Spidroin Silk Fibers with Bioactive Motifs of Extracellular Proteins for Neural Tissue Engineering

Veronica A Revkova; Konstantin V Sidoruk; Vladimir A Kalsin; Pavel A Melnikov; Mikhail A Konoplyannikov; Svetlana Kotova; Anastasia A Frolova; Sergey A Rodionov; Mikhail M Smorchkov; Alexey V Kovalev; Alexander V Troitskiy; Peter S Timashev; Vladimir P Chekhonin; Vladimir G Bogush; Vladimir P Baklaushev

doi:10.1021/acsomega.1c01576

Spidroin Silk Fibers with Bioactive Motifs of Extracellular Proteins for Neural Tissue Engineering

ACS Omega. 2021 May 30;6(23):15264-15273. doi: 10.1021/acsomega.1c01576. eCollection 2021 Jun 15.

Authors

Veronica A Revkova¹, Konstantin V Sidoruk², Vladimir A Kalsin¹, Pavel A Melnikov³, Mikhail A Konoplyannikov^{1

4}, Svetlana Kotova^{4

5}, Anastasia A Frolova⁴, Sergey A Rodionov⁶, Mikhail M Smorchkov⁶, Alexey V Kovalev⁶, Alexander V Troitskiy¹, Peter S Timashev^{4

5

7}, Vladimir P Chekhonin³, Vladimir G Bogush², Vladimir P Baklaushev¹

Affiliations

¹ Federal Research and Clinical Center of Specialized Medical Care and Medical Technologies FMBA of Russia, Moscow 115682, Russia.
² National Research Center "Kurchatov Institute", Moscow 123182, Russia.
³ Serbsky National Medical Research Center for Psychiatry and Narcology, Moscow 119034, Russia.
⁴ Institute for Regenerative Medicine, Sechenov First Moscow State Medical University, Moscow 119048, Russia.
⁵ N. N. Semenov Federal Research Center for Chemical Physics, Russian Academy of Sciences, Moscow 119991, Russia.
⁶ N. N. Priorov National Medical Research Center of Traumatology and Orthopedics, Moscow 127299, Russia.
⁷ Chemistry Department, Lomonosov Moscow State University, Moscow 119991, Russia.

Abstract

The interaction of neural progenitor cells (NPCs) with the extracellular matrix (ECM) plays an important role in neural tissue regeneration. Understanding which motifs of the ECM proteins are crucial for normal NPC adhesion, proliferation, and differentiation is important in order to create more adequate tissue engineered models of neural tissue and to efficiently study the central nervous system regeneration mechanisms. We have shown earlier that anisotropic matrices prepared from a mixture of recombinant dragline silk proteins, such as spidroin 1 and spidroin 2, by electrospinning are biocompatible with NPCs and provide good proliferation and oriented growth of neurites. This study objective was to find the effects of spidroin-based electrospun materials, modified with peptide motifs of the extracellular matrix proteins (RGD, IKVAV, and VAEIDGIEL) on adhesion, proliferation, and differentiation of directly reprogrammed neural precursor cells (drNPCs). The structural and biomechanical studies have shown that spidroin-based electrospun mats (SBEM), modified with ECM peptides, are characterized by a uniaxial orientation and elastic moduli in the swollen state, comparable to those of the dura mater. It has been found for the first time that drNPCs on SBEM mostly preserve their stemness in the growth medium and even in the differentiation medium with brain-derived neurotrophic factor and glial cell line-derived neurotrophic factor, while addition of the mentioned ECM-peptide motifs may shift the balance toward neuroglial differentiation. We have demonstrated that the RGD motif promotes formation of a lower number of neurons with longer neurites, while the IKVAV motif is characterized by formation of a greater number of NF200-positive neurons with shorter neurites. At the same time, all the studied matrices preserve up to 30% of neuroglial progenitor cells, phenotypically similar to radial glia derived from the subventricular zone. We believe that, by using this approach and modifying spidroin by various ECM-motifs or other substances, one may create an in vitro model for the neuroglial stem cell niche with the potential control of their differentiation.