中国组织工程研究 ›› 2014, Vol. 18 ›› Issue (47): 7585-7590.doi: 10.3969/j.issn.2095-4344.2014.47.008

• 纳米生物材料 nanobiomaterials • 上一篇    下一篇

静电纺丝聚乳酸复合物纳米纤维材料与小鼠神经干细胞的生物相容性

刘  畅,戎利民,李尚福,庞  卯,杨  阳,刘  斌   

  1. 中山大学附属第三医院脊柱外科,广东省广州市  510630
  • 修回日期:2014-10-10 出版日期:2014-11-19 发布日期:2014-11-19
  • 通讯作者: 刘斌,博士,副主任医师。中山大学附属第三医院脊柱外科,广东省广州市 510630
  • 作者简介:刘畅,男,安徽省人,汉族,1990年生,2012年皖南医学院毕业,硕士。
  • 基金资助:

    国家自然科学基金(31170947,31470949,81472122);广东省自然科学基金(S2012020011099,S2013010016413);广东省科技计划项目(2012B060300008);广州市科技计划项目(2013J4100062);教育部博士点新教师基金(20100171120088)

Biocompatibility of electrospun poly(lactide-co-glycolide)/polyethylene glycol nanofibrous scaffold with mouse neural stem cells

Liu Chang, Rong Li-min, Li Shang-fu, Pang Mao, Yang Yang, Liu Bin   

  1. Department of Spinal Surgery, Third Affiliated Hospital of Sun Yat-sen University, Guangzhou 510630, Guangdong Province, China
  • Revised:2014-10-10 Online:2014-11-19 Published:2014-11-19
  • Contact: Liu Bin, M.D., Associate chief physician, Department of Spinal Surgery, Third Affiliated Hospital of Sun Yat-sen University, Guangzhou 510630, Guangdong Province, China
  • About author:Liu Chang, Master, Department of Spinal Surgery, Third Affiliated Hospital of Sun Yat-sen University, Guangzhou 510630, Guangdong Province, China
  • Supported by:

    the National Natural Science Foundation of China, No. 31170947, 31470949, 81472122; Guangdong Natural Science Foundation, No. S2012020011099, S2013010016413; Guangdong Science and Technology Planning Project, No. 2012B060300008; Guangzhou Science and Technology Planning Project, No. 2013J4100062; New Teachers’ Fund for Doctor Stations of Ministry of Education of China, No. 20100171120088

摘要:

背景:聚乳酸聚乙醇酸支架材料广泛应用于组织工程学领域,但其细胞黏附性较差、缺乏活性功能基团以及疏水性较强等缺点限制了其进一步的发展和应用。
目的:观察小鼠神经干细胞与静电纺丝聚乳酸聚乙醇酸/聚乙二醇共聚物纳米纤维支架材料的体外相容性。
方法:自孕15 d CD-1小鼠胚胎大脑皮质分离培养小鼠神经干细胞。静电纺丝法制备聚乳酸聚乙醇酸和聚乳酸聚乙醇酸/聚乙二醇纳米纤维支架材料,扫描电镜观察材料结构;取第5代神经干细胞分别接种于聚乳酸聚乙醇酸和静电纺丝聚乳酸聚乙醇酸/聚乙二醇纳米纤维支架材料上,进行体外培养。
结果与结论:扫描电镜检测显示,两种支架材料呈现相互交联的多孔网状结构。聚乳酸聚乙醇酸组和静电纺丝聚乳酸聚乙醇酸/聚乙二醇组纤维直径和孔隙率差异无显著性意义(P > 0.05)。CCK-8检测显示,两种材料无明显细胞毒性。神经干细胞在支架材料中生长良好,两组吸光度值均随培养时间延长而增大,两组在培养  1,3,5,7,9,11 d 吸光度值差异均有显著性意义(P < 0.05)。两组材料培养3,6,9 h,静电纺丝聚乳酸聚乙醇酸/聚乙二醇组的细胞黏附率明显高于聚乳酸聚乙醇酸组(P < 0.05)。Hoechst染色显示两组细胞核质均染,形态正常,静电纺丝聚乳酸聚乙醇酸/聚乙二醇组细胞数量明显多于聚乳酸聚乙醇酸组(P < 0.05)。扫描电镜观察显示,与聚乳酸聚乙醇酸组相比,静电纺丝聚乳酸聚乙醇酸/聚乙二醇组神经干细胞在支架上的生长情况和基质分泌更好。结果说明,静电纺丝法制备的静电纺丝聚乳酸聚乙醇酸/聚乙二醇纳米纤维支架细胞生物相容性良好,安全无毒,具备合适的孔径和孔隙率,适宜神经干细胞生长,是一种适用于组织工程优质的支架载体。


中国组织工程研究杂志出版内容重点:生物材料;骨生物材料; 口腔生物材料; 纳米材料; 缓释材料; 材料相容性;组织工程


全文链接:

关键词: 生物材料, 材料相容性, 纤维纳米支架材料, 静电纺丝, 聚乳酸聚乙醇酸, 聚乙二醇, 复合支架, 生物相容性, 小鼠神经干细胞, 细胞毒性, 细胞增殖, 细胞黏附率, 国家自然科学基金

Abstract:

BACKGROUND: Poly(lactide-co-glycolide) (PLGA) scaffold is widely used in tissue engineering, but its poor cell adhesion ability and strong hydrophobicity limit its further development and application.
OBJECTIVE: To study the biocompatibility of electrospun poly (lactide-co-glycolide)/polyethylene glycol (PLGA-PEG) nanofibrous scaffolds with mouse neural stem cells in vitro.
METHODS: Neural stem cells were isolated from embryos of CD-1 mice at 15 embryonic days. Electrospinning was used to prepare PLGA and PLGA-PEG nanofibrous scaffolds. Scanning electron microscope was used for scanning observation of scaffolds. The 5th passage neural stem cells were seeded onto PLGA and PLGA-PEG scaffolds respectively, and cultured in vitro.
RESULTS AND CONCLUSION: Interconnected porous network structure was observed in both two kinds of scaffolds under the scanning electron microscope. Fiber diameters and porosities of PLGA and PLGA-PEG scaffolds showed no significant differences (P > 0.05). Cell Counting Kit-8 detection showed neural stem cells grew well on both two kinds of scaffolds and the absorbance value of two groups increased continuously with incubation time (1, 3, 5, 7, 9, 11 days). And there were statistically significant differences in the absorbance values between two groups at each time point (P < 0.05). Moreover, the cell adhesion rate was significantly higher in the PLGA-PEG group than in the PLGA group at 3, 6, 9 hours of culture (P < 0.05). Hoechst 33342 staining showed normal morphology and quality of the nuclei, and significantly more cells were observed in the PLGA-PEG group than the PLGA group (P < 0.05). Under the scanning electron microscope, compared with the PLGA scaffold, the PLGA-PEG scaffold was better for growth and matrix secretion of neural stem cells. In conclusion, PLGA-PEG nanofibrous scaffolds prepared by electrospinning are safe, non-toxic and suitable for neural stem cells growth with well biocompatibility, appropriate aperture and porosity.


中国组织工程研究杂志出版内容重点:生物材料;骨生物材料; 口腔生物材料; 纳米材料; 缓释材料; 材料相容性;组织工程


全文链接:

Key words: biocompatible materials, neural stem cells, mice, cell proliferation

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