Potential of Plant-derived Exosome-like Nanoparticles from Physalis peruviana Fruit for Human Dermal Fibroblast Regeneration and Remodeling.

Q2 Pharmacology, Toxicology and Pharmaceutics

Pharmaceutical nanotechnology Pub Date : 2025-01-01 DOI:10.2174/0122117385281838240105110106

Filia Natania, Iriawati Iriawati, Fitria Dwi Ayuningtyas, Anggraini Barlian

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引用次数: 0

Abstract

Aims: This research aimed to study the potential of PDEN from P. peruviana fruits (PENC) for regenerating and remodeling HDF.

Background: Large wounds are dangerous and require prompt and effective healing. Various efforts have been undertaken, but have been somewhat ineffective. Plant-derived exosome-like nanoparticles (PDEN) are easily sampled, relatively cost-effective, exhibit high yields, and are nonimmunogenic.

Objectives: The objective of the study was to isolate and characterize PDEN from Physalis peruviana (PENC), and determine PENC's internalization and toxicity on HDF cells, PENC's ability to regenerate HDF (proliferation and migration), and PENC ability's to remodel HDF (collagen I and MMP-1 production).

Methods: PENC was isolated using gradual filtration and centrifugation, followed by sedimentation using PEG6000. Characterization was done using a particle size analyzer, zeta potential analyzer, TEM, and BCA assay. Internalization was done using PKH67 staining. Toxicity and proliferation assays were conducted using MTT assay; meanwhile, migration assay was carried out by employing the scratch assay. Collagen I production was performed using immunocytochemistry and MMP-1 production was conducted using ELISA.

Results: MTT assay showed a PENC concentration of 2.5 until 500 μg/mL and being non-toxic to cells. PENC has been found to induce cell proliferation in 1, 3, 5, and 7 days. PENC at a concentration of 2.5, 5, and 7.5 μg/mL, also accelerated HDF migration using the scratch assay in two days. In remodeling, PENC upregulated collagen-1 expression from day 7 to 14 compared to control. MMP-1 declined from day 2 to 7 in every PENC concentration and increased on day 14. Overall, PENC at concentrations of 2.5, 5, and 7.5 μg/mL induced HDF proliferation and migration, upregulated collagen I production, and decreased MMP-1 levels.

Conclusion: Isolated PENC was 190-220 nm in size, circular, covered with membrane, and its zeta potential was -6.7 mV; it could also be stored at 4°C for up to 2 weeks in aqua bidest. Protein concentration ranged between 170-1,395 μg/mL. Using PKH67, PENC could enter HDF within 6 hours. PENC was non-toxic up to a concentration of 500 μg/mL. Using MTT and scratch assay, PENC was found to elevate HDF proliferation and migration, and reorganize actin. Using immunocytochemistry, collagen I was upregulated by PENC, whereas MMP-1 concentration was reduced.

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从秘鲁苦蘵果实中提取的植物外泌体纳米颗粒对人类真皮成纤维细胞再生和重塑的潜力

目的：本研究旨在研究来自秘鲁果实（PENC）的 PDEN 在再生和重塑 HDF 方面的潜力：背景：大面积伤口非常危险，需要及时有效的愈合。背景：大面积伤口非常危险，需要及时、有效地愈合。人们已经做出了各种努力，但效果不佳。植物外泌体纳米颗粒（PDEN）易于取样、成本效益相对较高、产量高且无免疫原性：本研究的目的是从秘鲁苦蘵（Physalis peruviana，PENC）中分离和鉴定 PDEN，并确定 PENC 对 HDF 细胞的内化和毒性、PENC 再生 HDF 的能力（增殖和迁移）以及 PENC 重塑 HDF 的能力（胶原蛋白 I 和 MMP- 1 的产生）：方法：使用渐进过滤和离心法分离 PENC，然后使用 PEG6000 进行沉淀。使用粒度分析仪、ZETA 电位分析仪、TEM 和 BCA 分析法进行表征。内化是通过 PKH67 染色法完成的。毒性和增殖试验采用 MTT 法进行；迁移试验采用划痕法进行。胶原蛋白 I 的产生采用免疫细胞化学法，MMP-1 的产生采用酶联免疫吸附法：MTT 检测显示，PENC 的浓度为 2.5 至 500 μg/mL，对细胞无毒性。研究发现，PENC 可在 1、3、5 和 7 天内诱导细胞增殖。浓度为 2.5、5 和 7.5 μg/mL 的 PENC 还能在两天内通过划痕试验加速 HDF 的迁移。在重塑过程中，与对照组相比，PENC 可在第 7 到 14 天内上调胶原蛋白-1 的表达。从第 2 天到第 7 天，MMP-1 在每个 PENC 浓度下都会下降，并在第 14 天上升。同时，与对照组相比，ROS 在第 2 天至第 7 天和第 14 天均有所下降，而 PENC 则会诱导 ROS 水平上升。总之，浓度为 2.5、5 和 7.5 μg/mL 的 PENC 可诱导 HDF 增殖和迁移，上调胶原蛋白 I 的生成，并降低 MMP-1 的水平：分离出的PENC大小为190-220 nm，呈圆形，被膜覆盖，其zeta电位为-6.7 mV；在4°C水溶液中可保存2周。蛋白质浓度在 170-1,395 μg/mL 之间。使用 PKH67，PENC 可在 6 小时内进入 HDF。五氯苯甲醚在浓度为 500 μg/mL 时无毒。通过 MTT 和划痕试验发现，五氯苯甲醚能促进 HDF 的增殖和迁移，并使肌动蛋白重组。通过免疫细胞化学，PENC 上调了胶原蛋白 I 的浓度，而 MMP-1 的浓度则有所降低。

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来源期刊

Pharmaceutical nanotechnology Pharmacology, Toxicology and Pharmaceutics-Pharmaceutical Science

CiteScore

4.20

自引率

0.00%

发文量

期刊介绍： Pharmaceutical Nanotechnology publishes original manuscripts, full-length/mini reviews, thematic issues, rapid technical notes and commentaries that provide insights into the synthesis, characterisation and pharmaceutical (or diagnostic) application of materials at the nanoscale. The nanoscale is defined as a size range of below 1 µm. Scientific findings related to micro and macro systems with functionality residing within features defined at the nanoscale are also within the scope of the journal. Manuscripts detailing the synthesis, exhaustive characterisation, biological evaluation, clinical testing and/ or toxicological assessment of nanomaterials are of particular interest to the journal’s readership. Articles should be self contained, centred around a well founded hypothesis and should aim to showcase the pharmaceutical/ diagnostic implications of the nanotechnology approach. Manuscripts should aim, wherever possible, to demonstrate the in vivo impact of any nanotechnological intervention. As reducing a material to the nanoscale is capable of fundamentally altering the material’s properties, the journal’s readership is particularly interested in new characterisation techniques and the advanced properties that originate from this size reduction. Both bottom up and top down approaches to the realisation of nanomaterials lie within the scope of the journal.