TY - JOUR

T1 - Biofilm development of Porphyromonas gingivalis on titanium surfaces in response to 1,4-dihydroxy-2-naphthoic acid-a hybrid in vitro-in silico approach

AU - Mukherjee, Rumjhum

AU - Klempt, Felix

AU - Fuchs, Florian

AU - Doll-Nikutta, Katharina

AU - Soleimani, Meisam

AU - Wriggers, Peter

AU - Junker, Philipp

AU - Stiesch, Meike

AU - Szafrański, Szymon P.

N1 - Publisher Copyright: © 2025 Mukherjee et al. This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International license.

PY - 2025/10/7

Y1 - 2025/10/7

N2 - Colonization of titanium dental implants by the oral pathogen Porphyromonas gingivalis can lead to peri-implant diseases and, ultimately, implant failure. P. gingivalis growth can be stimulated by 1,4-dihydroxy-2-naphthoic acid (DHNA), a menaquinone precursor from various oral bacteria; however, its impact on biofilm formation remains unclear. The aim of the study was to evaluate P. gingivalis growth and metabolic activity over 6 days in response to DHNA on two titanium grade IV surfaces with different roughness using a hybrid in vitro-in silico approach. P. gingivalis growth was modestly stimulated by DHNA and exhibited an inverse correlation with ammonia concentration in culture medium. Notably, this growth pattern transitioned from an initial linear phase to a later exponential phase, with DHNA-treated biofilms reaching this exponential shift at an earlier stage than untreated controls. Confocal microscopy revealed that DHNA-treated biofilms exhibited surface-dependent growth patterns, with larger biofilm volumes observed on rougher surfaces in later biofilm stages, compared with smoother surfaces. Regardless of surface characteristics, the area occupied by biofilms and the size of the aggregates exhibited a consistent and progressive increase over time and were larger in late DHNA-treated biofilms. The experimental data were used to calibrate a coupled finite element method (FEM)-based model that simulated P. gingivalis biofilm dynamics and nutrient utilization. Summarizing, DHNA moderately stimulated P. gingivalis growth, accelerated its transition to ammonia-independent growth, and promoted an increase in biofilm area and aggregate size. Our coupled approach offers significant potential for advancing in vitro biofilm research.IMPORTANCEThe results of our hybrid in vitro-in silico experiments advance the research on P. gingivalis physiology and its DHNA-dependent colonization of implant surfaces. Our findings reveal that DHNA accelerates P. gingivalis growth, induces aggregation, and promotes colonization of titanium surfaces. For the first time, DHNA-induced P. gingivalis growth acceleration and an earlier shift away from ammonia dependency were observed fluorometrically, highlighting ammonia assimilation as a promising marker of P. gingivalis physiology during early biofilm expansion. Understanding how growth factors together with surface properties influence P. gingivalis colonization offers a basis for future preventive strategies. Our study's stringent characterization of 3D surface texture parameters is expected to improve the reproducibility of biofilm-surface interactions experiments. The findings were validated using a continuum-based in silico model, initiating a hybrid approach where computational models complement in vitro research. Our interdisciplinary approach offers a versatile framework for investigating additional aspects of oral biofilms on titanium.

AB - Colonization of titanium dental implants by the oral pathogen Porphyromonas gingivalis can lead to peri-implant diseases and, ultimately, implant failure. P. gingivalis growth can be stimulated by 1,4-dihydroxy-2-naphthoic acid (DHNA), a menaquinone precursor from various oral bacteria; however, its impact on biofilm formation remains unclear. The aim of the study was to evaluate P. gingivalis growth and metabolic activity over 6 days in response to DHNA on two titanium grade IV surfaces with different roughness using a hybrid in vitro-in silico approach. P. gingivalis growth was modestly stimulated by DHNA and exhibited an inverse correlation with ammonia concentration in culture medium. Notably, this growth pattern transitioned from an initial linear phase to a later exponential phase, with DHNA-treated biofilms reaching this exponential shift at an earlier stage than untreated controls. Confocal microscopy revealed that DHNA-treated biofilms exhibited surface-dependent growth patterns, with larger biofilm volumes observed on rougher surfaces in later biofilm stages, compared with smoother surfaces. Regardless of surface characteristics, the area occupied by biofilms and the size of the aggregates exhibited a consistent and progressive increase over time and were larger in late DHNA-treated biofilms. The experimental data were used to calibrate a coupled finite element method (FEM)-based model that simulated P. gingivalis biofilm dynamics and nutrient utilization. Summarizing, DHNA moderately stimulated P. gingivalis growth, accelerated its transition to ammonia-independent growth, and promoted an increase in biofilm area and aggregate size. Our coupled approach offers significant potential for advancing in vitro biofilm research.IMPORTANCEThe results of our hybrid in vitro-in silico experiments advance the research on P. gingivalis physiology and its DHNA-dependent colonization of implant surfaces. Our findings reveal that DHNA accelerates P. gingivalis growth, induces aggregation, and promotes colonization of titanium surfaces. For the first time, DHNA-induced P. gingivalis growth acceleration and an earlier shift away from ammonia dependency were observed fluorometrically, highlighting ammonia assimilation as a promising marker of P. gingivalis physiology during early biofilm expansion. Understanding how growth factors together with surface properties influence P. gingivalis colonization offers a basis for future preventive strategies. Our study's stringent characterization of 3D surface texture parameters is expected to improve the reproducibility of biofilm-surface interactions experiments. The findings were validated using a continuum-based in silico model, initiating a hybrid approach where computational models complement in vitro research. Our interdisciplinary approach offers a versatile framework for investigating additional aspects of oral biofilms on titanium.

KW - ammonia

KW - biofilms

KW - confocal microscopy

KW - DHNA

KW - implanted devices

KW - mathematical modeling

KW - Porphyromonas gingivalis

KW - surface structures

KW - vitamin biosynthesis

UR - http://www.scopus.com/inward/record.url?scp=105017999133&partnerID=8YFLogxK

U2 - 10.1128/spectrum.00410-25

DO - 10.1128/spectrum.00410-25

M3 - Article

C2 - 40815221

AN - SCOPUS:105017999133

VL - 13

SP - e00410-25

JO - Microbiology spectrum

JF - Microbiology spectrum

SN - 2165-0497

IS - 10

ER -