Keratinocyte-Specific Peptide-Based Surfaces for Hemidesmosome Upregulation and Prevention of Bacterial Colonization.

Percutaneous devices like orthopedic prosthetic implants for amputees, catheters, and dental implants suffer from high infection rates. A critical aspect mediating peri-implant infection of dental implants is the lack of a structural barrier between the soft tissue and the implant surface which could impede bacteria access and colonization of exposed implant surfaces. Parafunctional soft tissue regeneration around dental implants is marked by a lack of hemidesmosome formation and thereby weakened mechanical attachment. In response to this healthcare burden, a simultaneously hemidesmosome-inducing, antimicrobial, multifunctional implant surface was engineered. A designer antimicrobial peptide, GL13K, and a laminin-derived peptide, LamLG3, were coimmobilized with two different surface fractional areas. The coimmobilized peptide surfaces showed antibiofilm activity against Streptococcus gordonii while enhancing proliferation, hemidesmosome formation, and mechanical attachment of orally derived keratinocytes. Notably, the coatings demonstrated specific activation of keratinocytes: the coatings showed no effects on gingival fibroblasts which are known to impede the quality of soft tissue attachment to dental implants. These coatings demonstrated stability and retained activity against mechanical and thermochemical challenges, suggesting their intraoral durability. Overall, these multifunctional surfaces may be able to reduce peri-implantitis rates and enhance the success rates of all percutaneous devices via strong antimicrobial activity and enhanced soft tissue attachment to implants.

Physical-chemical interactions between dental materials surface, salivary pellicle and Streptococcus gordonii.

Dental materials are susceptible to dental plaque formation, which increases the risk of biofilm-associated oral diseases. Physical-chemical properties of dental material surfaces can affect salivary pellicle formation and bacteria attachment, but relationships between these properties have been understudied. We aimed to assess the effects of surface properties and adsorbed salivary pellicle on Streptococcus gordonii adhesion to traditional dental materials. Adsorption of salivary pellicle from one donor on gold, stainless steel, alumina and zirconia was monitored with a quartz crystal microbalance with dissipation monitoring (QCM-D). Surfaces were characterized by X-ray photoelectron spectroscopy, atomic force microscopy and water contact angles measurement before and after pellicle adsorption. Visualization and quantification of Live/Dead stained bacteria and scanning electron microscopy were used to study S. gordonii attachment to materials with and without pellicle. The work of adhesion between surfaces and bacteria was also determined. Adsorption kinetics and the final thickness of pellicle formed on the four materials were similar. Pellicle deposition on all materials increased surface hydrophilicity, surface energy and work of adhesion with bacteria. Surfaces with pellicle had significantly more attached bacteria than surfaces without pellicle, but the physical-chemical properties of the dental material did not significantly alter bacteria attachment. Our findings suggested that the critical factor increasing S. gordonii attachment was the salivary pellicle formed on dental materials. This is attributed to increased work of adhesion between bacteria and substrates with pellicle. New dental materials should be designed for controlling bacteria attachment by tuning thickness, composition and structure of the adsorbed salivary pellicle.

Peptide coatings enhance keratinocyte attachment towards improving the peri-implant mucosal seal.

There is a critical need for preventing peri-implantitis as its prevalence has increased and dental implants lack features to prevent it. Research strategies to prevent peri-implantitis have focused on modifying dental implants to incorporate different antimicrobial agents. An alternative strategy consists of barring the expansion of the biofilm subgingivally by forming a long-lasting permucosal seal between the soft tissue and the implant surface. Here, we innovatively biofunctionalized titanium with bioinspired peptide coatings to strengthen biological interactions between epithelial cells and the titanium surface. We selected laminin 332- and ameloblastin-derived peptides (Lam, Ambn). Laminin 332 participates in the formation of hemidesmosomes by keratinocytes and promotes epithelial attachment around teeth; and ameloblastin, an enamel derived protein, is involved in tissue regeneration events following disruption of the periodontium. Lam, Ambn or combinations of both peptides were covalently immobilized on titanium discs. Successful immobilization of the peptides was confirmed by contact angle goniometry, X-ray photoelectron spectroscopy and fluorescent labelling of the peptides. Additionally, we confirmed the mechanical and thermochemical stability of the peptides on Ti substrates. Proliferation and hemidesmosome formation of human keratinocytes (TERT-2/OKF-6) were assessed by immunofluorescence labelling. The peptide-coated surfaces increased cell proliferation for up to 48 h in culture compared to control surfaces. Most importantly, formation of hemidesmosomes by keratinocytes was significantly increased on surfaces coated with Ambn + Lam peptides compared to control (p < 0.01) and monopeptide coatings (p < 0.005). Together, these results support the Ambn + Lam multipeptide coating as a promising candidate for inducing a permucosal seal around dental implants.