组织工程与重建外科杂志

组织工程与重建外科杂志 ›› 2025, Vol. 21 ›› Issue (6): 561-.

• • 上一篇    下一篇

基于DLP的3D打印仿生支架负载ADSC-EVs修复长骨缺损的实验研究

  

  • 出版日期:2025-12-25 发布日期:2025-12-30

Experimental research of long bone defect repair using biomimetic  scaffold constructed via DLP 3D printing loaded with ADSC extracellular vesicles#br#

  • Online:2025-12-25 Published:2025-12-30

PDF (PC)

7

摘要：

目的 探究负载脂肪干细胞细胞外囊泡（Adipose stem cell extracellular vesicles，ADSC-EVs）的3D打印仿生骨
支架对长骨缺损的修复作用。方法 通过数字光处理（Digital light processing，DLP）3D打印机，制备聚己内酯三丙烯酸
酯（Polycaprolactone triacrylate，PCLTA）高精度仿生骨支架，滴加ADSC-EVs于支架表面，扫描电镜下观察支架上的囊泡
形态，鬼笔环肽与茜素红染色检测支架的细胞相容性与成骨能力。兔双侧前肢桡骨制备长15 mm的临界骨缺损，随机
分3组：空白组不植入支架，对照组植入纯材料支架，实验组植入滴覆ADSC-EVs的支架。植入后12周取材，通过X线
片、免疫组化切片染色，观察成骨与成血管效果。结果 鬼笔环肽染色可见细胞在支架表面均匀生长，细胞形态正常。
茜素红染色显示滴覆囊泡的支架具有更强的成骨诱导能力（P<0.05）。扫描电镜下见囊泡均匀负载于支架表面及内部。
X线片显示实验组缺损区域新骨形成速度与成骨量均优于对照组（P<0.05）与空白组（P<0.01），HE染色进一步证实了X线
片的结果。免疫组化染色显示，实验组血管内皮细胞特异性标记CD31阳性表达高于对照组（P<0.05）与空白组（P<0.01）。
结论 负载ADSC-EVs的PCLTA高仿生骨支架具有良好的促成骨与促血管化能力，可有效促进长骨缺损的修复。

关键词:

Abstract:

Objective To investigate the reparative effect of 3D printed biomimetic bone scaffolds loaded with adipose stem cell-derived extracellular vesicles （ADSC-EVs） on long bone defects. Methods High-precision biomimetic bone
scaffolds were fabricated using digital light processing （DLP）3D printing with polycaprolactone triacrylate （PCLTA）. ADSCEVs were drop-loaded onto the scaffold surface. Scanning electron microscopy was used to observe the morphology of the vesicles on the scaffold. Cell compatibility and osteogenic ability of the scaffold were evaluated using immunofluorescence staining with phalloidin and Alizarin Red. Critical bone defects （15 mm） were created in the bilateral radius of New Zealand  white rabbits. Rabbits were randomly assigned to 3 groups： blank control group without scaffold implantation， control group with pure material scaffold implantation， and experimental group with scaffold loaded with ADSC-EVs. Samples were harvested at 12 weeks post-implantation for evaluation of osteogenesis and angiogenesis using X-ray， immunohistochemical staining， and histological analysis. Results Phalloidin staining showed uniform cell growth on the scaffold surface with normal cell morphology. Alizarin Red staining revealed that the vesicle-loaded scaffold had stronger osteoinductive ability （P< 0.05）. Scanning electron microscopy demonstrated uniform distribution of vesicles on the scaffold surface and within the scaffold. X-ray images indicated that the experimental group had faster new bone formation and higher bone mass than the
control group （P<0.05） and blank control group （P<0.01）， which was further confirmed by histological analysis with  Hematoxylin-Eosin staining. Immunohistochemical staining of tissue sections showed that the expression of the endothelial cell-specific marker CD31 was higher in the experimental group than in the control group （P<0.05） and blank control group （P<0.01）. Conclusion PCLTA biomimetic bone scaffolds loaded with ADSC-EVs exhibit excellent osteogenic and angiogenic abilities， effectively promoting the repair of long bone defects.

Key words: