Transglutaminase enables highly hydrolytically and proteolytically stable crosslinking of collagen on titanium surfaces and promotes osteogenic differentiation of human mesenchymal stem cells.

Collagen with its bioactive ligand motives would be predestined as coating on bone implant surfaces like titanium hip stems to facilitate receptor-mediated cell adhesion and thereby improve early osseointegration. Unfortunately, collagen as coating exhibits very low proteolytic resistance in vivo. To overcome this limitation, different crosslinking methods of collagen (transglutaminase, GTA, EDC/NHS, riboflavin, and lysyl oxidase) with silanized titanium alloy (Ti6Al4V) were investigated in terms of degradation resistance, hydrolysis stability, tensile strength, and metabolic cell activity. The in vitro osteogenic differentiation ability of human mesenchymal stem cells (hMSCs) induced by the surface modification was evaluated by immunofluorescence of early osteogenic markers, Alizarin red staining, and energy dispersive X-ray spectroscopy. The expression of the adhesion-related protein vinculin was analyzed on the different functionalized surfaces. The results revealed that the enzymatic crosslinker transglutaminase offered high degradation resistance, tensile strength, and hydrolysis stability compared to the other crosslinking reagents tested. Remarkably, the adhesion sequences within the collagen were accessible to the hMSCs despite the transglutaminase crosslinking procedure. In conclusion, the organochemical functionalization of Ti6Al4V surfaces with collagen using transglutaminase holds great potential to facilitate an enhanced interaction with attached bone cells and thereby could potentially improve and accelerate osseointegration of a titanium-based bone implant in vivo.