Congenital microtia patients: the genetically engineered exosomes released from porous gelatin methacryloyl hydrogel for downstream small RNA profiling, functional modulation of microtia chondrocytes and tissue-engineered ear cartilage regeneration
Abstract Background Mesenchymal stem cells (MSCs) exosomes were previously shown to be effective in articular cartilage repairing. However, whether MSCs exosomes promote mature cartilage formation of microtia chondrocytes and the underlying mechanism of action remains unknown. Additionally, some hur...
| Main Authors: | , , , , , |
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| Format: | Article |
| Language: | English |
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BMC
2022-03-01
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| Series: | Journal of Nanobiotechnology |
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| Online Access: | https://doi.org/10.1186/s12951-022-01352-6 |
| _version_ | 1798005811207733248 |
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| author | Jianguo Chen Tianyu Huang Ruiquan Liu Chenyu Wang Haiyue Jiang Hengyun Sun |
| author_facet | Jianguo Chen Tianyu Huang Ruiquan Liu Chenyu Wang Haiyue Jiang Hengyun Sun |
| author_sort | Jianguo Chen |
| collection | DOAJ |
| description | Abstract Background Mesenchymal stem cells (MSCs) exosomes were previously shown to be effective in articular cartilage repairing. However, whether MSCs exosomes promote mature cartilage formation of microtia chondrocytes and the underlying mechanism of action remains unknown. Additionally, some hurdles, such as the low yield and unsatisfactory therapeutic effects of natural exosomes have emerged when considering the translation of exosomes-therapeutics to clinical practices or industrial production. Herein, we investigated the roles of human adipose-derived stem cells (ADSCs) exosomes in modulating microtia chondrocytes and the underlying mechanism of action. Special attention was also paid to the mass production and functional modification of ADSCs exosomes. Results We firstly used porous gelatin methacryloyl (Porous Gelma) hydrogel with pores size of 100 to 200 μm for 3D culture of passage 2, 4 and 6 ADSCs (P2, P4 and P6 ADSCs, respectively), and obtained their corresponding exosomes (Exo 2, Exo 4 and Exo 6, respectively). In vitro results showed Exo 2 outperformed both Exo 4 and Exo 6 in enhancing cell proliferation and attenuating apoptosis. However, both Exo 4 and Exo 6 promoted chondrogenesis more than Exo 2 did. Small RNA sequencing results indicated Exo 4 was similar to Exo 6 in small RNA profiles and consistently upregulated PI3K/AKT/mTOR signaling pathway. Notably, we found hsa-miR-23a-3p was highly expressed in Exo 4 and Exo 6 compared to Exo 2, and they modulated microtia chondrocytes by transferring hsa-miR-23a-3p to suppress PTEN expression, and consequently to activate PI3K/AKT/mTOR signaling pathway. Then, we designed genetically engineered exosomes by directly transfecting agomir-23a-3p into parent P4 ADSCs and isolated hsa-miR-23a-3p-rich exosomes for optimizing favorable effects on cell viability and new cartilage formation. Subsequently, we applied the engineered exosomes to in vitro and in vivo tissue-engineered cartilage culture and consistently found that the engineered exosomes could enhance cell proliferation, attenuate apoptosis and promote cartilage regeneration. Conclusions Taken together, the porous Gelma hydrogel could be applied to exosomes mass production, and functional modification could be achieved by selecting P4 ADSCs as parent cells and genetically modifying ADSCs. Our engineered exosomes are a promising candidate for tissue-engineered ear cartilage regeneration. Graphical Abstract |
| first_indexed | 2024-04-11T12:45:00Z |
| format | Article |
| id | doaj.art-377bd0ac51924be684b6f90d139735b5 |
| institution | Directory Open Access Journal |
| issn | 1477-3155 |
| language | English |
| last_indexed | 2024-04-11T12:45:00Z |
| publishDate | 2022-03-01 |
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| series | Journal of Nanobiotechnology |
| spelling | doaj.art-377bd0ac51924be684b6f90d139735b52022-12-22T04:23:23ZengBMCJournal of Nanobiotechnology1477-31552022-03-0120112310.1186/s12951-022-01352-6Congenital microtia patients: the genetically engineered exosomes released from porous gelatin methacryloyl hydrogel for downstream small RNA profiling, functional modulation of microtia chondrocytes and tissue-engineered ear cartilage regenerationJianguo Chen0Tianyu Huang1Ruiquan Liu2Chenyu Wang3Haiyue Jiang4Hengyun Sun5Plastic Surgery Hospital, Chinese Academy of Medical Sciences and Peking Union Medical CollegePlastic Surgery Hospital, Chinese Academy of Medical Sciences and Peking Union Medical CollegePlastic Surgery Hospital, Chinese Academy of Medical Sciences and Peking Union Medical CollegePlastic Surgery Hospital, Chinese Academy of Medical Sciences and Peking Union Medical CollegePlastic Surgery Hospital, Chinese Academy of Medical Sciences and Peking Union Medical CollegePlastic Surgery Hospital, Chinese Academy of Medical Sciences and Peking Union Medical CollegeAbstract Background Mesenchymal stem cells (MSCs) exosomes were previously shown to be effective in articular cartilage repairing. However, whether MSCs exosomes promote mature cartilage formation of microtia chondrocytes and the underlying mechanism of action remains unknown. Additionally, some hurdles, such as the low yield and unsatisfactory therapeutic effects of natural exosomes have emerged when considering the translation of exosomes-therapeutics to clinical practices or industrial production. Herein, we investigated the roles of human adipose-derived stem cells (ADSCs) exosomes in modulating microtia chondrocytes and the underlying mechanism of action. Special attention was also paid to the mass production and functional modification of ADSCs exosomes. Results We firstly used porous gelatin methacryloyl (Porous Gelma) hydrogel with pores size of 100 to 200 μm for 3D culture of passage 2, 4 and 6 ADSCs (P2, P4 and P6 ADSCs, respectively), and obtained their corresponding exosomes (Exo 2, Exo 4 and Exo 6, respectively). In vitro results showed Exo 2 outperformed both Exo 4 and Exo 6 in enhancing cell proliferation and attenuating apoptosis. However, both Exo 4 and Exo 6 promoted chondrogenesis more than Exo 2 did. Small RNA sequencing results indicated Exo 4 was similar to Exo 6 in small RNA profiles and consistently upregulated PI3K/AKT/mTOR signaling pathway. Notably, we found hsa-miR-23a-3p was highly expressed in Exo 4 and Exo 6 compared to Exo 2, and they modulated microtia chondrocytes by transferring hsa-miR-23a-3p to suppress PTEN expression, and consequently to activate PI3K/AKT/mTOR signaling pathway. Then, we designed genetically engineered exosomes by directly transfecting agomir-23a-3p into parent P4 ADSCs and isolated hsa-miR-23a-3p-rich exosomes for optimizing favorable effects on cell viability and new cartilage formation. Subsequently, we applied the engineered exosomes to in vitro and in vivo tissue-engineered cartilage culture and consistently found that the engineered exosomes could enhance cell proliferation, attenuate apoptosis and promote cartilage regeneration. Conclusions Taken together, the porous Gelma hydrogel could be applied to exosomes mass production, and functional modification could be achieved by selecting P4 ADSCs as parent cells and genetically modifying ADSCs. Our engineered exosomes are a promising candidate for tissue-engineered ear cartilage regeneration. Graphical Abstracthttps://doi.org/10.1186/s12951-022-01352-6Adipose-derived stem cellsEngineered exosomesMicrotiaPorous gelatin methacryloylTissue-engineered ear cartilage |
| spellingShingle | Jianguo Chen Tianyu Huang Ruiquan Liu Chenyu Wang Haiyue Jiang Hengyun Sun Congenital microtia patients: the genetically engineered exosomes released from porous gelatin methacryloyl hydrogel for downstream small RNA profiling, functional modulation of microtia chondrocytes and tissue-engineered ear cartilage regeneration Journal of Nanobiotechnology Adipose-derived stem cells Engineered exosomes Microtia Porous gelatin methacryloyl Tissue-engineered ear cartilage |
| title | Congenital microtia patients: the genetically engineered exosomes released from porous gelatin methacryloyl hydrogel for downstream small RNA profiling, functional modulation of microtia chondrocytes and tissue-engineered ear cartilage regeneration |
| title_full | Congenital microtia patients: the genetically engineered exosomes released from porous gelatin methacryloyl hydrogel for downstream small RNA profiling, functional modulation of microtia chondrocytes and tissue-engineered ear cartilage regeneration |
| title_fullStr | Congenital microtia patients: the genetically engineered exosomes released from porous gelatin methacryloyl hydrogel for downstream small RNA profiling, functional modulation of microtia chondrocytes and tissue-engineered ear cartilage regeneration |
| title_full_unstemmed | Congenital microtia patients: the genetically engineered exosomes released from porous gelatin methacryloyl hydrogel for downstream small RNA profiling, functional modulation of microtia chondrocytes and tissue-engineered ear cartilage regeneration |
| title_short | Congenital microtia patients: the genetically engineered exosomes released from porous gelatin methacryloyl hydrogel for downstream small RNA profiling, functional modulation of microtia chondrocytes and tissue-engineered ear cartilage regeneration |
| title_sort | congenital microtia patients the genetically engineered exosomes released from porous gelatin methacryloyl hydrogel for downstream small rna profiling functional modulation of microtia chondrocytes and tissue engineered ear cartilage regeneration |
| topic | Adipose-derived stem cells Engineered exosomes Microtia Porous gelatin methacryloyl Tissue-engineered ear cartilage |
| url | https://doi.org/10.1186/s12951-022-01352-6 |
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