Department of Pharmaceutical Sciences, University of Perugia, Perugia, Italy.
Department of Experimental Medicine, University of Perugia, Perugia, Italy.
J Control Release. 2014 Oct 28;192:249-61. doi: 10.1016/j.jconrel.2014.08.001. Epub 2014 Aug 8.
Neonatal porcine Sertoli cells (NPSC) are immune privileged cells showing innate phagocytic and antibacterial activities. NPSC have been shown capable of immunoaltering the body's response and possess lung homing capacity. These properties encourage investigation of NPSC as functional components of cell-based therapeutic protocols to treat lung infections and related complications. In this work, for the first time, NPSC were tailored to carry an antibiotic drug loaded into poly(d,l lactic) acid microparticles (MP). A loading protocol was developed, which afforded 30% drug uptake and high stability over time, with little or no effects on NPSC viability, morphology, reactive oxygen species production and DNA integrity. FSH receptor integrity, and TGFβ (transforming growth factor β) and AMH (anti-Müllerian hormone) expressions were unchanged after 1month of cryopreservation. Protein tyrosine kinase activation due to phagocytosis may have had resulted in changes in inhibin B expression. The activity of MP-loaded or NPSC alone against Pseudomonas aeruginosa was maintained throughout 1month of storage. NPSC couple an innate antibacterial activity with the capacity to embody drug loaded MP. We showed for the first time that engineered NPSC can be cryopreserved with no loss of their basic properties, thereby possibly representing a novel approach for cell-based therapeutic and drug delivery system.
新生猪睾丸支持细胞(NPSC)是具有先天吞噬和抗菌活性的免疫特权细胞。已证明 NPSC 能够改变机体的免疫反应,并具有肺部归巢能力。这些特性鼓励研究 NPSC 作为基于细胞的治疗方案的功能成分,以治疗肺部感染和相关并发症。在这项工作中,NPSC 首次被设计用于携带负载于聚(D,L 乳酸)微球(MP)中的抗生素药物。开发了一种载药方案,可实现 30%的药物摄取,并且随着时间的推移具有很高的稳定性,对 NPSC 的活力、形态、活性氧产生和 DNA 完整性几乎没有或没有影响。FSH 受体完整性以及 TGFβ(转化生长因子β)和 AMH(抗苗勒管激素)的表达在 1 个月的冷冻保存后保持不变。吞噬作用引起的蛋白酪氨酸激酶的激活可能导致抑制素 B 表达的改变。负载 MP 或单独的 NPSC 对铜绿假单胞菌的活性在 1 个月的储存期间得以维持。NPSC 将先天抗菌活性与负载药物的 MP 结合在一起。我们首次表明,经工程改造的 NPSC 可以冷冻保存而不会丧失其基本特性,从而可能为基于细胞的治疗和药物输送系统提供一种新方法。