掺钆羟基磷灰石复合兔脂肪间充质干细胞修复膝关节软骨缺损

doi:10.16098/j.issn.0529-1356.2025.03.013

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解剖学报 ›› 2025, Vol. 56 ›› Issue (3) : 342-350. DOI: 10.16098/j.issn.0529-1356.2025.03.013

生物工程学

掺钆羟基磷灰石复合兔脂肪间充质干细胞修复膝关节软骨缺损

鲍瑛¹孔维丽² 杨钰³ 申福国¹ 张帅³ 孙文才^1*

作者信息 +

Repair of knee joint cartilage defects in rabbits using Gd-HA composite with adipose-derived mesenchymal stem cells

BAO Ying¹ KONG Wei-li² YANG Yu³ SHEN Fu-guo¹ ZHANG Shuai³ SUN Wen-cai^1*

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摘要

目的探讨掺钆羟基磷灰石 (Gd-HA) 支架材料复合脂肪间充质干细胞 (ADSCs) 对膝关节软骨缺损的修复作用。方法分离、培养和鉴定兔ADSCs，建立兔膝关节全层软骨缺损模型，18只兔随机分成空白对照组、Gd-HA支架组和ADSCs+Gd-HA支架组。术后12、24周取材行大体观察及评分（ICRS），HE、甲苯胺蓝、改良番红O亮绿和Ⅱ型胶原(ColⅡ)免疫组织化学染色分析及组织学染色评分（O’Driscoll），Real-time PCR检测修复组织ColⅡ与糖胺聚糖 (GAG) mRNA表达量，Western blotting检测修复组织ColⅡ蛋白表达，二甲基亚甲蓝比色法(DMMB)检测GAG含量，压痕实验检测生物力学强度，PKH26标记的ADSCs示踪，评价Gd-HA复合ADSCs的组织工程支架对兔膝关节软骨缺损的修复效果。结果体外分离培养的ADSCs形态学观察生长良好，表型稳定，定向分化良好。大体观察及组织学染色可见，植入Gd-HA支架组的膝关节全层软骨缺损模型修复程度及修复效果均优于空白对照组，而ADSCs+Gd-HA支架组的软骨修复情况优于Gd-HA支架 (P<0.05)；ICRS和改良O’Driscoll评分均高于其他两组 (P<0.05)。与Gd-HA组相比，ADSCs+Gd-HA组关节软骨修复过程中可产生更多的ColⅡ、GAG，其修复组织的力学强度表现更强 (P<0.05) ；ADSCs+Gd-HA组修复组织内发现PKH26标记的ADSCs，且参与新生组织的组成。结论 Gd-HA支架材料复合ADSCs对膝关节全层软骨缺损具有良好的修复效果，可能成为生物学修复关节软骨缺损的新型材料。

Abstract

Objective To investigate the effect of Gd-hydroxyapatite(Gd-HA) stents with adipose mesenchymal cells (ADSCs) on the repair of knee articular cartilage defects. Methods To isolate, culture, and identify rabbit ADSCs by establishing a rabbit knee joint full-thickness cartilage defect model, a total of 18 rabbits were randomly divided into blank control group, Gd-HA scaffold group, and ADSCs+Gd-HA scaffold group. At week 12 and 24 after surgery, International Curtilage Repair Society(ICRS) score, HE, toluidine blue, modified red O bright green and ColⅡ were detected by immunohistochemical staining, then ColⅡ and GAG mRNA expression levels were detected by O’Driscoll and Realtime PCR. ColⅡ protein expression was detected by Western blotting, GAG content was detected by DMMB, biomechanical strength was detected by indentation test, and PKH26 labeled ADSCs was used to trace the tissue engineering scaffold with Gd-HA composite ADSCs to evaluate the repair effect of rabbit knee cartilage defects. Results The ADSCs isolated and cultured in vitro showed good growth, stable phenotype and good directional differentiation through macroscopic observation and histological staining, it could be seen that the repair degree and effect of the knee joint fullthickness cartilage defect model implanted with Gd-HA scaffold group were better than those of the blank control group, while the cartilage repair situation of the ADSCs+Gd-HA scaffold group was better than that of the Gd-HA scaffold group (P<0.05); The ICRS and improved O’Driscoll scores were higher than the other two groups (P<0.05). Compared with the Gd-HA group, the ADSCs+Gd-HA group could produce ColⅡ and GAG during the process of cartilage repair, with stronger mechanical strength of the repaired tissue (P<0.05); PKH26 labeled ADSCs were found in the repaired tissues of the ADSCs+Gd-HA group, and they were involved in the composition of newly formed tissues. Conclusion Gd-HA scaffold material combined with ADSCs has a good repair effect on full-thickness cartilage defects in the knee joint as a new type of biological material for repairing joint cartilage defects.

关键词

软骨修复

/ 软骨组织缺损 / 生物材料 / 免疫组织化学 / 实时定量聚合酶链反应 / 免疫印迹法 / 兔

Key words

Cartilage repair

/ Cartilage tissue defect / Biomaterial / Immunohistochemistry / Real-time PCR / Western blotting / Rabbit

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鲍瑛孔维丽杨钰申福国张帅孙文才. 掺钆羟基磷灰石复合兔脂肪间充质干细胞修复膝关节软骨缺损[J]. 解剖学报. 2025, 56(3): 342-350 https://doi.org/10.16098/j.issn.0529-1356.2025.03.013

BAO Ying KONG Wei-li YANG Yu SHEN Fu-guo ZHANG Shuai SUN Wen-cai. Repair of knee joint cartilage defects in rabbits using Gd-HA composite with adipose-derived mesenchymal stem cells[J]. Acta Anatomica Sinica. 2025, 56(3): 342-350 https://doi.org/10.16098/j.issn.0529-1356.2025.03.013

中图分类号： R322 R681.3

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