Elsevier

Biomaterials

Volume 192, February 2019, Pages 51-61
Biomaterials

Native nanodiscs from blood inhibit pulmonary fibrosis

https://doi.org/10.1016/j.biomaterials.2018.10.045Get rights and content

Abstract

Blood is a treasure trove whose constituents have attracted increasing attention for use in understanding and controlling disease. However, the functions of blood, especially with regard to its composition at the nanoscale, remain largely unknown. Inspired by exosomes and lipoproteins, the present work isolated and characterized biotic nanodiscs from human blood (BNHBs) using multiple techniques. The isolated BNHBs had diameters of 10–30 nm and a thicknesses of approximately 2.9 nm. The BNHB concentration in blood peaked at 34.5 ± 5.19 mg/mL (20-fold higher than that of high-density lipoproteins and exosomes). BNHBs had high biocompatibility, facile cell internalization and strong biological control of pulmonary fibrosis. The BNHBs were hybrids of many metalloproteins and metabolites and contained a few functional proteins similar to lipoproteins or exosomal proteins. BNHBs inhibited transforming growth factor-beta 1 (TGF-β1)-induced fibrosis damage in human embryonic lung fibroblasts (HELFs) by inhibiting the expression of α-smooth muscle actin and collagen-1 protein. BNHBs also intensively bound TGF-β1 to inhibit TGF-β1 activity in fibrogenesis. BNHBs successfully reduced pulmonary inflammation and collagen deposition in a mouse model, preventing pulmonary fibrosis. Applying the protective properties of nanodiscs may be a novel therapeutic approach for pulmonary and other diseases.

Introduction

An increasing number of recent studies support the idea that blood from healthy humans promotes patient recovery [[1], [2], [3], [4], [5]], suggesting that something in the blood protects our health. However, the specific blood constituents that promote patient recovery or protect human health are unknown. With the development of nanotechnology, the nanoscale composition (e.g., extracellular vesicle and lipoprotein (LP) levels) of blood has attracted much attention worldwide [6,7]. Low-density LPs (LDLs) and high-density LPs (HDLs) with sizes of approximately 5–12 nm, determined by transmission electron microscopy (TEM) images, modulated thrombosis by preventing von Willebrand factor self-association and subsequent platelet adhesion [8,9]. Exosomes are 40-100-nm double-membrane extracellular vesicles produced by nearly every cell type that mediate intercellular communication and affects human health and diseases [[10], [11], [12]]. The properties and functions of native materials with sizes (e.g., approximately 20–30 nm) between those of exosomes and LPs are unclear, but these species likely also affect human health and disease.

Idiopathic pulmonary fibrosis (IPF) is a chronic, progressive lung disease characterized by progressive lung scarring and a histological pattern of usual interstitial pneumonia (UIP). IPF affects almost 3 million people worldwide, with incidence increasing dramatically with age [13]. Many therapies, such as nintedanib, have been identified in clinical trials as harmful and ineffective in the treatment of IPF, causing gastrointestinal (diarrhea and nausea) side effects [14,15]. Although blood pressure, red blood cells and white blood cells have been linked to pulmonary fibrosis [16,17], whether IPF is triggered or inhibited by blood is unclear.

In this work, biotic nanodiscs from human blood (BNHBs) were isolated, and they exhibited clear differences in concentration, morphology and composition from LPs and exosomes. Furthermore, the protective effects of isolated BNHBs against transforming growth factor-beta 1 (TGF-β1)-induced fibrosis damage in human embryonic lung fibroblasts (HELFs) and zeocin-induced pulmonary fibrosis in mice and the related specific signaling pathways involved were assessed. BNHBs may benefit by imitating LPs and exosomes by applying the protective properties of nanodiscs as a novel therapeutic approach for pulmonary and other diseases.

Section snippets

Isolation of nanoparticles from human blood

Whole-blood samples (n = 28) were collected from healthy volunteers from Shenyang, China. All experiments related to human blood were handled humanely and conducted in compliance with the guidelines approved by the Human & Animal Experiments Ethical Committee of Nankai University. All participants, 18 males and 10 females aged 18–76, provided informed consent prior to blood sampling. Venous blood samples were taken by venipuncture, collected in citrate tubes, quickly placed in a cooling box at

Nanodiscs are hybrids of proteins and metabolites

Nanoparticles isolated from human blood had a regular round morphology, and the diameters of the nanoparticles ranged from 10 to 30 nm, as shown by the TEM image in Fig. 1A. However, AFM images showed that the nanoparticles were nanodiscs with thicknesses of approximate 2.9 nm (Fig. 1B). The mass concentrations of the isolated nanoparticles were 34.5 ± 5.19 mg/mL, accounting for approximately 3.3% of the content of whole blood, and clearly higher than those of general LPs (less than 2 mg/mL)

Discussion

Recent work supports the idea that blood from healthy human bodies can control disease [[1], [2], [3], [4]], but the specific constituents of blood that promote the recovery of patients or protect human health are unknown. In the present work, BNHBs isolated from blood had different characteristics from those of LPs and exosomes but inhibited TGF-β1-induced damage in HELFs and zeocin-induced pulmonary fibrosis in mice. Based on TEM images, the diameters of HDLs and LDLs are approximately 5 and

Conclusions

The functions of the specific nanoscale constituents (e.g., LP and exosome levels) of blood have attracted much attention but remain largely unknown. In the present work, isolated nanodiscs (i.e., BNHBs) with a lateral size of 10–30 nm and a 2.9-nm thickness had high concentrations in blood, up to 34.5 ± 5.19 mg/mL (20-fold higher than HDL, LDL and exosome concentrations), and exhibited high biological inhibition of pulmonary fibrosis. BNHBs were hybrids of metabolites and a few functional

Ethics statement

All experiments related to human blood were performed humanely and conducted in compliance with the guidelines approved by the Human & Animal Experiments Ethical Committee of Nankai University. All experiments related to mice were performed humanely and conducted in compliance with the guidelines approved by the Human & Animal Experiments Ethical Committee of Nankai University.

Conflicts of interest

The authors declare no competing financial interests.

Acknowledgments

This work was financially supported by the National Natural Science Foundation of China (grant nos. 21722703, 31770550, 21577070 and 21876092), the Tianjin Natural Science Foundation (grant no. 16JCQNJC08400) and a 111 program (grant no. T2017002).

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    Anqi Sun and Ziyang Lai contributed equally to this work.

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