Dendrimer-Mediated Intracellular Delivery of Fibronectin Guides Macrophage Polarization to Alleviate Acute Lung Injury

Abstract

Fibronectin (FN) is an essential glycoprotein in the extracellular matrix with favorable biological functions for potential applications in various biomedical fields including wound healing, regenerative medicine, tissue engineering, as well as diagnosis and treatment of cancer and inflammatory diseases. Herein, we aim to explore the influence of intracellular FN delivery on macrophage functions and its possible therapeutic applications. We prepared phenylboronic acid (PBA)-functionalized generation 5 (G5) poly(amidoamine) dendrimers (G5.NH 2 -PBA) as a nanocarrier to load FN, and reveal that the obtained dendrimers enable efficient intracellular delivery of FN at an optimized dendrimer-to-FN weight ratio of 8, which guides macrophages toward antiinflammatory M2 phenotype polarization. Studies on action mechanisms show that the dendrimer-mediated FN intracellular delivery acts strongly on suppressing the nuclear factor-κB pathway, leading to reduced pro-inflammatory cytokine secretion and enhanced reactive oxygen species depletion in lipopolysaccharide (LPS)-activated macrophages. Further investigation in vivo using an LPSinduced mouse model of acute lung injury (ALI) shows that the dendrimer-mediated FN delivery can effectively alleviate the ALI symptoms through alleviation of lung inflammation and oxidation stress. Our work suggests a general approach to using dendrimers for mediating intracellular delivery of FN, thereby offering many opportunities to explore the biological functions of FN for different therapeutic applications toward inflammation-associated diseases.

Bioactive Phosphorus Dendrimers as a Universal Protein Delivery System for Enhanced Anti-inflammation Therapy

“…FN, a noncollagenous glycoprotein in the extracellular matrix, is known to play a crucial role in mediating cell surface interactions including cell differentiation, growth, adhesion, and other biological processes. Importantly, FN can be used as a therapeutic protein for the treatment of inflammatory diseases by inducing macrophage polarization and reducing the expression of pro-inflammatory factors. , However, the low intracellular delivery efficiency of FN quite limits its biological activity. Therefore, we utilized the AK-137 dendrimer with intrinsic anti-inflammatory activity for intracellular FN delivery to exert enhanced synergistic anti-inflammatory effects.…”

“…FN has been reported to increase the intracellular antioxidant enzyme expression to alleviate oxidative stress from inflammatory responses. , We next investigated the ability of AK-137@FN NCs to scavenge reactive oxygen species (ROS) in macrophage cells. The fluorescence intensity of NCs-treated macrophages was detected by flow cytometry after coincubation with an ROS fluorescence probe DCFH-DA (Figure E,F).…”

“…In this context, noncovalent nanocarriers, such as liposomes, nanogels, exosomes, , and polymers , have been developed to provide more options for efficient intracellular protein delivery. It has been demonstrated that poly­(amidoamine) (PAMAM) dendrimers or polyethylenimines (PEI) modified with phenylboronic acid, − guanidyl ligand, , fluorous chains, , and natural polyphenols can bind proteins through physical interactions and operate as a universal vehicle for the intracellular transport of various proteins. However, these polymer-based protein delivery systems rely on the chemical modification of functional molecules and are frequently endowed with a high surface positive potential to promote their binding to proteins, and hence this protein delivery technology is time-consuming and accompanied by biosafety concerns raised by the positive charges of vectors .…”

Nanoparticle-Mediated Multiple Modulation of Bone Microenvironment To Tackle Osteoarthritis

“…G4-TBP, a white solid, C 2064 H 3024 N 279 O 762 P 267 S 90 , M w = 55058.13 g/ mol, and yield = 42%. 31 Synthesis of G4-TBP NPs. In brief, G4-TBP (1.50 mg) and HOOC-PEG-TK-PLGA (1.05 mg, obtained from Xi'an Ruixi Biological Technology, Shanghai, China) were co-dissolved in DMSO (500 μL), dropped into water (5 mL) under sonication and stirred for 24 h at room temperature.…”

Self-Assembly of Epitope-Tagged Proteins and Antibodies for Delivering Biologics to Antigen Presenting Cells

“…Antigen-presenting cells (APCs) have gained significant attention as therapeutic targets due to their role in the immune system and various diseases including arthritis, Gaucher, and cancer. − Therefore, APC-targeting nanocarriers have been of recent interest. − Most of these studies mainly focus on hydrophobic drugs and very little attention has been given to biologics delivery, ,− despite the enormous impact potential in gene-editing, drugging the undruggable, growth factor-based therapeutics, and treating genetic diseases (e.g., Gaucher). Designing delivery vehicles for biologics is challenging because of their large size, sensitivity to non-native environments, and inherently lower operating concentrations.…”