by Keyword: hMSC
Canal, C., Fontelo, R., Hamouda, I., Guillem-Marti, J., Cvelbar, U., Ginebra, M. P., (2017). Plasma-induced selectivity in bone cancer cells death Free Radical Biology and Medicine 110, 72-80
Background: Current therapies for bone cancers - either primary or metastatic – are difficult to implement and
unfortunately not completely effective. An alternative therapy could be found in cold plasmas generated at
atmospheric pressure which have already demonstrated selective anti-tumor action in a number of carcinomas
and in more relatively rare brain tumors. However, its effects on bone cancer are still unknown.
Methods: Herein, we employed an atmospheric pressure plasma jet (APPJ) to validate its selectivity towards
osteosarcoma cell line vs. osteoblasts & human mesenchymal stem cells.
Results: Cytotoxicity following direct interaction of APPJ with cells is comparable to indirect interaction when
only liquid medium is treated and subsequently added to the cells, especially on the long-term (72 h of cell
culture). Moreover, following contact of the APPJ treated medium with cells, delayed effects are observed which
lead to 100% bone cancer cell death through apoptosis (decreased cell viability with incubation time in contact
with APPJ treated medium from 24 h to 72 h), while healthy cells remain fully viable and unaffected by the
Conclusions: The high efficiency of the indirect treatment indicates that an important role is played by the
reactive oxygen species (ROS) and reactive nitrogen species (RNS) in the gaseous plasma stage and then
transmitted to the liquid phase, which overall lead to lethal and selective action towards osteosarcoma cells.
These findings open new pathways for treatment of metastatic bone disease with a minimally invasive approach.
Keywords: Atmospheric pressure plasma jet, Bone cancer, hMSC, HOb, Liquids, Osteoblasts, Osteosarcoma, SaOS-2
Fraioli, R., Dashnyam, K., Kim, J. H., Perez, R. A., Kim, H. W., Gil, J., Ginebra, M. P., Manero, J. M., Mas-Moruno, C., (2016). Surface guidance of stem cell behavior: Chemically tailored co-presentation of integrin-binding peptides stimulates osteogenic differentiation in vitro and bone formation in vivo Acta Biomaterialia 43, 269-281
Surface modification stands out as a versatile technique to create instructive biomaterials that are able to actively direct stem cell fate. Chemical functionalization of titanium has been used in this work to stimulate the differentiation of human mesenchymal stem cells (hMSCs) into the osteoblastic lineage, by covalently anchoring a synthetic double-branched molecule (PTF) to the metal that allows a finely controlled presentation of peptidic motifs. In detail, the effect of the RGD adhesive peptide and its synergy motif PHSRN is studied, comparing a random distribution of the two peptides with the chemically-tailored disposition within the custom made synthetic platform, which mimics the interspacing between the motifs observed in fibronectin. Contact angle measurement and XPS analysis are used to prove the efficiency of functionalization. We demonstrate that, by rationally designing ligands, stem cell response can be efficiently guided towards the osteogenic phenotype: In vitro, PTF-functionalized surfaces support hMSCs adhesion, with higher cell area and formation of focal contacts, expression of the integrin receptor Î±5Î²1 and the osteogenic marker Runx2, and deposition a highly mineralized matrix, reaching values of mineralization comparable to fibronectin. Our strategy is also demonstrated to be efficient in promoting new bone growth in vivo in a rat calvarial defect. These results highlight the efficacy of chemical control over the presentation of bioactive peptides; such systems may be used to engineer bioactive surfaces with improved osseointegrative properties, or can be easily tuned to generate multi-functional coatings requiring a tailored disposition of the peptidic motifs. Statement of significance Organic coatings have been proposed as a solution to foster osseointegration of orthopedic implants. Among them, extracellular matrix-derived peptide motifs are an interesting biomimetic strategy to harness cell-surface interactions. Nonetheless, the combination of multiple peptide motifs in a controlled manner is essential to achieve receptor specificity and fully exploit the potentiality of synthetic peptides. Herein, we covalently graft to titanium a double branched molecule to guide stem cell fate in vitro and generate an osseoinductive titanium surface in vivo. Such synthetic ligand allows for the simultaneous presentation of two bioactive motifs, thus is ideal to test the effect of synergic sequences, such as RGD and PHSRN, and is a clear example of the versatility and feasibility of rationally designed biomolecules.
Keywords: hMSCs, Integrin-binding peptides, Osseointegration, RGD-PHSRN, Titanium