Matrilin-2 within a three-dimensional lysine-modified chitosan porous scaffold enhances Schwann cell migration and axonal outgrowth for peripheral nerve regeneration
Background: Matrilin-2 is a key extracellular matrix protein involved in peripheral nerve regeneration. We sought to develop a biomimetic scaffold to enhance peripheral nerve regeneration by incorporating matrilin-2 within a chitosan-derived porous scaffold. We hypothesized that the use of such a no...
| Main Authors: | , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Frontiers Media S.A.
2023-05-01
|
| Series: | Frontiers in Bioengineering and Biotechnology |
| Subjects: | |
| Online Access: | https://www.frontiersin.org/articles/10.3389/fbioe.2023.1142610/full |
| _version_ | 1797831629645807616 |
|---|---|
| author | Neill Y. Li Neill Y. Li Brandon Vorrius Jonathan Ge Zhen Qiao Shuang Zhu Julia Katarincic Qian Chen |
| author_facet | Neill Y. Li Neill Y. Li Brandon Vorrius Jonathan Ge Zhen Qiao Shuang Zhu Julia Katarincic Qian Chen |
| author_sort | Neill Y. Li |
| collection | DOAJ |
| description | Background: Matrilin-2 is a key extracellular matrix protein involved in peripheral nerve regeneration. We sought to develop a biomimetic scaffold to enhance peripheral nerve regeneration by incorporating matrilin-2 within a chitosan-derived porous scaffold. We hypothesized that the use of such a novel biomaterial delivers microenvironmental cues to facilitate Schwann cell (SC) migration and enhance axonal outgrowth during peripheral nerve regeneration.Materials and Methods: The effect of matrilin-2 on SC migration was evaluated with agarose drop migration assay on matrilin-2 coated dishes. SC adhesion was determined with SCs cultured atop tissue culture dishes coated with matrilin-2. Various formulations of chitosan vs matrilin-2 in scaffold constructs were examined with scanning electron microscopy. The effect of the matrilin-2/chitosan scaffold on SC migration in the collagen conduits was determined by capillary migration assays. Neuronal adhesion and axonal outgrowth were evaluated with three-dimensional (3D) organotypic assay of dorsal root ganglions (DRG). DRG axonal outgrowth within the scaffolds was determined by immunofluorescence staining of neurofilaments.Results: Matrilin-2 induced SC migration and enhanced its adhesion. A formulation of 2% chitosan with matrilin-2 demonstrated an optimal 3D porous architecture for SC interaction. Matrilin-2/chitosan scaffold enabled SCs to migrate against gravity within conduits. Chemical modification of chitosan with lysine (K-chitosan) further improved DRG adhesion and axonal outgrowth than the matrilin-2/chitosan scaffold without lysine modification.Conclusion: We developed a matrilin-2/K-chitosan scaffold to mimic extracellular matrix cues and provide a porous matrix to enhance peripheral nerve regeneration. Taking advantage of matrilin-2’s capability to stimulate SC migration and adhesion, we formulated a porous matrilin-2/chitosan scaffold to support axongal outgrowth. Chemical modification of chitosan with lysine further improved matrilin-2 bioactivity in the 3D scaffold. The 3D porous matrilin-2/K-chitosan scaffolds have high potential for enhancing nerve repair by stimulating SC migration, neuronal adhesion, and axonal outgrowth. |
| first_indexed | 2024-04-09T13:54:45Z |
| format | Article |
| id | doaj.art-87ff68b6ee6d4744b799feaf3d9b80eb |
| institution | Directory Open Access Journal |
| issn | 2296-4185 |
| language | English |
| last_indexed | 2024-04-09T13:54:45Z |
| publishDate | 2023-05-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Bioengineering and Biotechnology |
| spelling | doaj.art-87ff68b6ee6d4744b799feaf3d9b80eb2023-05-08T10:28:05ZengFrontiers Media S.A.Frontiers in Bioengineering and Biotechnology2296-41852023-05-011110.3389/fbioe.2023.11426101142610Matrilin-2 within a three-dimensional lysine-modified chitosan porous scaffold enhances Schwann cell migration and axonal outgrowth for peripheral nerve regenerationNeill Y. Li0Neill Y. Li1Brandon Vorrius2Jonathan Ge3Zhen Qiao4Shuang Zhu5Julia Katarincic6Qian Chen7Laboratory of Molecular Biology and Nanomedicine, Department of Orthopaedics, Warren Alpert Medical School of Brown University, Providence, RI, United StatesDepartment of Orthopaedics, Duke University School of Medicine, Durham, NC, United StatesLaboratory of Molecular Biology and Nanomedicine, Department of Orthopaedics, Warren Alpert Medical School of Brown University, Providence, RI, United StatesLaboratory of Molecular Biology and Nanomedicine, Department of Orthopaedics, Warren Alpert Medical School of Brown University, Providence, RI, United StatesLaboratory of Molecular Biology and Nanomedicine, Department of Orthopaedics, Warren Alpert Medical School of Brown University, Providence, RI, United StatesLaboratory of Molecular Biology and Nanomedicine, Department of Orthopaedics, Warren Alpert Medical School of Brown University, Providence, RI, United StatesLaboratory of Molecular Biology and Nanomedicine, Department of Orthopaedics, Warren Alpert Medical School of Brown University, Providence, RI, United StatesLaboratory of Molecular Biology and Nanomedicine, Department of Orthopaedics, Warren Alpert Medical School of Brown University, Providence, RI, United StatesBackground: Matrilin-2 is a key extracellular matrix protein involved in peripheral nerve regeneration. We sought to develop a biomimetic scaffold to enhance peripheral nerve regeneration by incorporating matrilin-2 within a chitosan-derived porous scaffold. We hypothesized that the use of such a novel biomaterial delivers microenvironmental cues to facilitate Schwann cell (SC) migration and enhance axonal outgrowth during peripheral nerve regeneration.Materials and Methods: The effect of matrilin-2 on SC migration was evaluated with agarose drop migration assay on matrilin-2 coated dishes. SC adhesion was determined with SCs cultured atop tissue culture dishes coated with matrilin-2. Various formulations of chitosan vs matrilin-2 in scaffold constructs were examined with scanning electron microscopy. The effect of the matrilin-2/chitosan scaffold on SC migration in the collagen conduits was determined by capillary migration assays. Neuronal adhesion and axonal outgrowth were evaluated with three-dimensional (3D) organotypic assay of dorsal root ganglions (DRG). DRG axonal outgrowth within the scaffolds was determined by immunofluorescence staining of neurofilaments.Results: Matrilin-2 induced SC migration and enhanced its adhesion. A formulation of 2% chitosan with matrilin-2 demonstrated an optimal 3D porous architecture for SC interaction. Matrilin-2/chitosan scaffold enabled SCs to migrate against gravity within conduits. Chemical modification of chitosan with lysine (K-chitosan) further improved DRG adhesion and axonal outgrowth than the matrilin-2/chitosan scaffold without lysine modification.Conclusion: We developed a matrilin-2/K-chitosan scaffold to mimic extracellular matrix cues and provide a porous matrix to enhance peripheral nerve regeneration. Taking advantage of matrilin-2’s capability to stimulate SC migration and adhesion, we formulated a porous matrilin-2/chitosan scaffold to support axongal outgrowth. Chemical modification of chitosan with lysine further improved matrilin-2 bioactivity in the 3D scaffold. The 3D porous matrilin-2/K-chitosan scaffolds have high potential for enhancing nerve repair by stimulating SC migration, neuronal adhesion, and axonal outgrowth.https://www.frontiersin.org/articles/10.3389/fbioe.2023.1142610/fullnerve regenerationscaffoldchitosanmatrilin-2schwann cellaxon |
| spellingShingle | Neill Y. Li Neill Y. Li Brandon Vorrius Jonathan Ge Zhen Qiao Shuang Zhu Julia Katarincic Qian Chen Matrilin-2 within a three-dimensional lysine-modified chitosan porous scaffold enhances Schwann cell migration and axonal outgrowth for peripheral nerve regeneration Frontiers in Bioengineering and Biotechnology nerve regeneration scaffold chitosan matrilin-2 schwann cell axon |
| title | Matrilin-2 within a three-dimensional lysine-modified chitosan porous scaffold enhances Schwann cell migration and axonal outgrowth for peripheral nerve regeneration |
| title_full | Matrilin-2 within a three-dimensional lysine-modified chitosan porous scaffold enhances Schwann cell migration and axonal outgrowth for peripheral nerve regeneration |
| title_fullStr | Matrilin-2 within a three-dimensional lysine-modified chitosan porous scaffold enhances Schwann cell migration and axonal outgrowth for peripheral nerve regeneration |
| title_full_unstemmed | Matrilin-2 within a three-dimensional lysine-modified chitosan porous scaffold enhances Schwann cell migration and axonal outgrowth for peripheral nerve regeneration |
| title_short | Matrilin-2 within a three-dimensional lysine-modified chitosan porous scaffold enhances Schwann cell migration and axonal outgrowth for peripheral nerve regeneration |
| title_sort | matrilin 2 within a three dimensional lysine modified chitosan porous scaffold enhances schwann cell migration and axonal outgrowth for peripheral nerve regeneration |
| topic | nerve regeneration scaffold chitosan matrilin-2 schwann cell axon |
| url | https://www.frontiersin.org/articles/10.3389/fbioe.2023.1142610/full |
| work_keys_str_mv | AT neillyli matrilin2withinathreedimensionallysinemodifiedchitosanporousscaffoldenhancesschwanncellmigrationandaxonaloutgrowthforperipheralnerveregeneration AT neillyli matrilin2withinathreedimensionallysinemodifiedchitosanporousscaffoldenhancesschwanncellmigrationandaxonaloutgrowthforperipheralnerveregeneration AT brandonvorrius matrilin2withinathreedimensionallysinemodifiedchitosanporousscaffoldenhancesschwanncellmigrationandaxonaloutgrowthforperipheralnerveregeneration AT jonathange matrilin2withinathreedimensionallysinemodifiedchitosanporousscaffoldenhancesschwanncellmigrationandaxonaloutgrowthforperipheralnerveregeneration AT zhenqiao matrilin2withinathreedimensionallysinemodifiedchitosanporousscaffoldenhancesschwanncellmigrationandaxonaloutgrowthforperipheralnerveregeneration AT shuangzhu matrilin2withinathreedimensionallysinemodifiedchitosanporousscaffoldenhancesschwanncellmigrationandaxonaloutgrowthforperipheralnerveregeneration AT juliakatarincic matrilin2withinathreedimensionallysinemodifiedchitosanporousscaffoldenhancesschwanncellmigrationandaxonaloutgrowthforperipheralnerveregeneration AT qianchen matrilin2withinathreedimensionallysinemodifiedchitosanporousscaffoldenhancesschwanncellmigrationandaxonaloutgrowthforperipheralnerveregeneration |