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Dendronized chitosan hydrogel with GIT1 to accelerate bone defect repair through increasing local neovascular amount
Lin Cheng1,2, Zhimin Zhou2, Qingqing Li2
1Department of Orthopedics, The Affiliated Hospital of Xuzhou Medical University, Huaihai West Road 99, Xuzhou, Jiangsu Province 221000, China.
Bone Reports
|September 25, 2023
Summary
Injectable hydrogels loaded with G protein-coupled receptor kinase 2 interacting protein 1 (GIT1) plasmids significantly enhance bone defect healing and neovascularization. This strategy promotes mesenchymal stem cell differentiation into endothelial cells, aiding bone tissue engineering.
Area of Science:
- Biomaterials Science
- Regenerative Medicine
- Orthopedic Research
Background:
- Bone defects present significant healthcare challenges due to complex regeneration and high treatment costs.
- G protein-coupled receptor kinase 2 interacting protein 1 (GIT1) is crucial for vascular development and bone fracture healing.
- Current bone regeneration strategies often face limitations in efficacy and cost-effectiveness.
Purpose of the Study:
- To develop an injectable hydrogel system for delivering GIT1 to enhance bone defect repair.
- To investigate the role of GIT1 in promoting mesenchymal stem cell (MSC) differentiation and neovascularization.
- To evaluate the efficacy of a combined hydrogel-GIT1 approach in a bone defect model.
Main Methods:
- Fabrication of thermoresponsive injectable hydrogel from oligoethylene glycol-based dendronized chitosan (G1-CS).
- Loading of GIT1-plasmids into G1-CS hydrogel (G1-CS/GIT1) for delivery.
- In vitro and in vivo assessment of DNA transfection, bone healing, and neovascularization using RT-PCR, micro-CT, and histological analysis.
- Construction of adeno-associated virus (AAV)-GIT1 for MSC transfection and analysis of endothelial differentiation via capillary tube formation assay, immunofluorescence, and Western blot.
Main Results:
- G1-CS/GIT1 hydrogel demonstrated enhanced DNA transfection in MSCs both in vitro and in vivo.
- Micro-CT, RT-PCR, and histological analyses confirmed accelerated bone healing and increased neovascularization around bone defects treated with G1-CS/GIT1.
- High GIT1 expression induced MSC differentiation into endothelial cells, evidenced by capillary tube formation, immunofluorescence, and Western blot.
- The Notch signaling pathway was confirmed to be activated during this differentiation process.
Conclusions:
- The injectable G1-CS/GIT1 hydrogel is an effective strategy for enhancing bone defect regeneration.
- GIT1 plays a key role in promoting MSC differentiation into endothelial cells, contributing to neovascularization.
- This combined approach of injectable hydrogel and GIT1 holds significant promise for bone tissue engineering applications.
