Adipose-Derived Stem-Cell-Membrane-Coated PLGA-PEI Nanoparticles Promote Wound Healing via Efficient Delivery of miR-21
Abstract
:1. Introduction
2. Materials and Methods
2.1. Cell Culture
2.2. Preparation of Nanocarriers
2.3. Characterization of Nanocarriers
2.4. Cell Uptake
2.5. Scratch Wound Assay
2.6. Tube Formation Assay
2.7. qRT-PCR
2.8. Western Blot
2.9. Mouse Skin Wound Model and Treatment
2.10. Histochemistry and Immunofluorescence
2.11. Statistical Analysis
3. Results
3.1. Preparation and Characteristics of Nanocarriers
3.2. Cellular Uptake Behavior of m/NP/miR-21
3.3. m/NP/miR-21 Accelerates Mouse Skin Wound Repair
3.4. m/NP/miR-21 Promotes Re-Epithelialization by Facilitating Keratinocyte Migration
3.5. m/NP/miR-21 Accelerates Angiogenesis
3.6. m/NP/miR-21 Promotes Dermal Reconstruction by Enhancing Fibroblast Function
3.7. m/NP/miR-21 Regulates Skin Repair via the PTEN/Akt Axis
4. Discussion
5. Conclusions
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
Abbreviations
ADSCs | Adipose mesenchymal stem cells |
PTEN | Phosphate and tension homology deleted on chromosome ten |
PLGA-PEI | Poly(d,l-lactide-co-glycolide)-polyethyleneimine |
NPs | Nanoparticles |
MSCs | Mesenchymal stem cells |
RES | Reticuloendothelial system |
D/H | DMEM high glucose medium |
D/F | DMEM/F12 |
FBS | Fetal bovine serum |
HUVECs | Human umbilical vein endothelial cells |
ECM | Endothelial culture medium and additive kit |
hDFs | Human fibroblasts |
Hacats | Human immortalized keratinocytes |
m/NP/miR | Membrane nanovesicles coated PLGA-PEI loaded with miRNA |
NP/miR | PLGA-PEI loaded with miRNA |
m/miR | Membrane nanovesicles loaded with miRNA |
mNVs | Membrane nanovesicles |
DLS | Dynamic light scattering |
NTA | Nanoparticle tracking analysis |
TEM | Transmission electron microscopy |
GAPDH | Glyceraldehyde-3-phosphate dehydrogenase |
MMP1 | Matrix metallopeptidase 1 |
MMP3 | Matrix metallopeptidase 3 |
CDH2 | Chromodomain helicase DNA binding protein 2 |
COL1A1 | Collagen type I alpha 1 chain |
VEGFA | Vascular endothelial growth factor a |
COL1 | Collagen type I alpha 1 |
K14 | Keratin 14 |
K15 | Keratin 15 |
SCD | Stearoyl-CoA desaturase |
α-SMA | Actin Alpha 2, smooth muscle |
EG | Epithelial gap |
SGs | Sebaceous glands |
HFs | Hair follicles |
RBCs | Red blood cells |
PI3K | Phosphatidylinositol-4,5-bisphosphate 3-kinase |
Arranged in order of first appearance in manuscripts. |
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Gene Names | Primer Sequences |
---|---|
MMP1 | forward: 5′-ATGAAGCAGCCCAGATGTGGAG-3′ |
reverse: 5′-TGGTCCACATCTGCTCTTGGCA-3′ | |
MMP3 | forward: 5′-CACTCACAGACCTGACTCGGTT-3′ |
reverse: 5′-AAGCAGGATCACAGTTGGCTGG-3′ | |
CDH2 | forward: 5′-CGAAAACAGGCACTGGACCACT-3′ |
reverse: 5′-GATGACGACTGTGTCCGCTGAA-3′ | |
COL1A1 | forward: 5′-GATTCCCTGGACCTAAAGGTGC-3′ |
reverse: 5′-AGCCTCTCCATCTTTGCCAGCA-3′ | |
GAPDH | forward: 5′-GTCTCCTCTGACTTCAACAGCG-3′ |
reverse: 5′-ACCACCCTGTTGCTGTAGCCAA-3′ | |
Vegfa | forward: 5′-CTGCTGTAACGATGAAGCCCTG-3′ |
reverse: 5′-GCTGTAGGAAGCTCATCTCTCC-3′ | |
pten | forward: 5′-TGAGTTCCCTCAGCCGTTACCT-3′ |
reverse: 5′-GAGGTTTCCTCTGGTCCTGGTA-3′ | |
U6 | forward: 5′-CTCGCTTCGGCAGCACAT-3′ |
reverse: 5′-TTTGCGTGTCATCCTTGCG-3′ |
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Peng, H.; Du, F.; Wang, J.; Wu, Y.; Wei, Q.; Chen, A.; Duan, Y.; Shi, S.; Zhang, J.; Yu, S. Adipose-Derived Stem-Cell-Membrane-Coated PLGA-PEI Nanoparticles Promote Wound Healing via Efficient Delivery of miR-21. Pharmaceutics 2024, 16, 1113. https://doi.org/10.3390/pharmaceutics16091113
Peng H, Du F, Wang J, Wu Y, Wei Q, Chen A, Duan Y, Shi S, Zhang J, Yu S. Adipose-Derived Stem-Cell-Membrane-Coated PLGA-PEI Nanoparticles Promote Wound Healing via Efficient Delivery of miR-21. Pharmaceutics. 2024; 16(9):1113. https://doi.org/10.3390/pharmaceutics16091113
Chicago/Turabian StylePeng, Huiyu, Fangzhou Du, Jingwen Wang, Yue Wu, Qian Wei, Aoying Chen, Yuhan Duan, Shuaiguang Shi, Jingzhong Zhang, and Shuang Yu. 2024. "Adipose-Derived Stem-Cell-Membrane-Coated PLGA-PEI Nanoparticles Promote Wound Healing via Efficient Delivery of miR-21" Pharmaceutics 16, no. 9: 1113. https://doi.org/10.3390/pharmaceutics16091113