新型亲水涂层冠脉支架的制备及相关研究
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摘要
目的：制备并表征具有不同亲水程度涂层的冠脉支架,探讨改性材料亲水程度对冠脉支架血液及组织生物相容性的影响,并明确其在生物体内形成血栓的风险及可能机制,为新型支架的研究探索新的方向。
方法：在不锈钢表面进行羟基化,然后进行硅烷化处理,与不同的硅烷偶联剂进行反应,从而在冠脉支架表面形成接合了不同亲水基团的硅烷衍生材料,对这些不同亲水程度的材料表面进行表征；对这些材料进行体外相容性试验,通过溶血实验,动态凝血实验,血小板吸附实验,细胞毒实验检测不同亲水程度材料的生物相容性差别；用前述亲水涂层制备方法制备不同亲水程度的冠脉支架,植入大白兔腹主动脉,通过造影,血管内超声及最后的支架切片免疫组化研究,观察不同亲水程度的支架抗血栓及内皮增生程度及凝血因子等激活的程度指标。多组间计量资料比较采用方差分析,计数资料采用X2检验,P<0.05为差异有统计学意义。
结果：含二氧化钛亲水涂层支架,3-巯基丙基三甲氧基硅烷支架及3-脲基丙基三甲氧基硅烷支架虽然亲水程度较高,但支架表面比较容易吸附血小板,并激活凝血系统,内皮增生情况不良,血中各种凝血及炎症因子水平较高；N-(2-氨乙基)-3-氨丙基三甲氧基硅烷支架、3-氨丙基三乙氧基硅烷(KH550)具有适度的亲水性,血小板吸附较少,不易激活凝血系统,血中各种凝血及炎症因子水平低。
结论：1、可以通过对不锈钢表面羟基化并硅烷处理,然后与硅烷偶联剂共反应的方法制备不同亲水程度的冠脉支架。
2、冠脉支架的生物相容性并非与支架的亲水程度成正比,亲水程度过高的改性材料抗凝血性并不理想。
3、N-(2-氨乙基)-3-氨丙基三甲氧基硅烷支架对血小板的吸附与激活最少,生物相容性好,可以继续接枝各种基团进一步加以改善,是支架改性材料研究的一个新方向。
Fabrication and Study of New Type Hydrophilic Coronary Stent
At present, clinical coronary stents are usually covered by some antiproliferative medicine, such as Rapamycin, Paclitaxel and so on to be a slow-release system in body. But drugs on stents make delayed endothelialization and activate all kinds of cells release cytokine to cause blood clotting. Moreover, it needs a long time to release drugs from stents into blood. So for preventing thrombosis with stents, patients have to take anticoagulants in long term. In order to enhance anticoagulant effect and biocompatibility, researchers are working on changing physical and chemical characters of stents on surface by methods of chemical reaction, graft polymer, spray coating process, electrodeposit, et al.
Our study mainly utilizated advanced chemical technology to make stainless steel surface hydroxylation and silanization. After responding with different silane coupling agents, different hydrophilic groups of the silane-derived materials are fabricated. Biocompatibility and risks of thrombosis of different degree of hydrophilic-coated coronary stents were tested by experiments in vivo and in vitro. The results of our study provide important theoretical and experimental bases for new coronary stents.
The investigation indicated:1. By hydroxylation and silanization, different hydrophilic groups of the silane-derived materials are fabricated.2. The ability of absorbing platelets is not dependent in degree of hydrophilic materials. N-(2-aminoethyl)-3-Ammonia propyl trimethoxysilane stents absorb less platelets on its surface than other hydrophilic materials.3. The haematolysis rate of all hydrophilic materials are all less than 5% and have little toxicity to red blood cells(RBC). N-(2-aminoethyl)-3-Ammonia propyl trimethoxysilane stent and 3-aminopropyl triethoxysilane (KH550) have similar least toxicity to RBC and are better than other hydrophilic materials.4. Assay of cytotoxicity showed that N-(2-aminoethyl)-3 Ammonia propyl trimethoxysilane stent and 3-aminopropyl triethoxysilane (KH550) have little toxicity, which belong to 0 grade:316L stainless steel belongs to 1 grade; the other three materials belong to 2 grade.5. Endothelial cells covered well and less thrombsis and angiostegnosis with N-(2-aminoethyl)-3-Ammoniapropyl trimethoxysilane stent after coronary stent implantation for one month. It has better bioactivity than others.
In this study.we utilizate advanced chemical technology to fabricate different hydrophilic stents. Results of experiments show that biocompatibility of different stents is not directly with their hydrophilic degree. N-(2-aminoethyl)-3-Ammonia propyl trimethoxysilane stent has better biocompatibility with other hydrophilic stents, which may be developed for a novel coronary stent. However, many problems still need to be solved. There is a long way to go before practical use in clinic.
方法：在不锈钢表面进行羟基化,然后进行硅烷化处理,与不同的硅烷偶联剂进行反应,从而在冠脉支架表面形成接合了不同亲水基团的硅烷衍生材料,对这些不同亲水程度的材料表面进行表征；对这些材料进行体外相容性试验,通过溶血实验,动态凝血实验,血小板吸附实验,细胞毒实验检测不同亲水程度材料的生物相容性差别；用前述亲水涂层制备方法制备不同亲水程度的冠脉支架,植入大白兔腹主动脉,通过造影,血管内超声及最后的支架切片免疫组化研究,观察不同亲水程度的支架抗血栓及内皮增生程度及凝血因子等激活的程度指标。多组间计量资料比较采用方差分析,计数资料采用X2检验,P<0.05为差异有统计学意义。
结果：含二氧化钛亲水涂层支架,3-巯基丙基三甲氧基硅烷支架及3-脲基丙基三甲氧基硅烷支架虽然亲水程度较高,但支架表面比较容易吸附血小板,并激活凝血系统,内皮增生情况不良,血中各种凝血及炎症因子水平较高；N-(2-氨乙基)-3-氨丙基三甲氧基硅烷支架、3-氨丙基三乙氧基硅烷(KH550)具有适度的亲水性,血小板吸附较少,不易激活凝血系统,血中各种凝血及炎症因子水平低。
结论：1、可以通过对不锈钢表面羟基化并硅烷处理,然后与硅烷偶联剂共反应的方法制备不同亲水程度的冠脉支架。
2、冠脉支架的生物相容性并非与支架的亲水程度成正比,亲水程度过高的改性材料抗凝血性并不理想。
3、N-(2-氨乙基)-3-氨丙基三甲氧基硅烷支架对血小板的吸附与激活最少,生物相容性好,可以继续接枝各种基团进一步加以改善,是支架改性材料研究的一个新方向。
Fabrication and Study of New Type Hydrophilic Coronary Stent
At present, clinical coronary stents are usually covered by some antiproliferative medicine, such as Rapamycin, Paclitaxel and so on to be a slow-release system in body. But drugs on stents make delayed endothelialization and activate all kinds of cells release cytokine to cause blood clotting. Moreover, it needs a long time to release drugs from stents into blood. So for preventing thrombosis with stents, patients have to take anticoagulants in long term. In order to enhance anticoagulant effect and biocompatibility, researchers are working on changing physical and chemical characters of stents on surface by methods of chemical reaction, graft polymer, spray coating process, electrodeposit, et al.
Our study mainly utilizated advanced chemical technology to make stainless steel surface hydroxylation and silanization. After responding with different silane coupling agents, different hydrophilic groups of the silane-derived materials are fabricated. Biocompatibility and risks of thrombosis of different degree of hydrophilic-coated coronary stents were tested by experiments in vivo and in vitro. The results of our study provide important theoretical and experimental bases for new coronary stents.
The investigation indicated:1. By hydroxylation and silanization, different hydrophilic groups of the silane-derived materials are fabricated.2. The ability of absorbing platelets is not dependent in degree of hydrophilic materials. N-(2-aminoethyl)-3-Ammonia propyl trimethoxysilane stents absorb less platelets on its surface than other hydrophilic materials.3. The haematolysis rate of all hydrophilic materials are all less than 5% and have little toxicity to red blood cells(RBC). N-(2-aminoethyl)-3-Ammonia propyl trimethoxysilane stent and 3-aminopropyl triethoxysilane (KH550) have similar least toxicity to RBC and are better than other hydrophilic materials.4. Assay of cytotoxicity showed that N-(2-aminoethyl)-3 Ammonia propyl trimethoxysilane stent and 3-aminopropyl triethoxysilane (KH550) have little toxicity, which belong to 0 grade:316L stainless steel belongs to 1 grade; the other three materials belong to 2 grade.5. Endothelial cells covered well and less thrombsis and angiostegnosis with N-(2-aminoethyl)-3-Ammoniapropyl trimethoxysilane stent after coronary stent implantation for one month. It has better bioactivity than others.
In this study.we utilizate advanced chemical technology to fabricate different hydrophilic stents. Results of experiments show that biocompatibility of different stents is not directly with their hydrophilic degree. N-(2-aminoethyl)-3-Ammonia propyl trimethoxysilane stent has better biocompatibility with other hydrophilic stents, which may be developed for a novel coronary stent. However, many problems still need to be solved. There is a long way to go before practical use in clinic.
引文
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