Speaker
Description
"Introduction
Adipose-derived mesenchymal stem cells (hADMSCs) are a promising resource for regenerative medicine. They can be harvested directly from the patient with moderately invasive procedures and re-implanted without eliciting immune response. To date, several clinical trials are active in phase II-III, but new technologies are needed to make these therapies more efficient. In particular, the physiologic niche microenvironment of stem cells, characterized by a unique combination of biophysical, biochemical and biomechanical properties, is a challenging research key. In this context, the micro-scaffold “Nichoid” has shown to induce pluripotency in stem cells thanks to its peculiar 3D structure inducing isotropic mechanical stimuli (1,2,3). The aim of this study is to fabricate and validate a new geometry of the Nichoid optimized for the expansion of hADMSCs and investigate the mechanotransduction process of cells in different scaffold pore configurations, using a combined computational and experimental approach
Materials and Methods
Nichoids were fabricated by an innovative two-photon polymerization technique using a biocompatible photoresist (4). To investigate the biomechanical stimuli of cells, we developed a computational model of the structural interaction between cells and the scaffold pore, using COMSOL Multiphysics. Then, we compared these mechanical parameters with several biomolecular and bioinformatics analyses performed on hADMSCs expanded on Nichoid scaffolds, as RNA-seq, Proteomics, Real Time RT-PCR, Western Blot and immunofluorescence assays.
Results
We compared different pore dimensions (10x10um, 20x20um and 30x30um) and we identified the most suitable structure for hADMSCs. Our hypothesis is based on how geometries of the scaffold influence the isotropy of the cells. This phenomenon is strictly connected to a roundish nucleus which corresponds to a specific accessibility of chromatin and higher expression of pluripotency genes (Figure 1). Indeed, the cubic conformation of the scaffold is connected to increased pluripotency, verified by the up-regulation of the stemness transcriptional core.
Discussion
The cubic Nichoid recreated the ex-vivo physiological environment optimized for MSCs. This new scaffold demonstrates how engineering technologies can support regenerative medicine and in particular stem cells therapies. Indeed, cells expanded in this new configuration of the scaffold could be easily collected in view of an autologous transplantation and they represent a valid therapeutic stem cell approach, in particular in Spinal Cord Injury.
References
- Raimondi et al., Acta Biomat, 4579-4584, 2013 Carelli et al., Nanothno, 5.1: 8, 2021 Rey et al., IJMS, 8: 6775, 2020 Zandrini et al., Sci Rep, 1:9, 2019
Acknowledgements
ERC projects NICHOID G.A. 646990, NICHOIDS G.A. 754467, and MOAB G.A. 825159; FET-OPEN project IN2SIGHT G.A. 964481; ESA project NICHOID-ET G.A. 4000133244/20/NL/GLC; NC3Rs projects MOAB, G.A. NC/C01903/1 and NC/C019201/1; MIUR-FARE project BEYOND, G.A. R16ZNN2R9K; FISR-COVID project LUNGCHILDMSC G.A. FISR2020IP_02959; Fondazione Giordano Dell’Amore, Cariplo Factory, Politecnico di Milano, Fondazione Bassetti and Fondazione Triulza (S2P project) and Fondazione Romeo ed Enrica Invernizzi.
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