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Research
Jie Zhao
Shanghai Jiao Tong University School of Medicine
Corresponding Author
ORCiD: https://orcid.org/0000-0003-1000-5641
License:
This work is licensed under a CC BY 4.0 License. Read Full License
Background: Lower back pain is often accredited to loss of intervertebral disc (IVD) height and compromised spine stability as a result of intervertebral disc degeneration (IVDD). We aim to locally use Bleomycin to induce the fibrotic transformation of bone marrow stromal cells (BMSCs) as a means to induce reparative fibrosis to slow down the height loss.
Methods: IVD from patients were gathered for histological examination. The expression of transforming growth factor beta 1 (TGF-β) signaling pathway was determined by qPCR and western blotting. Nucleus pulposus (NP) cells, annulus fibrosus (AF) cells and the Rats’ bone marrow stromal cells (BMSC)were cultured and their responsiveness to Bleomycin was evaluated by Cell Counting Kit-8, comet assay, transwell migration and wound healing assays. Rat IVDD models were created by puncture, rescued by Bleomycin injection and the effectiveness was evaluated by images (X-ray and MRI) and atomic force microscope.
Results: Histological examination showed increased levels of pro-fibrotic markers in IVDD tissues from patients. AF cells and BMSC cells was induced to adopt to a pro-fibrotic phenotype with increased expression fibrotic markers Col1a1, Col3a1, FSP1. The pro-fibrotic effect of Bleomycin on AF cells and BMSCs was in part due to the activation of the TGFβ-TGFβR1-SMAD2/3 signaling pathway. Pharmacological inhibition or gene knock-down of TGFβR1 could mitigate the pro-fibrotic effects.
Conclusion: Locally injection of Bleomycin in rats’ IVD induced rapid fibrosis and maintained its height through TGFβ-TGFβR1-SMAD2/3 signaling pathway.
Keywords
Intervertebral Disc Degeneration, Bleomycin, Bone Marrow Stromal Cells, Annulus Fibrosus Cells, Reparative Fibrosis, TGFβ-TGFβR1-SMAD2/3 Signaling Pathway
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CURRENT STATUS: Published
View the published article at Stem Cell Research & Therapy.
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Review #2 received
Received 18 Nov, 2020
Editorial decision: Accept
On 18 Nov, 2020
Reviewer #2 agreed
On 17 Nov, 2020
Reviewers invited
Invitations sent on 16 Nov, 2020
Reviewer #1 agreed
On 16 Nov, 2020
Review #1 received
Received 16 Nov, 2020
Editor assigned
On 15 Nov, 2020
Submission checks complete
On 15 Nov, 2020
Editor invited
On 15 Nov, 2020
Version 1
Posted 04 Sep, 2020
No comments provided
Review #3 received
Received 12 Oct, 2020
Editorial decision: Major Revision
On 12 Oct, 2020
Review #2 received
Received 06 Oct, 2020
Reviewer #4 agreed
On 04 Oct, 2020
Review #1 received
Received 30 Sep, 2020
Reviewer #2 agreed
On 28 Sep, 2020
Reviewer #3 agreed
On 28 Sep, 2020
Editor assigned
On 07 Sep, 2020
Reviewers invited
Invitations sent on 07 Sep, 2020
Reviewer #1 agreed
On 07 Sep, 2020
Editor invited
On 06 Sep, 2020
Submission checks complete
On 03 Sep, 2020
First submitted
On 02 Sep, 2020
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Subject Areas
Stem Cell & Developmental Cell Biology
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A new preprint design is coming!
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See the published version of this preprint at Stem Cell Research & Therapy.
This is a preprint, a preliminary version of a manuscript that has not completed peer review at a journal. Research Square does not conduct peer review prior to posting preprints. The posting of a preprint on this server should not be interpreted as an endorsement of its validity or suitability for dissemination as established information or for guiding clinical practice.
Learn more about In Review
Research
Jie Zhao
Shanghai Jiao Tong University School of Medicine
Corresponding Author
ORCiD: https://orcid.org/0000-0003-1000-5641
Badges
Prescreen
This manuscript passed our Prescreen assessment
Complete author information
Appropriate declaration statements
Potential risks to human health
Prescreen
Peer Review Timeline
CURRENT STATUS: Published
View the published article at Stem Cell Research & Therapy.
No community comments so far
Review #2 received
Received 18 Nov, 2020
Editorial decision: Accept
On 18 Nov, 2020
Reviewer #2 agreed
On 17 Nov, 2020
Reviewers invited
Invitations sent on 16 Nov, 2020
Reviewer #1 agreed
On 16 Nov, 2020
Review #1 received
Received 16 Nov, 2020
Editor assigned
On 15 Nov, 2020
Submission checks complete
On 15 Nov, 2020
Editor invited
On 15 Nov, 2020
Version 1
Posted 04 Sep, 2020
No comments provided
Review #3 received
Received 12 Oct, 2020
Editorial decision: Major Revision
On 12 Oct, 2020
Review #2 received
Received 06 Oct, 2020
Reviewer #4 agreed
On 04 Oct, 2020
Review #1 received
Received 30 Sep, 2020
Reviewer #2 agreed
On 28 Sep, 2020
Reviewer #3 agreed
On 28 Sep, 2020
Editor assigned
On 07 Sep, 2020
Reviewers invited
Invitations sent on 07 Sep, 2020
Reviewer #1 agreed
On 07 Sep, 2020
Editor invited
On 06 Sep, 2020
Submission checks complete
On 03 Sep, 2020
First submitted
On 02 Sep, 2020
Metrics
Comments: 0
PDF Downloads: ...
HTML Views: ...
Subject Areas
Stem Cell & Developmental Cell Biology
License:
This work is licensed under a CC BY 4.0 License. Read Full License
View the published article at Stem Cell Research & Therapy.
Background: Lower back pain is often accredited to loss of intervertebral disc (IVD) height and compromised spine stability as a result of intervertebral disc degeneration (IVDD). We aim to locally use Bleomycin to induce the fibrotic transformation of bone marrow stromal cells (BMSCs) as a means to induce reparative fibrosis to slow down the height loss.
Methods: IVD from patients were gathered for histological examination. The expression of transforming growth factor beta 1 (TGF-β) signaling pathway was determined by qPCR and western blotting. Nucleus pulposus (NP) cells, annulus fibrosus (AF) cells and the Rats’ bone marrow stromal cells (BMSC)were cultured and their responsiveness to Bleomycin was evaluated by Cell Counting Kit-8, comet assay, transwell migration and wound healing assays. Rat IVDD models were created by puncture, rescued by Bleomycin injection and the effectiveness was evaluated by images (X-ray and MRI) and atomic force microscope.
Results: Histological examination showed increased levels of pro-fibrotic markers in IVDD tissues from patients. AF cells and BMSC cells was induced to adopt to a pro-fibrotic phenotype with increased expression fibrotic markers Col1a1, Col3a1, FSP1. The pro-fibrotic effect of Bleomycin on AF cells and BMSCs was in part due to the activation of the TGFβ-TGFβR1-SMAD2/3 signaling pathway. Pharmacological inhibition or gene knock-down of TGFβR1 could mitigate the pro-fibrotic effects.
Conclusion: Locally injection of Bleomycin in rats’ IVD induced rapid fibrosis and maintained its height through TGFβ-TGFβR1-SMAD2/3 signaling pathway.
Figure 1
Figure 2
Figure 3
Figure 4
Figure 5
Figure 6
Figure 7
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