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Research

PULSED ELECTRON DEPOSITION OF BONE-LIKE APATITE THIN FILMS FROM A BIOGENIC SOURCE: FROM MATERIAL CHARACTERIZATION TO IN VITRO STEM CELL DIFFERENTIATION

The European Orthopaedic Research Society (EORS) 25th Annual and Anniversary Meeting, Munich, Germany, September 2017. Part 2 of 2.



Abstract

Fabrication of biogenic coatings with suitable mechanical properties is a key goal in orthopedics, to overcome the limitations of currently available coatings and improve the clinical results of coated implants compared to uncoated ones. In this paper, biological-like apatite coatings were deposited from a natural bone-apatite source by a pulsed electron deposition technique (PED).

Bone apatite-like (BAL) films were deposited directly from bone targets, obtained by standard deproteinization of bovine tibial cortical shafts and compared to films deposited by sintered stoichiometric-hydroxyapatite targets (HA). Deposition was performed at room temperature by PED in the Ionized Jet Deposition (IJD) version. Half of the samples was annealed at 400°C for 1h (BAL_400 and HA_400). As-deposited and annealed coatings were characterized in terms of composition and crystallinity (XRD, FT-IR), microstructure and morphology (SEM-EDS, AFM) and mechanical properties (nanoindentation and micro-scratch). For the biological tests, human dental pulp stem cells (hDPSCs) were isolated from dental pulp from patients undergoing a routine tooth extraction, plated on the samples (2500 cells/cm2) and cultured for 3 weeks, when the expression of typical osteogenic markers Runx-2, osteopontin, Osx and Osteocalcin in hDPSCs was evaluated.

Results showed that deposition by PED allows for a close transfer of the targets” composition. As-deposited coatings exhibited low cristallinity, that was significantly increased by post-deposition annealing, up to resembling that of biogenic apatite target. As a result of annealing, mechanical properties increased up to values comparable to those of commercial plasma-sprayed HA-coatings.

In vitro biological tests indicated that BAL_400 promotes hDPSCs proliferation to a higher extent compared to non-annealed bone coating and HA-references. Furher, immunofluorescence and western blot analyses revealed that the typical osteogenic markers were expressed, indicating that BAL_400 alone can efficiently promote the osteogenic commitment of the cells, even in absence of an osteogenic medium.

In conclusion, bone-like apatite coatings were deposited by PED, which closely resembled composition and structure of natural-apatite. Upon annealing at 400°C, the coatings exhibited satisfactory mechanical properties and were capable of providing a suitable microenvironment for hDPSCs adherence and proliferation and for them to reach osteogenic commitment.

These results suggest that bone apatite-like thin films obtained by biogenic source may represent an innovative platform to boost bone regeneration in the orthopedic, maxillofacial and odontoiatric field.


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