Modified amelogenin is a new and versatile nanomaterial for biomedical applications.
Amelogenin self-assembly is crucial for tooth biomineralization and crystallite enamel orientation. Amelogenin forms stable nanoparticles under physiological conditions. Here, we tested whether the surface properties and binding characteristics of these particles could be modified to enhance amelogenin function as a biomaterial. We evaluated different amelogenin fusion proteins for their ability to form hybrid nanoparticles. As a proof-of-concept, the integrin-binding tripeptide Arg-Gly-Asp (RGD) sequence from fibronectin was integrated into mouse amelogenin (rM179) at three different positions. Dynamic light scattering (DLS) measurements revealed that these amelogenin fusion proteins still form nanospheres. Additional DLS and isothermal titration calorimetry measurements showed that the mixtures of RGD-modified amelogenin and wild-type amelogenin form stable particles. We determined that insertion of the RGD-loop at the amelogenin C-terminus converts the nanoparticle into a cell-binding substrate. Calvarial osteoblasts efficiently attached and spread on modified amelogenin, whereas almost no binding was observed on wild-type amelogenin. These results establish amelogenin as a new versatile biomaterial that can be easily modified to add additional functions.
Institute for Dental Research and Oral Musculoskeletal Biology, Center for Biochemistry, Medical Faculty, University of Cologne, Kerpener-Str. 32, Cologne, 50931, Germany.
Thomas
T
Imhof
Department of Chemistry, Institute of Biochemistry, Center for Molecular Medicine, University of Cologne, Zuelpicher Str. 47, Cologne, 50674, Germany.
Nora
N
Gruenewald
Department of Chemistry, Institute of Biochemistry, Center for Molecular Medicine, University of Cologne, Zuelpicher Str. 47, Cologne, 50674, Germany.
Guenter
G
Schwarz
Department of Operative Dentistry, School of Dentistry, University of Cologne, Cologne, Germany.
Michael J
MJ
Noack
Institute for Dental Research and Oral Musculoskeletal Biology, Center for Biochemistry, Medical Faculty, University of Cologne, Kerpener-Str. 32, Cologne, 50931, Germany. Manuel.Koch@uni-koeln.de.|Centre for Ecology, Evolution and Environmental Changes, Faculdade de Ciências, Universidade de Lisboa, Lisbon, 1749-016, Portugal. Manuel.Koch@uni-koeln.de.
Manuel
M
Koch
D053523:Amelogenin / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D000818:Animals; D001672:Biocompatible Materials / Q000737:chemistry* / Q000378:metabolism*; D002151:Calorimetry; D002448:Cell Adhesion; D000067493:Dynamic Light Scattering; D051379:Mice; D049329:Nanostructures / Q000737:chemistry*; D010006:Osteoblasts / Q000502:physiology; D055503:Protein Multimerization; D055550:Protein Stability; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D013499:Surface Properties
D053523:Amelogenin; D001672:Biocompatible Materials; D011993:Recombinant Fusion Proteins
RGD tripeptide; amelogenin; functionalization; fusion protein; nanoparticle