Speaker
Description
INTRODUCTION
In recent years, regenerative medicine is becoming more and more important, especially the field related to stem cell transplantations. In the body, the physiological niche microenvironment of stem cells (SCs), characterized by a unique combination of biomechanical, biochemical and biophysical properties, is a challenging research key. In this context, the micro-scaffold “Nichoid” has shown to induce pluripotency in several SCs thanks to its peculiar 3D structure inducing isotropic mechanical stimuli (1). Indeed, the Nichoid induces the expression of pluripotency genes and potentiate the therapeutic potential of neural precursor SCs (NPCs). In our work we investigated the therapeutic effects of cell expanded inside the Nichoid in a preclinical animal model of Parkinson’s disease (2,3). Meanwhile, we investigate the potential of human adipose-derived mesenchymal stem cells (hADMSCs) for the treatment of Spinal Cord Injury (4).
METHODS
Nichoids were fabricated by an innovative two-photon polymerization technique using a biocompatible photoresist. The biological validation of the scaffold was started by expanding NPCs and hADMSCs for 7 days. Then, cells expanded inside the Nichoid have been characterized by viability assay, Real Time PCR, Western Blot and immunofluorescence analysis. Furthermore, neural differentiation was also investigated.
Moreover, RNA-seq analysis of cell grown inside the scaffold was performed to evaluate the gene expression profiles. For preclinical animal tests, parkinsonism was induced by the administration of neurotoxin MPTP in C57/black mice by a consolidated protocol, The recovery of motor functions was investigated with vertical and horizontal grid behavioural tests, and, after the animals’ sacrifice, observations were corroborated by immunofluorescence studies performed on brain sections.
RESULTS
SCs expanded in 3D Nichoid scaffold presented a specific organization and morphology. 7 days after plating, both NPCs and hADMSCs show an increase in pluripotency potential and mechanotransduction features. To evaluate the in vivo efficacy, after intra-striatal infusion in PD mice, NPCs expanded inside Nichoid showed an increase in their therapeutic features by counteracting neurodegeneration and promoting recovery of function. Specifically, these cells favored the recovery of dopaminergic markers in brain area affected by the neurodegeneration.
DISCUSSION AND CONCLUSIONS
These results represent an incredible improvement needed to reach the biological and clinical demands. Here we provide evidence that the expansion of stem cells inside the Nichoid both favours their pluripotency and potentiates their efficacy showing great promise in the field of regenerative medicine.
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 and “Neurogel-en-Marche” Foundation.
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