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Preprints

Bioactivation of 3D Cell-imprinted Polydimethylsiloxane Surface by Bone Proteins Nanocoating for Osteogenic Differentiation

Babaei M, Nasernejad B, Sharifikolouei E, Shokrgozar MA, Bonakdar S.
Preprint from
Research Square
18 January 2022
PPR
PPR444009
Abstract
Physical and chemical parameters that mimic the physiological niche of the human body have an influence on stem cell fate by creating directional signals to cells. Micro/nano cell patterned polydimethylsiloxane (PDMS) substrates, due to their ability to mimic the physiological niche, have been widely used in surface modifications. Integration of other factors such as the biochemical coating on the surface can achieve more similar microenvironmental conditions and promote stem cell differentiation to the target cell line. Herein, we investigated the effect of physical topography, chemical functionalization by acid bone lysate (ABL) nanocoating, and the combined functionalization of bone proteins nanocoated surface and topographically edited surface. We have prepared four distinguishing surfaces: plain PDMS, physically modified PDMS by 3D cell topography pattern, chemically modified PDMS with bone proteins nanocoating, and chemically modified nano 3D cell-imprinted PDMS by bone proteins (ABL). Characterization of ABL was carried out by Bradford staining and SDS page analysis followed by the MTT assay for evaluation of cell viability on ABL coated PDMS. Moreover, FESEM and Profilometry for determination of optimal coating thickness were utilized and the best coating concentration was identified and selected. The binding and retention of ABL to PDMS were confirmed by FTIR and BCA analysis. Sessile drop static water contact angle measurements on substrates have shown that the combined chemical functionalization and nano 3D cell-imprinting on PDMS surface leads to better surface wettability by 68% compared to plain PDMS. Eventually, the results of ALP measurement, Alizarin red S staining, Immunofluorescence staining, and real-time PCR have shown that nano 3D cell-imprinted PDMS surface functionalized by extracted bone proteins, ABL, is able to guide the fate of ADSCs toward osteogenic differentiation. Eventually, chemical modification of the cell-imprinted PDMS substrate by bone proteins extraction, not only improved the cell adhesion and proliferation but also contributed to the topographical effect itself and caused a significantly synergistic influence on the process of osteogenic differentiation.