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TGF-βl Suppresses Inflammation in Cell Therapy for Intervertebral Disc Degeneration.

Yang H, Cao C, Wu C, Yuan C, Gu Q, Shi Q, Zou J - Sci Rep (2015)

Bottom Line: As a strong immune suppressor, TGF-β1 has been shown to inhibit inflammation respond effectively.In vitro assays demonstrated that co-culturing of nucleus pulposus cells with bone marrow mesenchymal stem cells resulted in significantly higher levels of TGF-βl secretion.However, the NF-κB positive cells were significantly less than other two control groups.

View Article: PubMed Central - PubMed

Affiliation: Department of Orthopaedic Surgery, The First Affiliated Hospital of Soochow University, Suzhou, Jiangsu 215006, China.

ABSTRACT
Recent studies suggest that cell therapy may be an effective way to repair intervertebral disc degeneration. As a strong immune suppressor, TGF-β1 has been shown to inhibit inflammation respond effectively. The objective of this study was to explore the effects of TGF-β1 during bone marrow mesenchymal stem cells-based therapy for disc degeneration. In vitro assays demonstrated that co-culturing of nucleus pulposus cells with bone marrow mesenchymal stem cells resulted in significantly higher levels of TGF-βl secretion. This increase inhibited IκB phosphorylation and NF-κB activation, detected by western blot analysis. Meanwhile, in a rabbit model, MRI analysis revealed significant recovery of signal intensity in the degenerative discs of rabbits receiving cells transplantation, than receiving cells treated with a TGF-β1 inhibitor or saline. These findings indicated that enhanced TGF-β1 production recovered the degeneration of intervertebral disc. And also immunohistochemical staining detected enhanced collagen II expression in the rabbits treated with cell transplantation. However, the NF-κB positive cells were significantly less than other two control groups. Thus, cell therapy promoted TGF-β1 expression in nucleus pulposus, leading to anti-inflammatory effects via the inhibition of NF-κB, and the amelioration of disc degradation due to increased expression of collagen II and aggrecan in degenerative intervertebral disc.

No MeSH data available.


Related in: MedlinePlus

The supernatant media collected from different groups were analyzed for TGF-β1 concentration measured by ELISA.ELISA assay showed that the expression of TGF-β1 in NPCs increased gradually after co-cultured with BMSCs, compared to single culture group. However, there was no significant difference between the co-culture group and the TGF-βl-induced group.
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f1: The supernatant media collected from different groups were analyzed for TGF-β1 concentration measured by ELISA.ELISA assay showed that the expression of TGF-β1 in NPCs increased gradually after co-cultured with BMSCs, compared to single culture group. However, there was no significant difference between the co-culture group and the TGF-βl-induced group.

Mentions: To evaluate the BMSCs’ capability of TGF-βl induction on NPCs, we co-cultured these two cells in six-well Transwell cell culture plates. About 1 × 106 BMSCs were used to coat the bottom chamber of the Transwell system, while 1 × 106 NPCs were added to the upper chamber. About 1 × 106 NPCs treated with recombinant TGF-β1 proteins (at a final concentration of 30 ng/mL) were used as a positive control group, while a single culture containing 1 × 106 NPCs served as the negative control. At 1, 2 and 3 days after co-culture, cells were harvested and seeded in at 1 × 106 cells/well. The supernatants from different populations were then harvested for ELISA experiments. The levels of TGF-βl protein secretion by NPCs increased from 555.16 ± 133.64 to 805.14 ± 212.84 pg/mL between days 1 and 2 of co-culturing with BMSCs, as determined by ELISA analyses. Conversely, there were relatively low concentrations of TGF-βl at days 1 (315.74 ± 71.42 pg/mL) and 2 (342.33 ± 148.49 pg/mL) in the control group (NPCs single culture). At the day 3 time point, there was a significant increase in the levels of TGF-βl secretion by the co-culture group compared with control group (1648.75 ± 155.56 vs. 555.16 ± 133.64 pg/mL, p < 0.05). Meanwhile, there was no significant difference in cytokine secretion between the co-culture group and the recombinant TGF-βl-induced group (1761.25 ± 165.46 pg/mL, p > 0.05). (Fig. 1).


TGF-βl Suppresses Inflammation in Cell Therapy for Intervertebral Disc Degeneration.

Yang H, Cao C, Wu C, Yuan C, Gu Q, Shi Q, Zou J - Sci Rep (2015)

The supernatant media collected from different groups were analyzed for TGF-β1 concentration measured by ELISA.ELISA assay showed that the expression of TGF-β1 in NPCs increased gradually after co-cultured with BMSCs, compared to single culture group. However, there was no significant difference between the co-culture group and the TGF-βl-induced group.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4542522&req=5

f1: The supernatant media collected from different groups were analyzed for TGF-β1 concentration measured by ELISA.ELISA assay showed that the expression of TGF-β1 in NPCs increased gradually after co-cultured with BMSCs, compared to single culture group. However, there was no significant difference between the co-culture group and the TGF-βl-induced group.
Mentions: To evaluate the BMSCs’ capability of TGF-βl induction on NPCs, we co-cultured these two cells in six-well Transwell cell culture plates. About 1 × 106 BMSCs were used to coat the bottom chamber of the Transwell system, while 1 × 106 NPCs were added to the upper chamber. About 1 × 106 NPCs treated with recombinant TGF-β1 proteins (at a final concentration of 30 ng/mL) were used as a positive control group, while a single culture containing 1 × 106 NPCs served as the negative control. At 1, 2 and 3 days after co-culture, cells were harvested and seeded in at 1 × 106 cells/well. The supernatants from different populations were then harvested for ELISA experiments. The levels of TGF-βl protein secretion by NPCs increased from 555.16 ± 133.64 to 805.14 ± 212.84 pg/mL between days 1 and 2 of co-culturing with BMSCs, as determined by ELISA analyses. Conversely, there were relatively low concentrations of TGF-βl at days 1 (315.74 ± 71.42 pg/mL) and 2 (342.33 ± 148.49 pg/mL) in the control group (NPCs single culture). At the day 3 time point, there was a significant increase in the levels of TGF-βl secretion by the co-culture group compared with control group (1648.75 ± 155.56 vs. 555.16 ± 133.64 pg/mL, p < 0.05). Meanwhile, there was no significant difference in cytokine secretion between the co-culture group and the recombinant TGF-βl-induced group (1761.25 ± 165.46 pg/mL, p > 0.05). (Fig. 1).

Bottom Line: As a strong immune suppressor, TGF-β1 has been shown to inhibit inflammation respond effectively.In vitro assays demonstrated that co-culturing of nucleus pulposus cells with bone marrow mesenchymal stem cells resulted in significantly higher levels of TGF-βl secretion.However, the NF-κB positive cells were significantly less than other two control groups.

View Article: PubMed Central - PubMed

Affiliation: Department of Orthopaedic Surgery, The First Affiliated Hospital of Soochow University, Suzhou, Jiangsu 215006, China.

ABSTRACT
Recent studies suggest that cell therapy may be an effective way to repair intervertebral disc degeneration. As a strong immune suppressor, TGF-β1 has been shown to inhibit inflammation respond effectively. The objective of this study was to explore the effects of TGF-β1 during bone marrow mesenchymal stem cells-based therapy for disc degeneration. In vitro assays demonstrated that co-culturing of nucleus pulposus cells with bone marrow mesenchymal stem cells resulted in significantly higher levels of TGF-βl secretion. This increase inhibited IκB phosphorylation and NF-κB activation, detected by western blot analysis. Meanwhile, in a rabbit model, MRI analysis revealed significant recovery of signal intensity in the degenerative discs of rabbits receiving cells transplantation, than receiving cells treated with a TGF-β1 inhibitor or saline. These findings indicated that enhanced TGF-β1 production recovered the degeneration of intervertebral disc. And also immunohistochemical staining detected enhanced collagen II expression in the rabbits treated with cell transplantation. However, the NF-κB positive cells were significantly less than other two control groups. Thus, cell therapy promoted TGF-β1 expression in nucleus pulposus, leading to anti-inflammatory effects via the inhibition of NF-κB, and the amelioration of disc degradation due to increased expression of collagen II and aggrecan in degenerative intervertebral disc.

No MeSH data available.


Related in: MedlinePlus