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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.


Related in: MedlinePlus

Real-time RT-PCR analysis. Relative mRNA expression levels for COL1A1, ALPL, BMP2, VEGFA, TNFRSF11B and BGLAP are shown in osteoblasts cultured on scaffolds with 10 nM and 10 µM SIM compared to scaffolds without SIM and normalized to reference gene GAPDH at 7, 14 and 21 days. Values represent the mean ± SD. Statistical analysis: (a) p ≤ 0.05 versus alginate-coated scaffold without SIM and (b) p ≤ 0.05 versus alginate-coated scaffold with SIM 10 µM.SIM: simvastatin; RT-PCR: reverse transcriptase–polymerase chain reaction; mRNA: messenger RNA; GAPDH: glyceraldehyde-3-phosphate dehydrogenase; COL1A1: collagen type I alpha 1; ALPL: alkaline phosphatase; BMP2: bone morphogenetic protein 2; VEGFA: vascular endothelial growth factor A; TNFRSF11B: osteoprotegerin; BGLAP: osteocalcin; SD: standard deviation.
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fig7-2041731413515670: Real-time RT-PCR analysis. Relative mRNA expression levels for COL1A1, ALPL, BMP2, VEGFA, TNFRSF11B and BGLAP are shown in osteoblasts cultured on scaffolds with 10 nM and 10 µM SIM compared to scaffolds without SIM and normalized to reference gene GAPDH at 7, 14 and 21 days. Values represent the mean ± SD. Statistical analysis: (a) p ≤ 0.05 versus alginate-coated scaffold without SIM and (b) p ≤ 0.05 versus alginate-coated scaffold with SIM 10 µM.SIM: simvastatin; RT-PCR: reverse transcriptase–polymerase chain reaction; mRNA: messenger RNA; GAPDH: glyceraldehyde-3-phosphate dehydrogenase; COL1A1: collagen type I alpha 1; ALPL: alkaline phosphatase; BMP2: bone morphogenetic protein 2; VEGFA: vascular endothelial growth factor A; TNFRSF11B: osteoprotegerin; BGLAP: osteocalcin; SD: standard deviation.

Mentions: After 21 days of culture, the relative expression of BGLAP was significantly increased in osteoblasts cultured on scaffolds with 10 µM SIM when compared to scaffolds without SIM and normalized to GAPDH (p = 0.012). Furthermore, the relative expression of BMP2 was significantly enhanced in osteoblasts cultured on scaffolds with 10 nM SIM when compared to scaffolds with 10 µM SIM and normalized to GAPDH (p = 0.038). No significant differences were observed in the expression of ALPL, COL1A1, TNFRSF11B or VEGFA messenger RNA (mRNA) levels among experimental groups at any of the time points studied (Figure 7).


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)

Real-time RT-PCR analysis. Relative mRNA expression levels for COL1A1, ALPL, BMP2, VEGFA, TNFRSF11B and BGLAP are shown in osteoblasts cultured on scaffolds with 10 nM and 10 µM SIM compared to scaffolds without SIM and normalized to reference gene GAPDH at 7, 14 and 21 days. Values represent the mean ± SD. Statistical analysis: (a) p ≤ 0.05 versus alginate-coated scaffold without SIM and (b) p ≤ 0.05 versus alginate-coated scaffold with SIM 10 µM.SIM: simvastatin; RT-PCR: reverse transcriptase–polymerase chain reaction; mRNA: messenger RNA; GAPDH: glyceraldehyde-3-phosphate dehydrogenase; COL1A1: collagen type I alpha 1; ALPL: alkaline phosphatase; BMP2: bone morphogenetic protein 2; VEGFA: vascular endothelial growth factor A; TNFRSF11B: osteoprotegerin; BGLAP: osteocalcin; 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

fig7-2041731413515670: Real-time RT-PCR analysis. Relative mRNA expression levels for COL1A1, ALPL, BMP2, VEGFA, TNFRSF11B and BGLAP are shown in osteoblasts cultured on scaffolds with 10 nM and 10 µM SIM compared to scaffolds without SIM and normalized to reference gene GAPDH at 7, 14 and 21 days. Values represent the mean ± SD. Statistical analysis: (a) p ≤ 0.05 versus alginate-coated scaffold without SIM and (b) p ≤ 0.05 versus alginate-coated scaffold with SIM 10 µM.SIM: simvastatin; RT-PCR: reverse transcriptase–polymerase chain reaction; mRNA: messenger RNA; GAPDH: glyceraldehyde-3-phosphate dehydrogenase; COL1A1: collagen type I alpha 1; ALPL: alkaline phosphatase; BMP2: bone morphogenetic protein 2; VEGFA: vascular endothelial growth factor A; TNFRSF11B: osteoprotegerin; BGLAP: osteocalcin; SD: standard deviation.
Mentions: After 21 days of culture, the relative expression of BGLAP was significantly increased in osteoblasts cultured on scaffolds with 10 µM SIM when compared to scaffolds without SIM and normalized to GAPDH (p = 0.012). Furthermore, the relative expression of BMP2 was significantly enhanced in osteoblasts cultured on scaffolds with 10 nM SIM when compared to scaffolds with 10 µM SIM and normalized to GAPDH (p = 0.038). No significant differences were observed in the expression of ALPL, COL1A1, TNFRSF11B or VEGFA messenger RNA (mRNA) levels among experimental groups at any of the time points studied (Figure 7).

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.


Related in: MedlinePlus