[1]李旭,刘征宇,赵宇,等.三维打印人工骨支架结合BMP-7和VEGF双重转染干细胞修复兔桡骨缺损[J].中国中医骨伤科杂志,2025,33(12):8-14+21.[doi:10.20085/j.cnki.issn1005-0205.251202]
　LI Xu,LIU Zhengyu,ZHAO Yu,et al.3D Printing of Artificial Bone Scaffold and Dual Transfection of BMP-7/VEGF Stem Cells for Repairing Rabbit Radius Defects[J].Chinese Journal of Traditional Medical Traumatology & Orthopedics,2025,33(12):8-14+21.[doi:10.20085/j.cnki.issn1005-0205.251202]
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三维打印人工骨支架结合BMP-7和VEGF双重转染干细胞修复兔桡骨缺损()

《中国中医骨伤科杂志》[ISSN:1005-0205/CN:42-1340/R]

卷:
第33卷
期数:
2025年12期
页码:
8-14+21
栏目:
实验研究
出版日期:
2025-12-15

文章信息/Info

Title:
3D Printing of Artificial Bone Scaffold and Dual Transfection of BMP-7/VEGF Stem Cells for Repairing Rabbit Radius Defects
文章编号:
1005-0205(2025)12-0008-07
作者:
李旭1; 刘征宇1; 赵宇1; 王楠1; 崔军1; 郑海峰1△
1沈阳医学院附属中心医院(沈阳,110024)
Author(s):
LI Xu1; LIU Zhengyu1; ZHAO Yu1; WANG Nan1; CUI Jun1; ZHENG Haifeng1△
1Affiliated Central Hospital of Shenyang Medical College,Shenyang 110024,China.
关键词:
三维打印;  骨组织工程;  羟基磷灰石;  丝素蛋白;  聚乳酸;  间充质干细胞;  骨缺损修复
Keywords:
3D printing;  bone tissue engineering;  hydroxyapatite;  silk fibroin;  polylactic acid;  mesenchymal stem cells;  bone defect repair
分类号:
R687
DOI:
10.20085/j.cnki.issn1005-0205.251202
文献标志码:
A
摘要:
目的:通过3D打印羟基磷灰石(HA)、丝素蛋白(SF)和聚乳酸(PLA)三元复合材料人工骨支架,结合双重转染骨形态发生蛋白-7(BMP-7)和血管内皮生长因子(VEGF)间充质干细胞(MSCs),构建骨组织工程材料用于修复兔桡骨骨缺损模型,并观察其骨组织修复效果。方法:利用3D打印技术制作羟基磷灰石/丝素蛋白/聚乳酸复合支架,通过扫描电子显微镜(SEM)观察其微观结构,确定其多孔性和抗压强度。培养兔间充质干细胞,随后采用双重基因转染技术,将BMP-7和VEGF导入细胞内,并检测转染后的基因表达水平。将转染后的干细胞与羟基磷灰石/丝素蛋白/聚乳酸支架共培养,并移植至兔桡骨骨缺损模型中。本实验设四组:对照组、单独支架组、单独干细胞组、支架与转染干细胞组。通过碱性磷酸酶(ALP)活性检测、茜素红染色、Micro-CT扫描、组织切片染色等方法评估各组成骨效果。结果:成骨活性(碱性磷酸酶活性):支架+MSC组的碱性磷酸酶活性显著高于其他组,差异有统计学意义(P<0.01)。支架+MSC组随着时间推移,成骨活性不断增强。矿化水平:支架+MSC组矿化面积占比在术后第4,8,12周均显著高于其他组(P<0.01),尤其在术后第12周时达到71.9%,显示出较强骨矿化能力。骨密度:术后第12周,支架+MSC组骨密度为0.85 g/cm3,显著高于其他组,差异有统计学意义(P<0.01),表明其成骨效果最优。血管生成:支架+MSC组血管密度显著高于其他组,差异有统计学意义(P<0.01),VEGF基因转染显著促进血管生成,增强新生骨血供。成骨基因表达:术后第8周时,支架+MSC组成骨相关基因(RUNX2、ALP、OCN)表达显著高于其他组,差异有统计学意义(P<0.01),证实双重基因转染对成骨过程有促进作用。结论:成功构建基于羟基磷灰石/丝素蛋白/聚乳酸复合材料的骨组织工程支架,并通过BMP-7和VEGF双重基因转染技术促进间充质干细胞的成骨和血管生成,可为临床开展大段骨缺损修复提供理论依据。
Abstract:
Objective:To construct a bone tissue engineering material using a 3D-printed artificial bone scaffold composed of hydroxyapatite(HA),silk fibroin(SF),and polylactic acid(PLA),combined with bone morphogenetic protein-7(BMP-7)and vascular endothelial growth factor(VEGF)dual-transfected mesenchymal stem cells(MSCs),to repair rabbit radial bone defects and observe its bone tissue repair efficacy.Methods:A HA/SF/PLA composite scaffold was fabricated using 3D printing technology,and its microstructure was observed via scanning electron microscopy(SEM)to assess porosity and compressive strength.Rabbit MSCs were cultured,and dual-gene transfection of BMP-7 and VEGF was performed,with gene expression levels monitored post-transfection.The transfected MSCs were co-cultured with the HA/SF/PLA scaffold and implanted into a rabbit radial bone defect model.The study included four groups:control group with no cells or scaffold,scaffold-only group,MSC-only group,and scaffold + transfected MSC group.The osteogenic effects were evaluated using alkaline phosphatase(ALP)activity,Alizarin Red staining,Micro-CT scanning,and histological staining.Results:Osteogenic activity(ALP activity):At all time points,the ALP activity in the scaffold+MSC group was significantly higher than that in the other groups(P<0.01).Over time,the ALP activity gradually increased,the scaffold+MSC group indicating a continuous enhancement of osteogenic activity.Mineralization Level:The scaffold+MSC group showed a significantly higher percentage of mineralized area at 4,8,and 12 weeks compared to the other groups(P<0.01),reaching 71.9% at 12 weeks,demonstrating a strong bone mineralization capacity.Bone Density:At 12 weeks post-surgery,the bone mineral density(BMD)in the scaffold+MSC group was 0.85 g/cm3,significantly higher than that in the other groups(P<0.01),indicating optimal osteogenic performance.Vascularization:The vascular density in the scaffold+MSC group was significantly higher than that in the other groups(P<0.01).VEGF gene transfection significantly promoted vascular formation,enhancing the blood supply to the newly formed bone.Osteogenic Gene Expression:At 8 weeks,the expression levels of osteogenic-related genes(RUNX2,ALP,OCN)in the scaffold+MSC group were significantly higher than those in the other groups(P<0.01),further confirming the promoting effect of dual gene transfection on the osteogenesis process.Conclusion:This study successfully constructed a bone tissue engineering scaffold based on HA/SF/PLA composite material and demonstrated that BMP-7 and VEGF dual-gene transfection effectively promoted MSC osteogenesis and angiogenesis.This provides a theoretical foundation for the clinical repair of large bone defects.

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(收稿日期:2025-04-11)

备注/Memo

备注/Memo:
基金项目:沈阳市中青年科技创新人才支持计划项目(RC210304)
△通信作者 E-mail:18618464483@163.com
更新日期/Last Update: 2025-12-15