Limlawan Pirawish, Insin Numpon, Marger Laurine, Freudenreich Mélanie, Durual Stéphane, Vacharaksa Anjalee
Department of Oral Medicine, Faculty of Dentistry, Chulalongkorn University, Pathumwan, Bangkok, 10330, Thailand.
Research Unit on Oral Microbiology and Immunology, Faculty of Dentistry, Chulalongkorn University, Pathumwan, Bangkok, 10330, Thailand.
BDJ Open. 2023 Nov 24;9(1):50. doi: 10.1038/s41405-023-00177-1.
To demonstrate hydroxyapatite nanoparticles modified with cationic functional molecules. 3-aminopropyltriethoxysilane (HA-NPs-APTES) carrying microRNA-302a-3p (miR) in the 3D-printed tricalcium phosphate/Hydroxyapatite (TCP/HA) scaffold can increase healing of the critical-sized bone defect.
3D-printed TCP/HA were modified with HA-NPs-APTES by two methods (M1, M2). The dispersion of particles was visualized by fluorescent microscopy. Biocompatibility of the scaffolds was tested by alizarin assay. Delivery of miR to the cells and osteogenic gene expression were evaluated by qPCR. After selecting best method (M2), scaffolds, scaffolds+HA-NPs-APTES with or without miR were implanted in 4 mm mouse calvarium defect (n = 4 per group). After 2,4 and 6 weeks, bone regeneration were evaluated by microCT and histology sections.
Both M1 and M2 scaffolds were biocompatible with cell adhesion on its surface. M2 scaffold showed significant increase of miR, suggesting successful delivery, resulted in downregulation of its target mRNA COUP-TFII, and upregulation of RUNX2 mRNA. Calvarium defect with M2 scaffold also showed significantly higher BV/TV and higher number of filled spaces at all time points. Histomorphometry demonstrated new bone formed at the center of the HA-NPs-APTES-miR scaffold earlier than controls.
TCP/HA scaffold modified with HA-NPs-APTES facilitated delivery of miR and enhanced bone regeneration.
展示用阳离子功能分子修饰的羟基磷灰石纳米颗粒。在3D打印的磷酸三钙/羟基磷灰石（TCP/HA）支架中携带微小RNA-302a-3p（miR）的3-氨丙基三乙氧基硅烷（HA-NPs-APTES）可促进临界尺寸骨缺损的愈合。
采用两种方法（M1、M2）用HA-NPs-APTES对3D打印的TCP/HA进行修饰。通过荧光显微镜观察颗粒的分散情况。通过茜素试验测试支架的生物相容性。通过qPCR评估miR向细胞的递送和骨生成基因表达。在选择最佳方法（M2）后，将支架、含或不含miR的支架+HA-NPs-APTES植入4毫米的小鼠颅骨缺损处（每组n = 4）。在2、4和6周后，通过显微CT和组织学切片评估骨再生情况。
M1和M2支架均具有生物相容性，细胞可在其表面黏附。M2支架显示miR显著增加，表明递送成功，导致其靶mRNA COUP-TFII下调，RUNX2 mRNA上调。植入M2支架后的颅骨缺损在所有时间点的骨体积分数（BV/TV）也显著更高，填充空间数量更多。组织形态计量学显示，HA-NPs-APTES-miR支架中心形成新骨的时间早于对照组。
用HA-NPs-APTES修饰的TCP/HA支架促进了miR的递送并增强了骨再生。
