Micropatterning of Physisorbed Laminin-1 Physiological Matrices

Nicoletta Sgarbi1, Dario Pisignano1, Francesca Di Benedetto1, Alessandra Aloisi2, Giuseppe Nicolardi2, Roberto Cingolani1, and Ross Rinaldi1

1National Nanotechnology Laboratory of Istituto Nazionale di Fisica della Materia, Dipartimento di Ingegneria dell'Innovazione, Università di Lecce, Via per Arnesano, 73100 Lecce, Italy, and
2Dipartimento di Scienze e Tecnologie Biologiche ed Ambientali, Università di Lecce, via Monteroni, I-73100 Lecce, Italy

e-mail: nicoletta.sgarbi@unile.it

URL: http://www.nnl.it


Laminin-1 (LN-1) is an extracellular matrix (ECM) protein playing an important role for many cellular functions. LN-1 is able to self-assembly in network of polygonal structures by a well defined process of polymerization, forming matrices whose physiological properties strongly depend on their morphologies.

We exploited microcontact printing (µCP) to pattern regions of physisorbed LN-1 onto glass substrates. Indirect immunofluorescence assays with anti-laminin antibody show well-defined features on the whole printed area and the recognizing of the LN-1 molecules epitopes from the primary antibody, which can indicate negligible conformational changes of the proteins.

Atomic Force Microscopy (AFM) investigations show the inner structure of the printed laminin films revealing matrices with polygonal features with recognizable single laminin molecules, having physiological domain interactions.

The formation of these physiological mesh-like protein matrices by the microcontact printing technique was found to be strongly dependent on the LN-1 initial concentration, and it was achieved by physisorption without any previous chemical functionalization of the glass surface. The investigation of the influence of these patterns on the behaviour of adhered cells is currently in progress in our laboratory, aiming to assess the best culture conditions in view of the application in hybrid cell-based electronic chips and tissue engineering.