Surface modified biphasic calcium phosphate-particles enhance expression of bone markers in osteoblast-like cells
(English)Manuscript (preprint) (Other academic)
Objectives: The aim of the present in vitro study was to evaluate the response of human osteoblast-like cells (HOB) to nano-crystalline-diamond-particle-modified (nDP-modified) and un-modified (control) deproteinized bovine bone (DBB) and biphasic calcium phosphate (BCP) scaffolds.
Materials and methods: nDP-modification of DBB and BCP- particles was carried out through different steps of preparation including grinding and ultrasonic technique. In each experiment, 100 mg materials from each of the 4 groups (nDP-modified DBB, nDP-modified BCP, un-modified DBB and un-modified BCP) were plated into a 48 well cell culture plate and 200.000 cells/well were seeded onto the materials. Scanning electron microscopy (SEM) was carried out after 24 hours and 3 days. Real time-polymerase chain reaction (PCR) was carried out after 3 days, 1 week and 2 weeks of incubation. The following markers were analyzed; alkaline phosphatase (ALP), osteocalcin (OC), bone morphogenetic protein type 2 (BMP-2) and integrin alpha 10 (ITGA 10). The general tendency of DBB and BCP was compared.
Results: Cellular responses were evaluated in terms of attachment and differentiation. SEM after 24 hours and 3 days of incubation disclosed similar cell attachment and spreading for nDP-modified and non-modified DBB and BCP particles. Real-time PCR revealed significant up-regulation of mRNA expression of ALP and OC by HOB grown on nDP-modified DBB and BCP-particles after 1 and 2 weeks compared to non-modified particles. A significant down-regulation of BMP-2 on nDP-modified DBB and a significant up-regulation of BMP-2 on nDP-modified BCP were disclosed for HOB in relation to un-modified particles. Cell adhesion marker ITGA 10 showed significant down-regulated in the mRNA level for both nDP-modified groups after 2 weeks of incubation (nDP-BCP (p<.01) and nDP-DBB (p<.05) compared to the non-modified materials.
Conclusion: nDP-modified BCP-particles seem to enhance the expression of osteogenic markers in HOB in vitro. The results indicate that nDP-modified BCP enhance the osteoblast phenotype and suggest that these scaffolds might be appropriate cell carriers, superior to nonmodified BCP-particles. The osteogenic markers of OC and ALP were increased for nDPmodified DBB particles but BMP-2 was down regulated in relation to non-modified DBB particles.
Tissue engineering, scaffolds, gene expression, human osteoblast-like cells, biphasic calcium phosphate, deproteinized bovine bone
Medical and Health Sciences
IdentifiersURN: urn:nbn:se:liu:diva-72937OAI: oai:DiVA.org:liu-72937DiVA: diva2:463799