IRCCS SDN, Naples, 80143, Italy.
Department of Pharmacy and Research Centre on Bioactive Peptides (CIRPeB), University of Naples "Federico II", Naples, 80134, Italy.
Int J Nanomedicine. 2021 Mar 1;16:1617-1630. doi: 10.2147/IJN.S296272. eCollection 2021.
The clinical use of the antitumoral drug doxorubicin (Dox) is reduced by its dose-limiting toxicity, related to cardiotoxic side effects and myelosuppression. In order to overcome these drawbacks, here we describe the synthesis, the structural characterization and the in vitro cytotoxicity assays of hydrogels (HGs) and nanogels (NGs) based on short peptide sequences loaded with Dox or with its liposomal formulation, Doxil.
Fmoc-FF alone or in combination with (FY)3 or PEG8-(FY)3 peptides, at two different ratios (1/1 and 2/1 v/v), were used for HGs and NGs formulations. HGs were prepared according to the "solvent-switch" method, whereas NGs were obtained through HG submicronition by the top-down methodology in presence of TWEEN60 and SPAN60 as stabilizing agents. HGs gelation kinetics were assessed by Circular Dichroism (CD). Stability and size of NGs were studied using Dynamic Light Scattering (DLS) measurements. Cell viability of empty and filled Dox HGs and NGs was evaluated on MDA-MB-231 breast cancer cells. Moreover, cell internalization of the drug was evaluated using immunofluorescence assays.
Dox filled hydrogels exhibit a high drug loading content (DLC=0.440), without syneresis after 10 days. Gelation kinetics (20-40 min) and the drug release (16-28%) over time of HGs were found dependent on relative peptide composition. Dox filled NGs exhibit a DLC of 0.137 and a low drug release (20-40%) after 72 h. Empty HGs and NGs show a high cell viability (>95%), whereas Dox loaded ones significantly reduce cell viability after 24 h (49-57%) and 72 h (7-25%) of incubation, respectively. Immunofluorescence assays evidenced a different cell localization for Dox delivered through HGs and NGs with respect to the free drug.
A modulation of the Dox release can be obtained by changing the ratios of the peptide components. The different cellular localization of the drug loaded into HGs and NGs suggests an alternative internalization mechanism. The high DLC, the low drug release and preliminary in vitro results suggest a potential employment of peptide-based HGs and NGs as drug delivery tools.
抗肿瘤药物阿霉素（Dox）的临床应用受到其剂量限制毒性的限制，这与心脏毒性副作用和骨髓抑制有关。为了克服这些缺点，我们在这里描述了基于短肽序列的水凝胶（HG）和纳米凝胶（NG）的合成、结构表征和体外细胞毒性试验，这些短肽序列负载有 Dox 或其脂质体制剂 Doxil。
单独使用 Fmoc-FF 或与（FY）3 或 PEG8-（FY）3 肽组合，以两种不同的比例（1/1 和 2/1 v/v），用于 HG 和 NG 的配方。HG 是根据“溶剂切换”方法制备的，而 NG 是通过在 TWEEN60 和 SPAN60 作为稳定剂存在的情况下通过自上而下的方法使 HG 亚微米化得到的。通过圆二色性（CD）评估 HG 的凝胶化动力学。通过动态光散射（DLS）测量研究 NG 的稳定性和粒径。在 MDA-MB-231 乳腺癌细胞上评估空和填充 Dox HG 和 NG 的细胞活力。此外，使用免疫荧光测定法评估药物的细胞内化。
负载 Dox 的水凝胶表现出高药物负载含量（DLC=0.440），10 天后无收缩。凝胶化动力学（20-40 分钟）和 HG 随时间的药物释放（16-28%）发现取决于相对肽组成。负载 Dox 的 NG 的 DLC 为 0.137，72 小时后药物释放率（20-40%）较低。空 HG 和 NG 的细胞活力>95%，而负载 Dox 的 HG 和 NG 在孵育 24 小时（49-57%）和 72 小时（7-25%）后显著降低细胞活力。免疫荧光测定表明，与游离药物相比，通过 HG 和 NG 递送的 Dox 具有不同的细胞定位。
通过改变肽成分的比例可以调节 Dox 的释放。负载 Dox 的 HG 和 NG 的药物不同的细胞定位表明存在替代的内化机制。高 DLC、低药物释放和初步的体外结果表明，基于肽的 HG 和 NG 作为药物递送工具具有潜在的应用前景。
