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Extracellular Matrix of Current Biological Scaffolds Promotes the Differentiation Potential of Mesenchymal Stem Cells.
Arthroscopy 2016 November
PURPOSE: The purpose of this study was to quantitatively assess the ability of bone marrow-derived mesenchymal stem cells (bMSC) to differentiate toward bone, fat, cartilage, and tendon lineages when grown on commercially available scaffolds compared with control and native tendon tissue.
METHODS: BMSCs were cultured and analyzed by fluorescent automated cells sorting for surface markers CD73, -90, and -105. BMSCs were grown on rotator cuff tendon (RCT), decellularized human dermis patch (DDP), bilayer collagen matrix, and fibrin matrix (FM) to test their differentiation potential using quantitative polymerase chain reaction and establish markers for osteogenic, adipogenic, chondrogenic, and tenogenic lineages. Immunocytochemical testing was used to determine the specific proteins present on the scaffolds.
RESULTS: Alkaline phosphatase and osteocalcin gene expression was significantly higher on RCT (P < .001) and collagen scaffold (CS) (P < .001) compared with DDP and FM scaffolds (P < .001, P < .001). When differentiated toward a cartilage lineage, bMSCs grown on CS had significantly more type II collagen and aggrecan compared with DDP (P < .001, P < .001), FM (P < .001, P < .001), and RCT (P < .001, P < .001). Differentiated bMSCs grown on the CS had a significant increase in PPARγ and FABP4 gene expression compared with bMSCs grown on all other scaffolds (all P < .001). The differentiation of bMSCs into tendon on CSs had significantly more tenacin C, decorin, and type III collagen gene expression when compared with RCT, DDP, and FM (all P < .001). Decorin gene expression in the control undifferentiated CS was also significantly increased, suggesting that the matrix alone may promote a tenogenic lineage (P = .637).
CONCLUSIONS: Differences in the extracellular matrix composition of scaffolds significantly impact their potential to promote differentiation of bMSCs. Comparing the native RCT to the tested scaffolds showed that a high content of type I and III collagen significantly increased the potential of bMSCs to differentiate toward bone, tendon, fat, and cartilage lineages.
CLINICAL RELEVANCE: This in vitro study shows the differences between commercially available scaffolds for rotator cuff repairs. Therefore, these results support clinical use depending on the surgical intention and the potential of bMSCs to differentiate into bone, tendon, cartilage, and fat tissue.
METHODS: BMSCs were cultured and analyzed by fluorescent automated cells sorting for surface markers CD73, -90, and -105. BMSCs were grown on rotator cuff tendon (RCT), decellularized human dermis patch (DDP), bilayer collagen matrix, and fibrin matrix (FM) to test their differentiation potential using quantitative polymerase chain reaction and establish markers for osteogenic, adipogenic, chondrogenic, and tenogenic lineages. Immunocytochemical testing was used to determine the specific proteins present on the scaffolds.
RESULTS: Alkaline phosphatase and osteocalcin gene expression was significantly higher on RCT (P < .001) and collagen scaffold (CS) (P < .001) compared with DDP and FM scaffolds (P < .001, P < .001). When differentiated toward a cartilage lineage, bMSCs grown on CS had significantly more type II collagen and aggrecan compared with DDP (P < .001, P < .001), FM (P < .001, P < .001), and RCT (P < .001, P < .001). Differentiated bMSCs grown on the CS had a significant increase in PPARγ and FABP4 gene expression compared with bMSCs grown on all other scaffolds (all P < .001). The differentiation of bMSCs into tendon on CSs had significantly more tenacin C, decorin, and type III collagen gene expression when compared with RCT, DDP, and FM (all P < .001). Decorin gene expression in the control undifferentiated CS was also significantly increased, suggesting that the matrix alone may promote a tenogenic lineage (P = .637).
CONCLUSIONS: Differences in the extracellular matrix composition of scaffolds significantly impact their potential to promote differentiation of bMSCs. Comparing the native RCT to the tested scaffolds showed that a high content of type I and III collagen significantly increased the potential of bMSCs to differentiate toward bone, tendon, fat, and cartilage lineages.
CLINICAL RELEVANCE: This in vitro study shows the differences between commercially available scaffolds for rotator cuff repairs. Therefore, these results support clinical use depending on the surgical intention and the potential of bMSCs to differentiate into bone, tendon, cartilage, and fat tissue.
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