| 吕兰欣,杨红宁,颜晓庆,胡书群,燕宪亮,许 铁.氨基修饰的纳米纤维对人和大鼠 MSCs增殖分化的影响[J].现代生物医学进展英文版,2018,(18):3420-3424. |
| 氨基修饰的纳米纤维对人和大鼠 MSCs增殖分化的影响 |
| The Effect of Amino-group Modified Nanofibers on Proliferation and Differentiation of MSCs derived from Human and Rat Bone Marrow |
| Received:April 22, 2018 Revised:May 18, 2018 |
| DOI：10.13241/j.cnki.pmb.2018.18.004 |
| 中文关键词: 纳米纤维 氨基修饰 成骨分化 间充质干细胞 |
| 英文关键词: Nanofibers Amino-group modification Osteogenesis Mesenchymal stem cells |
| 基金项目:国家自然科学基金项目（81271267）；江苏省高校自然科学研究面上项目（16KJB310018）；徐州市科技计划项目（KC16SY156） |
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| 中文摘要: |
| 摘要 目的：探讨氨基修饰后的静电纺丝纳米纤维对大鼠和人骨髓来源的间充质干细胞（Rat and human bone marrow mesenchymal stem cells, rMSCs and hMSCs）增殖及成骨分化的影响。方法：采用静电纺丝法制备聚乳酸 -羟基乙酸共聚物(poly(lactic-co-glycolic acid)，PLGA)纳米纤维，用氨气等离子体处理其表面来接枝氨基；通过测量 PLGA纳米纤维（NF）及氨基修饰后的纳米纤维（NF-NH2）接触角来证明修饰效果；将 rMSCs和 hMSCs分别接种于 NF和 NF-NH2，用 CCK-8试剂盒检测接种后 1, 3 (4), 7天的细胞增殖；接种后的 21天，用茜素红 S染色 (ARS)法检测细胞成骨分化情况。结果：氨气等离子体处理后纳米纤维接触角从81.28 ± 0.33降低至 53.99 ± 0.79，说明氨基修饰后的 PLGA NF亲水性增加；CCK-8结果显示氨基修饰增加了 rMSCs的黏附，接种 24 h后 rMSCs在 NF和 NF-NH2上的检测吸光值分别为 0.096 ± 0.011和 0.175 ± 0.014（ P<0.001），而对 hMSCs黏附和增殖没有影响，接种 24 h后 hMSCs在 NF和 NF-NH2上的检测吸光值分别为 0.237 ± 0.004和 0.238 ± 0.006（ P>0.05）；ARS染色结果显示氨基修饰后 rMSCs成骨分化增多（在 NF和 NF-NH2表面 ARS染色区域比例分别 13.147 ± 3.223%和 36.677 ± 5.230%），而hMSCs在修饰前后的纳米纤维上均有表达（修饰前后 ARS染色比例分别为 50.283 ± 2.942%和 38.254 ± 3.272%）。结论：氨基修饰的 NF可以促进大鼠来源的 MSCs黏附增殖以及成骨分化，而对人骨髓来源的 MSCs没有显著影响，这提示我们 MSCs的增殖分化行为可能具有种属依赖性。 |
| 英文摘要: |
| ABSTRACT Objective: To investigate the effects of ammine group modified electrospinning nanofibers on the proliferation and osteogenic differentiation of rat and human bone marrow derived mesenchymal stem cells (rMSCs, hMSCs). Methods:Electrospun technique was employed to fabricate poly(lactic-co-glycolic acid) (PLGA) nanofibers, whose surface was grafted with amino-group through ammonia plasma treatment; Contact angle was measured on the PLGA nanofibers (NF) surface and amino-group modified (NF-NH2) surface; Cell Counting Kit-8 (CCK-8) was used to test the proliferation of rMSCs and hMSCs after seeded onto NF and NF-NH2 for 1, 3(4), and 7 days; Alizarin Red S (ARS) was used to detect the osteogenesis of rMSCs and hMSCs after seeded 21 days. Results:The contact angle was reduced from 81.28 ± 0.33 to 53.99 ± 0.79 by ammonia plasma treatment, which indicated that after modified by amino-group the surface of NF became hydrophilic. CCK-8 results showed that more rMSCs attached to NF-NH2 surface. After 24 h seeded onto surfaces, the absorbance data of rMSCs on PLGA NF and NF-NH2 was 0.096 ± 0.011 and 0.175 ± 0.014 respectively (P <0.001). However the NF-NH2 had no effects on the adhesion and proliferation of hMSCs. After 24 h seeded onto surfaces, the absorbance data of hMSCs on PLGA NF and NF-NH2 was 0.237 ± 0.004 and 0.238 ± 0.006 respectively (P >0.05). It can be seen from the ARS staining results that hMSCs produced calcium deposit equally on both NF and NF-NH2 surfaces, while NF-NH2 enhanced osteogenic differentiation of rMSCs (the ARS staining area was increased from 13.147 ± 3.223% to 36.677 ± 5.230%). Conclusion:NF modified by amino-groups can enhance rMSCs' adhesion, proliferation and osteogenic differentiation. The same phenomenon didn't happened on hMSCs, indicating that MSCs have a species dependent response. |
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