[1]钟宗雨,黄应攀,王旭明.负载硬骨化蛋白沉默分子的同轴纤维膜促进大鼠骨髓间充质干细胞募集与骨缺损修复[J].陕西医学杂志,2026,(7):902-911.[doi:DOI:10.3969/j.issn.1000-7377.2026.07.006]
　ZHONG Zongyu,HUANG Yingpan,WANG Xuming.SOST-silencing molecule-loaded coaxial fibrous membranes promote the recruitment of rat bone marrow mesenchymal stem cells and cranial bone defect repair[J].,2026,(7):902-911.[doi:DOI:10.3969/j.issn.1000-7377.2026.07.006]
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负载硬骨化蛋白沉默分子的同轴纤维膜促进大鼠骨髓间充质干细胞募集与骨缺损修复

《陕西医学杂志》[ISSN:1000-7377/CN:61-1281/TN]

卷:
期数:
2026年7期
页码:
902-911
栏目:
基础研究
出版日期:
2026-07-05

文章信息/Info

Title:
SOST-silencing molecule-loaded coaxial fibrous membranes promote the recruitment of rat bone marrow mesenchymal stem cells and cranial bone defect repair
作者:
钟宗雨; 黄应攀; 王旭明
（昆明医科大学第一附属医院创伤中心，云南 昆明 650000）
Author(s):
ZHONG Zongyu; HUANG Yingpan; WANG Xuming
(Trauma Center,The First Affiliated Hospital of Kunming Medical University,Kunming 650000,China)
关键词:
SOST沉默分子; 同轴静电纺丝纤维膜; 骨髓间充质干细胞; 细胞募集; 颅骨骨缺损修复; 成骨分化; 骨形成蛋白 2
Keywords:
SOST-silencing molecule; Coaxial electrospun fibrous membrane; Bone marrow mesenchymal stem cells; Cell recruitment; Cranial bone defect repair; Osteogenic differentiation; Morphogenetic protein 2
分类号:
R 681
DOI:
DOI:10.3969/j.issn.1000-7377.2026.07.006
文献标志码:
A
摘要:
目的：构建负载硬骨化蛋白（SOST）沉默分子（沉默SOST的短发夹RNA慢病毒，SOST-shRNA）的同轴静电纺丝纤维膜，探讨其对大鼠骨髓间充质干细胞（BMSCs）募集及颅骨骨缺损修复的作用及分子机制。方法：采用同轴静电纺丝技术制备明胶为芯材（负载SOST-shRNA慢病毒或空载体慢病毒）、聚乳酸-羟基乙酸共聚物（PLGA）为壳材的同轴纤维膜，以空白明胶/PLGA纤维膜为对照，通过扫描电子显微镜（SEM）、粒径分析仪及酶联免疫吸附试验（ELISA）表征纤维膜。分离培养并鉴定大鼠骨髓间充质干细胞（BMSCs）；将48只Sprague-Dawley（SD）大鼠随机分为正常组（Normal组）、骨缺损组（Defect组）、空白纤维膜组（Blank组）、空载体纤维膜组（SOST-shRNA-NC组）、SOST沉默纤维膜组（SOST-shRNA组）及β-磷酸三钙组（β-TCP组），除Normal组外均建立颅骨8 mm骨缺损模型，对应植入相应材料。术后采用5-溴脱氧尿苷（BrdU）免疫组化检测BMSCs募集，显微计算机断层扫描（Micro-CT）分析骨修复参数，苏木精-伊红染色（HE染色）、马松染色（Masson染色）观察组织愈合及胶原沉积，蛋白质印迹法（Western blot）检测SOST、Runt相关转录因子2（Runx2）、骨桥蛋白（OPN）、骨形态发生蛋白2（BMP-2）表达。结果：成功制备负载SOST-shRNA的同轴纤维膜，纤维直径均匀（208±32）nm，平均孔径（801±96）nm，具有持续释放特性，21 d累计释放率达75.32%±3.15%。分离的BMSCs呈成纤维样形态，高表达CD29、CD44、CD90，低表达CD45，具备成骨分化能力。动物实验显示，与Normal组比较，Defect组4周骨缺损区域BrdU阳性细胞数显著减少（P＜0.05）；术后4、8周骨体积分数（BV/TV）、骨密度（BMD）等骨修复参数显著降低，术后8周缺损区域SOST蛋白表达上调（均P＜0.05）。与SOST-shRNA-NC组相比较，SOST-shRNA组以及β-TCP组术后4周骨缺损区域BrdU阳性细胞数显著增加（均P＜0.05）；术后4、8周骨体积分数（BV/TV）、骨密度（BMD）等骨修复参数显著升高（均P＜0.05）；且术后8周SOST-shRNA组的 BV/TV、BMD 显著高于β-TCP 组（P＜0.05）；术后8周两组缺损区域均有大量新生骨形成、胶原沉积丰富，SOST 蛋白表达下调，Runx2、OPN、BMP-2 蛋白表达上调（均 P＜0.05），且SOST-shRNA组的SOST蛋白表达显著低于β-TCP组，Runx2、OPN、BMP-2 蛋白表达显著高于β-TCP 组（均P＜0.05）。结论：负载SOST沉默分子的同轴纤维膜可有效募集BMSCs，通过调控SOST及成骨相关蛋白表达显著促进大鼠颅骨骨缺损修复，为骨缺损修复提供新型生物材料策略。
Abstract:
Objective:To fabricate coaxial electrospun fibrous membranes loaded with a sclerostin (SOST)-silencing molecule (short hairpin RNA lentivirus targeting SOST,SOST-shRNA),and to investigate their effects and the underlying molecular mechanisms on the recruitment of rat bone marrow mesenchymal stem cells (BMSCs) and the repair of cranial bone defects.Methods：Coaxial fibrous membranes were prepared using coaxial electrospinning technology,with gelatin as the core material (loaded with either SOST-shRNA lentivirus or empty vector lentivirus) and poly(lactic-co-glycolic acid) (PLGA) as the shell material.Blank gelatin/PLGA fibrous membranes served as the control.The membranes were characterized by scanning electron microscopy (SEM),a particle size analyzer,and enzyme-linked immunosorbent assay (ELISA).Rat BMSCs were isolated,cultured,and identified.Forty-eight Sprague-Dawley (SD) rats were randomly divided into the following groups：Normal group,Bone defect group (Defect group),Blank membrane group (Blank group),Empty vector membrane group (SOST-shRNA-NC group),SOST-silencing membrane group (SOST-shRNA group),and β-tricalcium phosphate group (β-TCP group).Except for the Normal group,critical-sized (8 mm diameter) cranial bone defect models were established in all rats,followed by the implantation of the corresponding materials.Postoperatively,the recruitment of BMSCs was detected by 5-bromo-2’-deoxyuridine (BrdU) immunohistochemistry.Bone repair parameters were analyzed by micro-computed tomography (Micro-CT).Tissue healing and collagen deposition were observed by hematoxylin-eosin (HE) staining and Masson’s trichrome staining.The protein expressions of sclerostin (SOST),runt-related transcription factor 2 (Runx2),osteopontin (OPN),and bone morphogenetic protein 2 (BMP-2) were detected by Western blot.Results:The SOST-shRNA-loaded coaxial fibrous membranes were successfully fabricated,exhibiting uniform fiber diameter (208±32)nm,an average pore size of (801±96) nm,and a sustained release profile,with a cumulative release rate reaching 75.32%±3.15% at 21 days.The isolated BMSCs displayed a fibroblast-like morphology,highly expressed CD29,CD44,and CD90,weakly expressed CD45,and possessed osteogenic differentiation potential.Animal experiments showed that compared with the Normal group,the Defect group had a significantly reduced number of BrdU-positive cells in the bone defect area at 4 weeks (P<0.05).The bone repair parameters such as bone volume/total volume (BV/TV) and bone mineral density (BMD) were significantly decreased at 4 and 8 weeks postoperatively,and the protein expression of SOST in the defect area was upregulated at 8 weeks postoperatively (all P<0.05).Compared with the SOST-shRNA-NC group,both the SOST-shRNA group and the β-TCP group showed a significant increase in the number of BrdU-positive cells in the bone defect area at 4 weeks postoperatively (all P<0.05).The bone repair parameters including BV/TV and BMD were significantly elevated at 4 and 8 weeks postoperatively (all P<0.05),and the BV/TV and BMD in the SOST-shRNA group were significantly higher than those in the β-TCP group at 8 weeks postoperatively (all P<0.05).At 8 weeks postoperatively,abundant new bone formation and rich collagen deposition were observed in the defect areas of these two groups,accompanied by downregulated SOST protein expression and upregulated expressions of Runx2,OPN,and BMP-2 proteins (all P<0.05).Furthermore,the SOST protein expression in the SOST-shRNA group was significantly lower than that in the β-TCP group,while the expressions of Runx2,OPN and BMP-2 proteins were significantly higher (all P<0.05).Conclusion:The SOST-silencing molecule-loaded coaxial fibrous membranes can effectively recruit BMSCs and significantly promote the repair of cranial bone defects in rats by regulating the expressions of SOST and osteogenesis-related proteins,providing a novel biomaterial strategy for bone defect repair.

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备注/Memo

备注/Memo:
云南省教育厅科学研究基金资助项目(2025J0190)
更新日期/Last Update: 2026-07-10