Y. Dong (Shreveport, US)

LSU Health-Shreveport
Dr. Yufeng Dong; MD & PhD; a tenured Associate professor; is the Director of Translational Research in the Department of Orthopaedic Surgery. Dr. Dong received his PhD degree from Shanghai Jiaotong University and finished his postdoctoral training in University of Rochester. He received a significant funding from the NIH; Orthopaedic Research and Education foundation; Airlift Research Foundation and Lonza research Foundation to study the critical effects of stem cells on cartilage and bone regeneration.

Presenter Of 1 Presentation

Poster Cartilage /Cell Transplantation

P071 - Inhibition of Transcription Factor Twist1 Promotes Stromal Cell-Based Cartilage Repair

Presentation Topic
Cartilage /Cell Transplantation
Date
13.04.2022
Lecture Time
09:30 - 09:30
Room
Exhibition Foyer
Session Name
7.3 - Poster Viewing / Coffee Break / Exhibition
Session Type
Poster Session
Disclosure
No Significant Commercial Relationship

Abstract

Purpose

Placenta-derived Mesenchymal stromal cells (PMSCs) have been widely explored for tissue engineering applications and demonstrated capacity for chondrogenic differentiation. However, rapid induction of PMSC chondrogenic differentiation during therapeutic transplantation remains extremely challenging. This study is to determine if Twist1 inhibition could be utilized to accelerate PMSC-mediated cartilage repair in a cartilage defect model.

Methods and Materials

PMSCs were isolated from human placenta delivered from normal term. Cell phenotype was evaluated by flow cytometry, in vitro osteogenic, chondrogenic and adipogenic differentiation assays. Lentivirus-mediated gene silence was used to inhibit Twist1 in PMSC pellet culture. A mouse knee joint cartilage defect model was used for in vivo study. Wild type and Twist1 deficient PMSC pellets were generated and inserted to fill the cartilage defects. After 4 weeks postoperatively, osteochondral units containing the defects were harvested for Alcian Blue/Orange G staining, Immunohistochemical staining (IHC) and RT-PCR analysis.

Results

The flow cytometry results indicated that human stromal cell markers CD29, CD73, CD90 and proliferation marker Ki-67 were highly expressed in PMSCs, and these cells could be induced into osteoblasts, adipocytes and chondrocytes in vitro. Particularly, silencing Twist1 significantly enhanced chondrogenesis in pellet culture by showing increased Alcian blue staining, enhanced type II collagen (Col-II) expression when compared to control PMSCs. Importantly, the in vivo transplantation of Twist1-deficient PMSCs into knee joint cartilage defects had a significantly enhanced cartilage formation when compared with wild type PMSCs at 4 weeks after operation. Finally, our PCR data showed an increased expression of chondrogenic markers Sox9, Col-II and aggrecan in knee joint tissue with transplantation of Twist1 deficient PMSCs.

Conclusion

These findings demonstrate that 1) PMSC is a favorable cell source for cartilage repair. 2) Silencing Twist1 could accelerate PMSC differentiation into chondrocyte in vivo. Our results strongly suggest that targeting Twist1 in stromal cells may be a promising strategy for cartilage regeneration.

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Presenter Of 1 Presentation

Cartilage /Cell Transplantation

P071 - Inhibition of Transcription Factor Twist1 Promotes Stromal Cell-Based Cartilage Repair

Abstract

Purpose

Placenta-derived Mesenchymal stromal cells (PMSCs) have been widely explored for tissue engineering applications and demonstrated capacity for chondrogenic differentiation. However, rapid induction of PMSC chondrogenic differentiation during therapeutic transplantation remains extremely challenging. This study is to determine if Twist1 inhibition could be utilized to accelerate PMSC-mediated cartilage repair in a cartilage defect model.

Methods and Materials

PMSCs were isolated from human placenta delivered from normal term. Cell phenotype was evaluated by flow cytometry, in vitro osteogenic, chondrogenic and adipogenic differentiation assays. Lentivirus-mediated gene silence was used to inhibit Twist1 in PMSC pellet culture. A mouse knee joint cartilage defect model was used for in vivo study. Wild type and Twist1 deficient PMSC pellets were generated and inserted to fill the cartilage defects. After 4 weeks postoperatively, osteochondral units containing the defects were harvested for Alcian Blue/Orange G staining, Immunohistochemical staining (IHC) and RT-PCR analysis.

Results

The flow cytometry results indicated that human stromal cell markers CD29, CD73, CD90 and proliferation marker Ki-67 were highly expressed in PMSCs, and these cells could be induced into osteoblasts, adipocytes and chondrocytes in vitro. Particularly, silencing Twist1 significantly enhanced chondrogenesis in pellet culture by showing increased Alcian blue staining, enhanced type II collagen (Col-II) expression when compared to control PMSCs. Importantly, the in vivo transplantation of Twist1-deficient PMSCs into knee joint cartilage defects had a significantly enhanced cartilage formation when compared with wild type PMSCs at 4 weeks after operation. Finally, our PCR data showed an increased expression of chondrogenic markers Sox9, Col-II and aggrecan in knee joint tissue with transplantation of Twist1 deficient PMSCs.

Conclusion

These findings demonstrate that 1) PMSC is a favorable cell source for cartilage repair. 2) Silencing Twist1 could accelerate PMSC differentiation into chondrocyte in vivo. Our results strongly suggest that targeting Twist1 in stromal cells may be a promising strategy for cartilage regeneration.

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