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Cells inside a 3D matrix (such as tissue extracellular matrix or biomaterials) sense their insoluble environment through specific binding interactions between their adhesion receptors and ligands present on the matrix surface. Despite the critical role of the insoluble matrix in cell regulation, there exist no widely-applicable methods for quantifying the chemical stimuli provided by a matrix to cells. Here, we describe a general-purpose technique for quantifying in situ the density of ligands for specific cell adhesion receptors of interest on the surface of a 3D matrix. This paper improves significantly the accuracy of the procedure introduced in a previous publication by detailed marker characterization, optimized staining, and improved data interpretation. The optimized methodology is utilized to quantify the ligands of integrins α1β1, α2β1 on two kinds of matched porous collagen scaffolds, which are shown to possess significantly different ligand density, and significantly different ability to induce peripheral nerve regeneration in vivo. Data support the hypothesis that cell adhesion regulates contractile cell phenotypes, recently shown to be inversely related to organ regeneration. The technique provides a standardized way to quantify the surface chemistry of 3D matrices, and a means for introducing matrix effects in quantitative biological models.

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The use of indocyanine green-doped albumin protein solders has been shown to vastly improve the anastomotic strength that can be achieved by laser tissue repair techniques, while at the same time minimizing collateral thermal tissue damage. However, the safety of the degradation products of the chromophore following laser irradiation is uncertain. Therefore, we studied the feasibility of using alternative chromophores in terms of temperature rise at the solder/tissue interface, the extent of thermal damage in the surrounding tissue, and the tensile strength of repairs. Biodegradable polymer scaffolds of controlled porosity were fabricated with poly(L-lactic-co-glycolic acid), using a solvent-casting and particulate-leaching technique. The porous scaffold acted as a carrier to the traditional protein solder composition of serum albumin and an absorbing chromophore mixed in deionized water. Two commonly used chromophores, indocyanine green and methylene blue were investigated, as well as blue and green food colorings.Temperature rise at the solder surface and at the interface between the solder and tissue were monitored by an IR temperature monitoring system and a type-K thermocouple, respectively, and the extent of thermal damage in the underlying tissue was determined using light microscopy. As expected, temperature rise at the solder/tissue interface, and consequently the degree of collateral thermal tissue damage, was directly related to the penetration depth of the laser light in the protein solder. Optimal tensile strength of repairs was achieved by selecting a chromophore concentration that resulted in a temperature of 66 ± 3 °C at the solder/tissue interface.