Ergün Fazilet Canatan, Kars Meltem Demirel, Kars Gökhan
Department of Biomedical Engineering, Faculty of Engineering, Necmettin Erbakan University, Konya 42140, Turkey.
Department of Molecular Biology and Genetics, Faculty of Science, Necmettin Erbakan University, Konya 42140, Turkey.
Polymers (Basel). 2025 Jul 7;17(13):1884. doi: 10.3390/polym17131884.
CSNPs synthesized via the ionic gelation method have emerged as a promising nanoplatform in diverse fields such as pharmaceuticals, nanotechnology, and polymer science due to their biocompatibility, ease of fabrication, and tunable properties. This study focuses on the development and characterization of LL37-loaded CSNPs, designed to enhance antibacterial efficacy while maintaining biocompatibility. This study pioneers a systematic loading optimization approach by evaluating the encapsulation efficiency (%EE) of antimicrobial peptide LL37 across multiple concentrations (7.5, 15, and 30 µg/mL), thereby identifying the formulation that maximizes peptide incorporation while preserving controlled release characteristics. The multi-concentration analysis establishes a new methodological benchmark for peptide delivery system development. To achieve this, CSNPs were optimized for size and stability by adjusting parameters such as the chitosan concentration, pH, and stabilizer. LL37, a potent antimicrobial peptide, was successfully encapsulated into CSNPs at concentrations of 7.5, 15, and 30 µg/mL, yielding formulations with favorable physicochemical properties. Dynamic light scattering (DLS) and Zeta sizer analyses revealed that blank CSNPs exhibited an average particle size of 180.40 ± 2.16 nm, a zeta potential (ZP) of +40.57 ± 1.82 mV, and a polydispersity index (PDI) of 0.289. In contrast, 15-LL37-CSNPs demonstrated an increased size of 210.9 ± 2.59 nm with an enhanced zeta potential of +51.21 ± 0.93 mV, indicating an improved stability and interaction potential. Field emission scanning electron microscopy (FE-SEM) analyses exhibited the round shaped morphology of nanoparticles. The release profile of LL37 exhibited a concentration-dependent rate and showed the best fit with the first-order kinetic model. Cytocompatibility assessments using the XTT assay confirmed that both blank and LL37-loaded CSNPs did not exhibit cytotoxicity on keratinocyte cells across a range of concentrations (150 µg/mL to 0.29 µg/mL). Notably, LL37-loaded CSNPs demonstrated significant antibacterial activity against and , with the 15-LL37-CSNP formulation exhibiting superior efficacy. Overall, these findings highlight the potential of LL37-CSNPs as a versatile antibacterial delivery system with applications in drug delivery, wound healing, and tissue engineering.
通过离子凝胶法合成的壳聚糖纳米粒（CSNPs）因其生物相容性、易于制备和可调节的性质，已成为制药、纳米技术和聚合物科学等不同领域中一种很有前景的纳米平台。本研究聚焦于负载LL37的CSNPs的开发与表征，旨在提高抗菌效果同时保持生物相容性。本研究开创了一种系统的负载优化方法，通过评估抗菌肽LL37在多个浓度（7.5、15和30 µg/mL）下的包封率（%EE），从而确定在保持控释特性的同时能使肽掺入最大化的配方。多浓度分析为肽递送系统的开发建立了一个新的方法学基准。为此，通过调整壳聚糖浓度、pH值和稳定剂等参数对CSNPs的尺寸和稳定性进行了优化。强效抗菌肽LL37成功以7.5、15和30 µg/mL的浓度包封入CSNPs中，得到具有良好理化性质的配方。动态光散射（DLS）和Zeta粒度分析仪分析表明，空白CSNPs的平均粒径为180.40 ± 2.16 nm，zeta电位（ZP）为 +40.57 ± 1.82 mV，多分散指数（PDI）为0.289。相比之下，15-LL37-CSNPs的粒径增大至210.9 ± 2.59 nm，zeta电位增强至 +51.21 ± 0.93 mV，表明稳定性和相互作用潜力得到改善。场发射扫描电子显微镜（FE-SEM）分析显示纳米颗粒呈圆形形态。LL37的释放曲线呈现浓度依赖性速率，并且与一级动力学模型拟合最佳。使用XTT法进行的细胞相容性评估证实，空白和负载LL37的CSNPs在一系列浓度（150 µg/mL至0.29 µg/mL）下对角质形成细胞均未表现出细胞毒性。值得注意的是，负载LL37的CSNPs对[具体细菌名称1]和[具体细菌名称2]表现出显著的抗菌活性，其中15-LL37-CSNP配方表现出更优异的效果。总体而言，这些发现突出了LL37-CSNPs作为一种多功能抗菌递送系统在药物递送、伤口愈合和组织工程中的应用潜力。
