CoChat
Sign Up
Cited by 111Tissue Engineering and Regenerative MedicineMesenchymal stem cell researchMuscle Physiology and Disorders

The DOI matches its Crossref record; 64 of the 64 references it cites were found in a registry or an index.

Mesenchymal stem cells and extracellular matrix scaffold promote muscle regeneration by synergistically regulating macrophage polarization toward the M2 phenotype

Open access
Qiu, Xinyu, Liu, Shiyu, Zhang, Hao, Zhu, Bin, Su, Yuting, Zheng, Chenxi, Tian, Rong, Wang, Miao, Kuang, Huijuan, Zhao, Xinyi, Jin, Yan·2018·Stem Cell Research & Therapy
DOI 10.1186/s13287-018-0821-5PMC PMC5883419
Open this paper in CoChat — free accountOpen PDF

Abstract

BACKGROUND: Skeletal muscle plays an important role in the body's physiology but there are still no effective treatments for volumetric muscle loss (VML) resulting from severe traumatic injury or tumor excision. Recent studies show that a tissue engineering strategy using a compound containing mesenchymal stem cells (MSCs) and decellularized extracellular matrix (ECM) scaffold generates significant regenerative effects on VML injury, but the underlying mechanisms are not fully understood. METHODS: The characteristics of human umbilical cord MSCs, including multiplication capacity and multidifferentiation ability, were determined. We constructed a compound containing MSCs and decellularized ECM scaffold which was used for tissue regeneration in a VML model. RESULTS: We found that MSCs and decellularized ECM scaffold generated synergistic effects on promoting skeletal muscle tissue regeneration. Interestingly, both MSCs and decellularized ECM scaffold could promote macrophage polarization toward the M2 phenotype and suppress macrophage polarization toward the M1 phenotype, which is widely regarded as an important promoting factor in tissue regeneration. More importantly, MSCs and decellularized ECM scaffold generate synergistic promoting effects on macrophage polarization toward the M2 phenotype, not just an additive effect. CONCLUSIONS: Our findings uncover a previously unknown mechanism that MSCs and decellularized ECM scaffold promote tissue regeneration via collaboratively regulating macrophage polarization.

Overview

The claims and statistics of this paper haven’t been checked yet.

Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.

A few free checks a day · papers already verified open instantly · results are public

Integrity signals

Registry

Crossref · 10.1186/s13287-018-0821-5

Cites

64 of 64 references verified0 citation errors · 0 not checkable

Statistics

Figures

Fig. 1 Decellularization of native heart tissue and biochemical analysis. a Representative optical images of native and decellularized heart tissue. b H&E staining images, Masson’s Trichrome staining images and SEM images of native and decellularized heart tissue. Scale bar, 100 μm. c Representative images of immunofluorescence staining for collagen I, fibronectin and laminin of native and decellularized heart tissue. Scale bar, 200 μm. d ECM components (COL and GAGs) and DNA contents of native and decellularized heart tissue. All experiments performed in triplicate. Data shown as mean ± SD. *P < 0.05; NS, not significant (P > 0.05). H&E hematoxylin and eosin, SEM scanning electron microscopy, H-dECM heart decellularized extracellular matrix, GAG glycosaminoglycan
Fig. 1 Decellularization of native heart tissue and biochemical analysis. a Representative optical images of native and decellularized heart tissue. b H&E staining images, Masson’s Trichrome staining images and SEM images of native and decellularized heart tissue. Scale bar, 100 μm. c Representative images of immunofluorescence staining for collagen I, fibronectin and laminin of native and decellularized heart tissue. Scale bar, 200 μm. d ECM components (COL and GAGs) and DNA contents of native and decellularized heart tissue. All experiments performed in triplicate. Data shown as mean ± SD. *P < 0.05; NS, not significant (P > 0.05). H&E hematoxylin and eosin, SEM scanning electron microscopy, H-dECM heart decellularized extracellular matrix, GAG glycosaminoglycanCC BY · PubMed Central
Fig. 2 Characterization of human umbilical cord mesenchymal stem cells (hUCMSCs). a Flow cytometric analysis of ex-vivo-expanded hUCMSCs revealed positive expression of CD29, CD44, CD90 and CD105, and negative expression of CD31, CD34 and CD45. b Representative percentage of cell markers for identification. c Representative proliferation of single clone of hUCMSCs. d hUCMSCs seeded in plates induced with osteogenic medium for 10 days. Activity of ALP detected by ALP staining. e hUCMSCs cultured in osteogenic inductive conditions for 28 days, mineralized nodules found by Alizarin Red staining. f Cultured hUCMSCs formed Oil Red O-positive lipid cluster following 14 days of adipogenic induction. g–j Cell aggregate (g control) positively expressed fibronectin (h), collagen I (i) and integrin-β1 (j). k Arrangement of cells in cell aggregate observed by SEM. l H&E staining showing cell aggregate was dense and contained plenty of cells. n = 6 per group. Scale bar, 100 μm. PE phycoerythrin, CFU-F fibroblastic colony-forming unit, ALP alkaline phosphatase, Con control, SEM scanning electron microscopy, H&E hematoxylin and eosin
Fig. 2 Characterization of human umbilical cord mesenchymal stem cells (hUCMSCs). a Flow cytometric analysis of ex-vivo-expanded hUCMSCs revealed positive expression of CD29, CD44, CD90 and CD105, and negative expression of CD31, CD34 and CD45. b Representative percentage of cell markers for identification. c Representative proliferation of single clone of hUCMSCs. d hUCMSCs seeded in plates induced with osteogenic medium for 10 days. Activity of ALP detected by ALP staining. e hUCMSCs cultured in osteogenic inductive conditions for 28 days, mineralized nodules found by Alizarin Red staining. f Cultured hUCMSCs formed Oil Red O-positive lipid cluster following 14 days of adipogenic induction. g–j Cell aggregate (g control) positively expressed fibronectin (h), collagen I (i) and integrin-β1 (j). k Arrangement of cells in cell aggregate observed by SEM. l H&E staining showing cell aggregate was dense and contained plenty of cells. n = 6 per group. Scale bar, 100 μm. PE phycoerythrin, CFU-F fibroblastic colony-forming unit, ALP alkaline phosphatase, Con control, SEM scanning electron microscopy, H&E hematoxylin and eosinCC BY · PubMed Central
Fig. 3 Gross morphology and functional evaluation of TA muscle after VML injury and repair. a Gross appearance of VML-injured muscle in four groups. b–d Representative central thickness of TA muscle (b) and widths of the distal (c) and proximal (d) thirds of the TA muscle in each group. ECM-treated group, MSC-treated group and compound-treated group can all partially recover central thickness and distal width of TA muscle. e, f Mechanical function evaluation by ES-induced contractions observed in each group presented at 1 month (e) and 2 months (f). g Mean body weight of four groups over the course of 8 weeks. No differences observed among all groups at each time point. n= 6 per group. Data shown as mean ± SD. *P < 0.05; **P < 0.01; NS, not significant (P > 0.05). PBS phosphate buffer saline, ECM extracellular matrix, MSC mesenchymal stem cell, TA tibialis anterior
Fig. 3 Gross morphology and functional evaluation of TA muscle after VML injury and repair. a Gross appearance of VML-injured muscle in four groups. b–d Representative central thickness of TA muscle (b) and widths of the distal (c) and proximal (d) thirds of the TA muscle in each group. ECM-treated group, MSC-treated group and compound-treated group can all partially recover central thickness and distal width of TA muscle. e, f Mechanical function evaluation by ES-induced contractions observed in each group presented at 1 month (e) and 2 months (f). g Mean body weight of four groups over the course of 8 weeks. No differences observed among all groups at each time point. n= 6 per group. Data shown as mean ± SD. *P < 0.05; **P < 0.01; NS, not significant (P > 0.05). PBS phosphate buffer saline, ECM extracellular matrix, MSC mesenchymal stem cell, TA tibialis anteriorCC BY · PubMed Central
Fig. 4 TA muscle histological study following repair in four groups after transplantation for 8 weeks. a TA muscles harvested 8 weeks after VML injury. Longitudinal and cross-sections from four groups stained with H&E (D–K) and Masson’s Trichrome (L–S). b, c Surface ratio of new muscle fibers (b) and collagen fibers (c) formed in each group analyzed by Image-Pro Plus 6.0. n = 6 per group. #Area of the defect. Dashed line represents separation between two areas. Scale bar, 200 μm. Data shown as mean ± SD. **P < 0.01; ***P < 0.001; NS, not significant (P > 0.05). PBS phosphate buffer saline, ECM extracellular matrix, MSC mesenchymal stem cell
Fig. 4 TA muscle histological study following repair in four groups after transplantation for 8 weeks. a TA muscles harvested 8 weeks after VML injury. Longitudinal and cross-sections from four groups stained with H&E (D–K) and Masson’s Trichrome (L–S). b, c Surface ratio of new muscle fibers (b) and collagen fibers (c) formed in each group analyzed by Image-Pro Plus 6.0. n = 6 per group. #Area of the defect. Dashed line represents separation between two areas. Scale bar, 200 μm. Data shown as mean ± SD. **P < 0.01; ***P < 0.001; NS, not significant (P > 0.05). PBS phosphate buffer saline, ECM extracellular matrix, MSC mesenchymal stem cellCC BY · PubMed Central
Fig. 5 Macrophage polarization toward M2 phenotype after transplantation for 2 weeks. a Only a small amount of CD206 (red)-positive macrophage appeared in the PBS-treated group, few in the ECM-treated group and MSC-treated group, and plenty of CD206-positive macrophages detected in the compound treated group. Nuclei stained with Hoechst 33342 (blue). b Field of view for CD206-positive macrophages in each group analyzed by Image-Pro Plus 6.0. c Plenty of iNOS (red)-positive macrophages showed in the PBS-treated group, meanwhile, only few can be detected in the compound-treated group. Nuclei were stained with Hoechst 33342 (blue). d Field of view for iNOS-positive macrophages in each group analyzed by Image-Pro Plus 6.0. n = 6 per group. #Area of defect. Dashed line represents separation between two areas. Scale bar, 200 μm. Data shown as mean ± SD. **P < 0.01; ***P < 0.001; ****P < 0.0001; NS, not significant (P > 0.05). PBS phosphate buffer saline, ECM extracellular matrix, MSC mesenchymal stem cell, iNOS inducible nitric oxide synthase
Fig. 5 Macrophage polarization toward M2 phenotype after transplantation for 2 weeks. a Only a small amount of CD206 (red)-positive macrophage appeared in the PBS-treated group, few in the ECM-treated group and MSC-treated group, and plenty of CD206-positive macrophages detected in the compound treated group. Nuclei stained with Hoechst 33342 (blue). b Field of view for CD206-positive macrophages in each group analyzed by Image-Pro Plus 6.0. c Plenty of iNOS (red)-positive macrophages showed in the PBS-treated group, meanwhile, only few can be detected in the compound-treated group. Nuclei were stained with Hoechst 33342 (blue). d Field of view for iNOS-positive macrophages in each group analyzed by Image-Pro Plus 6.0. n = 6 per group. #Area of defect. Dashed line represents separation between two areas. Scale bar, 200 μm. Data shown as mean ± SD. **P < 0.01; ***P < 0.001; ****P < 0.0001; NS, not significant (P > 0.05). PBS phosphate buffer saline, ECM extracellular matrix, MSC mesenchymal stem cell, iNOS inducible nitric oxide synthaseCC BY · PubMed Central
Fig. 6 Gene expression in inguinal lymph node after 2 weeks post operation. Gene expression measured in local (inguinal) lymph nodes at 2 weeks after transplantation to measure M1 phenotype-related (TNF-α, IL-1β, IL-6) (a–c) and M2 phenotype-related (IL-4, IL-10, IL-13) (d–f) cytokine changes dependent on ECM and MSCs application. n = 6 per group. Data shown as mean ± SD. **P < 0.01; ****P < 0.0001; NS, not significant (P > 0.05). IL, interleukin, PBS phosphate buffer saline, ECM extracellular matrix, MSC mesenchymal stem cell, TNF-α tumor necrosis factor alpha
Fig. 6 Gene expression in inguinal lymph node after 2 weeks post operation. Gene expression measured in local (inguinal) lymph nodes at 2 weeks after transplantation to measure M1 phenotype-related (TNF-α, IL-1β, IL-6) (a–c) and M2 phenotype-related (IL-4, IL-10, IL-13) (d–f) cytokine changes dependent on ECM and MSCs application. n = 6 per group. Data shown as mean ± SD. **P < 0.01; ****P < 0.0001; NS, not significant (P > 0.05). IL, interleukin, PBS phosphate buffer saline, ECM extracellular matrix, MSC mesenchymal stem cell, TNF-α tumor necrosis factor alphaCC BY · PubMed Central

Verification

Claims the paper makes

The claims and statistics of this paper haven’t been checked yet.

Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.

A few free checks a day · papers already verified open instantly · results are public

Claims other papers make about this one

Each is checked against this paper’s own text: does it say what the citing paper says it does?

Self-check

Percentages, sums and reported differences re-computed from the paper’s own numbers.

The self-check needs the paper’s full text.

Citations

Cites

Cited by

Sorted by citations

See all 111

Related

See all

Metadata

Authors
Qiu, X., Liu, S., Zhang, H., Zhu, B., Su, Y., Zheng, C., Tian, R., Wang, M., Kuang, H., Zhao, X., & Jin, Y.
Affiliations
  • Air Force Medical University (CN)
  • Chinese People's Liberation Army (CN)
Corresponding author
  • Zhao, Xinyi
  • Jin, Yan
ORCID
Venue
Stem Cell Research & Therapy
Publisher
Springer Science and Business Media LLC
Published
2018-04-03
Volume
9
Issue
1
Pages
88-88
ISSN
1757-6512 (ISSN-L)
DOI
10.1186/s13287-018-0821-5
PMC
PMC5883419
OpenAlex
W2800380986
Cited by
111
References
64
Citations by year
2026: 5 · 2025: 12 · 2024: 13 · 2023: 7 · 2022: 17 · 2021: 17 · 2020: 21 · 2019: 13 · 2018: 6
FWCI
14.81
Type
journal article
Language
en
Open access
Licence
Locations
  • Stem Cell Research & Therapy (publishedVersion, cc-by)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
Funders
  • National Natural Science Foundation of China (31670995, 81470679)
  • National Key Research and Development Program of China (2016YFC1101400, 2016TFC1102903)
Topics
  • Health Sciences › Medicine › Surgery › Tissue Engineering and Regenerative Medicine
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Mesenchymal stem cell research
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Muscle Physiology and Disorders
Keywords
mesenchymal stem cells, skeletal muscle regeneration, volumetric muscle loss, macrophage polarization, M2 macrophages, M1 macrophages, human umbilical cord mesenchymal stem cells, synergistic effect, tissue engineering, traumatic muscle injury
MeSH terms
Animals, Cell Differentiation, Cells, Cultured, Extracellular Matrix, Humans, Macrophages, Regeneration, Rats, Sprague-Dawley, Muscle, Skeletal, Mesenchymal Stem Cell Transplantation, Rats, Mesenchymal Stem Cells

About these checks

Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.

Statistics and arithmetic are recomputed, not judged.

Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.

What counts as a claim ›How we measure confidence ›How we check this ›

Are you an author, or do you see a problem with this page? Request a review