Abstract
Tissue engineering (TE) and regenerative medicine offer strategies to improve damaged tissues by using scaffolds and cells. The use of collagen-based biomaterials in the field of TE has been intensively growing over the past decades. Mesenchymal stromal cells (MSCs) and dental pulp stem cells (DPSCs) are promising cell candidates for development of clinical composites. In this study, we proposed the development of a bovine collagen type I: chondroitin-6-sulphate (CG) scaffold, obtained from Uruguayan raw material (certified as free bovine spongiform encephalopathy), with CG crosslinking enhancement using different gamma radiation doses. Structural, biomechanical and chemical characteristics of the scaffolds were assessed by Scanning Electron Microscopy, axial tensile tests, FT-IR and Raman Spectroscopy, respectively. Once we selected the most appropriate scaffold for future use as a TE product, we studied the behavior of MSCs and DPSCs cultured on the scaffold by cytotoxicity, proliferation and differentiation assays. Among the diverse porous scaffolds obtained, the one with the most adequate properties was the one exposed to 15 kGy of gamma radiation. This radiation dose contributed to the crosslinking of molecules, to the formation of new bonds and/or to the reorganization of the collagen fibers. The selected scaffold was non-cytotoxic for the tested cells and a suitable substrate for cell proliferation. Furthermore, the scaffold allowed MSCs differentiation to osteogenic, chondrogenic, and adipogenic lineages. Thus, this work shows a promising approach to the synthesis of a collagen-scaffold suitable for TE.
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The authors confirm that the data supporting the findings of this study are available within the article, but if additional data is required are available from the corresponding author, Pardo, H., upon reasonable request.
Abbreviations
- TE:
-
Tissue engineering
- INDT:
-
Instituto Nacional de Donación y Trasplante
- MSCs:
-
Mesenchymal stromal cells
- DPSCs:
-
Dental pulp stem cells
- GAG:
-
Chondroitin-6-sulphate
- CG:
-
Collagen type I: chondroitin-6-sulphate
- Col 1:
-
Collagen type I
- TS:
-
Tensile strength
- %EX:
-
Percentage extension
- SEM:
-
Scanning electron microscopy
- No-GAG:
-
Scaffolds without GAG
- 0 kGy:
-
Scaffolds with GAG and non-irradiated
- 5 kGy, 15 kGy and 25 kGy:
-
Scaffolds with GAG irradiated at different radiation doses
- FT-IR:
-
Fourier transform infrared spectrometry analysis
- SD:
-
Standard deviation
- DPBS:
-
Dulbecco's phosphate-buffered saline
- FBS:
-
Fetal bovine serum
- DAPI:
-
4′,6-Diamidino-2-phenylindole
- CFSE:
-
Carboxyfluorescein succinimidyl ester dye
- PR:
-
Proliferation rate
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Acknowledgements
This research was supported by: IAEA Research Contract No.18282, CRP Code: E 31007; IAEA contribution with the National Project URU 6/036, Proyectos I+D 2014, Comisión Sectorial de Investigación Científica (CSIC), UDELAR (Universidad de la República), INDT-Secretary of Public Health, Agencia Nacional de Investigación e Innovación (ANII), Centro Argentino Brasileño de Biotecnología (CABBIO).
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We express our gratitude to the following researchers for their contribution to the experiments: Dr. PL, QFSC, MSR, Br. JPV, QFPM, LAC, Dr. PZ, LMDD, LNM, Dr. MB, LSR.
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Pereira, L., Echarte, L., Romero, M. et al. Synthesis and characterization of a bovine collagen: GAG scaffold with Uruguayan raw material for tissue engineering. Cell Tissue Bank 25, 123–142 (2024). https://doi.org/10.1007/s10561-021-09960-6
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DOI: https://doi.org/10.1007/s10561-021-09960-6