Chondrogenic Commitment of Human Bone Marrow Mesenchymal Stem Cells in a Perfused Collagen Hydrogel Functionalized with hTGF-β1-Releasing PLGA Microcarrier
Abstract
:1. Introduction
2. Materials and Methods
2.1. hBM-MSCs Isolation and Harvesting
2.2. PLGA-MCs Fabrication by Supercritical Emulsion Extraction Technology
2.3. Carrier Size Distribution and Morphological Analyses
2.4. hTGF-β1 Release Study
2.5. 3D Collagen Scaffolds Preparation and Characterization
2.5.1. Static and Dynamic Culture
2.5.2. Hematoxylin and Eosin and Sirius Red Staining
2.5.3. Immunofluorescence Assay
2.6. RNA Isolation and Gene Expression Profile
2.7. FEM Modeling
2.8. Statistical Analysis
3. Result and Discussion
3.1. PLGA Carriers Characterization and hTGF-β1 Release Profile
3.2. 3D Scaffold Environment Assembly and Characterization
3.3. Dynamic Culture by Perfusion Bioreactor
3.4. 3D Microenvironment and hBM-MSC Chondrogenic Commitment: Gene Expression and Immunofluorescence Assay
4. Conclusions and Perspectives
5. Patents
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
Abbreviations
hTGF-β1 | Human transforming growth factor β1 |
PLGA-MCs | Poly-lactic-co-glycolic acid microcarriers |
hBM-MSCs | Human bone marrow mesenchymal stem cells |
hASC | Human adipose stem cells |
COL1A1 | Type I collagen |
COL2A1 | Type II collagen |
COL3A1 | Type III collagen |
SOX9 | SRY-Box transcription factor 9 |
ACAN | Aggrecan |
RT-qPCR | Reverse transcription quantitative polymerase chain reaction |
q-IF | Semiquantitative immunofluorescence |
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FEM Modeling Parameters | Values | Units |
---|---|---|
Glucose at time zero | 0 | mol/m3 |
Waste at time zero | 0 | mol/m3 |
Nutrient consumption | 4.74·10−12 | mol/s |
Waste production | 3.4·10−12 | mol/s |
D nutrients coefficient | 3.9·10−9 | m2/s |
D waste coefficient | 1.6·10−9 | m2/s |
κ collagen hydrogel | 2·10−9 | m2 |
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Lamparelli, E.P.; Lovecchio, J.; Ciardulli, M.C.; Giudice, V.; Dale, T.P.; Selleri, C.; Forsyth, N.; Giordano, E.; Maffulli, N.; Della Porta, G. Chondrogenic Commitment of Human Bone Marrow Mesenchymal Stem Cells in a Perfused Collagen Hydrogel Functionalized with hTGF-β1-Releasing PLGA Microcarrier. Pharmaceutics 2021, 13, 399. https://doi.org/10.3390/pharmaceutics13030399
Lamparelli EP, Lovecchio J, Ciardulli MC, Giudice V, Dale TP, Selleri C, Forsyth N, Giordano E, Maffulli N, Della Porta G. Chondrogenic Commitment of Human Bone Marrow Mesenchymal Stem Cells in a Perfused Collagen Hydrogel Functionalized with hTGF-β1-Releasing PLGA Microcarrier. Pharmaceutics. 2021; 13(3):399. https://doi.org/10.3390/pharmaceutics13030399
Chicago/Turabian StyleLamparelli, Erwin Pavel, Joseph Lovecchio, Maria Camilla Ciardulli, Valentina Giudice, Tina P. Dale, Carmine Selleri, Nicholas Forsyth, Emanuele Giordano, Nicola Maffulli, and Giovanna Della Porta. 2021. "Chondrogenic Commitment of Human Bone Marrow Mesenchymal Stem Cells in a Perfused Collagen Hydrogel Functionalized with hTGF-β1-Releasing PLGA Microcarrier" Pharmaceutics 13, no. 3: 399. https://doi.org/10.3390/pharmaceutics13030399