An MMP-degradable and conductive hydrogel to stabilize HIF-1α for recovering cardiac functions

Xiaojuan Wei, Si Chen, Tian Xie, Hongchi Chen, Xin Jin, Jumin Yang, Shafaq Sahar, Huanlei Huang, Shuoji Zhu, Nanbo Liu, Changjiang Yu, Ping Zhu, Wei Wang, Wei Zhang, Xiaojuan Wei, Si Chen, Tian Xie, Hongchi Chen, Xin Jin, Jumin Yang, Shafaq Sahar, Huanlei Huang, Shuoji Zhu, Nanbo Liu, Changjiang Yu, Ping Zhu, Wei Wang, Wei Zhang

Abstract

Rationale: Although a few injectable hydrogels have shown a reliable biosafety and a moderate promise in treating myocardial infarction (MI), the updated hydrogel systems with an on-demand biodegradation and multi-biofunctions to deliver therapeutic drug would achieve more prominent efficacy in the future applications. In this report, a conductive and injectable hydrogel crosslinked by matrix metalloproteinase-sensitive peptides (MMP-SP) was rationally constructed to stabilize hypoxia-inducible factor-1α (HIF-1α) to recover heart functions after MI. Methods: Firstly, tetraaniline (TA) was incorporated into partially oxidized alginate (ALG-CHO) to endow the hydrogels with conductivity. The 1,4-dihydrophenonthrolin-4-one-3-carboxylic acid (DPCA) nanodrug was manufactured with high drug loading capacity and decorated with polymerized dopamine (PDA) to achieve a stable release of the drug. Both ALG-CHO and DPCA@PDA can be cross-linked by thiolated hyaluronic acid (HA-SH) and thiolated MMP-SP to construct a MMP-degradable and conductive hydrogel. After administration in the infarcted heart of rats, echocardiographic assessments, histological evaluation, and RT-PCR were used to evaluate therapeutic effects of hydrogels. Results: The cell viability and the results of subcutaneous implantation verify a good cytocompatibility and biocompatibility of the resulting hydrogels. The hydrogel shows remarkable strength in decreasing the expression of inflammatory factors, maintaining a high level of HIF-1α to promote the vascularization, and promoting the expression of junctional protein connexin 43. Meanwhile, the multifunctional hydrogels greatly reduce the infarcted area (by 33.8%) and improve cardiac functions dramatically with ejection fraction (EF) and fractional shortening (FS) being increased by 31.3% and 19.0%, respectively. Conclusion: The as-prepared hydrogels in this report achieve a favorable therapeutic effect, offering a promising therapeutic strategy for treating heart injury.

Trial registration: ClinicalTrials.gov NCT01226563.

Keywords: conductivity; hypoxia-inducible factor-1α; injectable hydrogel; matrix metalloproteinase; myocardial infarction.

Conflict of interest statement

Competing Interests: The authors have declared that no competing interest exists.

© The author(s).

Figures

Figure 1
Figure 1
The schematic design of an MMP-degradable and conductive hydrogel to stabilize HIF-1α for recovering heart functions after MI. The hydrogel was fabricated based on functionalized HA and ALG. The conductive TA was incorporated into ALG-CHO to endow electric conductivity on the hydrogels. DPCA nanodrug was coated with PDA to achieve a high drug loading amount and maintain a stable releasing manner. Both ALG-CHO and DPCA@PDA are crosslinked by HA-SH and thiolated MMP-SP to construct the multifunctional hydrogels. The resulted hydrogels were administrated by intramyocardial injection into the infarcted rat hearts to evaluate therapy effects.
Figure 2
Figure 2
The synthesis and characterization of DPCA@PDA NPs. (A) The synthesis process of DPCA@PDA. (B) The size distribution of DPCA and DPCA@PDA NPs assessed by DLS. TEM images of DPCA NPs (C) and DPCA@PDA NPs (D).
Figure 3
Figure 3
Properties of the as-prepared ALG-CHO-TA/DPCA@PDA/MMP-SP/HA-SH hydrogels. (A) Rheological analysis based on a time-sweep model. (B) The macro picture to demonstrate the injectable capability of the hydrogels. (C) The shear-thinning behavior and (D) the fatigue resistance of ALG-CHO-TA/DPCA@PDA/MMP-SP/HA-SH hydrogel. (E) The macro picture indicates that the conductive hydrogel can act as a wire to light up LED. (F) The conductivity of the hydrogels. (G) The DPCA releasing behavior in vitro. (H) The macro pictures illustrate the subcutaneous injection of hydrogels within 7 d. (I) H&E staining of the tissue around the hydrogels (black zone in the images) after 7-day subcutaneous injection. (III: ALG-CHO/HA-SH; IV: ALG-CHO-TA/HA-SH; V: ALG-CHO-TA/DPCA@PDA/HA-SH; VI: ALG-CHO-TA/DPCA@PDA/MMP-SP/HA-SH.)
Figure 4
Figure 4
Evaluation of the cardiac functions after various treatments at different time points. (A) The representative echocardiographic images for various groups at 28 d. The statistical values of (B) EF, (C) FS, (D) LVID; d, and (E) LVID; s. (I: Sham; II: MI; III: ALG-CHO/HA-SH; IV: ALG-CHO-TA/HA-SH; V: ALG-CHO-TA/DPCA@PDA/HA-SH; VI: ALG-CHO-TA/DPCA@PDA/MMP-SP/HA-SH.)
Figure 5
Figure 5
Histological analysis of the cardiac tissue at 28 d. (A) TTC staining. (B) Masson staining. (C) Sirius red staining. Statistical data on LV wall thickness (D), the infarcted area (E), and the fibrosis area (F). (I: Sham; II: MI; III: ALG-CHO/HA-SH; IV: ALG-CHO-TA/HA-SH; V: ALG-CHO-TA/DPCA@PDA/HA-SH; VI: ALG-CHO-TA/DPCA@PDA/MMP-SP/HA-SH.)
Figure 6
Figure 6
Protein expression level of TNF-α (A) and Cas-3 (B) determined by immunofluorescent staining at 28 d. (I: Sham; II: MI; III: ALG-CHO/HA-SH; IV: ALG-CHO-TA/HA-SH; V: ALG-CHO-TA/DPCA@PDA/HA-SH; VI: ALG-CHO-TA/DPCA@PDA/MMP-SP/HA-SH.)
Figure 7
Figure 7
Immunofluorescent staining of cTnT (A, B) and Cx43 (C, D) at 28 d. B, D are the statistical data quantified and normalized to the number of nuclei. (I: Sham; II: MI; III: ALG-CHO/HA-SH; IV: ALG-CHO-TA/HA-SH; V: ALG-CHO-TA/DPCA@PDA/HA-SH; VI: ALG-CHO-TA/DPCA@PDA/MMP-SP/HA-SH.)
Figure 8
Figure 8
Protein expression level of HIF-1α (A, B), VEGFA (C, D), and α-SMA (E, F) at 28 d measured by immunofluorescent staining. B, D, and F are the statistical data quantified and normalized to the number of nuclei. (I: Sham; II: MI; III: ALG-CHO/HA-SH; IV: ALG-CHO-TA/HA-SH; V: ALG-CHO-TA/DPCA@PDA/HA-SH; VI: ALG-CHO-TA/DPCA@PDA/MMP-SP/HA-SH.)
Figure 9
Figure 9
Gene expression level at 28 d determined by RT-PCR. A-F indicate TNF-α, IL-1β, Ang-1, HIF-1α, α-Actinin, and cTnT. (I: Sham; II: MI; III: ALG-CHO/HA-SH; IV: ALG-CHO-TA/HA-SH; V: ALG-CHO-TA/DPCA@PDA/HA-SH; VI: ALG-CHO-TA/DPCA@PDA/MMP-SP/HA-SH.)

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