Abstract
Decellularized extracellular matrix in the form of patches and locally injected hydrogels has long been used as therapies in animal models of disease. Here we report the safety and feasibility of an intravascularly infused extracellular matrix as a biomaterial for the repair of tissue in animal models of acute myocardial infarction, traumatic brain injury and pulmonary arterial hypertension. The biomaterial consists of decellularized, enzymatically digested and fractionated ventricular myocardium, localizes to injured tissues by binding to leaky microvasculature, and is largely degraded in about 3 d. In rats and pigs with induced acute myocardial infarction followed by intracoronary infusion of the biomaterial, we observed substantially reduced left ventricular volumes and improved wall-motion scores, as well as differential expression of genes associated with tissue repair and inflammation. Delivering pro-healing extracellular matrix by intravascular infusion post injury may provide translational advantages for the healing of inflamed tissues ‘from the inside out’.
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Data availability
The main data supporting the findings of this study are available within the Article and its Supplementary Information. Nanostring data are available from the NCBI GEO database via the identifier LINK. Source data are provided with this paper.
Code availability
Custom Matlab script for cell counting is available at https://doi.org/10.5281/zenodo.7196555. Custom R script for NanoString analysis is available at https://doi.org/10.5281/zenodo.7190262.
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Acknowledgements
We thank J. Placone for confocal imaging assistance, M. Davis for providing the rat cardiac endothelial cells, P. Duran for helpful comments during the manuscript-editing process, and the veterinary staff at the Institutional Animal Care and Use Program of the University of California, San Diego, for assistance with large-animal procedures and safety. K.L.C. and O.E.-A. acknowledge funding support for the research described in this study from the NIH NHLBI (grant numbers R01HL113468, 1R01HL165232 and R43HL150917 to K.L.C. and R01HL145709 to O.E.-A). M.T.S., M.D., J.H. and R.W. acknowledge support from the NIH NHLBI (grant number T32HL105373). M.D., J.H., H.S. and R.W. acknowledge support from the NIH NHLBI (grant numbers F31HL152686, F31HL158212, F31HL152610 and F31HL137347). M.T.S. acknowledges support from an A.H.A. predoctoral fellowship. M.A.V. acknowledges support from the N.S.F. (grant number DGE-1842165) and the Dr. John N. Nicholson Fellowship, which was awarded by Northwestern University. This work made use of the San Diego Nanotechnology Infrastructure (SDNI) of UCSD, a member of the National Nanotechnology Coordinated Infrastructure, which is supported by the National Science Foundation (Grant ECCS-2025752), and the EPIC facility of Northwestern University’s NUANCE Center, which has received support from the SHyNE Resource (NSF ECCS-2025633), the IIN, and Northwestern’s MRSEC program (NSF DMR-1720139).
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K.L.C. obtained the funding. M.T.S., K.L.C., R.R.R. and A.N.D. conceptually designed the studies. M.T.S., K.L.C. and A.N.D. interpreted the results. M.T.S., M.D., J.H. and J.M. performed the in vitro characterization and analyses. C.L. performed small-animal surgeries. S.I. performed large-animal echocardiography. R.R.R. performed large-animal surgeries. K.G.O. performed large-animal necropsy and histopathology analyses. M.T.S., R.M., M.D., R.W., J.H., J.M., H.S., T.S.L., S.B., J.C., R.K., G.D. and G.S.-G. processed tissue samples and performed analyses. R.M., R.W., J.H., J.M., T.K. and C.L. assisted with large-animal surgeries. A.B. and O.E.-A. designed and analysed the in vitro flow-adhesion assay. M.D. and R.W. assisted with gene-expression analyses. M.A.V., K.G. and N.G. designed and performed cryo-TEM imaging and analysed the data. P.C. designed and performed haemocompatibility analyses. E.K. and F.C. provided consultation, experimental design and data evaluation expertise related to imaging. M.T.S. and K.L.C. drafted the manuscript. All authors edited the manuscript.
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K.L.C. and A.N.D. hold equity in Ventrix Bio, Inc. K.L.C. is a co-founder, consultant and board member of Ventrix Bio, Inc. The other authors declare no competing interests.
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Extended data
Extended Data Fig. 1 Histological measurements post-infusion show no significant differences in infarct size or fibrosis between iECM and saline infused rats.
Infarct area at 5 weeks (a,b) and 3 days (c,d) post-infusion, reported as area (a,c) and percentage of the LV (b,d). Infarct fibrosis reported as area (e) and percentage of infarct area (f). g, Interstitial fibrosis of the remote myocardium reported as a percentage of area. N = 5 for saline and n = 6 for iECM for day 3 measurements, and n = 10 for both groups for 5 week measurements. Data are mean ± SEM. Add data are biological replicates.
Extended Data Fig. 2 Correlation matrices for the NanoString nCounter data.
Correlation matrix for day 1 post-infusion significantly differentially expressed genes only (a) and all genes in the Nanostring nCounter custom cardiac codeset (b). Correlation matrix for day 3 post-infusion significantly differentially expressed genes (c) and all genes in the Nanostring nCounter custom cardiac codeset (d). The Nanostring panel used was a 380 gene custom panel designed to probe for differences in gene expression across a wide range of myocardial injury models.
Extended Data Fig. 3 Effect of iECM on metabolic activity and viability of rat cardiac endothelial cells, ROS scavenging, and thiol content.
Percent difference in reduction in alamarBlue activity of rat cardiac endothelial cells in response to hydrogen peroxide with and without iECM, relative to controls with no hydrogen peroxide (a, n = 6 both groups, **p = 0.006). Percent viability as measured by Calcein-AM staining of rat cardiac endothelial cells in response to hydrogen peroxide with and without iECM, relative to controls with no hydrogen peroxide (b, n = 4 both groups, ***p < 0.001). Concentration of hydrogen peroxide was monitored following incubation with either PBS or iECM, showing a continuing decrease with iECM (c, *p = 0.01, **p = 0.003, n = 3 for both groups at 1 and 24 hrs, n = 2 at 6 hrs). Thiol content was determined compared to N-acetylcysteine standard in iECM per mg of material (d, n = 3). Data are mean ± SEM. All data are biological replicates and were evaluated with a two tailed unpaired t-test.
Extended Data Fig. 4 Additional echocardiography results showing that iECM infusions mitigate negative LV remodelling in a pig acute MI model.
a, Representative M-mode echocardiographic images showing that iECM mitigates negative LV remodelling. Yellow arrows represent LV diastolic dimension, red arrows represent wall thickness, and white arrows represent wall thinning. b-g, LV diastolic dimension (LVDd, #p = 0.08), LV systolic dimension (LVDs), and fractional shortening (FS) over time (b,d,f) and changes from post-MI to 8 weeks post-MI (c, e (*p = 0.03), g (*p = 0.047)). h, Diagram demonstrating how infarct angle was measured. N = 10 all groups. Data are mean ± SEM. All data are biological replicates and were evaluated with a two tailed unpaired t-test.
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Spang, M.T., Middleton, R., Diaz, M. et al. Intravascularly infused extracellular matrix as a biomaterial for targeting and treating inflamed tissues. Nat. Biomed. Eng 7, 94–109 (2023). https://doi.org/10.1038/s41551-022-00964-5
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DOI: https://doi.org/10.1038/s41551-022-00964-5
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