Osteogenic differentiation of adipose-derived canine mesenchymal stem cells seeded in porous calcium-phosphate scaffolds
Por:
Herrera, David, Lodoso-Torrecilla I, Ginebra MP, Rappe, Katrin and Franch, Jordi
Publicada:
2 jun 2023
Categoría:
Veterinary (miscellaneous)
Resumen:
Introduction: Engineered bone graft substitutes are a promising
alternative and supplement to autologous bone grafts as treatments for
bone healing impairment. Advances in human medicine extend an invitation
to pursue these biomimetic strategies in animal patients, substantiated
by the theory that specialized scaffolds, multipotent cells, and
biological cues may be combined into a bioactive implant intended for
the enhancement of tissue regeneration.
Methods: This proof-of-concept study was designed to evaluate and
validate the feasibility of beta-tricalcium phosphate foam scaffolds
seeded with canine mesenchymal stem cells derived from adipose tissue.
Cell-inoculated samples and sham controls were cultured statically for
72 hours in complete growth medium to evaluate seeding capacity, while a
subset of loaded scaffolds was further induced with osteogenic culture
medium for 21 days. Produced implants were characterized and validated
with a combination of immunofluorescence and reflection confocal
microscopy, scanning electron microscopy, and polymerase chain reaction
to confirm osteogenic differentiation in tridimensional-induced samples.
Results: After 72 hours of culture, all inoculated scaffolds presented
widespread yet heterogeneous surface seeding, distinctively congregating
stem cells around pore openings. Furthermore, at 21 days of osteogenic
culture conditions, robust osteoblastic differentiation of the seeded
cells was confirmed by the change of cell morphology and evident
deposition of extra-cellular matrix, accompanied by mineralization and
scaffold remodeling; furthermore, all induced cell-loaded implants lost
specific stemness immunophenotype expression and simultaneously
upregulated genomic expression of osteogenic genes Osterix and
Ostecalcin.
Conclusions: ss-TCP bio-ceramic foam scaffolds proved to be suitable
carriers and hosts of canine adipose-derived MSCs, promoting not only
surface attachment and proliferation, but also demonstrating strong
in-vitro osteogenic potential. Although this research provides
satisfactory in-vitro validation for the conceptualization and
feasibility of a canine bio-active bone implant, further testing such as
patient safety, large-scale reproducibility, and quality assessment are
needed for regulatory compliance in future commercial clinical
applications.
Filiaciones:
Herrera, David:
Autonomous Univ Barcelona, Vet Fac, Dept Anim Med & Surg, Bone Regenerat Res Grp, Cerdanyola Del Valles, Spain
Lodoso-Torrecilla I:
Univ Politecn Cataluna, Dept Mat Sci & Engn, Biomat Biomech & Tissue Engn Grp, Barcelona, Spain
Ginebra MP:
Univ Politecn Cataluna, Dept Mat Sci & Engn, Biomat Biomech & Tissue Engn Grp, Barcelona, Spain
Rappe, Katrin:
Autonomous Univ Barcelona, Vet Fac, Dept Anim Med & Surg, Bone Regenerat Res Grp, Cerdanyola Del Valles, Spain
Franch, Jordi:
Autonomous Univ Barcelona, Vet Fac, Dept Anim Med & Surg, Bone Regenerat Res Grp, Cerdanyola Del Valles, Spain
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