Monascus pigment-protected bone marrow-derived stem cells for heart failure treatment

Abstract

Mesenchymal stem cells (MSCs) have demonstrated significant therapeutic potential in heart failure (HF) treatment. However, their clinical application is impeded by low retention rate and low cellular activity of MSCs caused by high inflammatory and reactive oxygen species (ROS) microenvironment. In this study, monascus pigment (MP) nanoparticle (PPM) was proposed for improving adverse microenvironment and assisting in transplantation of bone marrow-derived MSCs (BMSCs). Meanwhile, in order to load PPM and reduce the mechanical damage of BMSCs, injectable hydrogels based on Schiff base cross-linking were prepared. The PPM displays ROS-scavenging and macrophage phenotype-regulating capabilities, significantly enhancing BMSCs survival and activity in HF microenvironment. This hydrogel demonstrates superior biocompatibility, injectability, and tissue adhesion. With the synergistic effects of injectable, adhesive hydrogel and the microenvironment-modulating properties of MP, cardiac function was effectively improved in the pericardial sac of rats. Our results offer insights into advancing BMSCs-based HF therapies and their clinical applications.

Keywords: BMSCs; Heart failure; Hydrogel; Microenvironment; Monascus pigment.

Graphical abstract

Fig. 1

Schematic design of the monascus pigments-encapsulated HY for BMSCs-based HF treatment.

Fig. 2

(A) Synthesis pathway of PEG-PCL. (B) FT-IR spectra of COOH-PEG-COOH, PEG, PCL, and PEG-PCL. Particle size of (C) PP and (D) PPM. (E) Zeta potential of PP and PPM. TEM image of (E) PP and (F) PPM. (H) MP release profile of PPM and HY@PPM; (I–J) Free radical scavenging ability of MP particles at different concentrations.

Fig. 3

(A) Synthesis pathways of OHA, HADop, and HY. (B) FT-IR spectra of HADop, OHA, CMC, and HY. (C) Gelation behavior, injectability and tissue adhesion of HY. SEM images of (D) HY and (E) HY@PPM. (F–G) Rheological properties of HY. (H) Swelling rate of HY. (I) Degradation behavior of HY.

Fig. 4

(A) Macrophage phagocytosis according to the MP fluorogram. (B–C) FC results and statistics of macrophage phagocytosis of MP. (D–E) FC results and statistics of macrophage phenotypes. (F) ELISA analysis of TNF-α and IL-10 secretion by macrophages.

Fig. 5

(A) DCFH-DA fluorescence assay of ROS in H9c2 cells. (B–C) FC assay results and statistics of ROS in H9c2 cells. (D) Results of WB analysis of HO-1 protein expression in H9c2 cells. (E) Statistics of HO-1 protein expression in H9c2 cells (quantitative statistics using tubulin as a control). (F) Fluorescence 3-D modeling of the distribution of HY@BMSCs and HY@PPM&BMSCs under normal and ROS conditions. (G) Activity of HY@BMSCs as well as HY@PPM in normal and ROS environments for 1–7 days. (H) Effect of PPM on the VEGF secretion of BMSCs in adverse microenvironments. (I) Expression of the BAX gene after 1–7 days of coculture of BMSCs with H9c2 cells.

Fig. 6

(A–B) DCFH-DA FC assay and statistics of intracellular ROS in the cardiomyocytes of treated HF rats. (C) ELISA results for ROS in the ECM. (D) Neutrophil percentage in routine blood tests in HF rats after treatment. (E–F) Distribution and statistics of CD86⁺ cells in the heart tissue of HF rats after treatment. (G) Expression levels of TNF-α and IL-10 in cardiac tissues of HF rats after treatment. (H–I) Retention of BMSCs in the pericardium for 1–4 days and statistical analysis. Unless otherwise specified, the relative quantities in the above figures were calculated and statistically analyzed using normal rats as controls.

Fig. 7

(A) H&E staining of heart tissue from treated HF rats. (B) Statistical analysis of ventricular wall thickness. (C) CD31 (red) staining of rat heart tissue slices after treatment. (D) Random statistics of the number of new blood vessels (n = 6). (E) Masson staining of cardiac tissue. (F) Statistics of the relative fibrotic area (calculated using normal rats as a control).

Fig. 8

(A–B) Annexin-FITC (Green) staining of cardiac tissue slices and statistical analysis of myocardial cell apoptosis. (C) Echocardiographic evaluation of left ventricular function. (D) The LVEF of normal rats and treated HF rats. (E) The FS of normal rats and treated HF rats.