Application of Epigallocatechin-3-gallate (EGCG) Modified 1-Ethyl-3-(3-dimethylaminopropylcarbodiimide hydrochloride/N-hydroxy-succinimide (EDC/NHS) Cross-Linked Collagen Membrane to Promote Macrophage Adhesion
Abstract
:1. Introduction
2. Materials and Methods
2.1. Materials
2.2. Surface Morphology
2.3. Mechanical and Chemical Properties
- a.
- Mechanical properties measurement
- b.
- Chemical properties measurement
- c.
- Release profile of EGCG-EDC/NHS-Col in vitro
2.4. Cell Viability
2.5. Cell Adhesion
2.6. Surgical Procedures
2.7. HE Staining
- (a)
- Dehydration with ethanol;
- (b)
- Stain for 5 min using hematoxylin and differentiate for 2 s in 1% hydrochloric acid alcohol;
- (c)
- Incubate for 2 min in 0.2% ammonia water and stain for 1 min using eosin;
- (d)
- Gradually dehydrate using 95% ethanol, then clear and mount the sections with neutral resin. These sections were observed with light microscopy (Olympus, Tokyo, Japan) and scanned by the Digital Slide Scanning System (PRECICE, Beijing, China).
2.8. Promotion of Revascularization
2.9. Statistical Analysis
3. Results
3.1. Surface Morphology
3.2. Mechanical and Chemical Properties
3.3. Cell Viability
3.4. Cell Adhesion
3.5. Promotion of Revascularization
3.6. Recruitment of Monocyte/Macrophage
4. Discussion
5. Conclusions
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
References
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Genes | Oligonucleotide Sequence |
---|---|
GAPDH | Forward: TGAAGCAGGCATCTGAGGG Reverse: TGAAGTCGCAGGAGACAACC |
Integrin b1 | Forward: CCAAGTGGGACACGGGTGA Reverse: CTGCTGCTGTGAGCTTGGTG |
Integrin b2 | Forward: GCAGCAGAAGGACGGAAACG Reverse: AGGGGGTTGTCGTTGTTCCA |
Integrin b3 | Forward: AGACAGCGCCCAGATCACTC Reverse: GCCAATCCGAAGGTTGCTGG |
F4/80 | Forward: ATTCCCGTGTTGTTGGTGG Reverse: TGCTTTGGCTGGATGTGC |
CD86 | Forward: TGTTTCCGTGGAGACGCAAG Reverse: TTGAGCCTTTGTAAATGGGCA |
CD206 | Forward: CTCTGTTCAGCTATTGGACGC Reverse: CGGAATTTCTGGGATTCAGCTTC |
VEGF | Forward: ACACGGGAGACAATGGGATG Reverse: GGCAGGCAAAAGGACTTCG |
Groups | Ultimate Stress (MPa) | Ultimate Elongation (%) | Young’s Modulus |
---|---|---|---|
EDC/NHS-Col | 18.32 ± 3.12 | 22.44 ± 3.12 | 0.77 ± 0.16 |
EGCG-EDC/NHS-Col | 18.18 ± 2.97 | 29.29 ± 2.78 | 0.57 ± 0.09 |
Groups | 1235/1450 |
---|---|
NHS/EDC-Col | 1.001 ± 0.002 |
EGCG-NHS/EDC-Col | 0.989 ± 0.003 |
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Rung, S.; Zhao, X.; Chu, C.; Yang, R.; Qu, Y.; Man, Y. Application of Epigallocatechin-3-gallate (EGCG) Modified 1-Ethyl-3-(3-dimethylaminopropylcarbodiimide hydrochloride/N-hydroxy-succinimide (EDC/NHS) Cross-Linked Collagen Membrane to Promote Macrophage Adhesion. Materials 2021, 14, 4660. https://doi.org/10.3390/ma14164660
Rung S, Zhao X, Chu C, Yang R, Qu Y, Man Y. Application of Epigallocatechin-3-gallate (EGCG) Modified 1-Ethyl-3-(3-dimethylaminopropylcarbodiimide hydrochloride/N-hydroxy-succinimide (EDC/NHS) Cross-Linked Collagen Membrane to Promote Macrophage Adhesion. Materials. 2021; 14(16):4660. https://doi.org/10.3390/ma14164660
Chicago/Turabian StyleRung, Shengan, Xiwen Zhao, Chenyu Chu, Renli Yang, Yili Qu, and Yi Man. 2021. "Application of Epigallocatechin-3-gallate (EGCG) Modified 1-Ethyl-3-(3-dimethylaminopropylcarbodiimide hydrochloride/N-hydroxy-succinimide (EDC/NHS) Cross-Linked Collagen Membrane to Promote Macrophage Adhesion" Materials 14, no. 16: 4660. https://doi.org/10.3390/ma14164660