that the PAP fragments and PMA in the degradation products of the PAP-PLA material had no obvious cytotoxicity, and had good cytocompatibility.
(5) the detection of cell adhesion and spreading morphology on the PAP-PLA material membrane showed that the overly hydrophilic or hydrophobic surfaces were not conducive to cell adhesion, of which the two materials of PAPD4/20 and PAPD4/25 were moderately hydrophobic, the cell adhesion and spreading state were better, while PAPD4/15 and PLA materials were relatively poor.
6. The proliferation of cells on the PAP-PLA membrane showed that the proliferation rate of the cells on PLA was lower than that of PAPD4/20 and PAPD4/25, and the proliferation of the cells in PAP-PLA material was good, and the proliferation rate of the cells on the surface of each group showed an upward trend.
The test of cell differentiation and mineralization on the PAP-PLA material membrane showed that the differentiation, mineralization and collagen secretion on the PAP-PLA copolymer were better than that of the PLA material.
The results of the above tests show that the PAP-PLA copolymer has good cytocompatibility, which provides a good reference for the biocompatibility evaluation of the modified polylactic acid materials and other potential PAP modified materials.
【學(xué)位授予單位】：重慶大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2012
【分類號】：R318.08

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