Conditioned bio-interfaces of silicon/porous silicon micro-patterns lead to the chondrogenesis of hMSCs

Abstract

aThe interaction between cells and materials is of scientific and technological interest for the development of new biomaterials with improved functional properties. In this work the chondrogenesis (CG) of human mesenchymal stem cells (hMSCs) derived from the bone marrow of healthy donors has been achieved on Si/PSi surfaces after pre-treatment with a conditioned medium derived from hMSCs (CM). The chondrogenic process was analyzed in fixed-cell preparations by immune fluorescence, determining the cellular localization of b-catenin and the transcription factors STAT-5, Runx2 and vitamin D receptor (VDR). In a week of CG on Si/PSi or CM-Si/PSi, b-catenin showed cell-cell contacts priming a preferential chondrogenic process on a PSi surface, in contrast with the preferential proliferation and migration processes observed on hexagonal/triangular micro-designed Si areas. Interestingly, the presence of a CM bio-interface improves this differentiation process with respect to a control PSi surface. The chondroinductive effect was also observed in STAT-5, which was highly expressed and is involved in intracellular signaling of many differentiation receptors related to development and tissue repair. In good agreement with the above results, the co-localization of transcription factors Runx2/VDR at nuclei supports active transcription of chondrogenic genes. Considering these observations, we propose that the combination of a rough PSi surface and the components of the CM-hMSCs biointerface boosts the chondrogenesis of hMSCs. These findings suggest that the tailoring of Si/PSi scaffolds with appropriate bio-interface combinations improves their therapeutic potential.

3D Printed Structures Filled with Carbon Fibers and Functionalized with Mesenchymal Stem Cell Conditioned Media as In Vitro Cell Niches for Promoting Chondrogenesis

“…As described in an earlier work [ 21 , 22 ], the clean supernatant was cooled down on ice for 30 min, centrifuged at 9000 rpm in a Sorvall centrifuge (Thermo Fisher Scientific, Waltham, MA USA) for a duration of 30 min to remove salt precipitations and, then, the clean supernatant was kept in 2 mL aliquots at −30 °C until use. We avoided repeatedly freeze–thawing the samples.…”

Research on the Methods for the Mass Production of Multi-Scale Organs-On-Chips

“…Cells were isolated, and culture expanded following processes described also in previous research with minor modifications [18,19,20]. The preparation of the hMSCs-CM has been also presented in previous studies by our team linked to biofunctionalization of biomedical microdevices [21,22].…”

Monolithic 3D labs- and organs-on-chips obtained by lithography-based ceramic manufacture

“…Then, cells were washed thoroughly with PBS, incubated in DMEM-LG, starved of FBS, complemented with 2 mM pyruvate and incubated during the following 24 h. Afterwards, the culture medium was collected, cleaned of any floating cells in a bench centrifuge at 1500 rpm for 5 min. As described in an early work [20,21], the clean supernatant was cooled down on ice for 30 min, centrifuged at 9000 rpm in a Sorvall centrifuge during 30 min to remove salt precipitations and then, the clean supernatant was kept in 2 mL aliquots at −30°C until use. We avoided repeatedly freeze-thawing the samples.…”