The Potential Selective Cytotoxicity of Poly (L- Lactic Acid)-Based Scaffolds Functionalized with Nanohydroxyapatite and Europium (III) Ions toward Osteosarcoma Cells

Abstract

Osteosarcoma (OSA) is malignant bone tumor, occurring in children and adults, characterized by poor prognosis. Despite advances in chemotherapy and surgical techniques, the survival of osteosarcoma patients is not improving significantly. Currently, great efforts are taken to identify novel selective strategies, distinguishing between cancer and normal cells. This includes development of biomimetic scaffolds with anticancer properties that can simultaneously support and modulate proper regeneration of bone tissue. In this study cytotoxicity of scaffolds composed from poly (L-lactic acid) functionalized with nanohydroxyapatite (nHAp) and doped with europium (III) ions—10 wt % 3 mol % Eu3+: nHAp@PLLA was tested using human osteosarcoma cells: U-2 OS, Saos-2 and MG-63. Human adipose tissue-derived stromal cells (HuASCs) were used as non-transformed cells to determine the selective cytotoxicity of the carrier. Analysis included evaluation of cells morphology (confocal/scanning electron microscopy (SEM)), metabolic activity and apoptosis profile in cultures on the scaffolds. Results obtained indicated on high cytotoxicity of scaffolds toward all OSA cell lines, associated with a decrease of cells’ viability, deterioration of metabolic activity and activation of apoptotic factors determined at mRNA and miRNA levels. Simultaneously, the biomaterials did not affect HuASCs’ viability and proliferation rate. Obtained scaffolds showed a bioimaging function, due to functionalization with luminescent europium ions, and thus may find application in theranostics treatment of OSA.

Nanohydroxyapatite (nHAp) Doped with Iron Oxide Nanoparticles (IO), miR-21 and miR-124 Under Magnetic Field Conditions Modulates Osteoblast Viability, Reduces Inflammation and Inhibits the Growth of Osteoclast – A Novel Concept for Osteoporosis Treatment: Part 1

“…The anti-inflammatory action of the static magnetic field and hydroxyapatites were described previously. 35,71 Here we showed that the anti-inflammatory potential of nHAp/IO composites could be enhanced by incorporating miR-21 and miR-124 molecules. The improved biological effects resulting from nHAp/IO functionalization by miRNAs was evidenced by the proinflammatory profiles of cytokines released by LPS-stimulated macrophages in experimental cultures.…”

“…The protocol was performed accordingly to manufacturers’ instruction, and it was shown in our previous studies. 35 , 36 After incubations with provided reagents, the samples were analysed using Muse TM Cell Analyzer (Merck/Sigma-Aldrich, Munich, Germany). Each measurement was performed in triplicate.…”

“…Additionally, the MTS assay (Abcam, Cambridge, UK) was performed using a well-known protocol performed according to the method described before. 35 …”

“…The procedures were performed accordingly to our previous studies. 33 , 35 , 36 Briefly, to isolate the RNA, cultures were homogenized by the addition of Extrazol ® (Blirt DNA, Gdańsk, Poland). The purity and the quantity of obtained RNA were measured by a spectrophotometer at 260 and 280 nm wavelength (Epoch, BioTek Instruments, Vermont, USA).…”

Bone marrow stromal cells (BMSCs CD45‐/CD44+/CD73+/CD90+) isolated from osteoporotic mice SAM/P6 as a novel model for osteoporosis investigation

“…Other reagents were prepared for the analysis as it was described elsewhere. 35 According to manufacturer instructions, the analysis of caspases activation was based on membrane permeable VAD-peptide that can detect multiple caspases for example caspase 1, −3, −4, −5, −6 −7, −8 and −9.…”