Human oral epithelial cell culture I. Improved conditions for reproducible culture in serum-free medium

Oda, Dolphine; Watson, Eileen L.

doi:10.1007/bf02624208

Cited by 115 publications

(84 citation statements)

References 18 publications

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“…After mincing 1-2 mm 2 pieces, the epithelium fragments were incubated in trypsin-EDTA at 37°C for 10 minutes to isolate HGECs. 29,30 Then, the cells were collected and fed with defined keratinocyte-serum-free medium (Thermo Fisher Scientific) supplemented with 1% penicillin/streptomycin, with culture media refreshed every 2 days. The remaining gingival connective tissues were placed in DMEM containing 10% FBS and 1% penicillin/ streptomycin, where they were cut into 2-3 mm 2 -sized pieces.…”

Section: Behaviors Of Hgecs and Hgfs On Different Samplesmentioning

confidence: 99%

The response of human osteoblasts, epithelial cells, fibroblasts, macrophages and oral bacteria to nanostructured titanium surfaces: a systematic study

Miao¹,

Wang²,

Xu³

et al. 2017

IJN

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Nanotopography modification is a major focus of interest in current titanium surface design; however, the influence of the nanostructured surface on human cell/bacterium behavior has rarely been systematically evaluated. In this study, a homogeneous nanofiber structure was prepared on a titanium surface (Nano) by alkali-hydrothermal treatment, and the effects of this Nano surface on the behaviors of human MG-63 osteoblasts, human gingival epithelial cells (HGECs) and human gingival fibroblasts (HGFs) were evaluated in comparison with a smooth titanium surface (Smooth) by polishing and a micro-rough titanium surface (Micro) by sandblasting and acid etching. In addition, the impacts of these different surface morphologies on human THP-1 macrophage polarization and Streptococcus mutans attachment were also assessed. Our findings showed that the nanostructured surface enhanced the osteogenic activity of MG-63 cells (Nano=Micro>Smooth) at the same time that it improved the attachment of HGECs (Nano>Smooth>Micro) and HGFs (Nano=Micro>Smooth). Furthermore, the surface with nanotexture did not affect macrophage polarization (Nano=Micro=Smooth), but did reduce initial bacterial adhesion (Nano

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Section: Behaviors Of Hgecs and Hgfs On Different Samplesmentioning

confidence: 99%

The response of human osteoblasts, epithelial cells, fibroblasts, macrophages and oral bacteria to nanostructured titanium surfaces: a systematic study

Miao¹,

Wang²,

Xu³

et al. 2017

IJN

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“…Using an informed consent protocol that was reviewed and approved by the Human Subjects Institutional Review Board of the University of Minnesota, healthy human gingiva was excised from sites overlying impacted third molar teeth. Gingival keratinocytes were isolated and cultured by the method of Oda and Watson (30) and passaged in KGM (Clonetics; BioWhittaker, Inc., Walkersville, Md.). The cells were propagated through successive passages to senescence.…”

Section: Methodsmentioning

confidence: 99%

Calprotectin Expression by Gingival Epithelial Cells

Ross

Herzberg

2001

Infect Immun

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Calprotectin, a heterodimer of MRP8 and MRP14 with antimicrobial properties, is found in the cytosol of neutrophils, monocytes, and human gingival keratinocytes. During inflammation of the oral mucosa, the expression of immunoreactive calprotectin appears upregulated. Given the possible cell sources, we sought to learn if epithelial cells upregulate calprotectin in response to proinflammmatory agents. First, human gingival keratinocytes were maintained in primary culture until senescence. At each passage, cells were harvested and analyzed for quantitative expression of MRP8 and MRP14 subunit mRNA by RNase protection assays and calprotectin complex by enzyme-linked immunosorbent assay. Calprotectin expression was constitutive in the primary gingival keratinocytes, but calprotectin-specific mRNA and protein tended to increase as the cells neared senescence. To test whether calprotectin expression was inducible, immortalized gingival keratinocyte cultures were treated for 2 to 4 h with lipopolysaccharide (LPS) or interleukin-1␤ (IL-1␤). As a positive control for inducible expression, immortalized keratinocytes were incubated with phorbol myristate acetate (PMA) (50 ng/ml) for 24 h. Incubation with PMA stimulated increased expression of MRP8 and MRP14 mRNA within 2 h, peaking within 5 h. MRP8-and MRP14-specific mRNA expression by immortalized keratinocytes appeared to be unaffected by LPS or IL-1␤. In contrast, LPS, IL-1␤, and PMA each upregulated IL-8. These data show that calprotectin mRNA is expressed constitutively in cultured keratinocytes, while expression by immortalized cells appears to be independent of the exogenous proinflammatory agents LPS and IL-1␤.Calprotectin appears to be constitutively expressed by squamous mucosal epithelia but is expressed by normal epidermis only in certain inflammatory skin diseases. During inflammation of the skin and mucosa, keratinocytes can actively contribute to immunological events that occur by expressing class II major histocompatibility complex (MHC) molecules (2), cell adhesion molecules (4, 14), and various cytokines (22,40). Concomitantly, calprotectin is induced in skin and may be upregulated in mucosal epithelium as detected immunohistochemically. A major constituent in the cytoplasm of granulocytes and monocytes, calprotectin constitutes up to 45 to 60% of the total cytosolic protein in neutrophils (9, 13) but is absent in resident tissue macrophages. The biological functions of the calprotectin complex are poorly defined.Calprotectin is a heterodimer of two noncovalently associated polypeptides, MRP8 and MRP14 (10.8 and 14 kDa, respectively). The calprotectin heterodimer is anionic and calcium binding. MRP8-MRP14 (31) synonyms include cystic fibrosis antigen (8), L1 antigen (1, 7), calgranulin A and B (42), and S100A8 and S100A9 (36, 43). MRP8 and MRP14 belong to the S100 protein family, whose members are involved in cell cycle progression, cell differentiation, and cytoskeletal membrane interactions with the plasma membrane (20). MRP14 phosphorylates in both mo...

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“…Primary cultures of GECs were generated as described previously (36,48). Briefly, healthy gingival tissue was collected from patients undergoing surgery for removal of impacted third molars, following Institutional Review Board guidelines.…”

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confidence: 99%

Role of the Clp System in Stress Tolerance, Biofilm Formation, and Intracellular Invasion inPorphyromonas gingivalis

et al. 2008

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Clp proteases and chaperones are ubiquitous among prokaryotes and eukaryotes, and in many pathogenic bacteria the Clp stress response system is also involved in regulation of virulence properties. In this study, the roles of ClpB, ClpC, and ClpXP in stress resistance, homotypic and heterotypic biofilm formation, and intracellular invasion in the oral opportunistic pathogen Porphyromonas gingivalis were investigated. Absence of ClpC and ClpXP, but not ClpB, resulted in diminished tolerance to high temperatures. Response to oxidative stress was not affected by the loss of any of the Clp proteins. The clpC and clpXP mutants demonstrated elevated monospecies biofilm formation, and the absence of ClpXP also enhanced heterotypic P. gingivalis-Streptococcus gordonii biofilm formation. All clp mutants adhered to gingival epithelial cells to the same level as the wild type; however, ClpC and ClpXP were found to be necessary for entry into host epithelial cells. ClpB did not play a role in entry but was required for intracellular replication and survival. ClpXP negatively regulated the surface exposure of the minor fimbrial (Mfa) protein subunit of P. gingivalis, which stimulates biofilm formation but interferes with epithelial cell entry. Collectively, these results show that the Clp protease complex and chaperones control several processes that are important for the colonization and survival of P. gingivalis in the oral cavity.

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Human oral epithelial cell culture I. Improved conditions for reproducible culture in serum-free medium

Cited by 115 publications

References 18 publications

The response of human osteoblasts, epithelial cells, fibroblasts, macrophages and oral bacteria to nanostructured titanium surfaces: a systematic study

The response of human osteoblasts, epithelial cells, fibroblasts, macrophages and oral bacteria to nanostructured titanium surfaces: a systematic study

Calprotectin Expression by Gingival Epithelial Cells

Role of the Clp System in Stress Tolerance, Biofilm Formation, and Intracellular Invasion inPorphyromonas gingivalis

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