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Article

In Vitro Biocompatibility Assessment of Bioengineered PLA-Hydrogel Core–Shell Scaffolds with Mesenchymal Stromal Cells for Bone Regeneration

1
Unit of Blood Diseases and Cell Therapies, Department of Clinical and Experimental Sciences, University of Brescia, “ASST-Spedali Civili” Hospital of Brescia, 25123 Brescia, Italy
2
Centro di Ricerca Emato-Oncologica AIL (CREA), ASST Spedali Civili, 25123 Brescia, Italy
3
University Center of Research “Stem Cells, bioENgineering and Regenerative MEDicine”—STENMED, University of Brescia, 25123 Brescia, Italy
4
Materials Science and Technology Laboratory, Department of Mechanical and Industrial Engineering, University of Brescia, 25123 Brescia, Italy
5
Department of Civil, Environmental, Architectural Engineering and Mathematics (DICATAM), University of Brescia, Via Valotti 9, 25123 Brescia, Italy
6
National Research Council (CNR)—Institute for Microelectronics and Microsystems, Via Gobetti 101, 40129 Bologna, Italy
7
Division of Anatomy and Physiopathology, Department of Clinical and Experimental Sciences, University of Brescia, 25123 Brescia, Italy
8
Interdepartmental University Center of Research “Adaption and Regeneration of Tissues and Organs (ARTO)”, University of Brescia, 25123 Brescia, Italy
9
Laboratory for Stem Cells Manipulation and Cryopreservation, Department of Transfusion Medicine, ASST Spedali Civili di Brescia, 25123 Brescia, Italy
10
Department of Molecular and Translational Medicine, University of Brescia, 25123 Brescia, Italy
11
Highly Specialized Laboratory, ASST Spedali Civili di Brescia, 25123 Brescia, Italy
12
National Center for Gene Therapy and Drugs based on RNA Technology—CN3, 35122 Padua, Italy
*
Author to whom correspondence should be addressed.
J. Funct. Biomater. 2024, 15(8), 217; https://doi.org/10.3390/jfb15080217 (registering DOI)
Submission received: 28 June 2024 / Revised: 22 July 2024 / Accepted: 25 July 2024 / Published: 31 July 2024
(This article belongs to the Special Issue Feature Papers in Bone Biomaterials)

Abstract

Human mesenchymal stromal cells (hMSCs), whether used alone or together with three-dimensional scaffolds, are the best-studied postnatal stem cells in regenerative medicine. In this study, innovative composite scaffolds consisting of a core–shell architecture were seeded with bone-marrow-derived hMSCs (BM-hMSCs) and tested for their biocompatibility and remarkable capacity to promote and support bone regeneration and mineralization. The scaffolds were prepared by grafting three different amounts of gelatin–chitosan (CH) hydrogel into a 3D-printed polylactic acid (PLA) core (PLA-CH), and the mechanical and degradation properties were analyzed. The BM-hMSCs were cultured in the scaffolds with the presence of growth medium (GM) or osteogenic medium (OM) with differentiation stimuli in combination with fetal bovine serum (FBS) or human platelet lysate (hPL). The primary objective was to determine the viability, proliferation, morphology, and spreading capacity of BM-hMSCs within the scaffolds, thereby confirming their biocompatibility. Secondly, the BM-hMSCs were shown to differentiate into osteoblasts and to facilitate scaffold mineralization. This was evinced by a positive Von Kossa result, the modulation of differentiation markers (osteocalcin and osteopontin), an expression of a marker of extracellular matrix remodeling (bone morphogenetic protein-2), and collagen I. The results of the energy-dispersive X-ray analysis (EDS) clearly demonstrate the presence of calcium and phosphorus in the samples that were incubated in OM, in the presence of FBS and hPL, but not in GM. The chemical distribution maps of calcium and phosphorus indicate that these elements are co-localized in the same areas of the sections, demonstrating the formation of hydroxyapatite. In conclusion, our findings show that the combination of BM-hMSCs and PLA-CH, regardless of the amount of hydrogel content, in the presence of differentiation stimuli, can provide a construct with enhanced osteogenicity for clinically relevant bone regeneration.
Keywords: scaffold design; PLA; human mesenchymal stromal cells; gelatin–chitosan hydrogel; human platelet lysate; bone regeneration; tissue engineering; 3D printing scaffold design; PLA; human mesenchymal stromal cells; gelatin–chitosan hydrogel; human platelet lysate; bone regeneration; tissue engineering; 3D printing

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MDPI and ACS Style

Re, F.; Sartore, L.; Pasini, C.; Ferroni, M.; Borsani, E.; Pandini, S.; Bianchetti, A.; Almici, C.; Giugno, L.; Bresciani, R.; et al. In Vitro Biocompatibility Assessment of Bioengineered PLA-Hydrogel Core–Shell Scaffolds with Mesenchymal Stromal Cells for Bone Regeneration. J. Funct. Biomater. 2024, 15, 217. https://doi.org/10.3390/jfb15080217

AMA Style

Re F, Sartore L, Pasini C, Ferroni M, Borsani E, Pandini S, Bianchetti A, Almici C, Giugno L, Bresciani R, et al. In Vitro Biocompatibility Assessment of Bioengineered PLA-Hydrogel Core–Shell Scaffolds with Mesenchymal Stromal Cells for Bone Regeneration. Journal of Functional Biomaterials. 2024; 15(8):217. https://doi.org/10.3390/jfb15080217

Chicago/Turabian Style

Re, Federica, Luciana Sartore, Chiara Pasini, Matteo Ferroni, Elisa Borsani, Stefano Pandini, Andrea Bianchetti, Camillo Almici, Lorena Giugno, Roberto Bresciani, and et al. 2024. "In Vitro Biocompatibility Assessment of Bioengineered PLA-Hydrogel Core–Shell Scaffolds with Mesenchymal Stromal Cells for Bone Regeneration" Journal of Functional Biomaterials 15, no. 8: 217. https://doi.org/10.3390/jfb15080217

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