Peng Yiyu, Li Lunhao, Yuan Qingyue, Gu Ping, You Zhengwei, Zhuang Ai, Bi Xiaoping
Department of Ophthalmology, Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine, 639 Zhizaoju Road, Shanghai 200011, P. R. China.
Shanghai Key Laboratory of Orbital Diseases and Ocular Oncology, Shanghai 200011, P. R. China.
ACS Omega. 2020 Feb 18;5(8):4302-4312. doi: 10.1021/acsomega.9b04249. eCollection 2020 Mar 3.
Bone tissue engineering has emerged as an effective alternative treatment to the problem of bone defect. To repair a bone defect, antibiosis and osteogenesis are two essential aspects of the repair process. By searching the literature and performing exploratory experiments, we found that β defensin 2 (BD2), with bifunctional properties of antibiosis and osteogenesis, was a feasible alternative for traditional growth factors. The antimicrobial ability of BD2 against and was studied by the spread plate and live/dead staining methods (low effective concentration of 20 ng/mL). BD2 was also demonstrated to enhance osteogenesis, with higher messenger RNA (mRNA) and protein expression of the osteogenic markers collagen I (Col1), runt-related transcription factor 2 (Runx2), osteopontin (Opn), and osteocalcin (Ocn) in vitro (1.5-2.5-fold increase compared with the control group in the most effective concentration group), which was consistent with the alkaline phosphatase (ALP) and alizarin red S (ARS) staining results. We implanted poly(sebacoyl diglyceride) (PSeD) combined with BD2 and rat bone tissue-derived mesenchymal stem cells (rBMSCs) under the back skin of rats and found that the inflammatory response was significantly lower with this combination than with the PSeD/rBMSCs scaffold without BD2 and the pure PSeD group and was similar to the control group. Importantly, when assessed in a critical-sized in vivo rat 8 m diameter calvaria defect model, a scaffold we developed combining bifunctional BD2 with porous organic polymer displayed an osteogenic effect that was 160-200% greater than the control group. The in vivo study results revealed a significant osteogenic response and antimicrobial effect and were consistent with the in vitro results. In summary, BD2 displayed a great potential of simultaneously promoting bone regeneration and preventing infection and could provide a viable alternative to traditional growth factors applied in bone defect repair.
骨组织工程已成为解决骨缺损问题的一种有效替代治疗方法。修复骨缺损时，抗菌和成骨是修复过程的两个重要方面。通过文献检索和探索性实验，我们发现具有抗菌和成骨双功能特性的β防御素2（BD2）是传统生长因子的可行替代品。采用平板涂布法和活/死染色法（最低有效浓度为20 ng/mL）研究了BD2对[具体细菌名称1]和[具体细菌名称2]的抗菌能力。体外实验还表明，BD2可促进成骨，成骨标志物I型胶原（Col1）、 runt相关转录因子2（Runx2）、骨桥蛋白（Opn）和骨钙素（Ocn）的信使核糖核酸（mRNA）和蛋白表达水平更高（在最有效浓度组中，与对照组相比增加了1.5至2.5倍），这与碱性磷酸酶（ALP）和茜素红S（ARS）染色结果一致。我们将聚（癸二酰甘油酯）（PSeD）与BD2及大鼠骨组织来源的间充质干细胞（rBMSCs）植入大鼠背部皮肤下，发现该组合的炎症反应明显低于不含BD2的PSeD/rBMSCs支架和纯PSeD组，且与对照组相似。重要的是，在直径8毫米的大鼠颅骨临界尺寸体内缺损模型中进行评估时，我们开发的一种将双功能BD2与多孔有机聚合物相结合的支架显示出比对照组高160%至200%的成骨效果。体内研究结果显示出显著的成骨反应和抗菌效果，与体外研究结果一致。总之，BD2在同时促进骨再生和预防感染方面具有巨大潜力，可为骨缺损修复中应用的传统生长因子提供可行的替代方案。
