IADR Abstract Archives
Potential Use of CLP Patient-Derived Fibroblasts for Alveolar Bone Reconstruction
Objectives: Autogenous bone graft is the gold standard for alveolar defects reconstruction in cleft lip and palate (CLP) affected patients. To relieve CLP individuals of the burden of another invasive surgery for harvesting bone, alternative strategies would be beneficial.
Cleft lips are surgically corrected at the age of 3-5 months, which requires the excision of superfluous lip tissue reaching into the cleft. This tissue remnant represents an optimal source for the isolation of primary cells, including fibroblasts (Fb), which display compelling similarities to mesenchymal stem cells (MSCs) known to have bone regenerative potential. These observations prompted us to investigate the in vitro characteristics of CLP-Fb in regard to their potential future clinical use for regenerative purposes.
Methods: We employed an explant culture approach to isolate CLP-Fb. Therefore, we thoroughly assess similarities of CLP-Fb with bone marrow-derived MSCs, including the expression of MSCs markers (CD73, CD90, CD105, CD106 and CD146) and the capacity to differentiate into fat- and bone-forming cells.
Results: Both CLP-Fb and MSCs were plastic adherent and showed a spindle-like morphology, with comparable levels of CD73, CD90 and CD105. However, CLP-Fb showed considerably lower CD106 and CD146 levels than MSCs. When induced to differentiate into fat- and bone-forming cells, both CLP-Fb and MSCs expressed lineage-specific transcription factors as well as distinct tissue markers. CLP-Fb also possess additional MSCs characteristics, such as the ability to form colonies from single cells and the capacity to modulate the immune response. Further analysis revealed the presence of specific CD106-positive as well as CD146-positive subpopulations within our CLP-Fb cultures. While the CD106- positive population could be enriched by serum starvation, the CD146-positive was reduced by passaging, resulting in a reduced bone-forming capacity.
Conclusions: Our data show that CLP-Fb significantly share MSC-like characteristics and their use might represent a promising alternative approach for the repair of bone defects.
Cleft lips are surgically corrected at the age of 3-5 months, which requires the excision of superfluous lip tissue reaching into the cleft. This tissue remnant represents an optimal source for the isolation of primary cells, including fibroblasts (Fb), which display compelling similarities to mesenchymal stem cells (MSCs) known to have bone regenerative potential. These observations prompted us to investigate the in vitro characteristics of CLP-Fb in regard to their potential future clinical use for regenerative purposes.
Methods: We employed an explant culture approach to isolate CLP-Fb. Therefore, we thoroughly assess similarities of CLP-Fb with bone marrow-derived MSCs, including the expression of MSCs markers (CD73, CD90, CD105, CD106 and CD146) and the capacity to differentiate into fat- and bone-forming cells.
Results: Both CLP-Fb and MSCs were plastic adherent and showed a spindle-like morphology, with comparable levels of CD73, CD90 and CD105. However, CLP-Fb showed considerably lower CD106 and CD146 levels than MSCs. When induced to differentiate into fat- and bone-forming cells, both CLP-Fb and MSCs expressed lineage-specific transcription factors as well as distinct tissue markers. CLP-Fb also possess additional MSCs characteristics, such as the ability to form colonies from single cells and the capacity to modulate the immune response. Further analysis revealed the presence of specific CD106-positive as well as CD146-positive subpopulations within our CLP-Fb cultures. While the CD106- positive population could be enriched by serum starvation, the CD146-positive was reduced by passaging, resulting in a reduced bone-forming capacity.
Conclusions: Our data show that CLP-Fb significantly share MSC-like characteristics and their use might represent a promising alternative approach for the repair of bone defects.