抗菌肽LL37固体脂质纳米粒的制备、优化和评价

Preparation,optimization and evaluation of antimicrobial peptides LL37-SLNs

目的制备搭载抗菌肽LL37的固体脂质纳米粒(LL37-SLNs),对配方进行优化、表征和评价。方法采用“乳化蒸发-低温固化”的方法制备17个批次的LL37-SLNs,以硬脂酸、软磷脂和表面活性剂Myrj52作为自变量,平均粒径和包封率作为因变量,通过Box-Behnken试验设计优化配方,并进行表征和评价。结果通过Design Expert 12.0软件对回归方程分析得到最佳配方为硬脂酸300 mg、软磷脂150 mg、Myrj52250 mg。对优化后的LL37-SLNs进行表征和评价,结果显示载药量为(2.92±0.06)%,包封率为(86.87±0.26)%,平均粒径为(221.7±1.3)nm,Zeta电位为(-30.4±0.2)mV,多分散指数为(0.233±0.012)。在透射电镜下观察其外观规则,粒径均一。LL37-SLNs的体外释药曲线表现出良好的缓释能力,体外抗菌活性研究显示出持久的抗菌特性,且在-20℃可稳定储存60 d。结论LL37-SLNs配方合理,颗粒分布均一、稳定,包封率佳,体外释放和抗菌活性良好。

Objective To prepare,optimize,characterize and evaluate the solid lipid nanoparticles carrying the antimicrobial peptide LL37(LL37-SLNs).Methods Totally 17 batches of LL37-SLNs were prepared by an emulsification and low-temperature solidification method.The LL37-SLNs formulations were optimized by Box-Behnken experiment design,with stearic acid,soft phospholipids and surfactant Myrj52 as the independent variables,while the average particle size and encapsulation efficiency as the dependent variables.LL37-SLNs were characterized and evaluated.Results The regression equation was obtanined by Design Expert 12.0 software and the optimized formulation was 300 mg stearic acid,150 mg soft phospholipids,and 250 mg surfactant(Myrj52).LL37-SLNs were prepared according to the optimized formulation.The drug loading rate was(2.92±0.06)%,the encapsulation rate was(86.87±0.26)%,the average particle size was(221.7±1.3)nm,the Zeta potential was(-30.4±0.2)mV,and the PDI was(0.233±0.012).The optimized formation was spherical with uniform size.In vitro,LL37-SLNs showed good slow release rate and sustained antibacterial activity,and were stable at-20℃for 60 days.Conclusion The formulation of LL37-SLNs is reasonable,with homogeneous and stable distribution,good release rate and antibacterial activity.