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首页|期刊导航|临床口腔医学杂志|氧化石墨烯气凝胶促进兔骨髓间充质干细胞增殖及成骨分化

氧化石墨烯气凝胶促进兔骨髓间充质干细胞增殖及成骨分化OACSTPCD

Graphene oxide aerogels promote proliferation and osteogenic differentiation of rabbit bone marrow mesenchy-mesenchymal stem cells

中文摘要英文摘要

目的:研究氧化石墨烯(graphene oxide,GO)气凝胶对兔骨髓间充质干细胞(bone marrow mesenchymal stem cells,BMSCs)增殖及成骨分化的影响.方法:组织块法培养新西兰大白兔下颌骨圆形骨片获得细胞,免疫荧光检测干细胞标记物CD44 以及BMSCs标记物CD90 表达.将培养所得细胞分别接种于 96 孔板、明胶海绵(gelatin sponge,GS)、GO气凝胶支架,通过CCK-8检测接种第 1~3 天 96 孔板(空白组)、GS组、GO组细胞OD值.经成骨诱导 7 d后,细胞碱性磷酸酶(alkaline phosphatase,ALP)活性检测.制备新西兰大白兔双侧下颌骨颊侧骨壁圆形骨缺损模型,分成空白组(单纯使缺损区内血液充盈)、骨粉组(缺损区充填Bio-Oss骨粉)、支架组(缺损区充填GO气凝胶支架).于 3 个月后,X线片及CBCT观察 3 组缺损区骨生成情况,制备组织切片经HE染色后进行组织学观察.结果:组织块法培养所得第 3 代细胞CD44 和CD90 均阳性,证实为BMSCs.GO支架和GS支架均可促进BMSCs增殖、向成骨细胞分化,且GO支架优于GS支架.动物实验结果显示,GO支架与骨粉组缺损区域的成骨效果均优于空白组,GO支架的成骨效果优于骨粉组.HE染色结果显示,GO支架组和骨粉组新生骨均能与缺损区边缘相连续,而GO支架对骨缺损边缘修复的效果更佳.结论:GO气凝胶可促进BMSCs的增殖和向成骨分化,能够促进骨缺损区新骨生成,是较好的组织工程支架材料.

Objective:To study the effects of graphene oxide(GO)aerogels on the proliferation and osteogenic differ-entiation of rabbit bone marrow mesenchymal stem cells(BMSCs).Methods:The cells were obtained by tissue culture from the round mandibular bone fragments of New Zealand white rabbits.The cells were identified by immunofluorescence staining of stem cell marker CD44 and BMSCs marker CD90.The identified cells were inoculated into 96 well plate pores,gelatin sponge(GS)scaffold,GO aerogel gel scaffold.Cell Counting Kit-8(CCK-8)was used to detect OD values of blank group,GS group and GO group cells on day 1~3.In addition,the alkaline phosphatase(ALP)activity of each group cells was detected by ALP kit post-induction 7 days.In vivo experiments were conducted in New Zealand white rabbits.Three circular bone de-fects were prepared on the buccal bone wall of bilateral mandibles in New Zealand white rabbits,and were filled with blood(blank group),Bio-Oss bone powder(bone meal group)and GO aerogel stent(scaffold group),separately.Three months lat-er,the bone formation in the defect area was observed by X-ray,cone-beam computed tomography(CBCT)and HE staining.Results:The third generation cells cultured by tissue block method were both CD44 and CD90 positive,which confirming that those were BMSCs.Both GO and GS scaffolds promoted the proliferation and differentiation of BMSCs into osteoblasts.The effect of GO scaffold on promoting osteogenic differentiation was better than that of GS scaffold.The animal experiment results showed that both GO and GS scaffolds promoted osteogenesis in the defect area,and the osteogenic effect of GO scaffold was better than that of bone powder group.The HE staining showed that both the GO scaffold group and the bone powder group showed continuity with the edge of the defect area,while the GO scaffold had a better effect.Conclusion:GO aerogel could promote the proliferation and osteogenic differentiation of BMSCs,as well as the formation of new bone in the bone defect area.It is a good scaffold material for tissue engineering.

杨莹;李玉凤;李林峰;赵谦;李健

唐山市人民医院口腔科,河北 唐山 063000唐山市人民医院中心实验室,河北 唐山 063000唐山博创口腔医院牙体牙髓科,河北 唐山 063000厦门大学附属翔安医院口腔科,福建 厦门 361000

口腔医学

组织工程细胞支架氧化石墨烯成骨骨髓间充质干细胞

Tissue engineeringCell scaffoldGraphene oxideOsteogenesisBone marrow mesenchymal stem cells

《临床口腔医学杂志》 2024 (002)

75-79 / 5

河北省医学科学研究课题计划项目(编号:20240104)

10.3969/j.issn.1003-1634.2024.02.003

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