Abstract
Objectives
We genetically modified dedifferentiated chondrocytes (DCs) using lentiviral vectors and adenoviral vectors encoding TGF-β3 (referred to as transgenic groups below) and encapsulated these DCs in the microcavitary hydrogel and investigated the combinational effect on redifferentiation of the genetically manipulated DCs.
Results
The Cell Counting Kit-8 data indicated that both transgenic groups exhibited significantly higher cell viability in the first week but inferior cell viability in the subsequent timepoints compared with those of the control group. Real-time polymerase chain reaction and western blot analysis results demonstrated that both transgenic groups had a better effect on redifferentiation to some extent, as evidenced by higher expression levels of chondrogenic genes, suggesting the validity of combination with transgenic DCs and the microcavitary hydrogel on redifferentiation. Although transgenic DCs with adenoviral vectors presented a superior extent of redifferentiation, they also expressed greater levels of the hypertrophic gene type X collagen. It is still worth further exploring how to deliver TGF-β3 more efficiently and optimizing the appropriate parameters, including concentration and duration.
Conclusions
The results demonstrated the better redifferentiation effect of DCs with the combinational use of transgenic TGF-β3 and a microcavitary alginate hydrogel and implied that DCs would be alternative seed cells for cartilage tissue engineering due to their easily achieved sufficient cell amounts through multiple passages and great potential to redifferentiate to produce cartilaginous extracellular matrix.
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Acknowledgements
This work was supported by the Science and Technology Program of Guangzhou (Dengfeng Hospital 2022) (Grant number 202201020460), Guangzhou Medical University Research Ability Enhancement Project (2024) (Grant number 2024SRP072) and Guangzhou Health Science and Technology Project (Grant number 20241A010112).
Funding
The authors have not disclosed any funding The Science and Technology Program of Guangzhou (Dengfeng Hospital 2022),202201020460,Yongchang Yao,Guangzhou Medical University Research Ability Enhancement Project,2024,Yongchang Yao,Guangzhou Health Science and Technology Project,Grant number 20241A010112.
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Yao, Y., Chen, K., Pan, Q. et al. Redifferentiation of genetically modified dedifferentiated chondrocytes in a microcavitary hydrogel. Biotechnol Lett 46, 483–495 (2024). https://doi.org/10.1007/s10529-024-03475-2
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DOI: https://doi.org/10.1007/s10529-024-03475-2