骨靶向脂质体包封丹酚酸 A 通过调控 HDAC3 介导的软骨内骨化促进骨不连愈合。
Bone-Targeting Liposome-Encapsulated Salvianic Acid A Improves Nonunion Healing Through the Regulation of HDAC3-Mediated Endochondral Ossification.
机构信息
Guangdong Key Laboratory for Research and Development of Natural Drugs, Department of Pharmacology, Guangdong Medical University, Zhanjiang, Guangdong 524023, People's Republic of China.
Department of Orthopaedics, Stem Cell Research and Cellular Therapy Center, The Affiliated Hospital of Guangdong Medical University, Zhanjiang, Guangdong 524001, People's Republic of China.
出版信息
Drug Des Devel Ther. 2020 Aug 26;14:3519-3533. doi: 10.2147/DDDT.S263787. eCollection 2020.
AIM
Nonunion is a major complication in fracture repair and remains a challenge in orthopaedics and trauma surgery. In this study, we aimed to evaluate the effectiveness of treatment of nonunion with a large radial defect using a bone-targeting liposome-encapsulated salvianic acid A (SAA-BTL)-incorporated collagen sponge and further elucidate whether the effects were closely related to histone deacetylase 3 (HDAC 3)-mediated endochondral ossification in nonunion healing process.
METHODS
Fifteen New Zealand female rabbits were randomly divided into three groups. Segmental radius critical size defects (15 mm) were created via surgery on both the forelimbs of the rabbits. The SAA-BTL/SAA/saline-incorporated collagen sponges were implanted into the defects in the three groups, respectively, for four weeks of treatment. X-ray imaging, micro-computed tomography (CT) analysis, histology, and immunofluorescence analysis (HDAC3, collagen II, VEGFA, and osteocalcin) were performed to determine the effects of the treatments. In addition, a short interfering RNA was applied to induce HDAC3 knockdown in the chondrogenic cell line ATDC5 to investigate the roles of HDAC3 and SAA intervention in endochondral ossification in nonunion healing.
RESULTS
X-ray imaging and micro-CT results revealed that SAA-BTL-incorporated collagen sponges significantly stimulated bone formation in the nonunion defect rabbit model. Furthermore, immunofluorescence double staining and histology analysis confirmed that SAA-BTL significantly increased the expression of P-HDAC3, collagen II, RUNX2, VEGFA, and osteocalcin in vivo; accelerated endochondral ossification turnover from cartilage to bone; and promoted long bone healing of nonunion defects. ATDC5 cells knocked down for HDAC3 showed significantly decreased expression of HDAC3, which resulted in reduced expression of chondrogenesis, osteogenesis, and angiogenesis biomarker genes (Sox9, Col10a1, VEGFA, RUNX2, and Col1a1), and increased expression of extracellular matrix degradation marker (MMP13). SAA treatment reversed these effects in the HDAC3 knockdown cell model.
CONCLUSION
SAA-BTL can improve nonunion healing through the regulation of HDAC3-mediated endochondral ossification.
目的
骨折修复后的不愈合是一个主要并发症，在矫形和创伤外科仍然是一个挑战。本研究旨在评估靶向骨的脂质体包载丹酚酸 A（SAA-BTL）结合胶原蛋白海绵治疗大桡骨缺损不愈合的效果，并进一步阐明这些效果是否与不愈合愈合过程中组蛋白去乙酰化酶 3（HDAC3）介导的软骨内骨化密切相关。
方法
15 只新西兰雌性兔随机分为 3 组。通过手术在兔的前肢上创建 15mm 节段性桡骨临界尺寸缺陷。将 SAA-BTL/SAA/生理盐水结合的胶原蛋白海绵分别植入这 3 组的缺陷部位，治疗 4 周。进行 X 射线成像、微计算机断层扫描（CT）分析、组织学和免疫荧光分析（HDAC3、胶原 II、VEGFA 和骨钙素），以确定治疗效果。此外，应用短干扰 RNA 诱导软骨细胞系 ATDC5 中的 HDAC3 敲低，以研究 HDAC3 和 SAA 干预在不愈合愈合中的软骨内骨化作用。
结果
X 射线成像和微 CT 结果显示，SAA-BTL 结合的胶原蛋白海绵显著刺激了兔不愈合缺陷模型中的骨形成。此外，免疫荧光双重染色和组织学分析证实，SAA-BTL 显著增加了 P-HDAC3、胶原 II、RUNX2、VEGFA 和骨钙素在体内的表达；加速了软骨内骨化从软骨到骨的转化；促进了不愈合缺陷处长骨的愈合。HDAC3 敲低的 ATDC5 细胞显示出 HDAC3 表达明显降低，导致软骨生成、成骨和血管生成生物标志物基因（Sox9、Col10a1、VEGFA、RUNX2 和 Col1a1）表达减少，细胞外基质降解标志物（MMP13）表达增加。SAA 处理逆转了 HDAC3 敲低细胞模型中的这些作用。
结论
SAA-BTL 可以通过调节 HDAC3 介导的软骨内骨化来改善不愈合的愈合。
Zotero 插件