Table 3.
Studies Considering Infection of Oral Mucosa Models with Microorganisms Other Than Candida and Porphyromonas
| Authors | Bacteria strains | Culture condition | Oral mucosa model | Time of contact between biofilm and mucosa | Assays | Results |
|---|---|---|---|---|---|---|
| Gursoy et al.23 | Two strains of F. nucleatum: ATCC25586 and AHN9508 (clinical oral isolate) | Two groups: anaerobically grown biofilm on a semipermeable membrane placed upside-down on OCC, 10 μL (3 × 106 CFUs/PBS) of planktonic bacteria | HaCaT epithelial cells grown on a fibroblast collagen matrix (OCC model) | 24 h | H&E, Ki-67, PASS, LDH release | Invasion of the collagen matrix by one of the strains; more cytotoxicity and invasiveness of biofilm in comparison to planktonic bacteria |
| Dabija-Wolter et al.62 | Four strains of F. nucleatum: ATCC 10953, ATCC 25586, and two other clinical isolates: AHN 8158 and MRC-23 | 5 × 107 unstained or FITC-labeled F. nucleatum in 20–30 μL FAD medium, in anaerobic atmosphere for 3 h and then at 37°C in aerobic conditions | 3D engineered models of human gingiva using primary gingival keratinocytes and fibroblasts | 24, 48 h | CLSM, IHC, qRT-PCR | Penetration of F. nucleatum to gingival epithelium without causing permanent damage |
| Pollanen et al.63 | F. nucleatum (ATCC) 25586 | Biofilm grown on semipermeable nitrocellulose membranes placed on OMM | HaCaT cells seeded on collagen fibroblast gels and a tooth piece placed on top | ≤24 h | IHC | Epithelial migration and altered epithelial proliferation pattern |
| De Ryck et al.22 | Microbiota derived from a swab of the inner cheek | Microbiota grown on an agar/mucin layer positioned on top of oral mucosa | TR146, HaCaT, or normal keratinocyte cells grown on collagen layer containing NIH-3T3 fibroblasts | 72 h | Oral scratch assay, Pyrosequencing, PCR-DGGE analysis, live/dead staining, flow cytometry, SCFA, MTT, SRB, LDH, Western blot, lactate analysis, Van Gieson, Alcian Blue, E-cadherin, Ki67, H&E | Reduced healing in the presence of microbiota, no reduction of the proliferation index, no increase of apoptotic or necrotic cells |
| Buskermolen et al.64 | Three biofilm types: commensal, gingivitis, and cariogenic | 10 μL of 105, 106, or 107 CFUs/equivalent diluted in HBSS | Immortalized human keratinocyte (KC-TERT) and fibroblast (Fib-TERT) embedded in collagen hydrogel | 24 h | IHC, FISH, fluorescence resonance energy transfer, ELISA | Increased expression of elafin, secretion of the antimicrobial cytokine and inflammatory cytokines in the gingiva epithelium |
| Shang et al.24 | From healthy human saliva, consists of typical commensal genera Granulicatella and major oral microbiota genera Veillonella and Streptococcus | 107 CFU of biofilm cells diluted in 10 μL HBSS, dripped onto the surface of the RHG | RHG: immortalized human keratinocyte (KC-TERT) and fibroblast (Fib-TERT)-populated hydrogel | 1, 2, 4, or 7 days | ELISA, RT-PCR, CFU count, H&E, FISH | Increased epithelial thickness, stratification, keratinocyte proliferation, and production of anti-microbial proteins in biofilm exposed RHG |
| Rahimi et al.65 | Streptococcus mutans (strain UA-159) | Injection of 2 μL of bacterial solution (with optical density between 0.2 and 0.3) into the keratinocyte-containing channel of the device | Microfluidic mucosal model-on-a-chip: fibroblast cell line-laden collagen, followed by a keratinocyte cell line (Gie-No3B11) layer | 24 h | DiI fluorescence staining, TEER | Some infiltration in collagen layer, lower TEER after bacterial exposure |
| Shang et al.66 | Commensal, gingivitis, or cariogenic biofilms from human healthy saliva | Biofilms cultured in the AAA model diluted as 1 × 107 CFU biofilm cells in 10 μL HBSS | RHG: keratinocyte (KC-TERT, OKG4/bmi1/TERT) on collagen-embedded fibroblast (Fib-TERT) | 24 h | FISH, H&E, RT-PCR, Western blotting | Upregulation of gene expression involved in TLR signaling by commensal biofilm, and suppression of some by cariogenic biofilm; no significant damaging effect on RHG morphology |
| Ingendoh-Tsakmakidis et al.67 | Biofilm of S. oralis (DSM 20627) on polyethersulfone membrane, biofilm of A. actinomycetemcomitans JP2 strain on coverslip | S. oralis or A. actinomycetemcomitans biofilm facing the peri-implant oral mucosa model with direct contact to titanium disk | Peri-implant oral mucosa model assembly: OKF6/TERT-2 seeded on titanium disks-HGF-collagen matrix | 24 h | Microarray data analysis, ELISA, IHC | Induction of a protective stress response by S. oralis. downregulation of genes involved in inflammatory response by A. actinomycetemcomitans |
| Beklen et al.68 | A. actinomycetemcomitans strain D7S | A. actinomycetemcomitans biofilm cultured on porous filter discs added on top of OMM | Immortalized human gingival keratinocyte cells seeded on fibroblast-collagen matrix | 24 h | IHC, TEM | Thick necrotic layer and decrease of keratin expression in epithelium following infection |
AAA-model, Amsterdam active attachment model; AHN, anaerobe Helsinki negative; DGGE, denaturing gradient gel electrophoresis; FITC, fluorescein-isothiocyanate; HaCaT, human adult low-calcium high-temperature; HBSS, Hank's Balanced Salt Solution; KC-TERT, telomerase reverse transcriptase-immortalized human keratinocyte; OCC, organotypic cell culture; RHG, reconstructed human gingiva; SCFA, short-chain fatty acid; SRB, sulforhodamine B colorimetric assay; TEER, transepithelial electrical resistance; TLR, toll-like receptor.