This study investigates the impact of hydroxyapatite (HA) nanoparticles (NPs) on the cellular responses of poly(L-lactide-co-ε-caprolactone) (PLCL) scaffolds in bone tissue engineering applications. Three types of PLCL scaffolds were fabricated, varying in HANPs content. Saos-2 osteoblast-like cells (OBs) and THP-1-derived osteoclast-like cells (OCs) were co-cultured on the scaffolds, and cell proliferation was assessed using the MTS assay. The amount of double-stranded DNA (dsDNA) was quantified to evaluate cell proliferation. Expression levels of OBs and OCs markers were analyzed via quantitative polymerase chain reaction (qPCR) and the production of Collagen type I was visualized using confocal microscopy. Additionally, enzymatic activity of alkaline phosphatase (ALP) and tartrate-resistant acid phosphatase (TRAP or ACP5) was measured to assess OB and OC function, respectively. Interestingly, despite the scaffold's structured character supporting the growth of the Saos-2 OBs and THP-1-derived OCs coculture, the incorporation of HANPs did not significantly enhance cellular responses compared to scaffolds without HANPs, except for collagen type I production. These findings suggest the need for further investigation into the potential benefits of HANPs in bone tissue engineering applications. Nevertheless, our study contributes valuable insights into optimizing biomaterial design for bone tissue regeneration, with implications for drug screening and material testing protocols.

• Klíčová slova
• PLCL, bone regeneration, hydroxyapatite, osteoblasts, osteoclasts, scaffold, tissue engineering,
• MeSH
• buněčné linie MeSH
• hydroxyapatit * chemie   MeSH
• kokultivační techniky MeSH
• lidé MeSH
• nanočástice * chemie   MeSH
• nanovlákna * chemie   MeSH
• osteoblasty cytologie   metabolismus   účinky léků   MeSH
• osteogeneze MeSH
• osteoklasty cytologie   metabolismus   účinky léků   MeSH
• polyestery * chemie   MeSH
• proliferace buněk účinky léků   MeSH
• regenerace kostí * účinky léků   MeSH
• THP-1 buňky MeSH
• tkáňové inženýrství metody   MeSH
• tkáňové podpůrné struktury chemie   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• hydroxyapatit * MeSH
• poly(lactic acid-co-epsilon-caprolactone) MeSH Prohlížeč
• polyestery * MeSH

Human dental pulp stem cells (hDPSCs) are mesenchymal stem cells that have been successfully used in human bone tissue engineering. To establish whether these cells can lead to a bone tissue ready to be grafted, we checked DPSCs for their osteogenic and angiogenic differentiation capabilities with the specific aim of obtaining a new tool for bone transplantation. Therefore, hDPSCs were specifically selected from the stromal-vascular dental pulp fraction, using appropriate markers, and cultured. Growth curves, expression of bone-related markers, calcification and angiogenesis as well as an in vivo transplantation assay were performed. We found that hDPSCs proliferate, differentiate into osteoblasts and express high levels of angiogenic genes, such as vascular endothelial growth factor and platelet-derived growth factor A. Human DPSCs, after 40 days of culture, give rise to a 3D structure resembling a woven fibrous bone. These woven bone (WB) samples were analysed using classic histology and synchrotron-based, X-ray phase-contrast microtomography and holotomography. WB showed histological and attractive physical qualities of bone with few areas of mineralization and neovessels. Such WB, when transplanted into rats, was remodelled into vascularized bone tissue. Taken together, our data lead to the assumption that WB samples, fabricated by DPSCs, constitute a noteworthy tool and do not need the use of scaffolds, and therefore they are ready for customized regeneration.

• Klíčová slova
• bone differentiation, bone regeneration, bone tissue engineering, hDPSCs, holotomography, human Dental Pulp Stem Cells, human serum, phc-microCT, woven bon, woven bone,
• MeSH
• buněčná diferenciace fyziologie   MeSH
• chemotaxe MeSH
• dospělí MeSH
• fyziologická kalcifikace fyziologie   MeSH
• fyziologická neovaskularizace fyziologie   MeSH
• kmenové buňky cytologie   MeSH
• kostní náhrady * MeSH
• kultivované buňky MeSH
• lidé MeSH
• mladý dospělý MeSH
• myši nahé MeSH
• osteogeneze fyziologie   MeSH
• osteokalcin metabolismus   MeSH
• proliferace buněk MeSH
• rentgenová mikrotomografie metody   MeSH
• separace buněk metody   MeSH
• tkáňové inženýrství metody   MeSH
• transplantace kostí metody   MeSH
• zubní dřeň cytologie   MeSH
• zvířata MeSH
• Check Tag
• dospělí MeSH
• lidé MeSH
• mladý dospělý MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• hodnotící studie MeSH
• Názvy látek
• kostní náhrady * MeSH
• osteokalcin MeSH