Engineered Protein Coatings to Improve the Osseointegration of Dental and

Orthopaedic Implants

Jordan Raphel1a, Johan Karlsson2a, Silvia Galli3, Ann Wennerberg3, Chris Lindsay1, Matthew
Haugh1, Jukka Pajarinen4, Stuart B. Goodman4, Ryo Jimbo3, Martin Andersson2, Sarah C.
Heilshorn1*
1
Department of Materials Science and Engineering, Stanford University, Stanford, CA, USA
2
Department of Chemistry and Chemical Engineering, Chalmers University of Technology,
Gothenburg, Sweden
3
Department of Prosthodontics, Faculty of Odontology, Malmö University, Malmö, Sweden
4
Department of Orthopaedic Surgery, Stanford University, Stanford, CA, USA
a
Authors contributed equally to this work

*Prof. S. Heilshorn, heilshorn@stanford.edu

Figure S1: Amino acid sequence of ELP. The bioactive domain consist of either a cell-
adhesive extended RGD sequence derived from fibronectin (“RGD”) or a non-adhesive
scrambled control sequence (“Scrambled”). The elastin-like structural domains consists of
repeating blocks of an elastin-like VPGIG pentapeptide sequence, with every fifth isoleucine
residue replaced by a lysine, K. The T7-lac promoter was used to initiate expression, the poly-
histadine (“His”) tag could be used for identification or purification, and the enterokinase
(“EK”) cleavage site could be used for cleaving upstream domains. The total length of both
the RGD and scrambled ELP variants is 417 amino acids.

1
Figure S2: Tunable thickness of ELP coatings. A) ELP film thickness on Ti6Al4V discs after
successive rounds of spin coating, photocrosslinking, and rinsing. B) Thickness of dip coated
scrambled and RGD ELP coated films on Ti6Al4V discs after photocrosslinking and rinsing.

Figure S3: Custom built dip coater. A) Rendering of 3D printed trough and sample holder
showing capability to hold six 12-mm discs. B) Rendering of 3D printed sample holder
showing attachment for holding twenty-six rods with diameter of 0.889-mm. C) Picture of
assembled dip coater, including linear actuator, sample holder, stepper motor, arduino, motor
shield, and framing.

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Figure S4: Endotoxin purification of ELP. Scrambled and RGD ELP were endotoxin purified
by heating in formic acid and the residual endotoxin levels were quantified (PyroGene rFC,
Lonza). Purification resulted in endotoxin levels of approximately 0.1 EU/mL, well below the
strictest FDA limit of 0.5 EU/mL. Endotoxin levels prior to purification in RGD ELP were
over 100 times higher than the FDA endotoxin limit.