Osteoregenerative Potential of 3D-Printed Poly ε-Caprolactone Tissue Scaffolds In Vitro Using Minimally Manipulative Expansion of Primary Human Bone Marrow Stem Cells
Abstract
:1. Introduction
1.1. Review of Literature
1.1.1. Human Bone Regeneration
1.1.2. 3D-Microfabrication
2. Results
2.1. Polycaprolactone (PCL) 3D-Printed Scaffold Fabrication
2.2. Human Bone Marrow (hBM) Sample Collection
2.3. Primary Whole Bone Marrow Progenitor Cell Line Isolation and In Vitro Expansion
2.4. Screening of Biocompatibility of Fabricated Polycaprolactone (PCL) Scaffolds via Cytotoxic and Cytostatic Analysis of PCL Extract Exposure and Histological Analysis of Established Osteosarcoma Cell Line SAOS-2
2.5. Investigation of Varying 3D-Printed PCL Scaffold Matrix Patterns on Cell Integration and Resulting Scaffold Biomass Increase In Vitro with Primary Human Bone Marrow Cell Culture as Compared to Standard-of-Care Allograft Cancellous Bone Cubes
2.6. Histologic Analysis of Human Bone Marrow (hBM) Stem Cell Viability, Integration, and Differentiation in 3D-Printed PCL Honeycomb Scaffold
3. Discussion
4. Materials and Methods
4.1. Human Bone Marrow (hBM) Sample Collection
4.2. Patient Samples of Primary Bone Marrow (hBM) Stem Cell Colonies
4.3. Cell Lines and Growth Medium
4.4. Cytotoxic and Cytostatic Analysis of PCL Extract on Established Osteosarcoma Cell Line SAOS-2 via MTT Analysis
4.5. hBM Stem-Cell Culture and Colony Expansion
4.6. Cell Counting
4.7. Scaffold Inoculation and Rotating Shaker Culture
4.8. PCL Scaffold Microfabrication and Specifications
4.9. In Vitro Biomass Increase
4.10. Histologic Processing: H&E and Immunohistochemical Evaluation of Cell Viability, Cell Integration, and Cell Differentiation of Primary (hBMs) in Fertilized Scaffolds
4.11. Sterilization of 3D-Printed PCL Scaffolds and Allowash® Cancellous Cubes
4.12. Statistical Analysis
5. Conclusions
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
Abbreviations
MTT | 4, 5-dimethylthiazol-2-yl]-2, 5 diphenyl tetrazolium bromide |
BVF | Bone Void Fillers |
Cub | Cubic |
DBM | Decellularized Bone Material |
DIW | Direct Ink Writing |
FFPE | Formalin Fixation and Paraffin Embedding |
FDM | Fused Deposition Modeling |
H&E | Hematoxylin & Eosin |
HC | Honeycomb |
hBM | Human Bone Marrow |
IC | Immunohistochemistry |
ICBG | Iliac Crest Bone Graft |
LGDW | Laser-Guided Direct Writing |
MSCs | Mesenchymal Stem Cells |
PME | Pneumatic Micro-Extrusion |
PCL | Polycaprolactone |
RL | Rectilinear |
3D | Three-Dimensional |
SLA | Stereolithography |
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Lawrence, L.M.; Salary, R.; Miller, V.; Valluri, A.; Denning, K.L.; Case-Perry, S.; Abdelgaber, K.; Smith, S.; Claudio, P.P.; Day, J.B. Osteoregenerative Potential of 3D-Printed Poly ε-Caprolactone Tissue Scaffolds In Vitro Using Minimally Manipulative Expansion of Primary Human Bone Marrow Stem Cells. Int. J. Mol. Sci. 2023, 24, 4940. https://doi.org/10.3390/ijms24054940
Lawrence LM, Salary R, Miller V, Valluri A, Denning KL, Case-Perry S, Abdelgaber K, Smith S, Claudio PP, Day JB. Osteoregenerative Potential of 3D-Printed Poly ε-Caprolactone Tissue Scaffolds In Vitro Using Minimally Manipulative Expansion of Primary Human Bone Marrow Stem Cells. International Journal of Molecular Sciences. 2023; 24(5):4940. https://doi.org/10.3390/ijms24054940
Chicago/Turabian StyleLawrence, Logan M., Roozbeh (Ross) Salary, Virginia Miller, Anisha Valluri, Krista L. Denning, Shannon Case-Perry, Karim Abdelgaber, Shannon Smith, Pier Paolo Claudio, and James B. Day. 2023. "Osteoregenerative Potential of 3D-Printed Poly ε-Caprolactone Tissue Scaffolds In Vitro Using Minimally Manipulative Expansion of Primary Human Bone Marrow Stem Cells" International Journal of Molecular Sciences 24, no. 5: 4940. https://doi.org/10.3390/ijms24054940