中国组织工程研究
中國組織工程研究
중국조직공정연구
Journal of Clinical Rehabilitative Tissue Engineering Research
2014年
30期
4851-4856
,共6页
生物材料%缓释材料%聚乙二醇水凝胶%生长因子%可控性释放%内皮祖细胞%细胞增殖%国家自然科学基金
生物材料%緩釋材料%聚乙二醇水凝膠%生長因子%可控性釋放%內皮祖細胞%細胞增殖%國傢自然科學基金
생물재료%완석재료%취을이순수응효%생장인자%가공성석방%내피조세포%세포증식%국가자연과학기금
polyethylene glycols%hydrogel%fibroblast growth factors%vascular endothelial growth factors
背景：大量研究表明，为了实现聚合物材料的内皮细胞化，向材料表面负载生物活性因子是一种重要的手段，并且在聚合物材料表面引入人体血管内皮细胞将有助于提高材料的生物相容性。目的：合成封装碱性成纤维细胞生长因子、血管内皮生长因子和内皮祖细胞的水凝胶，观察生长因子的缓释效果，以及内皮祖细胞在水凝胶中的培养状态。方法：合成含精氨酸-甘氨酸-天冬氨酸肽段的聚乙二醇水凝胶，先后加入大鼠内皮祖细胞、碱性成纤维细胞生长因子、血管内皮生长因子溶液、基质金属蛋白酶反应底物肽段溶液。将上述水凝胶浸泡于 PBS 中，每12 h用ELISA法检测上清液中生长因子的水平；72 h后在水凝胶PBS溶液中再分别加入不同质量的基质金属蛋白酶2(100，1000 ng)、基质金属蛋白酶9(100，1000 ng)，每12 h用ELISA法检测上清液中生长因子的水平。将封装内皮祖细胞的聚乙二醇水凝胶置于培养基中培养5 d，消化后用流式细胞仪检测存活细胞数。结果与结论：12-72 h内，碱性成纤维细胞生长因子、血管内皮生长因子的释放百分比一直维持在41%左右，72 h后分别加入不同质量的基质金属蛋白酶2、基质金属蛋白酶9，发现两种细胞因子的释放百分比呈稳步上升，72 h后已达95%以上，且基质金属蛋白酶质量越大两种因子的检测释放百分比越高。培养5 d后，在聚乙二醇水凝胶中仍有88.17%的内皮祖细胞存活。表明自组装聚乙二醇水凝胶既可以实现碱性成纤维细胞生长因子、血管内皮生长因子的可控性释放，又可以支持内皮祖细胞的增殖生长。
揹景：大量研究錶明，為瞭實現聚閤物材料的內皮細胞化，嚮材料錶麵負載生物活性因子是一種重要的手段，併且在聚閤物材料錶麵引入人體血管內皮細胞將有助于提高材料的生物相容性。目的：閤成封裝堿性成纖維細胞生長因子、血管內皮生長因子和內皮祖細胞的水凝膠，觀察生長因子的緩釋效果，以及內皮祖細胞在水凝膠中的培養狀態。方法：閤成含精氨痠-甘氨痠-天鼕氨痠肽段的聚乙二醇水凝膠，先後加入大鼠內皮祖細胞、堿性成纖維細胞生長因子、血管內皮生長因子溶液、基質金屬蛋白酶反應底物肽段溶液。將上述水凝膠浸泡于 PBS 中，每12 h用ELISA法檢測上清液中生長因子的水平；72 h後在水凝膠PBS溶液中再分彆加入不同質量的基質金屬蛋白酶2(100，1000 ng)、基質金屬蛋白酶9(100，1000 ng)，每12 h用ELISA法檢測上清液中生長因子的水平。將封裝內皮祖細胞的聚乙二醇水凝膠置于培養基中培養5 d，消化後用流式細胞儀檢測存活細胞數。結果與結論：12-72 h內，堿性成纖維細胞生長因子、血管內皮生長因子的釋放百分比一直維持在41%左右，72 h後分彆加入不同質量的基質金屬蛋白酶2、基質金屬蛋白酶9，髮現兩種細胞因子的釋放百分比呈穩步上升，72 h後已達95%以上，且基質金屬蛋白酶質量越大兩種因子的檢測釋放百分比越高。培養5 d後，在聚乙二醇水凝膠中仍有88.17%的內皮祖細胞存活。錶明自組裝聚乙二醇水凝膠既可以實現堿性成纖維細胞生長因子、血管內皮生長因子的可控性釋放，又可以支持內皮祖細胞的增殖生長。
배경：대량연구표명，위료실현취합물재료적내피세포화，향재료표면부재생물활성인자시일충중요적수단，병차재취합물재료표면인입인체혈관내피세포장유조우제고재료적생물상용성。목적：합성봉장감성성섬유세포생장인자、혈관내피생장인자화내피조세포적수응효，관찰생장인자적완석효과，이급내피조세포재수응효중적배양상태。방법：합성함정안산-감안산-천동안산태단적취을이순수응효，선후가입대서내피조세포、감성성섬유세포생장인자、혈관내피생장인자용액、기질금속단백매반응저물태단용액。장상술수응효침포우 PBS 중，매12 h용ELISA법검측상청액중생장인자적수평；72 h후재수응효PBS용액중재분별가입불동질량적기질금속단백매2(100，1000 ng)、기질금속단백매9(100，1000 ng)，매12 h용ELISA법검측상청액중생장인자적수평。장봉장내피조세포적취을이순수응효치우배양기중배양5 d，소화후용류식세포의검측존활세포수。결과여결론：12-72 h내，감성성섬유세포생장인자、혈관내피생장인자적석방백분비일직유지재41%좌우，72 h후분별가입불동질량적기질금속단백매2、기질금속단백매9，발현량충세포인자적석방백분비정은보상승，72 h후이체95%이상，차기질금속단백매질량월대량충인자적검측석방백분비월고。배양5 d후，재취을이순수응효중잉유88.17%적내피조세포존활。표명자조장취을이순수응효기가이실현감성성섬유세포생장인자、혈관내피생장인자적가공성석방，우가이지지내피조세포적증식생장。
BACKGROUND:Many studies have demonstrated that in order to achieve the endothelialization of the polymer materials, surface loading of bioactive factors is an important means and the introduction of human endothelial cells on the surface of the polymer materials wil contribute to increase the biocompatibility of materials. OBJECTIVE:To synthesize the polyethylene glycol hydrogel packaging vascular endothelial growth factor, basic fibroblast growth factor and endothelial progenitor cells, and to observe the sustained release of growth factors and culture status of endothelial progenitor cells. METHODS:The polyethylene glycol-modified hydrogel containing arginine-glycine-aspartic acid peptide was synthesized, and then, rat endothelial progenitor cells, basic fibroblast growth factor, vascular endothelial growth factor, matrix metal oproteinases were successively added. The hydrogel were immersed in PBS, and ELISA was used to test the levels of growth factors in supernatants every 12 hours. Matrix metal oproteinases-2 (100, 1 000 ng) and matrix metal oproteinases-9 (100, 1 000 ng) at different mass were added into the PBS containing hydrogel after 72 hours. ELISA was also used to test the levels of growth factors in supernatants every 12 hours. The hydrogel encapsulating endothelial progenitor cells was cultured in a culture medium for 5 days, and the number of survival cells was counted by a flow cytometer after digestion. RESULTS AND CONCLUSION:Within 12-72 hours, the release percentage of basic fibroblast growth factor and vascular endothelial growth factor remained at about 41%. After 72 hours, the release percentage of both growth factors was found to grow steadily when matrix metal oproteinases-2 (100, 1000 ng) and matrix metal oproteinases-9 (100, 1 000 ng) at different mass were added, which reached 95%. The release percentage was increased with the increasing mass of matrix metal oproteinases. After 5 days of culture, 88.17%cells stil survived in the hydrogel. These findings indicate that sel-assembly polyethylene glycol-modified hydrogel can realize the control ed release of basic fibroblast growth factor and vascular endothelial growth factor, and it can also support the proliferative growth of endothelial progenitor cells.
