Minimally invasive delivery of a hydrogel-based exosome patch to prevent heart failure
Graphical abstract
Introduction
Coronary heart disease (CHD) generally presents a high prevalence in adults, and the risks of CHD increase with age in both genders [1]. Although treatment strategies have improved in the past several decades, the loss of functional myocardium cannot be restored due to the lack of regenerative capability in the adult human heart [2]. Eventually, the patients may suffer from chronic and recurrent heart failure (HF) that contributes to death through alternations in calcium homeostasis, action potential prolongation, and HF-related lung complications [3]. Stem cell therapy has been studied for decades. It has been reported as a robust strategy that can promote proliferation in infarcted myocardium through paracrine cargos such as proteins [4,5], RNA species [6,7], and extracellular vesicles [7,8]. Among those different cell types, mesenchymal stem cells (MSCs) [9] have been reported as a promising therapeutic agent restoring cardiac function by multifaceted mechanisms.
Direct cardiac delivery of MSCs suffers from low cell retention and survival. Also, potential risks of cell transplantation include immunogenicity and tumorgenicity [10,11]. Therefore, MSC-derived exosomes, or nanosized extracellular vesicles, have been studied as an alternative for MSC therapy. MSC exosomes share similar protein factors, lipid messengers, and regulatory microRNAs (miRNAs) with MSCs and induce their treatment effects mainly by triggering pro-regenerative and anti-fibrotic signaling pathways mediated by different cargo miRNAs [12]. Our previous study showed that miR-21-5p from MSC exosomes regulated PI3K signaling pathway through Akt kinase activity inhibition [13]. Additionally, MSC exosomes play an essential role in the modulation of immune reactions [14] and the reduction of cardiomyocyte apoptosis through miR-199a-3p [15]. Such mechanisms of MSCs include enhanced proliferation and reduced apoptosis of cardiomyocytes [16,17]. However, intravenously injected exosomes could be cleared rapidly from the blood and accumulate in organs such as the brain, lung, and liver rather than in the heart [18]. Even though direct intramyocardial injection of exosomes has shown promising results, it is an invasive technique with restricted application scenarios.
Various bioengineering techniques have been designed to deliver exosomes to the heart efficiently. For example, exosomes were conjugated with cardiac homing peptide or hybridized with platelet membrane to enhance the exosome homing ability after intravenous injection [19,20]. Exosomes were also loaded in cardiac patches made of natural or synthetic materials to generate higher cardiac retention than intramyocardial injection [21]. However, transplantation of cardiac patches typically requires traumatic surgery, which is not acceptable for patients with mild-to-moderate heart diseases. We designed an injectable hyaluronic acid (HA) hydrogel patch loaded with MSC-derived exosomes (ExoGel) in the present study. We tested the feasibility of delivering ExoGel into the pericardial space of rodent or porcine hearts via a thoracoscope-guided minimally invasive procedure. The acute retention and inflammatory response to ExoGel delivery were determined in rodent models, while the safety assessments were performed in healthy pigs as a translational effort.
Section snippets
Fabrication and characterization of an injectable ExoGel
Exosomes were isolated from MSCs and concentrated using the ultrafiltration method as previously described [22]. HA Hydrogel has excellent gelling properties due to its capability to bind water quickly [6]. It is suitable to be delivered into the pericardial fluid environment for sustained drug release. The safety and efficacy of HA hydrogel in heart repair have been established. ExoGel was fabricated by embedding MSC exosomes in HA hydrogel at 1 × 109 exosomes/ml (Fig. 1A). Before ExoGel
Discussion
Although exosomes may vary among different cell types, cell culture conditions, exosome isolation, and purification methods [25], researchers are still enthusiastic about applying exosomes as therapeutics. Two major theories attribute to the mechanism of exosomes: possession of proteomic potency and release of regulating miRNAs [26]. Therefore, many different biotechnologies regarding exosomes are created based on these mechanisms to increase the delivery efficiency and therapeutic potency. For
Isolation and characterization of MSC exosomes
The MSC conditioned media was concentrated via centrifugal filter units Amicon Ultra-15 (3-kDa cutoff) and washed with PBS once at the beginning of exosome isolation. Then, we used the Ultra-15 centrifugal filter units (100 kDa cutoff) to concentrate the exosomes, which were washed with PBS twice. After the wash, NanoSight LM10 (Malvern Instruments Ltd., UK) was used to examine the concentration of exosomes after a 1000-fold dilution.
Preparation of HA hydrogel and ExoGel
The HA hydrogel was prepared to test the effectiveness of
Declaration of Competing Interest
The authors declare no competing financial interests.
Acknowledgments
GC, KH, and TGC conceived and designed the research, GC and DZ performed experiments, analyzed the data, and drafted the paper with guidance from KH and TGC. All authors have reviewed the final version and approved the content in this manuscript. This work was supported by the American Heart Association (21CDA855570 to KH).
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