Physicochemical properties of a novel chiral self-assembling peptide R-LIFE-1 and its controlled release to exosomes_Journal of Biomedical Engineering

Authors：

LUO Xinyi , SU Di , LU Na , WAN Yuan , LIU Guicen ,  LUO Zhongli

Molecular Medicine and Cancer Research Center, Collage of Basic Medicine, Chongqing Medical University, Chongqing 400016, P. R. China;

Corresponding author：

LUO Zhongli, Email: Zhongliluo@163.com

Keywords：

Chiral self-assembling peptide; Hydrogel; Exosomes; Controlled release

DOI：

10.7507/1001-5515.202207056

Video：

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Abstract Full text Figures/Tables Video References Cited by

This research aims to investigate the encapsulation and controlled release effect of the newly developed self-assembling peptide R-LIFE-1 on exosomes. The gelling ability and morphological structure of the chiral self-assembling peptide (CSAP) hydrogel were examined using advanced imaging techniques, including atomic force microscopy, transmission electron microscopy, and cryo-scanning electron microscopy. The biocompatibility of the CSAP hydrogel was assessed through optical microscopy and fluorescent staining. Exosomes were isolated via ultrafiltration, and their quality was evaluated using Western blot analysis, nanoparticle tracking analysis, and transmission electron microscopy. The controlled release effect of the CSAP hydrogel on exosomes was quantitatively analyzed using laser confocal microscopy and a BCA assay kit. The results revealed that the self-assembling peptide R-LIFE-1 exhibited spontaneous assembly in the presence of various ions, leading to the formation of nanofibers. These nanofibers were cross-linked, giving rise to a robust nanofiber network structure, which further underwent cross-linking to generate a laminated membrane structure. The nanofibers possessed a large surface area, allowing them to encapsulate a substantial number of water molecules, thereby forming a hydrogel material with high water content. This hydrogel served as a stable spatial scaffold and loading matrix for the three-dimensional culture of cells, as well as the encapsulation and controlled release of exosomes. Importantly, R-LIFE-1 demonstrated excellent biocompatibility, preserving the growth of cells and the biological activity of exosomes. It rapidly formed a three-dimensional network scaffold, enabling the stable loading of cells and exosomes, while exhibiting favorable biocompatibility and reduced cytotoxicity. In conclusion, the findings of this study support the notion that R-LIFE-1 holds significant promise as an ideal tissue engineering material for tissue repair applications.

Citation： LUO Xinyi, SU Di, LU Na, WAN Yuan, LIU Guicen, LUO Zhongli. Physicochemical properties of a novel chiral self-assembling peptide R-LIFE-1 and its controlled release to exosomes. Journal of Biomedical Engineering, 2023, 40(4): 770-777. doi: 10.7507/1001-5515.202207056 Copy

1.	Théry C, Zitvogel L, Amigorena S. Exosomes: Composition, biogenesis and function. Nat Rev Immunol, 2002: 569-579.
2.	Kowal J, Tkach M, Théry C. Biogenesis and secretion of exosomes. Curr Opin Cell Biol, 2014, 29: 116-125.
3.	Kalluri R, Lebleu V S. The biology, function, and biomedical applications of exosomes. Science, 2020, 367(6478): eaau6977.
4.	Kalluri R. The biology and function of exosomes in cancer. J Clin Invest, 2016, 126(4): 1208-1215.
5.	Zhang J, Li S, Li L, et al. Exosome and exosomal microRNA: Trafficking, sorting, and function. Genomics Proteomics Bioinformatics, 2015, 13(1): 17-24.
6.	Hessvik N P, Llorente A. Current knowledge on exosome biogenesis and release. Cell Mol Life Sci, 2018, 75(2): 193-208.
7.	Zhang L, Yu D. Exosomes in cancer development, metastasis, and immunity. Biochim Biophys Acta Rev Cancer, 2019, 1871(2): 455-468.
8.	Pishavar E, Luo H, Naserifar M, et al. Advanced hydrogels as exosome delivery systems for osteogenic differentiation of MSCs: Application in bone regeneration. Int J Mol Sci, 2021, 22(12): 6203.
9.	Rao F, Zhang D, Fang T, et al. Exosomes from human gingiva-derived mesenchymal stem cells combined with biodegradable chitin conduits promote rat sciatic nerve regeneration. Stem Cells Int, 2019, 2019: 2546367.
10.	Chen P, Zheng L, Wang Y, et al. Desktop-stereolithography 3D printing of a radially oriented extracellular matrix/mesenchymal stem cell exosome bioink for osteochondral defect regeneration. Theranostics, 2019, 9(9): 2439-2459.
11.	Zhang K, Zhao X, Chen X, et al. Enhanced therapeutic effects of mesenchymal stem cell-derived exosomes with an injectable hydrogel for hindlimb ischemia treatment. ACS Appl Mater Interfaces, 2018, 10(36): 30081-30091.
12.	朱耀斌, 张燕搏, 范祥明, 等. 外泌体在心肌梗死治疗中研究进展. 中华实用诊断与治疗杂志, 2017, 31(12): 1236-1238.
13.	Emanueli C, Shearn A I, Angelini G D, et al. Exosomes and exosomal miRNAs in cardiovascular protection and repair. Vascul Pharmacol, 2015, 71: 24-30.
14.	Basu J, Ludlow J W. Exosomes for repair, regeneration and rejuvenation. Expert Opin Biol Ther, 2016, 16(4): 489-506.
15.	Phinney D G, Pittenger M F. Concise review: MSC-derived exosomes for cell-free therapy. Stem Cells, 2017, 35(4): 851-858.
16.	Tan W S, Arulselvan P, Ng S F, et al. Healing effect of vicenin-2 (VCN-2) on human dermal fibroblast (HDF) and development VCN-2 hydrocolloid film based on alginate as potential wound dressing. Biomed Res Int, 2020, 2020: 4730858.
17.	Lago J C, Puzzi M B. The effect of aging in primary human dermal fibroblasts. PLoS One, 2019, 14(7): e0219165.
18.	Hu S, Li Z, Cores J, et al. Needle-free injection of exosomes derived from human dermal fibroblast spheroids ameliorates skin photoaging. ACS Nano, 2019, 13(10): 11273-11282.
19.	Han X, Wu P, Li L, et al. Exosomes derived from autologous dermal fibroblasts promote diabetic cutaneous wound healing through the Akt/β-catenin pathway. Cell Cycle, 2021, 20(5-6): 616-629.
20.	Shi A, Li J, Qiu X, et al. TGF-β loaded exosome enhances ischemic wound healing in vitro and in vivo. Theranostics, 2021, 11(13): 6616-6631.
21.	Luo Z, Zhao X, Zhang S. Self-organization of a chiral D-EAK16 designer peptide into a 3D nanofiber scaffold. Macromol Biosci, 2008, 8(8): 785-791.
22.	Hauser C A E, Zhang S. Designer self-assembling peptide nanofiber biological materials. Chem Soc Rev, 2010, 39(8): 2780.
23.	Gelain F, Horii A, Zhang S. Designer self-assembling peptide scaffolds for 3-D tissue cell cultures and regenerative medicine. Macromol Biosci, 2007, 7(5): 544-551.
24.	Zhang S, Holmes T, Lockshin C, et al. Spontaneous assembly of a self-complementary oligopeptide to form a stable macroscopic membrane. Proc Natl Acad Sci U S A, 1993, 90(8): 3334-3338.
25.	Vauthey S, Santoso S, Gong H, et al. Molecular self-assembly of surfactant-like peptides to form nanotubes and nanovesicles. Proc Natl Acad Sci U S A, 2002, 99(8): 5355-5360.
26.	Gelain F, Cigognini D, Caprini A, et al. New bioactive motifs and their use in functionalized self-assembling peptides for NSC differentiation and neural tissue engineering. Nanoscale, 2012, 4(9): 2946-2957.
27.	张慧楠, 李萌萌, 文静, 等. 自组装短肽R₂I₄R₂对皮肤创伤快速修复过程的研究. 中国生物工程杂志, 2018, 38(2): 7-12.
28.	吴书祎, 兰世建, 文静, 等. 手性自组装短肽对子宫创伤修复的影响. 生物工程学报, 2019, 35(6): 1079-1087.
29.	陈振银, 岳媛媛, 张慧楠, 等. 自组装短肽GFS-4作为细胞三维培养及心肌梗死修复支架材料的研究. 生物医学工程学杂志, 2017, 34(3): 388-393.
30.	文静, 李昭绪, 吴书祎, 等. 自组装短肽SciobioⅡ对关节软骨损伤修复过程的探究. 基因组学与应用生物学, 2020, 39(7): 3219-3224.
31.	Onak G, Gökmen O, Yaralı Z B, et al. Enhanced osteogenesis of human mesenchymal stem cells by self-assembled peptide hydrogel functionalized with glutamic acid templated peptides. J Tissue Eng Regen Med, 2020, 14(9): 1236-1249.
32.	Chow L W, Bitton R, Webber M J, et al. A bioactive self-assembled membrane to promote angiogenesis. Biomaterials, 2011, 32(6): 1574-1582.
33.	Bruggeman K F, Rodriguez A L, Parish C L, et al. Temporally controlled release of multiple growth factors from a self-assembling peptide hydrogel. Nanotechnology, 2016, 27(38): 385102.
34.	Ligorio C, Zhou M, Wychowaniec J K, et al. Graphene oxide containing self-assembling peptide hybrid hydrogels as a potential 3D injectable cell delivery platform for intervertebral disc repair applications. Acta Biomater, 2019, 92: 92-103.

1. Théry C, Zitvogel L, Amigorena S. Exosomes: Composition, biogenesis and function. Nat Rev Immunol, 2002: 569-579.
2. Kowal J, Tkach M, Théry C. Biogenesis and secretion of exosomes. Curr Opin Cell Biol, 2014, 29: 116-125.
3. Kalluri R, Lebleu V S. The biology, function, and biomedical applications of exosomes. Science, 2020, 367(6478): eaau6977.
4. Kalluri R. The biology and function of exosomes in cancer. J Clin Invest, 2016, 126(4): 1208-1215.
5. Zhang J, Li S, Li L, et al. Exosome and exosomal microRNA: Trafficking, sorting, and function. Genomics Proteomics Bioinformatics, 2015, 13(1): 17-24.
6. Hessvik N P, Llorente A. Current knowledge on exosome biogenesis and release. Cell Mol Life Sci, 2018, 75(2): 193-208.
7. Zhang L, Yu D. Exosomes in cancer development, metastasis, and immunity. Biochim Biophys Acta Rev Cancer, 2019, 1871(2): 455-468.
8. Pishavar E, Luo H, Naserifar M, et al. Advanced hydrogels as exosome delivery systems for osteogenic differentiation of MSCs: Application in bone regeneration. Int J Mol Sci, 2021, 22(12): 6203.
9. Rao F, Zhang D, Fang T, et al. Exosomes from human gingiva-derived mesenchymal stem cells combined with biodegradable chitin conduits promote rat sciatic nerve regeneration. Stem Cells Int, 2019, 2019: 2546367.
10. Chen P, Zheng L, Wang Y, et al. Desktop-stereolithography 3D printing of a radially oriented extracellular matrix/mesenchymal stem cell exosome bioink for osteochondral defect regeneration. Theranostics, 2019, 9(9): 2439-2459.
11. Zhang K, Zhao X, Chen X, et al. Enhanced therapeutic effects of mesenchymal stem cell-derived exosomes with an injectable hydrogel for hindlimb ischemia treatment. ACS Appl Mater Interfaces, 2018, 10(36): 30081-30091.
12. 朱耀斌, 张燕搏, 范祥明, 等. 外泌体在心肌梗死治疗中研究进展. 中华实用诊断与治疗杂志, 2017, 31(12): 1236-1238.
13. Emanueli C, Shearn A I, Angelini G D, et al. Exosomes and exosomal miRNAs in cardiovascular protection and repair. Vascul Pharmacol, 2015, 71: 24-30.
14. Basu J, Ludlow J W. Exosomes for repair, regeneration and rejuvenation. Expert Opin Biol Ther, 2016, 16(4): 489-506.
15. Phinney D G, Pittenger M F. Concise review: MSC-derived exosomes for cell-free therapy. Stem Cells, 2017, 35(4): 851-858.
16. Tan W S, Arulselvan P, Ng S F, et al. Healing effect of vicenin-2 (VCN-2) on human dermal fibroblast (HDF) and development VCN-2 hydrocolloid film based on alginate as potential wound dressing. Biomed Res Int, 2020, 2020: 4730858.
17. Lago J C, Puzzi M B. The effect of aging in primary human dermal fibroblasts. PLoS One, 2019, 14(7): e0219165.
18. Hu S, Li Z, Cores J, et al. Needle-free injection of exosomes derived from human dermal fibroblast spheroids ameliorates skin photoaging. ACS Nano, 2019, 13(10): 11273-11282.
19. Han X, Wu P, Li L, et al. Exosomes derived from autologous dermal fibroblasts promote diabetic cutaneous wound healing through the Akt/β-catenin pathway. Cell Cycle, 2021, 20(5-6): 616-629.
20. Shi A, Li J, Qiu X, et al. TGF-β loaded exosome enhances ischemic wound healing in vitro and in vivo. Theranostics, 2021, 11(13): 6616-6631.
21. Luo Z, Zhao X, Zhang S. Self-organization of a chiral D-EAK16 designer peptide into a 3D nanofiber scaffold. Macromol Biosci, 2008, 8(8): 785-791.
22. Hauser C A E, Zhang S. Designer self-assembling peptide nanofiber biological materials. Chem Soc Rev, 2010, 39(8): 2780.
23. Gelain F, Horii A, Zhang S. Designer self-assembling peptide scaffolds for 3-D tissue cell cultures and regenerative medicine. Macromol Biosci, 2007, 7(5): 544-551.
24. Zhang S, Holmes T, Lockshin C, et al. Spontaneous assembly of a self-complementary oligopeptide to form a stable macroscopic membrane. Proc Natl Acad Sci U S A, 1993, 90(8): 3334-3338.
25. Vauthey S, Santoso S, Gong H, et al. Molecular self-assembly of surfactant-like peptides to form nanotubes and nanovesicles. Proc Natl Acad Sci U S A, 2002, 99(8): 5355-5360.
26. Gelain F, Cigognini D, Caprini A, et al. New bioactive motifs and their use in functionalized self-assembling peptides for NSC differentiation and neural tissue engineering. Nanoscale, 2012, 4(9): 2946-2957.
27. 张慧楠, 李萌萌, 文静, 等. 自组装短肽R₂I₄R₂对皮肤创伤快速修复过程的研究. 中国生物工程杂志, 2018, 38(2): 7-12.
28. 吴书祎, 兰世建, 文静, 等. 手性自组装短肽对子宫创伤修复的影响. 生物工程学报, 2019, 35(6): 1079-1087.
29. 陈振银, 岳媛媛, 张慧楠, 等. 自组装短肽GFS-4作为细胞三维培养及心肌梗死修复支架材料的研究. 生物医学工程学杂志, 2017, 34(3): 388-393.
30. 文静, 李昭绪, 吴书祎, 等. 自组装短肽SciobioⅡ对关节软骨损伤修复过程的探究. 基因组学与应用生物学, 2020, 39(7): 3219-3224.
31. Onak G, Gökmen O, Yaralı Z B, et al. Enhanced osteogenesis of human mesenchymal stem cells by self-assembled peptide hydrogel functionalized with glutamic acid templated peptides. J Tissue Eng Regen Med, 2020, 14(9): 1236-1249.
32. Chow L W, Bitton R, Webber M J, et al. A bioactive self-assembled membrane to promote angiogenesis. Biomaterials, 2011, 32(6): 1574-1582.
33. Bruggeman K F, Rodriguez A L, Parish C L, et al. Temporally controlled release of multiple growth factors from a self-assembling peptide hydrogel. Nanotechnology, 2016, 27(38): 385102.
34. Ligorio C, Zhou M, Wychowaniec J K, et al. Graphene oxide containing self-assembling peptide hybrid hydrogels as a potential 3D injectable cell delivery platform for intervertebral disc repair applications. Acta Biomater, 2019, 92: 92-103.

Journal of Biomedical Engineering

Physicochemical properties of a novel chiral self-assembling peptide R-LIFE-1 and its controlled release to exosomes

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