Selective biofunctionalization of 3D cell-imprinted PDMS with collagen immobilization for targeted cell attachment

Abstract

Cell-imprinted polydimethylsiloxane substrates, in terms of their ability to mimic the physiological niche, low microfabrication cost, and excellent biocompatibility were widely used in tissue engineering. Cells inside the mature cells' cell-imprinted PDMS pattern have been shown in previous research to be capable of being differentiated into a specific mature cell line. On the other hand, the hydrophobicity of PDMS substrate leads to weak cell adhesion. Moreover, there was no guarantee that the cells would be exactly located in the cavities of the cells' pattern. In many studies, PDMS surface was modified by plasma treatment, chemical modification, and ECM coating. Hence, to increase the efficiency of cell-imprinting method, the concavity region created by the cell-imprinted pattern is conjugated with collagen. A simple and economical method of epoxy silane resin was applied for the selective protein immobilization on the desired regions of the PDMS substrate. This method could be paved to enhance the cell trapping into the cell-imprinted pattern, and it could be helpful for stem cell differentiation studies. The applied method for selective protein attachment, and as a consequence, selective cell integration was assessed on the aligned cell-imprinted PDMS. A microfluidic chip created the aligned cell pattern. After Ar+ plasma and APTES treatment of the PDMS substrate, collagen immobilization was performed. The immobilized collagen was removed by epoxy silane resin stamp from the ridge area where the substrate lacked cell pattern and leaving the collagen only within the patterned areas. Coomassie brilliant blue staining was evaluated for selective collagen immobilization, and the collagen-binding stability was assessed by BCA analysis. MTT assay for the evaluation of cell viability on the modified surface was further analyzed. Subsequently, the crystal violet staining has confirmed the selective cell integration to the collagen-immobilized site on the PDMS substrate. The results proved the successfully selective collagen immobilization on the cell-imprinted PDMS and showed that this method increased the affinity of cells to attach inside the cell pattern cavity.