Guozhu Yin, Yihua Wu, Zhu You, Li Dai, Tianqi Zhang, Jia Li, Shizhou Zhang
Department of Stomatology, Shandong Provincial Hospital Affiliated to Shandong First Medical University, 324 Jingwu Road, 250021, Jinan, China.
School of Material Science and Engineering, University of Jinan, Jinan 250022, China.
Regen Ther. 2025 Mar 14;29:140-147. doi: 10.1016/j.reth.2025.02.016. eCollection 2025 Jun.
Osteoporosis, which adversely affected osseointegration of dental implants, became prevalent with the entry of ageing era. Recent studies indicated that α-calcitonin gene-related peptide (α-CGRP) played a role in modulating osteoporosis. However, due to the plasma half-life of α-CGRP being estimated at merely 7-10 min, it was difficult to utilize a conventional method that administered sufficient α-CGRP to the implant site. This present study aimed to investigate the efficacy of an α-CGRP transgene in promoting implant osseointegration in ovariectomized (OVX) rats.
The osteoporosis rat model was established through bilateral ovariectomy, following which the subjects were categorized into three distinct groups: the α-CGRP transgene group, the empty virus vector group, and the blank control group. Ad-CGRP-EGFP was locally administered into the bone defect site prior to implant placement in the OVX rats. 7 and 28 days after implantation, the femurs were isolated for molecular and histological analyses, micro-CT analysis and biomechanical test.
Bone marrow stromal cells (BMSCs) transduced with Ad-CGRP-EGFP could continuously express α-CGRP more than 28 days in vitro. Successful transgene expression was confirmed through cryosectioning and Western blot analysis 7 days after implantation in vivo. The results indicated a substantial decrease in the quantity of TRAP + cells in the α-CGRP transgene group. Additionally, quantitative real-time RT-PCR and Western blot analysis revealed a significant elevation in the expression levels of Runx2 and ALP, coupled with a notable reduction in the expression of cathepsin K and RANKL . Moreover, the α-CGRP transgene group exhibited a significant enhancement in osteointegration and restoration of implant stability in OVX rats () 28 days after implantation.
This study implied a great therapeutic potential of α-CGRP in osseointegration of titanium implants in OVX-relative osteoporosis rats, offering valuable insights for guiding bone regeneration strategies under osteoporotic conditions.
随着老龄化时代的到来，骨质疏松症日益普遍，它对牙种植体的骨整合产生不利影响。最近的研究表明，α-降钙素基因相关肽（α-CGRP）在调节骨质疏松症中发挥作用。然而，由于α-CGRP的血浆半衰期估计仅为7 - 10分钟，很难采用常规方法向种植部位给予足够的α-CGRP。本研究旨在探讨α-CGRP转基因在促进去卵巢（OVX）大鼠种植体骨整合中的疗效。
通过双侧卵巢切除术建立骨质疏松大鼠模型，然后将实验对象分为三个不同的组：α-CGRP转基因组、空病毒载体组和空白对照组。在OVX大鼠植入种植体之前，将Ad-CGRP-EGFP局部注射到骨缺损部位。植入后7天和28天，分离股骨进行分子和组织学分析、显微CT分析和生物力学测试。
用Ad-CGRP-EGFP转导的骨髓间充质干细胞（BMSCs）在体外可连续表达α-CGRP超过28天。体内植入7天后，通过冰冻切片和蛋白质印迹分析证实了转基因的成功表达。结果表明，α-CGRP转基因组中抗酒石酸酸性磷酸酶（TRAP）+细胞数量显著减少。此外，定量实时逆转录聚合酶链反应（RT-PCR）和蛋白质印迹分析显示，Runx2和碱性磷酸酶（ALP）的表达水平显著升高，同时组织蛋白酶K和核因子κB受体活化因子配体（RANKL）的表达显著降低。此外，α-CGRP转基因组在植入后28天的OVX大鼠中骨整合显著增强，种植体稳定性恢复。
本研究表明α-CGRP在与OVX相关的骨质疏松大鼠钛种植体骨整合中具有巨大的治疗潜力，为指导骨质疏松条件下的骨再生策略提供了有价值的见解。
