Improved osteogenic differentiation of umbilical cord blood MSCs using custom made perfusion bioreactor
Background: 3D cell culture is an appropriate method to develop engineered bone tissue, where different bioreactors have been designed to mitigate the challenges in 3D culture. Currently, we tailored a perfusion reactor to witness human mesenchymal stem cells (MSCs) proliferation and differentiation...
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| Format: | Article |
| Language: | English |
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Elsevier
2018-10-01
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| Series: | Biomedical Journal |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2319417017303499 |
| _version_ | 1798024766877073408 |
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| author | Bhaskar Birru Naveen Kumar Mekala Sreenivasa Rao Parcha |
| author_facet | Bhaskar Birru Naveen Kumar Mekala Sreenivasa Rao Parcha |
| author_sort | Bhaskar Birru |
| collection | DOAJ |
| description | Background: 3D cell culture is an appropriate method to develop engineered bone tissue, where different bioreactors have been designed to mitigate the challenges in 3D culture. Currently, we tailored a perfusion reactor to witness human mesenchymal stem cells (MSCs) proliferation and differentiation over polylactic acid-polyethylene glycol (PLA/PEG) composite scaffolds. Methods: The composite scaffolds with different weight ratios of PLA and PEG were prepared using solvent casting-particulate leaching technique. Human umbilcal card blood MSCs were cultured under dynamic and static conditions to elucidate the role of dynamic fluid flow in osteogenesis of MSCs. Results: The human MSCs distribution over the scaffolds was confirmed with fluorescent microscopy. Alkaline phosphatase (ALP), calcium mineralization, and collagen formation were found to be higher in PLA90 scaffolds than PLA100 and PLA75. PLA90 scaffolds with better cell adhesion/proliferartion were considered for bioreactor studies and they exhibited enhanced ALP, Ca+2 mineralization and collagen formation under dynamic perfusion than static culture. We further confirmed our observation by looking at expression levels of osteogenic marker (Runx2 and osteonectin) in differentiated MSCs subjected to perfusion culture compared to static culture. Conclusion: The results of the current investigation once again proves that dynamic perfusion cultures improve the osteogenic differentiation of MSCs over hybrid polymer scaffolds (PLA90) for effective bone regeneration. Keywords: 3D cell culture, Mesenchymal stem cells (MSCs), Fluid flow, Osteogenic differentiation |
| first_indexed | 2024-04-11T18:07:55Z |
| format | Article |
| id | doaj.art-bdc8fc183f7942deb8479f2652337a01 |
| institution | Directory Open Access Journal |
| issn | 2319-4170 |
| language | English |
| last_indexed | 2024-04-11T18:07:55Z |
| publishDate | 2018-10-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Biomedical Journal |
| spelling | doaj.art-bdc8fc183f7942deb8479f2652337a012022-12-22T04:10:16ZengElsevierBiomedical Journal2319-41702018-10-01415290297Improved osteogenic differentiation of umbilical cord blood MSCs using custom made perfusion bioreactorBhaskar Birru0Naveen Kumar Mekala1Sreenivasa Rao Parcha2Department of Biotechnology, National Institute of Technology Warangal, TS, IndiaCollege of Medicine, Central Michigan University, Mt Pleasant, MI, USADepartment of Biotechnology, National Institute of Technology Warangal, TS, India; Corresponding author. Department of Biotechnology, National Institute of Technology Warangal, 506004, TS, India.Background: 3D cell culture is an appropriate method to develop engineered bone tissue, where different bioreactors have been designed to mitigate the challenges in 3D culture. Currently, we tailored a perfusion reactor to witness human mesenchymal stem cells (MSCs) proliferation and differentiation over polylactic acid-polyethylene glycol (PLA/PEG) composite scaffolds. Methods: The composite scaffolds with different weight ratios of PLA and PEG were prepared using solvent casting-particulate leaching technique. Human umbilcal card blood MSCs were cultured under dynamic and static conditions to elucidate the role of dynamic fluid flow in osteogenesis of MSCs. Results: The human MSCs distribution over the scaffolds was confirmed with fluorescent microscopy. Alkaline phosphatase (ALP), calcium mineralization, and collagen formation were found to be higher in PLA90 scaffolds than PLA100 and PLA75. PLA90 scaffolds with better cell adhesion/proliferartion were considered for bioreactor studies and they exhibited enhanced ALP, Ca+2 mineralization and collagen formation under dynamic perfusion than static culture. We further confirmed our observation by looking at expression levels of osteogenic marker (Runx2 and osteonectin) in differentiated MSCs subjected to perfusion culture compared to static culture. Conclusion: The results of the current investigation once again proves that dynamic perfusion cultures improve the osteogenic differentiation of MSCs over hybrid polymer scaffolds (PLA90) for effective bone regeneration. Keywords: 3D cell culture, Mesenchymal stem cells (MSCs), Fluid flow, Osteogenic differentiationhttp://www.sciencedirect.com/science/article/pii/S2319417017303499 |
| spellingShingle | Bhaskar Birru Naveen Kumar Mekala Sreenivasa Rao Parcha Improved osteogenic differentiation of umbilical cord blood MSCs using custom made perfusion bioreactor Biomedical Journal |
| title | Improved osteogenic differentiation of umbilical cord blood MSCs using custom made perfusion bioreactor |
| title_full | Improved osteogenic differentiation of umbilical cord blood MSCs using custom made perfusion bioreactor |
| title_fullStr | Improved osteogenic differentiation of umbilical cord blood MSCs using custom made perfusion bioreactor |
| title_full_unstemmed | Improved osteogenic differentiation of umbilical cord blood MSCs using custom made perfusion bioreactor |
| title_short | Improved osteogenic differentiation of umbilical cord blood MSCs using custom made perfusion bioreactor |
| title_sort | improved osteogenic differentiation of umbilical cord blood mscs using custom made perfusion bioreactor |
| url | http://www.sciencedirect.com/science/article/pii/S2319417017303499 |
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