Cargando…
Osteogenic Potential of Sheep Mesenchymal Stem Cells Preconditioned with BMP-2 and FGF-2 and Seeded on an nHAP-Coated PCL/HAP/β-TCP Scaffold
Mesenchymal stem cells (MSCs) attract interest in regenerative medicine for their potential application in bone regeneration. However, direct transplantation of cells into damaged tissue is not efficient enough to regenerate large bone defects. This problem could be solved with a biocompatible scaff...
Autores principales: | , , , , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2022
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9659177/ https://www.ncbi.nlm.nih.gov/pubmed/36359842 http://dx.doi.org/10.3390/cells11213446 |
_version_ | 1784830137231998976 |
---|---|
author | Stamnitz, Sandra Krawczenko, Agnieszka Szałaj, Urszula Górecka, Żaneta Antończyk, Agnieszka Kiełbowicz, Zdzisław Święszkowski, Wojciech Łojkowski, Witold Klimczak, Aleksandra |
author_facet | Stamnitz, Sandra Krawczenko, Agnieszka Szałaj, Urszula Górecka, Żaneta Antończyk, Agnieszka Kiełbowicz, Zdzisław Święszkowski, Wojciech Łojkowski, Witold Klimczak, Aleksandra |
author_sort | Stamnitz, Sandra |
collection | PubMed |
description | Mesenchymal stem cells (MSCs) attract interest in regenerative medicine for their potential application in bone regeneration. However, direct transplantation of cells into damaged tissue is not efficient enough to regenerate large bone defects. This problem could be solved with a biocompatible scaffold. Consequently, bone tissue engineering constructs based on biomaterial scaffolds, MSCs, and osteogenic cytokines are promising tools for bone regeneration. The aim of this study was to evaluate the effect of FGF-2 and BMP-2 on the osteogenic potential of ovine bone marrow-derived MSCs seeded onto an nHAP-coated PCL/HAP/β-TCP scaffold in vitro and its in vivo biocompatibility in a sheep model. In vitro analysis revealed that cells preconditioned with FGF-2 and BMP-2 showed a better capacity to adhere and proliferate on the scaffold than untreated cells. BM-MSCs cultured in an osteogenic medium supplemented with FGF-2 and BMP-2 had the highest osteogenic differentiation potential, as assessed based on Alizarin Red S staining and ALP activity. qRT-PCR analysis showed increased expression of osteogenic marker genes in FGF-2- and BMP-2-treated BM-MSCs. Our pilot in vivo research showed that the implantation of an nHAP-coated PCL/HAP/β-TCP scaffold with BM-MSCs preconditioned with FGF-2 and BMP-2 did not have an adverse effect in the sheep mandibular region and induced bone regeneration. The biocompatibility of the implanted scaffold-BM-MSC construct with sheep tissues was confirmed by the expression of early (collagen type I) and late (osteocalcin) osteogenic proteins and a lack of an elevated level of proinflammatory cytokines. These findings suggest that FGF-2 and BMP-2 enhance the osteogenic differentiation potential of MSCs grown on a scaffold, and that such a tissue engineering construct may be used to regenerate large bone defects. |
format | Online Article Text |
id | pubmed-9659177 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-96591772022-11-15 Osteogenic Potential of Sheep Mesenchymal Stem Cells Preconditioned with BMP-2 and FGF-2 and Seeded on an nHAP-Coated PCL/HAP/β-TCP Scaffold Stamnitz, Sandra Krawczenko, Agnieszka Szałaj, Urszula Górecka, Żaneta Antończyk, Agnieszka Kiełbowicz, Zdzisław Święszkowski, Wojciech Łojkowski, Witold Klimczak, Aleksandra Cells Article Mesenchymal stem cells (MSCs) attract interest in regenerative medicine for their potential application in bone regeneration. However, direct transplantation of cells into damaged tissue is not efficient enough to regenerate large bone defects. This problem could be solved with a biocompatible scaffold. Consequently, bone tissue engineering constructs based on biomaterial scaffolds, MSCs, and osteogenic cytokines are promising tools for bone regeneration. The aim of this study was to evaluate the effect of FGF-2 and BMP-2 on the osteogenic potential of ovine bone marrow-derived MSCs seeded onto an nHAP-coated PCL/HAP/β-TCP scaffold in vitro and its in vivo biocompatibility in a sheep model. In vitro analysis revealed that cells preconditioned with FGF-2 and BMP-2 showed a better capacity to adhere and proliferate on the scaffold than untreated cells. BM-MSCs cultured in an osteogenic medium supplemented with FGF-2 and BMP-2 had the highest osteogenic differentiation potential, as assessed based on Alizarin Red S staining and ALP activity. qRT-PCR analysis showed increased expression of osteogenic marker genes in FGF-2- and BMP-2-treated BM-MSCs. Our pilot in vivo research showed that the implantation of an nHAP-coated PCL/HAP/β-TCP scaffold with BM-MSCs preconditioned with FGF-2 and BMP-2 did not have an adverse effect in the sheep mandibular region and induced bone regeneration. The biocompatibility of the implanted scaffold-BM-MSC construct with sheep tissues was confirmed by the expression of early (collagen type I) and late (osteocalcin) osteogenic proteins and a lack of an elevated level of proinflammatory cytokines. These findings suggest that FGF-2 and BMP-2 enhance the osteogenic differentiation potential of MSCs grown on a scaffold, and that such a tissue engineering construct may be used to regenerate large bone defects. MDPI 2022-10-31 /pmc/articles/PMC9659177/ /pubmed/36359842 http://dx.doi.org/10.3390/cells11213446 Text en © 2022 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Article Stamnitz, Sandra Krawczenko, Agnieszka Szałaj, Urszula Górecka, Żaneta Antończyk, Agnieszka Kiełbowicz, Zdzisław Święszkowski, Wojciech Łojkowski, Witold Klimczak, Aleksandra Osteogenic Potential of Sheep Mesenchymal Stem Cells Preconditioned with BMP-2 and FGF-2 and Seeded on an nHAP-Coated PCL/HAP/β-TCP Scaffold |
title | Osteogenic Potential of Sheep Mesenchymal Stem Cells Preconditioned with BMP-2 and FGF-2 and Seeded on an nHAP-Coated PCL/HAP/β-TCP Scaffold |
title_full | Osteogenic Potential of Sheep Mesenchymal Stem Cells Preconditioned with BMP-2 and FGF-2 and Seeded on an nHAP-Coated PCL/HAP/β-TCP Scaffold |
title_fullStr | Osteogenic Potential of Sheep Mesenchymal Stem Cells Preconditioned with BMP-2 and FGF-2 and Seeded on an nHAP-Coated PCL/HAP/β-TCP Scaffold |
title_full_unstemmed | Osteogenic Potential of Sheep Mesenchymal Stem Cells Preconditioned with BMP-2 and FGF-2 and Seeded on an nHAP-Coated PCL/HAP/β-TCP Scaffold |
title_short | Osteogenic Potential of Sheep Mesenchymal Stem Cells Preconditioned with BMP-2 and FGF-2 and Seeded on an nHAP-Coated PCL/HAP/β-TCP Scaffold |
title_sort | osteogenic potential of sheep mesenchymal stem cells preconditioned with bmp-2 and fgf-2 and seeded on an nhap-coated pcl/hap/β-tcp scaffold |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9659177/ https://www.ncbi.nlm.nih.gov/pubmed/36359842 http://dx.doi.org/10.3390/cells11213446 |
work_keys_str_mv | AT stamnitzsandra osteogenicpotentialofsheepmesenchymalstemcellspreconditionedwithbmp2andfgf2andseededonannhapcoatedpclhapbtcpscaffold AT krawczenkoagnieszka osteogenicpotentialofsheepmesenchymalstemcellspreconditionedwithbmp2andfgf2andseededonannhapcoatedpclhapbtcpscaffold AT szałajurszula osteogenicpotentialofsheepmesenchymalstemcellspreconditionedwithbmp2andfgf2andseededonannhapcoatedpclhapbtcpscaffold AT goreckazaneta osteogenicpotentialofsheepmesenchymalstemcellspreconditionedwithbmp2andfgf2andseededonannhapcoatedpclhapbtcpscaffold AT antonczykagnieszka osteogenicpotentialofsheepmesenchymalstemcellspreconditionedwithbmp2andfgf2andseededonannhapcoatedpclhapbtcpscaffold AT kiełbowiczzdzisław osteogenicpotentialofsheepmesenchymalstemcellspreconditionedwithbmp2andfgf2andseededonannhapcoatedpclhapbtcpscaffold AT swieszkowskiwojciech osteogenicpotentialofsheepmesenchymalstemcellspreconditionedwithbmp2andfgf2andseededonannhapcoatedpclhapbtcpscaffold AT łojkowskiwitold osteogenicpotentialofsheepmesenchymalstemcellspreconditionedwithbmp2andfgf2andseededonannhapcoatedpclhapbtcpscaffold AT klimczakaleksandra osteogenicpotentialofsheepmesenchymalstemcellspreconditionedwithbmp2andfgf2andseededonannhapcoatedpclhapbtcpscaffold |