See the published version of this preprint at Stem Cell Research & Therapy.
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Research
Daniela Franco Bueno
Hospital Sírio-Libanês
Corresponding Author
ORCiD: https://orcid.org/0000-0001-5932-6134
License:
This work is licensed under a CC BY 4.0 License. Read Full License
Background. Bone reconstruction in congenital craniofacial differences, which affect about 2-3% of newborns, has long been the focus of intensive research in the field of bone tissue engineering. The possibility of using mesenchymal stem cells in regenerative medicine protocols has opened a new field of investigation aimed at finding optimal sources of multipotent stem cells that can be isolated via non-invasive procedures. Here we analysed whether levator veli palatini muscle fragments, which can be readily obtained in non-invasive manner during surgical rehabilitation of cleft patients during palatoplasty, represent a novel source of MSCs with osteogenic potential.
Methods. We obtained levator veli palatini muscle fragments, in non-invasive procedure during surgical rehabilitation of 5 unrelated cleft palate patients (palatoplasty surgery). The levator veli palatini muscle fragments was used to obtain the mesenchymal cells using pre-plating technique in a clean rooms infrastructure and all procedures were performed at good practices of manipulation conditions. To prove that levator veli palatini muscle are mesenchymal stem cells they were induced to flow cytometry analysis and to differentiation into bone, cartilage, fat and muscle. To demonstrate the osteogenic potential of these cells in vivo a bilateral full thickness calvarial defect model was made in immunocompentent rats.
Results. Flow cytometry analysis showed that the cells were positive for mesenchymal stem cell antigens (CD29, CD73, CD90), while negative for hematopoietic (CD45) or endothelial cell markers (CD31). Moreover, these cells were capable of undergoing chondrogenic, adipogenic, osteogenic and skeletal muscle cell differentiation under appropriate cell culture conditions characterizing them as mesenchymal stem cell. Defects treated with CellCeramTM scaffolds seeded with levator veli palatini muscle cells showed significantly greater bone healing compared to defects treated with acellular scaffolds.
Conclusion. We have demonstrated that cells derived from levator veli palatini muscle have phenotypic characteristics similar to other mesenchymal stem cells, both in vitro and in vivo. Our findings suggest that these cells may have clinical relevance in the rehabilitation of patients with cleft palate and other craniofacial anomalies characterized by significant bone deficit.
Keywords
Bone reconstruction, mesenchymal stem cells, levator veli palatini muscle, osteogenic differentiation, scaffold, craniofacial malformations
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View the published article at Stem Cell Research & Therapy.
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Editorial decision: Accept
On 26 Oct, 2020
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Received 09 Oct, 2020
Reviewer #2 agreed
On 09 Oct, 2020
Reviewer #3 agreed
On 09 Oct, 2020
Reviewer #4 agreed
On 09 Oct, 2020
Review #2 received
Received 09 Oct, 2020
Review #1 received
Received 09 Oct, 2020
Reviewer #1 agreed
On 07 Oct, 2020
Reviewers invited
Invitations sent on 06 Oct, 2020
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On 04 Oct, 2020
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On 03 Oct, 2020
Editor invited
On 03 Oct, 2020
Version 1
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Editorial decision: Major Revision
On 06 Aug, 2020
Review #3 received
Received 03 Aug, 2020
Review #1 received
Received 31 Jul, 2020
Review #4 received
Received 30 Jul, 2020
Reviewer #4 agreed
On 29 Jul, 2020
Review #2 received
Received 25 Jul, 2020
Reviewer #2 agreed
On 22 Jul, 2020
Reviewer #3 agreed
On 22 Jul, 2020
Reviewer #1 agreed
On 20 Jul, 2020
Reviewers invited
Invitations sent on 19 Jul, 2020
Editor assigned
On 16 Jul, 2020
Editor invited
On 15 Jul, 2020
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On 15 Jul, 2020
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On 13 Jul, 2020
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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
Daniela Franco Bueno
Hospital Sírio-Libanês
Corresponding Author
ORCiD: https://orcid.org/0000-0001-5932-6134
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
Editorial decision: Accept
On 26 Oct, 2020
Review #4 received
Received 21 Oct, 2020
Review #3 received
Received 09 Oct, 2020
Reviewer #2 agreed
On 09 Oct, 2020
Reviewer #3 agreed
On 09 Oct, 2020
Reviewer #4 agreed
On 09 Oct, 2020
Review #2 received
Received 09 Oct, 2020
Review #1 received
Received 09 Oct, 2020
Reviewer #1 agreed
On 07 Oct, 2020
Reviewers invited
Invitations sent on 06 Oct, 2020
Editor assigned
On 04 Oct, 2020
Submission checks complete
On 03 Oct, 2020
Editor invited
On 03 Oct, 2020
Version 1
Posted 17 Jul, 2020
No comments provided
Editorial decision: Major Revision
On 06 Aug, 2020
Review #3 received
Received 03 Aug, 2020
Review #1 received
Received 31 Jul, 2020
Review #4 received
Received 30 Jul, 2020
Reviewer #4 agreed
On 29 Jul, 2020
Review #2 received
Received 25 Jul, 2020
Reviewer #2 agreed
On 22 Jul, 2020
Reviewer #3 agreed
On 22 Jul, 2020
Reviewer #1 agreed
On 20 Jul, 2020
Reviewers invited
Invitations sent on 19 Jul, 2020
Editor assigned
On 16 Jul, 2020
Editor invited
On 15 Jul, 2020
Submission checks complete
On 15 Jul, 2020
First submitted
On 13 Jul, 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. Bone reconstruction in congenital craniofacial differences, which affect about 2-3% of newborns, has long been the focus of intensive research in the field of bone tissue engineering. The possibility of using mesenchymal stem cells in regenerative medicine protocols has opened a new field of investigation aimed at finding optimal sources of multipotent stem cells that can be isolated via non-invasive procedures. Here we analysed whether levator veli palatini muscle fragments, which can be readily obtained in non-invasive manner during surgical rehabilitation of cleft patients during palatoplasty, represent a novel source of MSCs with osteogenic potential.
Methods. We obtained levator veli palatini muscle fragments, in non-invasive procedure during surgical rehabilitation of 5 unrelated cleft palate patients (palatoplasty surgery). The levator veli palatini muscle fragments was used to obtain the mesenchymal cells using pre-plating technique in a clean rooms infrastructure and all procedures were performed at good practices of manipulation conditions. To prove that levator veli palatini muscle are mesenchymal stem cells they were induced to flow cytometry analysis and to differentiation into bone, cartilage, fat and muscle. To demonstrate the osteogenic potential of these cells in vivo a bilateral full thickness calvarial defect model was made in immunocompentent rats.
Results. Flow cytometry analysis showed that the cells were positive for mesenchymal stem cell antigens (CD29, CD73, CD90), while negative for hematopoietic (CD45) or endothelial cell markers (CD31). Moreover, these cells were capable of undergoing chondrogenic, adipogenic, osteogenic and skeletal muscle cell differentiation under appropriate cell culture conditions characterizing them as mesenchymal stem cell. Defects treated with CellCeramTM scaffolds seeded with levator veli palatini muscle cells showed significantly greater bone healing compared to defects treated with acellular scaffolds.
Conclusion. We have demonstrated that cells derived from levator veli palatini muscle have phenotypic characteristics similar to other mesenchymal stem cells, both in vitro and in vivo. Our findings suggest that these cells may have clinical relevance in the rehabilitation of patients with cleft palate and other craniofacial anomalies characterized by significant bone deficit.
Figure 1
Figure 2
Figure 3
Figure 4
Figure 5
This is a list of supplementary files associated with this preprint. Click to download.
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