Osteoregenerative Potential of 3D-Printed Poly <i>ε</i>-Caprolactone Tissue Scaffolds In Vitro Using Minimally Manipulative Expansion of Primary Human Bone Marrow Stem Cells
The repair of orthopedic and maxillofacial defects in modern medicine currently relies heavily on the use of autograft, allograft, void fillers, or other structural material composites. This study examines the in vitro osteo regenerative potential of polycaprolactone (PCL) tissue scaffolding, fabric...
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| Format: | Article |
| Language: | English |
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MDPI AG
2023-03-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/24/5/4940 |
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| author | Logan M. Lawrence Roozbeh (Ross) Salary Virginia Miller Anisha Valluri Krista L. Denning Shannon Case-Perry Karim Abdelgaber Shannon Smith Pier Paolo Claudio James B. Day |
| author_facet | Logan M. Lawrence Roozbeh (Ross) Salary Virginia Miller Anisha Valluri Krista L. Denning Shannon Case-Perry Karim Abdelgaber Shannon Smith Pier Paolo Claudio James B. Day |
| author_sort | Logan M. Lawrence |
| collection | DOAJ |
| description | The repair of orthopedic and maxillofacial defects in modern medicine currently relies heavily on the use of autograft, allograft, void fillers, or other structural material composites. This study examines the in vitro osteo regenerative potential of polycaprolactone (PCL) tissue scaffolding, fabricated via a three-dimensional (3D) additive manufacturing technology, i.e., a pneumatic micro extrusion (PME) process. The objectives of this study were: (i) To examine the innate osteoinductive and osteoconductive potential of 3D-printed PCL tissue scaffolding and (ii) To perform a direct in vitro comparison of 3D-printed PCL scaffolding with allograft Allowash<sup>®</sup> cancellous bone cubes with regards to cell-scaffold interactions and biocompatibility with three primary human bone marrow (hBM) stem cell lines. This study specifically examined cell survival, cell integration, intra-scaffold cell proliferation, and differentiation of progenitor cells to investigate the potential of 3D-printed PCL scaffolds as an alternative to allograft bone material for the repair of orthopedic injuries. We found that mechanically robust PCL bone scaffolds can be fabricated via the PME process and the resulting material did not elicit detectable cytotoxicity. When the widely used osteogenic model SAOS-2 was cultured in PCL extract medium, no detectable effect was observed on cell viability or proliferation with multiple test groups showing viability ranges of 92.2% to 100% relative to a control group with a standard deviation of ±10%. In addition, we found that the honeycomb infill pattern of the 3D-printed PCL scaffold allowed for superior mesenchymal stem-cell integration, proliferation, and biomass increase. When healthy and active primary hBM cell lines, having documented in vitro growth rates with doubling times of 23.9, 24.67, and 30.94 h, were cultured directly into 3D-printed PCL scaffolds, impressive biomass increase values were observed. It was found that the PCL scaffolding material allowed for biomass increase values of 17.17%, 17.14%, and 18.18%, compared to values of 4.29% for allograph material cultured under identical parameters. It was also found that the honeycomb scaffold infill pattern was superior to the cubic and rectangular matrix structures, and provided a superior microenvironment for osteogenic and hematopoietic progenitor cell activity and auto-differentiation of primary hBM stem cells. Histological and immunohistochemical studies performed in this work confirmed the regenerative potential of PCL matrices in the orthopedic setting by displaying the integration, self-organization, and auto-differentiation of hBM progenitor cells within the matrix. Differentiation products including mineralization, self-organizing “proto-osteon” structures, and in vitro erythropoiesis were observed in conjunction with the documented expression of expected bone marrow differentiative markers including CD-99 (>70%), CD-71 (>60%), and CD-61 (>5%). All of the studies were conducted without the addition of any exogenous chemical or hormonal stimulation and exclusively utilized the abiotic and inert material polycaprolactone; setting this work apart from the vast majority of contemporary investigations into synthetic bone scaffold fabrication In summary, this study demonstrates the unique clinical potential of 3D-printed PCL scaffolds for stem cell expansion and incorporation into advanced microstructures created via PME manufacturing to generate a physiologically inert temporary bony defect graft with significant autograft features for enhanced end-stage healing. |
| first_indexed | 2024-03-11T07:21:17Z |
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| institution | Directory Open Access Journal |
| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-11T07:21:17Z |
| publishDate | 2023-03-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | International Journal of Molecular Sciences |
| spelling | doaj.art-205d33ce019048c892f1549dbbfbd6242023-11-17T07:55:34ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672023-03-01245494010.3390/ijms24054940Osteoregenerative Potential of 3D-Printed Poly <i>ε</i>-Caprolactone Tissue Scaffolds In Vitro Using Minimally Manipulative Expansion of Primary Human Bone Marrow Stem CellsLogan M. Lawrence0Roozbeh (Ross) Salary1Virginia Miller2Anisha Valluri3Krista L. Denning4Shannon Case-Perry5Karim Abdelgaber6Shannon Smith7Pier Paolo Claudio8James B. Day9Department of Pathology, Joan C. Edwards School of Medicine, Cabell Huntington Hospital Laboratory, Marshall University, Huntington, WV 25701, USADepartment of Mechanical Engineering, Marshall University, Huntington, WV 25703, USADepartment of Pathology, Joan C. Edwards School of Medicine, Cabell Huntington Hospital Laboratory, Marshall University, Huntington, WV 25701, USAJoan C. Edwards School of Medicine, Marshall University, Huntington, WV 25701, USADepartment of Pathology, Joan C. Edwards School of Medicine, Cabell Huntington Hospital Laboratory, Marshall University, Huntington, WV 25701, USACabell Huntington Hospital Laboratory, Department of Histology, Mountain Health Network, Huntington, WV 25701, USAJoan C. Edwards School of Medicine, Marshall University, Huntington, WV 25701, USAJoan C. Edwards School of Medicine, Marshall University, Huntington, WV 25701, USADepartment of Pharmacology and Toxicology, University of Mississippi Medical Center, Jackson, MS 39216, USADepartment of Orthopaedic Surgery, Joan C. Edwards School of Medicine, Marshall University, Huntington, WV 25701, USAThe repair of orthopedic and maxillofacial defects in modern medicine currently relies heavily on the use of autograft, allograft, void fillers, or other structural material composites. This study examines the in vitro osteo regenerative potential of polycaprolactone (PCL) tissue scaffolding, fabricated via a three-dimensional (3D) additive manufacturing technology, i.e., a pneumatic micro extrusion (PME) process. The objectives of this study were: (i) To examine the innate osteoinductive and osteoconductive potential of 3D-printed PCL tissue scaffolding and (ii) To perform a direct in vitro comparison of 3D-printed PCL scaffolding with allograft Allowash<sup>®</sup> cancellous bone cubes with regards to cell-scaffold interactions and biocompatibility with three primary human bone marrow (hBM) stem cell lines. This study specifically examined cell survival, cell integration, intra-scaffold cell proliferation, and differentiation of progenitor cells to investigate the potential of 3D-printed PCL scaffolds as an alternative to allograft bone material for the repair of orthopedic injuries. We found that mechanically robust PCL bone scaffolds can be fabricated via the PME process and the resulting material did not elicit detectable cytotoxicity. When the widely used osteogenic model SAOS-2 was cultured in PCL extract medium, no detectable effect was observed on cell viability or proliferation with multiple test groups showing viability ranges of 92.2% to 100% relative to a control group with a standard deviation of ±10%. In addition, we found that the honeycomb infill pattern of the 3D-printed PCL scaffold allowed for superior mesenchymal stem-cell integration, proliferation, and biomass increase. When healthy and active primary hBM cell lines, having documented in vitro growth rates with doubling times of 23.9, 24.67, and 30.94 h, were cultured directly into 3D-printed PCL scaffolds, impressive biomass increase values were observed. It was found that the PCL scaffolding material allowed for biomass increase values of 17.17%, 17.14%, and 18.18%, compared to values of 4.29% for allograph material cultured under identical parameters. It was also found that the honeycomb scaffold infill pattern was superior to the cubic and rectangular matrix structures, and provided a superior microenvironment for osteogenic and hematopoietic progenitor cell activity and auto-differentiation of primary hBM stem cells. Histological and immunohistochemical studies performed in this work confirmed the regenerative potential of PCL matrices in the orthopedic setting by displaying the integration, self-organization, and auto-differentiation of hBM progenitor cells within the matrix. Differentiation products including mineralization, self-organizing “proto-osteon” structures, and in vitro erythropoiesis were observed in conjunction with the documented expression of expected bone marrow differentiative markers including CD-99 (>70%), CD-71 (>60%), and CD-61 (>5%). All of the studies were conducted without the addition of any exogenous chemical or hormonal stimulation and exclusively utilized the abiotic and inert material polycaprolactone; setting this work apart from the vast majority of contemporary investigations into synthetic bone scaffold fabrication In summary, this study demonstrates the unique clinical potential of 3D-printed PCL scaffolds for stem cell expansion and incorporation into advanced microstructures created via PME manufacturing to generate a physiologically inert temporary bony defect graft with significant autograft features for enhanced end-stage healing.https://www.mdpi.com/1422-0067/24/5/49403D microfabricationautograftallograftbone graftbone marrowbone regeneration |
| spellingShingle | Logan M. Lawrence Roozbeh (Ross) Salary Virginia Miller Anisha Valluri Krista L. Denning Shannon Case-Perry Karim Abdelgaber Shannon Smith Pier Paolo Claudio James B. Day Osteoregenerative Potential of 3D-Printed Poly <i>ε</i>-Caprolactone Tissue Scaffolds In Vitro Using Minimally Manipulative Expansion of Primary Human Bone Marrow Stem Cells International Journal of Molecular Sciences 3D microfabrication autograft allograft bone graft bone marrow bone regeneration |
| title | Osteoregenerative Potential of 3D-Printed Poly <i>ε</i>-Caprolactone Tissue Scaffolds In Vitro Using Minimally Manipulative Expansion of Primary Human Bone Marrow Stem Cells |
| title_full | Osteoregenerative Potential of 3D-Printed Poly <i>ε</i>-Caprolactone Tissue Scaffolds In Vitro Using Minimally Manipulative Expansion of Primary Human Bone Marrow Stem Cells |
| title_fullStr | Osteoregenerative Potential of 3D-Printed Poly <i>ε</i>-Caprolactone Tissue Scaffolds In Vitro Using Minimally Manipulative Expansion of Primary Human Bone Marrow Stem Cells |
| title_full_unstemmed | Osteoregenerative Potential of 3D-Printed Poly <i>ε</i>-Caprolactone Tissue Scaffolds In Vitro Using Minimally Manipulative Expansion of Primary Human Bone Marrow Stem Cells |
| title_short | Osteoregenerative Potential of 3D-Printed Poly <i>ε</i>-Caprolactone Tissue Scaffolds In Vitro Using Minimally Manipulative Expansion of Primary Human Bone Marrow Stem Cells |
| title_sort | osteoregenerative potential of 3d printed poly i ε i caprolactone tissue scaffolds in vitro using minimally manipulative expansion of primary human bone marrow stem cells |
| topic | 3D microfabrication autograft allograft bone graft bone marrow bone regeneration |
| url | https://www.mdpi.com/1422-0067/24/5/4940 |
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