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In this study, we aimed to compare how the microstructure and architecture of polymer supports influence adhesion, growth and differentiation of human mesenchymal stem cells (hMSC) in the context of bone tissue engineering. We manufactured poly(L-lactide-co-glycolide) (PLGA) three-dimensional supports in the form of microspheres by emulsification and porous scaffolds by solvent casting/ porogen leaching. HMSC were seeded on both materials and on control tissue culture polystyrene (TCPS, bottom of the wells) and cultured in basal or osteogenic medium for 1, 3, 7 and 14 days. HMSC proliferation and osteogenic differentiation were studied using lactate dehydrogenase (LDH) and alkaline phosphatase (ALP) assays, respectively. Furthermore, cell morphology and viability were analyzed after live/dead fluorescence staining. The results show that the optimized emulsification conditions allowed the production of PLGA microspheres with a median size of 95 µm. The PLGA scaffolds had a porosity of 82.1% ± 4.2% and a pore size of 360 µm ± 74 µm. HMSC cultured on control TCPS in osteogenic medium were more spread and polygonal than those in basal medium. They were characterized with a lower proliferation rate, as shown by the LDH results, but higher ALP activity. This suggests that hMSC osteogenic differentiation was achieved. The same tendency was observed for cells cultured on microspheres and scaffolds. Cell proliferation was more efficient on both materials and control in growth medium as compared to differentiation medium. The amount of ALP, i.e. a marker of osteogenic differentiation, was elevated, as expected, in differentiation medium. However, on day 14 cells cultured on the scaffolds in basal medium exhibited the same osteogenic potential as those cultured in differentiation medium. In general, both microspheres and scaffolds promoted hMSC adhesion, proliferation, and osteogenic differentiation and may be used for bone tissue engineering.
Czasopismo
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Tom
Strony
14--19
Opis fizyczny
Bibliogr. 15 poz., wykr., zdj.
Twórcy
autor
- AGH University of Krakow, Faculty of Materials Science and Ceramics, Department of Biomaterials and Composites, al. A. Mickiewicza 30, 30-059 Krakow, Poland
- AGH University of Krakow, Faculty of Materials Science and Ceramics, Department of Biomaterials and Composites, al. A. Mickiewicza 30, 30-059 Krakow, Poland
autor
- AGH University of Krakow, Faculty of Materials Science and Ceramics, Department of Biomaterials and Composites, al. A. Mickiewicza 30, 30-059 Krakow, Poland
Bibliografia
- [1] Ansari M.: Bone tissue regeneration: biology, strategies and interface studies. Progress in Biomaterials 8 (2019) 223-237.
- [2] Zhu G., Zhang T., Chen M., Yao K., Huang X., Zhang B., Li Y., Liu J., Wang Y., Zhao Z.: Bone physiological microenvironment and healing mechanism: Basis for future bone-tissue engineering scaffolds. Bioactive Materials 6 (2021) 4110-4140.
- [3] Henkel J., Woodruff M., Epari D., Steck R., Glatt V., Dickinson I., Choong P., Schuetz M., Hutmacher D.: Bone regeneration based on tissue engineering conceptions - a 21st century perspective. Bone Research 3 (2013) 216-248.
- [4] Asadi N., Del Bakhshayesh A., Davaran S., Akbarzadeh A.: Common biocompatible polymeric materials for tissue engineering and regenerative medicine. Materials Chemistry and Physics 242 (2020) 122528.
- [5] Kumar P., Dehiya B., Sindhu A.: Bioceramics for hard tissue engineering applications. International Journal for Applied Engineering Research 13 (2018) 2744-2752.
- [6] Turnbull G., Clarke J., Picard F., Riches P., Jia L., Han F., Li B., Shu W.: 3D bioactive composite scaffolds for bone tissue engineering. Bioactive Materials 3 (2018) 278-314.
- [7] Chan B., Leong K.: Scaffolding in tissue engineering: general approaches and tissue-specific considerations. European Spine Journal 17 (2008) 467-479.
- [8] Rumian Ł., Wojak I., Scharnweber D., Pamuła E.: Resorbable scaffolds modified with collagen type I or hydroxyapatite: in vitro studies on human mesenchymal stem cells. Acta of Bioengineering and Biomechanics 15 (2013) 61-67.
- [9] Gupta V., Khan Y., Berkland C., Laurencin C., Detamore M.: Microsphere-based scaffolds in regenerative engineering. Annual Review of Biomedical Engineering 19 (2017) 135-161.
- [10] Mielan B., Pamuła E.: Optimizing manufacturing conditions of polymer microspheres as cell carriers for modular tissue engineering. Engineering of Biomaterials 155 (2020) 2-9.
- [11] Liu S., Cai M., Deng R., Wang J., Liang R., Zhu J.: fabrication of porous polymer microparticles with tunable pore size and density through the combination of phase separation and emulsion-solvent evaporation approach. Korea-Australia Rheology Journal 26 (2014) 63-71.
- [12] Mielan B., Sousa D., Krok-Borkowicz M., Eloy P., Dupont C., Lamghari M., Pamuła E.: Polymeric microspheres/cells/extracellular matrix constructs produced by auto-assembly for bone modular tissue engineering. international Journal of Molecular Science 22 (2021) 7897.
- [13] Rougerie P., Silva dos Santos R., Farina M., Anselme K.: Molecular mechanisms of topography sensing by osteoblasts: An update. Applied Sciences 11 (2021) 1791.
- [14] Werner M., Blanquer S.B.G., Haimi S.P., Korus G., Dunlop J.W.C., Duda G.N., Grijpma D.W., Petersen A.: Surface curvature differentially regulates stem cell migration and differentiation via altered attachment morphology and nuclear deformation. Advanced Science News 4 (2017) 1600347.
- [15] Rutledge K. E., Cheng Q., Jabbarzadeh E.: Modulation of inflammatory response and induction of bone formation based on combinatorial effects of resveratrol. Journal of Nanomedicine and Nanotechnology 7 (2016) 350.
Typ dokumentu
Bibliografia
Identyfikator YADDA
bwmeta1.element.baztech-89fff05f-2030-47d6-80a5-503bf19ae339