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Enhanced in vitro osteoblast differentiation on TiO2 scaffold coated with alginate hydrogel containing simvastatin.

Pullisaar H, Tiainen H, Landin MA, Lyngstadaas SP, Haugen HJ, Reseland JE, Ostrup E - J Tissue Eng (2013)

Bottom Line: No cytotoxic effects on osteoblasts were observed by scaffolds with simvastatin when compared to scaffolds without simvastatin.The relative expression and secretion of osteocalcin was significantly increased by cells cultured on scaffolds with 10 µM simvastatin when compared to scaffolds without simvastatin after 21 days.In conclusion, the results indicate that simvastatin-coated TiO2 scaffolds can support a sustained release of simvastatin and induce osteoblast differentiation.

View Article: PubMed Central - PubMed

Affiliation: Department of Biomaterials, Institute of Clinical Dentistry, University of Oslo, Oslo, Norway.

ABSTRACT
The aim of this study was to develop a three-dimensional porous bone graft material as vehicle for simvastatin delivery and to investigate its effect on primary human osteoblasts from three donors. Highly porous titanium dioxide (TiO2) scaffolds were submerged into simvastatin containing alginate solution. Microstructure of scaffolds, visualized by scanning electron microscopy and micro-computed tomography, revealed an evenly distributed alginate layer covering the surface of TiO2 scaffold struts. Progressive and sustained simvastatin release was observed for up to 19 days. No cytotoxic effects on osteoblasts were observed by scaffolds with simvastatin when compared to scaffolds without simvastatin. Expression of osteoblast markers (collagen type I alpha 1, alkaline phosphatase, bone morphogenetic protein 2, osteoprotegerin, vascular endothelial growth factor A and osteocalcin) was quantified using real-time reverse transcriptase-polymerase chain reaction. Secretion of osteoprotegerin, vascular endothelial growth factor A and osteocalcin was analysed by multiplex immunoassay (Luminex). The relative expression and secretion of osteocalcin was significantly increased by cells cultured on scaffolds with 10 µM simvastatin when compared to scaffolds without simvastatin after 21 days. In addition, secretion of vascular endothelial growth factor A was significantly enhanced from cells cultured on scaffolds with both 10 nM and 10 µM simvastatin when compared to scaffolds without simvastatin at day 21. In conclusion, the results indicate that simvastatin-coated TiO2 scaffolds can support a sustained release of simvastatin and induce osteoblast differentiation. The combination of the physical properties of TiO2 scaffolds with the osteogenic effect of simvastatin may represent a new strategy for bone regeneration in defects where immediate load is wanted or unavailable.

No MeSH data available.


Lactate dehydrogenase (LDH) activity assay. LDH activity in culture medium from scaffolds with 10 nM and 10 µM SIM is shown compared to scaffolds without SIM for (a) donor 1, (b) donor 2 and (c) donor 3 measured every other day up till 14 days. Neither of the SIM concentrations caused a significant increase in LDH activity compared to the effect of alginate-coated scaffolds without SIM. Values represent the mean ± SD.SIM: simvastatin; SD: standard deviation.
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fig4-2041731413515670: Lactate dehydrogenase (LDH) activity assay. LDH activity in culture medium from scaffolds with 10 nM and 10 µM SIM is shown compared to scaffolds without SIM for (a) donor 1, (b) donor 2 and (c) donor 3 measured every other day up till 14 days. Neither of the SIM concentrations caused a significant increase in LDH activity compared to the effect of alginate-coated scaffolds without SIM. Values represent the mean ± SD.SIM: simvastatin; SD: standard deviation.

Mentions: The cytotoxic effect of SIM from alginate-coated scaffolds was tested for a wide range of concentrations (2.4 mM, 0.6 mM, 24 µM, 10 µM, 1 µM, 0.1 µM and 10 nM). SIM was found to be highly cytotoxic for osteoblasts at higher concentrations (above 10 µM), when cells were seeded on the scaffolds (data not shown). A 14-day cytotoxicity study was performed for lower concentrations of SIM (10 µM and 10 nM) to investigate the effect on osteoblast viability when exposed to SIM for a sustained time period. A higher LDH activity was generally detected in the medium from scaffolds with 10 µM SIM compared to scaffolds with 10 nM SIM throughout the 14-day period. Neither of the SIM concentrations caused a significant increase in LDH activity compared to the effect of alginate-coated scaffolds without SIM. Some variation was seen in the LDH activity profiles, indicating donor-dependent differences in the cellular response to SIM (Figure 4(a)–(c)).


Enhanced in vitro osteoblast differentiation on TiO2 scaffold coated with alginate hydrogel containing simvastatin.

Pullisaar H, Tiainen H, Landin MA, Lyngstadaas SP, Haugen HJ, Reseland JE, Ostrup E - J Tissue Eng (2013)

Lactate dehydrogenase (LDH) activity assay. LDH activity in culture medium from scaffolds with 10 nM and 10 µM SIM is shown compared to scaffolds without SIM for (a) donor 1, (b) donor 2 and (c) donor 3 measured every other day up till 14 days. Neither of the SIM concentrations caused a significant increase in LDH activity compared to the effect of alginate-coated scaffolds without SIM. Values represent the mean ± SD.SIM: simvastatin; SD: standard deviation.
© Copyright Policy - creative-commons
Related In: Results  -  Collection

License 1 - License 2 - License 3
Show All Figures
getmorefigures.php?uid=PMC3927861&req=5

fig4-2041731413515670: Lactate dehydrogenase (LDH) activity assay. LDH activity in culture medium from scaffolds with 10 nM and 10 µM SIM is shown compared to scaffolds without SIM for (a) donor 1, (b) donor 2 and (c) donor 3 measured every other day up till 14 days. Neither of the SIM concentrations caused a significant increase in LDH activity compared to the effect of alginate-coated scaffolds without SIM. Values represent the mean ± SD.SIM: simvastatin; SD: standard deviation.
Mentions: The cytotoxic effect of SIM from alginate-coated scaffolds was tested for a wide range of concentrations (2.4 mM, 0.6 mM, 24 µM, 10 µM, 1 µM, 0.1 µM and 10 nM). SIM was found to be highly cytotoxic for osteoblasts at higher concentrations (above 10 µM), when cells were seeded on the scaffolds (data not shown). A 14-day cytotoxicity study was performed for lower concentrations of SIM (10 µM and 10 nM) to investigate the effect on osteoblast viability when exposed to SIM for a sustained time period. A higher LDH activity was generally detected in the medium from scaffolds with 10 µM SIM compared to scaffolds with 10 nM SIM throughout the 14-day period. Neither of the SIM concentrations caused a significant increase in LDH activity compared to the effect of alginate-coated scaffolds without SIM. Some variation was seen in the LDH activity profiles, indicating donor-dependent differences in the cellular response to SIM (Figure 4(a)–(c)).

Bottom Line: No cytotoxic effects on osteoblasts were observed by scaffolds with simvastatin when compared to scaffolds without simvastatin.The relative expression and secretion of osteocalcin was significantly increased by cells cultured on scaffolds with 10 µM simvastatin when compared to scaffolds without simvastatin after 21 days.In conclusion, the results indicate that simvastatin-coated TiO2 scaffolds can support a sustained release of simvastatin and induce osteoblast differentiation.

View Article: PubMed Central - PubMed

Affiliation: Department of Biomaterials, Institute of Clinical Dentistry, University of Oslo, Oslo, Norway.

ABSTRACT
The aim of this study was to develop a three-dimensional porous bone graft material as vehicle for simvastatin delivery and to investigate its effect on primary human osteoblasts from three donors. Highly porous titanium dioxide (TiO2) scaffolds were submerged into simvastatin containing alginate solution. Microstructure of scaffolds, visualized by scanning electron microscopy and micro-computed tomography, revealed an evenly distributed alginate layer covering the surface of TiO2 scaffold struts. Progressive and sustained simvastatin release was observed for up to 19 days. No cytotoxic effects on osteoblasts were observed by scaffolds with simvastatin when compared to scaffolds without simvastatin. Expression of osteoblast markers (collagen type I alpha 1, alkaline phosphatase, bone morphogenetic protein 2, osteoprotegerin, vascular endothelial growth factor A and osteocalcin) was quantified using real-time reverse transcriptase-polymerase chain reaction. Secretion of osteoprotegerin, vascular endothelial growth factor A and osteocalcin was analysed by multiplex immunoassay (Luminex). The relative expression and secretion of osteocalcin was significantly increased by cells cultured on scaffolds with 10 µM simvastatin when compared to scaffolds without simvastatin after 21 days. In addition, secretion of vascular endothelial growth factor A was significantly enhanced from cells cultured on scaffolds with both 10 nM and 10 µM simvastatin when compared to scaffolds without simvastatin at day 21. In conclusion, the results indicate that simvastatin-coated TiO2 scaffolds can support a sustained release of simvastatin and induce osteoblast differentiation. The combination of the physical properties of TiO2 scaffolds with the osteogenic effect of simvastatin may represent a new strategy for bone regeneration in defects where immediate load is wanted or unavailable.

No MeSH data available.