鱼鳞骨粉及Bio-Oss骨粉对骨髓间充质干细胞成骨分化及体内骨再生作用的比较研究

doi:10.13591/j.cnki.kqyx.2026.02.005

摘要/Abstract

摘要：

目的观察并比较鱼鳞(fish scale,FS)骨粉及Bio-Oss骨粉的细胞毒性、促成骨分化能力及体内修复大鼠颅骨缺损作用。方法制备FS骨粉,Bio-Oss骨粉作为阳性对照,扫描电子显微镜观察FS骨粉微观结构,活/死细胞染色和CCK-8检测12.5~200.0 mg/mL Bio-Oss和FS浸提液的细胞毒性。Bio-Oss骨粉(Bio-Oss组)和FS骨粉(FS组)分别与大鼠骨髓间充质干细胞(bone marrow mesenchymal stem cells,BMSCs)共培养,RT-PCR检测细胞成骨相关基因ALP、RUNX2、OPN表达,茜素红染色观察成骨矿化能力;将两种骨粉分别植入大鼠颅骨临界缺损模型,通过Micro-CT、HE染色及Masson染色观察术后8周骨再生效果。结果 FS骨粉直径为200~800 μm,正面呈瓦楞状,背面较平滑,形态不规则。50 mg/mL及以下浓度的Bio-Oss和FS浸提液无细胞毒性;Bio-Oss及FS均可促进BMSCs钙化结节形成及成骨相关基因(ALP、RUNX2及OPN)表达(P<0.05),且FS组RUNX2、OPN表达水平显著高于Bio-Oss组(P<0.05)。植入Bio-Oss骨粉和FS骨粉后,Micro-CT分析发现,大鼠颅骨缺损处新生骨骨体积分数(bone volume tissue volume,BV/TV)明显增加(P<0.05);HE及Masson染色结果显示Bio-Oss组和FS组骨再生量也明显提高。结论 FS骨粉具有较好的细胞相容性及促成骨分化能力,在大鼠体内骨再生效果与Bio-Oss相似,应用前景广阔。

关键词: 鱼鳞骨粉, Bio-Oss骨粉, 成骨分化, 骨再生

Abstract:

Objective To evaluate and compare the cytotoxicity, osteogenic differentiation potential and cranial defect repair capacity of fish scale (FS) bone powder versus Bio-Oss bone powder. Methods FS bone powder was prepared, with Bio-Oss bone powder as the positive control. The microstructures of both materials were examined by scanning electron microscopy (SEM). Cytotoxicity of Bio-Oss and FS extracts (12.5-200.0 mg/mL) on rat bone marrow mesenchymal stem cells (BMSCs) was assessed using live/dead cll staining and CCK-8 assays. BMSCs were co-cultured with Bio-Oss and FS powder and osteogenic gene expression (ALP, RUNX2, OPN) was detected by RT-PCR. Osteogenic mineralization capacity was observed by alizarin red staining. Bio-Oss and FS powder was implanted into rat calvarial critical-sized defects. The bone regeneration was analyzed at 8 weeks post-surgery using Micro-CT, hematoxylin-eosin (HE), and Masson’s trichrome staining. Results FS bone powder particles ranged from 200 to 800 μm in diameter, featuring a corrugated front surface, a relatively smooth back surface, and an irregular morphology. Both Bio-Oss and FS extracts showed no cytotoxicity at concentrations ≤50 mg/mL. Bio-Oss and FS significantly promoted the formation of calcified nodules and upregulated osteogenic genes (ALP, RUNX2, OPN)(P<0.05), with FS exhibiting higher RUNX2 and OPN expression than Bio-Oss (P<0.05). Micro-CT analysis revealed that both the Bio-Oss and FS groups exhibited significantly higher bone volume/tissue volume (BV/TV) (P<0.05). Histological analysis with HE and Masson’s trichrome staining demonstrated greater bone regeneration in both the Bio-Oss and FS groups. Conclusion FS demonstrates excellent cytocompatibility and osteogenic differentiation capacity, achieving comparable bone regeneration to Bio-Oss in rats, suggesting potential applications in bone tissue engineering.

Key words: fish scale bone powder, Bio-Oss bone powder, osteogenic differentiation, bone regeneration

中图分类号:

R728.23

王晓蕾, 苏畅, 周忠政, 秦迪, 惠光艳, 王忠山, 吴广升. 鱼鳞骨粉及Bio-Oss骨粉对骨髓间充质干细胞成骨分化及体内骨再生作用的比较研究[J]. 口腔医学, 2026, 46(2): 112-117.

WANG Xiaolei, SU Chang, ZHOU Zhongzheng, QIN Di, HUI Guangyan, WANG Zhongshan, WU Guangsheng. A comparative study on fish scales and Bio-Oss bone powder on osteogenic differentiation of bone marrow mesenchymal stem cells and bone regeneration in vivo[J]. Stomatology, 2026, 46(2): 112-117.

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基因	引物序列(5'—3')
ALP	F:GCCTACACGGTCCTCCTATACGG
ALP	R:CACTGCTGACTGCTGCCGATAC
RUNX2	F:AGGCAGTTCCCAAGCATTTCATCC
RUNX2	R:TGGCAGGTAGGTGTGGTAGTGAG
OPN	F:CAGCCGTGGGAAGGACAGTTATG
OPN	R:TCACATCGGAATGCTCATTGCTCTC
GAPDH	F:TGAAGGTCGGAGTCAACGGATTTGGT
GAPDH	R:CATGTGGGCCATGAGGTCCACCAC