Table 2.

Studies Related to the Infection of Oral Mucosa Models with Porphyromonas Alone or in Association with Other Bacteria

Authors	Bacteria strain	Culture condition	Oral mucosa model	Time of contact between P. gingivalis and mucosa	Assays	Results
Andrian et al.5	P. gingivalis ATCC 33277 and the derivative gingipain-null mutant KDP128	106 and 109 bacteria (ATCC 33277 or KPD128)/mL in DMEH, incubated in an anaerobic chamber	EHOM (primary epithelial and fibroblasts cells in collagen)	24 h	TEM, ELISA	Higher penetration of nonmutant form in lamina propria; high secretion of cytokines from oral mucosa models after infection
Kimball et al.25	P. gingivalis (ATCC 33277 or strain 861), S. gordonii DL-1, and Fusobacterium nucleatum ATCC 25586	6 × 106 bacteria in 10–50 μL bacterial growth medium (MOI of 100:1 bacteria per surface cell)	EpiOralTM (MatTek Corporation, Ashland, MA)	24–72 h	H&E, IHC, qRT-PCR	Increase of hBD2 expression after infection
Andrian et al.53	P. gingivalis ATCC 33277 or its derivative gingipain-null mutant (KDP128)	100 μL of 109 bacteria/mL in DMEH, in an anaerobic chamber	EHOM (primary epithelial and fibroblasts cells in collagen)	4, 8, 24 h	RT-PCR, ELISA	Increase activation of TIMP-2 and expression of MMP-2 and MMP-9 by oral mucosa following infection
Wayakanon et al.54	Clinical strains (A245br) of P. gingivalis	MOI = 100	OMM (NOK or TR146 cells on collagen containing NOFs)	18 h	Bacteria count, IHC	Reduced number of intracellular P. gingivalis in presence of polymersome-encapsulated metronidazole or doxycycline
Belibasakis et al.55	P. gingivalis ATCC 33277T, Campylobacter rectus (OMZ 697), F. nucleatum (OMZ 596), Prevotella intermedia ATCC 25611T, Tannerella forsythia OMZ1047, Treponema denticola ATCC 35405T, Veillonella dispar ATCC 17748T, Actinomyces oris (OMZ 745), S. anginosus (OMZ 871), and S. oralis SK 248 (OMZ 607)	10-species “subgingival” biofilm model grown on sintered hydroxyapatite discs placed onto OMM	EpiGing, (MatTek, Ashland, MA)	3–24 h	qPCR, LDH activity, ELISA	Upregulation of IL-8 gene expression and secretion after 3 h in both biofilms, in the presence of the “red complex”
Pinnock et al.56	P. gingivalis strains NCTC 11834 and W50	MOI of 100 (monolayer) or 2 × 107 cells/300 mL	OMMs with either NOK or the H357 cell line on collagen containing NOFs	1.5 or 4 h	Antibiotic protection assay, IF, IHC, chemokine array	Higher intracellular survival of P. gingivalis in mucosal models compared with monolayer cultures
Thurnheer et al.57	P. gingivalis ATCC 33277T, S. oralis SK248 S. anginosus ATCC 9895, Actinomyces oris (OMZ 745), F. nucleatum subsp. Nucleatum OMZ 598, Veillonella dispar ATCC 17748T, Campylobacter rectus OMZ 698, Prevotella intermedia ATCC 25611T, T. forsythia OMZ 1047, and Treponema denticola ATCC 35405	Subgingival biofilm formed on hydroxyapatite discs put upside-down on the OMM	EpiGing (MatTek, Ashland, MA)	24, 48 h	IF, CLSM, SEM, histological staining	Colonization of OMM by “red-complex” species, a colonization of Streptococci on the gingival epithelia, in the absence of all three “red complex” bacteria from the biofilm
Bao et al.58	Porphyromonas gingivalis W50 (OMZ 308), Prevotella intermedia ATCC 25611T, A. actinomycetemcomitans JP2 (OMZ 295), Campylobacter rectus (OMZ 398), Veillonella dispar ATCC 17748T, F. nucleatum subsp. Nucleatum (OMZ 598), S. oralis SK248 (OMZ 607), Treponema denticola ATCC 35405T, Actinomyces oris (OMZ 745), S. anginosus ATCC 9895, and Tannerella forsythia (OMZ 1047)	11-species biofilm formed on hydroxyapatite discs co-cultured with the OMM in the bioreactor	Immortalized epithelial cells, fibroblasts, and a monocytic cell line perfused through 3D collagen scaffold into the bioreactor	24 h	Proteomic, LC-MS/MS analysis, gene ontology (GO) analysis	Identification of 896 proteins in the supernatant and 3363 proteins in the biofilm lysate, significant regulation of the levels of F. nucleatum, Actinomyces oris, and Campylobacter rectus proteins
Bao et al.59	Porphyromonas gingivalis W50 (OMZ 308), Prevotella intermedia ATCC 25611T, A. actinomycetemcomitans JP2 (OMZ 295), Campylobacter rectus (OMZ 398), Veillonella dispar ATCC 17748T, F. nucleatum subsp. Nucleatum (OMZ 598), S. oralis SK248 (OMZ 607), Treponema denticola ATCC 35405T, Actinomyces oris (OMZ 745), S. anginosus ATCC 9895, and Tannerella forsythia (OMZ 1047)	11-species biofilm formed on hydroxyapatite discs co-cultured with the OMM in the bioreactor (37°C, 2% O2 and 5% CO2)	Immortalized epithelial cells (HGEK-16), fibroblasts (GFB-16), and a monocytic cell line perfused through 3D collagen scaffold into the bioreactor	24 h	qPCR, quantification of cytokine secretion, Masson's Trichrome Staining, SEM	Reduced growth of Campylobacter rectus, Actinomyces oris, S. anginosus, Veillonella dispar, and P. gingivalis in the presence of OMM; upregulation of cytokine release in cell culture supernatants in presence of the biofilm
Bugueno et al.60	P. gingivalis strain 33277	MOI = 100	3D microtissue of TERT-2 OKF-6 cell line on 3D spheroid of NOFs	2–24 h	Antibiotic Protection Assay, qRT-PCR, IF, SEM, TEM	Invasion of the fibroblastic core and increased apoptosis after infection
Brown et al.61	P. gingivalis W83, S. mitis NCTC 12261, S. intermedius 20753, S. oralis NTCC 11427, F. nucleatum ATCC 10596, F. spp. vincentii DSM 19507, Act. naeslundii DSM 17233, Veillonella NCTC 11831, Prevotella intermedia DSM 20706, and A. actinomycetemcomitans ATCC 43718	Three multispecies oral biofilms representative of a “health associated” (3 species), “gingivitis-associated,” (7 species), and “periodontitis associated” (10 species) grown on coverslips attached to the underside of inserts, and then placed into inserts containing the HGE tissue	HGE (Episkin, Skinethic, Lyon, France) + PBMC/CD14 + monocytes	1–2 days	H&E, LDH assay, qRT-PCR, ELISA	High viability of HGE exposed to all multispecies biofilms, more differential inflammatory response in immune cells cultured with epithelium stimulated by “gingivitis-associated” biofilm

3D, three dimensional; PBMCs, peripheral blood mononuclear cells; HGE, human gingival epithelium; MS, mass spectrometry.