外源性bFGF对2型糖尿病大鼠钛种植体骨结合的影响及机制探讨

doi:10.19438/j.cjoms.2022.01.004

摘要/Abstract

摘要： 目的: 探讨外源性碱性成纤维细胞生长因子（bFGF）对2型糖尿病（T2DM）大鼠钛种植体骨结合的影响及机制。方法: 取SPF级雄性SD大鼠40只,随机分为对照组、T2DM组、bFGF-T2DM组、氯化锂（LiCl）-T2DM组。对照组正常饲料喂养,其余大鼠以高脂高糖喂养+腹腔注射链脲佐菌素（STZ）30 mg/kg建立T2DM模型。建模后,于右侧胫骨近骺端植入纯钛种植体。bFGF-T2DM组在植入种植体前以重组大鼠碱性成纤维细胞生长因子（rrbFGF）-聚乳酸-羟基乙酸共聚物（PLGA）微球处理种植体表面,其余大鼠以安慰剂-PLGA微球处理。LiCl-T2DM组种植体植入后皮下注射LiCl 60 mg·kg^-1·d^-1,连续8周,其余大鼠注射等体积生理盐水。观察大鼠术后一般情况,记录随机血糖。术后8周,采用丽春红染色观察骨组织学形态,计算骨组织形态学参数。利用Western免疫印迹检测种植体周围骨组织成骨相关指标骨钙素（OCN）、I型胶原蛋白（Col-1）、核心结合因子2（Runx2）、β连环蛋白（β-catenin）、反式激活活性蛋白（c-myc）的表达。采用SPSS 20.0软件包对数据进行统计学分析。结果: 注射STZ后1~8周,T2DM组、bFGF-T2DM组与LiCl-T2DM组随机血糖显著高于对照组（P<0.05）。种植体植入8周后,丽春红染色显示T2DM组无骨-种植体接触;bFGF-T2DM组骨-种植体接触范围广泛,与对照组接近;LiCl-T2DM组部分区域无骨组织接触。与对照组相比,T2DM组与LiCl-T2DM组骨-种植体接触率（BIC）、骨小梁容积率（TBV）、OCN、Col-1、Runx2蛋白相对表达量及T2DM组β-catenin、c-myc蛋白相对表达量显著降低,LiCl-T2DM组β-catenin、c-myc蛋白相对表达量显著增加（P<0.05）;对照组与bFGF-T2DM组BIC、TBV及OCN、Col-1、Runx2、β-catenin、c-myc蛋白相对表达量比较,差异无统计学意义（P>0.05）;与T2DM组相比,bFGF-T2DM组与LiCl-T2DM组BIC、TBV及OCN、Col-1、Runx2、β-catenin、c-myc蛋白相对表达量显著增加（P<0.05）;与bFGF-T2DM组相比,LiCl-T2DM组BIC、TBV与OCN、Col-1、Runx2蛋白相对表达量显著降低,β-catenin、c-myc蛋白相对表达量显著增加（P<0.05）。结论: 外源性bFGF能促进T2DM大鼠钛种植体骨组织愈合,促进骨形成;其机制可能与Wnt/β-catenin信号通路激活有关。

关键词: 2型糖尿病, 外源性碱性成纤维细胞生长因子, 植骨, 种植体, 骨结合

Abstract: PURPOSE: To investigate the effect and mechanism of exogenous basic fibroblast growth factor (bFGF) on osseointegration of titanium implants in type 2 diabetes mellitus (T2DM) rats. METHODS: Forty SPF male SD rats were randomly divided into control group, T2DM group, bFGF-T2DM group, and lithium chloride (LiCl)-T2DM group. The control group was fed with normal diet, and the remaining rats were fed with high-fat and high-sugar + intraperitoneal injection of streptozotocin (STZ) 30 mg/kg to establish a T2DM model. After modeling, pure titanium implants were implanted in the proximal epiphysis of the right tibia. The bFGF-T2DM group was treated with recombinant rat basic fibroblast growth factor (rrbFGF)-poly lactic-co-glycolic acid (PLGA) microspheres before implantation, and the remaining rats were treated with placebo- PLGA microsphere processing. After implantation of LiCl-T2DM group, LiCl 60 mg·kg^-1·d^-1 was injected subcutaneously for 8 weeks, and the remaining rats were injected with equal volume of normal saline. The general conditions of the rats after surgery were observed and random blood glucose was recorded. Eight weeks after operation, ponceau red staining was used to observe bone histological morphology and bone histological parameters were calculated. Western blotting was used to detect the expression of osteocalcin (OCN), type I collagen (collagen type I, Col-1), core binding factor 2 (runt-related transcription factor 2, Runx2), β-catenin and cell-myc(c-myc) in the bone tissue around the implants. SPSS 20.0 software package was used for data analysis. RESULTS: The random blood glucose of T2DM group, bFGF-T2DM group and LiCl-T2DM group was significantly higher than that of the control group (P<0.05). Eight weeks after implantation, no bone implant contact was found in T2DM group. bFGF-T2DM group had a wide range of bone implant contact, which was close to the control group. In LiCl-T2DM group, there was no bone tissue contact in some areas. Compared with the control group, the relative expression of β-catenin and c-myc protein in T2DM group, the relative expression of OCN, Col-1, Runx2 and bone-implant contact(BIC), trabecular bone volume(TBV) of LiCl-T2DM group and T2DM group decreased, and the relative expression of β-catenin and c-myc protein increased significantly in LiCl-T2DM group(P<0.05). There was no significant difference in BIC, TBV, relative expression of OCN, Col-1, Runx2, β-catenin and c-myc between the control group and bFGF-T2DM group(P>0.05). Compared with T2DM group, BIC, TBV, relative expressions of OCN, Col-1, Runx2, β-catenin and c-myc in bFGF-T2DM group and LiCl-T2DM group were increased significantly(P<0.05). Compared with bFGF-T2DM group, BIC, TBV and relative expressions of OCN, Col-1, Runx2 in LiCl-T2DM group were significantly decreased, while the relative expression levels of β-catenin and c-myc protein were significantly decreased (P<0.05). CONCLUSIONS: Exogenous bFGF can promote the healing of titanium implant bone tissue in T2DM rats and promote bone formation. The mechanism may be related to the activation of Wnt/β-catenin signaling pathway.

Key words: Type 2 diabetes mellitus, Exogenous basic fibroblast growth factor, Bone graft, Dental implant, Osseointegration

中图分类号:

R782.1

王慧丽, 谢冰, 魏育红. 外源性bFGF对2型糖尿病大鼠钛种植体骨结合的影响及机制探讨[J]. 中国口腔颌面外科杂志, 2022, 20(1): 22-27.

WANG Hui-li, XIE Bing, WEI Yu-hong. Effect of exogenous bFGF on osseointegration of titanium implants in type 2 diabetic rats and its possible mechanism[J]. China J Oral Maxillofac Surg, 2022, 20(1): 22-27.

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