The application of a 4D-printed chitosan-based stem cell carrier for the repair of corneal alkali burns
Abstract Background Corneal alkali burns can lead to ulceration, perforation, and even corneal blindness due to epithelial defects and extensive cell necrosis, resulting in poor healing outcomes. Previous studies have found that chitosan-based in situ hydrogel loaded with limbal epithelium stem cell...
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| Format: | Article |
| Language: | English |
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BMC
2024-02-01
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| Series: | Stem Cell Research & Therapy |
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| Online Access: | https://doi.org/10.1186/s13287-024-03653-z |
| _version_ | 1827328389692784640 |
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| author | Zibo Wang Changqing Jiang Yuqiao Fan Xiaodan Hao Yanhan Dong Xinjia He Jinning Gao Yongchun Zhang Meng Li Mengyuan Wang Yiming Liu Wenhua Xu |
| author_facet | Zibo Wang Changqing Jiang Yuqiao Fan Xiaodan Hao Yanhan Dong Xinjia He Jinning Gao Yongchun Zhang Meng Li Mengyuan Wang Yiming Liu Wenhua Xu |
| author_sort | Zibo Wang |
| collection | DOAJ |
| description | Abstract Background Corneal alkali burns can lead to ulceration, perforation, and even corneal blindness due to epithelial defects and extensive cell necrosis, resulting in poor healing outcomes. Previous studies have found that chitosan-based in situ hydrogel loaded with limbal epithelium stem cells (LESCs) has a certain reparative effect on corneal alkali burns. However, the inconsistent pore sizes of the carriers and low cell loading rates have resulted in suboptimal repair outcomes. In this study, 4D bioprinting technology was used to prepare a chitosan-based thermosensitive gel carrier (4D-CTH) with uniform pore size and adjustable shape to improve the transfer capacity of LESCs. Methods Prepare solutions of chitosan acetate, carboxymethyl chitosan, and β-glycerophosphate sodium at specific concentrations, and mix them in certain proportions to create a pore-size uniform scaffold using 4D bioprinting technology. Extract and culture rat LESCs (rLESCs) in vitro, perform immunofluorescence experiments to observe the positivity rate of deltaNp63 cells for cell identification. Conduct a series of experiments to validate the cell compatibility of 4D-CTH, including CCK-8 assay to assess cell toxicity, scratch assay to evaluate the effect of 4D-CTH on rLESCs migration, and Calcein-AM/PI cell staining experiment to examine the impact of 4D-CTH on rLESCs proliferation and morphology. Establish a severe alkali burn model in rat corneas, transplant rLESCs onto the injured cornea using 4D-CTH, periodically observe corneal opacity and neovascularization using a slit lamp, and evaluate epithelial healing by fluorescein sodium staining. Assess the therapeutic effect 4D-CTH-loaded rLESCs on corneal alkali burn through histological evaluation of corneal tissue paraffin sections stained with hematoxylin and eosin, as well as immunofluorescence staining of frozen sections. Results Using the 4D-CTH, rLESCs were transferred to the alkali burn wounds of rats. Compared with the traditional treatment group (chitosan in situ hydrogel encapsulating rLESCs), the 4D-CTH-rLESC group had significantly higher repair efficiency of corneal injury, such as lower corneal opacity score (1.2 ± 0.4472 vs 0.4 ± 0.5477, p < 0.05) and neovascularization score (5.5 ± 1.118 vs 2.6 ± 0.9618, p < 0.01), and significantly higher corneal epithelial wound healing rate (72.09 ± 3.568% vs 86.60 ± 5.004%, p < 0.01). Conclusion In summary, the corneas of the 4D-CTH-rLESC treatment group were similar to the normal corneas and had a complete corneal structure. These findings suggested that LESCs encapsulated by 4D-CTH significantly accelerated corneal wound healing after alkali burn and can be considered as a rapid and effective method for treating epithelial defects. Graphical abstract |
| first_indexed | 2024-03-07T15:17:15Z |
| format | Article |
| id | doaj.art-a3dc30a9a24c4184a169daa3a8c02805 |
| institution | Directory Open Access Journal |
| issn | 1757-6512 |
| language | English |
| last_indexed | 2024-03-07T15:17:15Z |
| publishDate | 2024-02-01 |
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| series | Stem Cell Research & Therapy |
| spelling | doaj.art-a3dc30a9a24c4184a169daa3a8c028052024-03-05T17:52:35ZengBMCStem Cell Research & Therapy1757-65122024-02-0115111710.1186/s13287-024-03653-zThe application of a 4D-printed chitosan-based stem cell carrier for the repair of corneal alkali burnsZibo Wang0Changqing Jiang1Yuqiao Fan2Xiaodan Hao3Yanhan Dong4Xinjia He5Jinning Gao6Yongchun Zhang7Meng Li8Mengyuan Wang9Yiming Liu10Wenhua Xu11Institute of Regenerative Medicine and Laboratory Technology Innovation, Qingdao UniversityDepartment of Pathology, Qingdao Municipal HospitalInstitute of Regenerative Medicine and Laboratory Technology Innovation, Qingdao UniversityInstitute of Translational Medicine, College of Medicine, Qingdao UniversityInstitute of Translational Medicine, College of Medicine, Qingdao UniversityDepartment of Oncology, Affiliated Hospital of Qingdao UniversityInstitute of Translational Medicine, College of Medicine, Qingdao UniversityDepartment of Oncology, Affiliated Hospital of Qingdao UniversityInstitute of Regenerative Medicine and Laboratory Technology Innovation, Qingdao UniversityInstitute of Regenerative Medicine and Laboratory Technology Innovation, Qingdao UniversityInstitute of Regenerative Medicine and Laboratory Technology Innovation, Qingdao UniversityInstitute of Regenerative Medicine and Laboratory Technology Innovation, Qingdao UniversityAbstract Background Corneal alkali burns can lead to ulceration, perforation, and even corneal blindness due to epithelial defects and extensive cell necrosis, resulting in poor healing outcomes. Previous studies have found that chitosan-based in situ hydrogel loaded with limbal epithelium stem cells (LESCs) has a certain reparative effect on corneal alkali burns. However, the inconsistent pore sizes of the carriers and low cell loading rates have resulted in suboptimal repair outcomes. In this study, 4D bioprinting technology was used to prepare a chitosan-based thermosensitive gel carrier (4D-CTH) with uniform pore size and adjustable shape to improve the transfer capacity of LESCs. Methods Prepare solutions of chitosan acetate, carboxymethyl chitosan, and β-glycerophosphate sodium at specific concentrations, and mix them in certain proportions to create a pore-size uniform scaffold using 4D bioprinting technology. Extract and culture rat LESCs (rLESCs) in vitro, perform immunofluorescence experiments to observe the positivity rate of deltaNp63 cells for cell identification. Conduct a series of experiments to validate the cell compatibility of 4D-CTH, including CCK-8 assay to assess cell toxicity, scratch assay to evaluate the effect of 4D-CTH on rLESCs migration, and Calcein-AM/PI cell staining experiment to examine the impact of 4D-CTH on rLESCs proliferation and morphology. Establish a severe alkali burn model in rat corneas, transplant rLESCs onto the injured cornea using 4D-CTH, periodically observe corneal opacity and neovascularization using a slit lamp, and evaluate epithelial healing by fluorescein sodium staining. Assess the therapeutic effect 4D-CTH-loaded rLESCs on corneal alkali burn through histological evaluation of corneal tissue paraffin sections stained with hematoxylin and eosin, as well as immunofluorescence staining of frozen sections. Results Using the 4D-CTH, rLESCs were transferred to the alkali burn wounds of rats. Compared with the traditional treatment group (chitosan in situ hydrogel encapsulating rLESCs), the 4D-CTH-rLESC group had significantly higher repair efficiency of corneal injury, such as lower corneal opacity score (1.2 ± 0.4472 vs 0.4 ± 0.5477, p < 0.05) and neovascularization score (5.5 ± 1.118 vs 2.6 ± 0.9618, p < 0.01), and significantly higher corneal epithelial wound healing rate (72.09 ± 3.568% vs 86.60 ± 5.004%, p < 0.01). Conclusion In summary, the corneas of the 4D-CTH-rLESC treatment group were similar to the normal corneas and had a complete corneal structure. These findings suggested that LESCs encapsulated by 4D-CTH significantly accelerated corneal wound healing after alkali burn and can be considered as a rapid and effective method for treating epithelial defects. Graphical abstracthttps://doi.org/10.1186/s13287-024-03653-zChitosan4D printing technologyCell scaffoldLimbal epithelium stem cellsCorneal wound healing |
| spellingShingle | Zibo Wang Changqing Jiang Yuqiao Fan Xiaodan Hao Yanhan Dong Xinjia He Jinning Gao Yongchun Zhang Meng Li Mengyuan Wang Yiming Liu Wenhua Xu The application of a 4D-printed chitosan-based stem cell carrier for the repair of corneal alkali burns Stem Cell Research & Therapy Chitosan 4D printing technology Cell scaffold Limbal epithelium stem cells Corneal wound healing |
| title | The application of a 4D-printed chitosan-based stem cell carrier for the repair of corneal alkali burns |
| title_full | The application of a 4D-printed chitosan-based stem cell carrier for the repair of corneal alkali burns |
| title_fullStr | The application of a 4D-printed chitosan-based stem cell carrier for the repair of corneal alkali burns |
| title_full_unstemmed | The application of a 4D-printed chitosan-based stem cell carrier for the repair of corneal alkali burns |
| title_short | The application of a 4D-printed chitosan-based stem cell carrier for the repair of corneal alkali burns |
| title_sort | application of a 4d printed chitosan based stem cell carrier for the repair of corneal alkali burns |
| topic | Chitosan 4D printing technology Cell scaffold Limbal epithelium stem cells Corneal wound healing |
| url | https://doi.org/10.1186/s13287-024-03653-z |
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