Abstract
In this study, the in vitro biocompatibility and osteoinductive ability of a recently developed biomorphic hydroxylapatite ceramic scaffold (B-HA) derived from transformation of wood structures were analyzed using human adipose stem cells (hASCs). Cell viability and metabolic activity were evaluated in hASCs, parental cells and in recombinant genetically engineered hASC-eGFP cells expressing the green fluorescence protein. B-HA osteoinductivity properties, such as differentially expressed genes (DEG) involved in the skeletal development pathway, osteocalcin (OCN) protein expression and mineral matrix deposition in hASCs, were evaluated. In vitro induction of osteoblastic genes, such as Alkaline phosphatase (ALPL), Bone gamma-carboxyglutamate (gla) protein (BGLAP), SMAD family member 3 (SMAD3), Sp7 transcription factor (SP7) and Transforming growth factor, beta 3 (TGFB3) and Tumor necrosis factor (ligand) superfamily, member 11 (TNFSF11)/Receptor activator of NF-κB (RANK) ligand (RANKL), involved in osteoclast differentiation, was undertaken in cells grown on B-HA. Chondrogenic transcription factor SRY (sex determining region Y)-box 9 (SOX9), tested up-regulated in hASCs grown on the B-HA scaffold. Gene expression enhancement in the skeletal development pathway was detected in hASCs using B-HA compared to sintered hydroxylapatite (S-HA). OCN protein expression and calcium deposition were increased in hASCs grown on B-HA in comparison with the control. This study demonstrates the biocompatibility of the novel biomorphic B-HA scaffold and its potential use in osteogenic differentiation for hASCs. Our data highlight the relevance of B-HA for bone regeneration purposes.
Highlights
In recent times, the steady rise in musculoskeletal disease, resulting from increases in life expectancy and lifestyle changes, has become a serious threat for the ever-increasing population and national health systems
We show that the biomorphic scaffold (B-HA) modulated Epidermal growth factor (EGF), GLI family zinc finger 1 (GLI1), Matrix metallopeptidase 8 (MMP8)-10, Sp7 transcription factor (SP7) and TNFSF11 gene expression with higher fold change values compared to sintered hydroxylapatite (S-HA) material
In vitro tests carried out with hydroxylapatite (HA) porous scaffolds, obtained using a recently developed biomorphic transformation process, reported enhanced osteoinductive ability, attested by overexpression of various genes involved in the osteogenesis process after 2 weeks, in comparison with a scaffold made of HA with similar porosity, obtained using the classical sintering process
Summary
The steady rise in musculoskeletal disease, resulting from increases in life expectancy and lifestyle changes, has become a serious threat for the ever-increasing population and national health systems. 20 million patients/year have been reported as suffering from loss of bone tissue due to trauma or diseases [1]. New knowledge about bone repair mechanism is essential to address the important steps required in translational and precise medicine to improve patients healing. Self-repair is known to be challenging when there are massive bone defects due to traumatic injury, tumor resection or congenital disease [2]. Autologous cancellous bone grafting is considered the gold standard for regeneration when dealing with bone defects [3], despite several limitations and drawbacks being frequently encountered, including low bone availability and surgery with its consequent pain and risk of infection [4]. There is growing demand for synthetic bone scaffolds, which are capable of imitating the extracellular matrix and provide an appropriate microenvironment for bone growth by supporting and accelerating cell migration, while facilitating osteogenic differentiation and the regenerative cascade, overall [5,6,7]
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