Background
Materials and methods
Protocol development
Information sources and search strategy
Eligibility criteria
PICOS | Inclusion Criteria | Exclusion Criteria |
---|---|---|
Population
| Studies assessing cultured macrophage cells | Studies assessing cells other than macrophages |
Intervention
| Studies evaluating the effect of P. gingivalis | Studies evaluating the effect of bacteria other than P. gingivalis |
Comparison
| Studies evaluating a group of macrophages without exposure to any bacterial species | - |
Outcome
| Studies assessing the rate of foam cell formation from macrophage cells | - |
Study Design
| In-vitro studies | Case reports, narrative reviews, systematic reviews with or without meta-analysis, letters to the editors, short communications, in-vivo studies, ex-vivo studies, animal studies, and non-comparative studies. |
Study selection
Data collection and data items
Risk of bias assessment
Criteria number | Criteria | Qi et al. (2003) [35] | Kuramitsu et al. (2003) [45] | Miyakawa et al. (2004) [36] | Giacona et al. (2004) [37] | Shaik-Dasthagirisaheb et al. (2013) [42] | Li et al. (2013) [47] | Shaik-Dasthagirisheb et al. (2016) [43] | Liang et al. (2016) [46] | Kim et al. (2018) [48] | Gupta et al. (2019) [44] | Yang et al. (2020) [41] |
---|---|---|---|---|---|---|---|---|---|---|---|---|
1 | Clearly stated aims/objectives | 2 | 1 | 2 | 2 | 2 | 2 | 1 | 2 | 2 | 2 | 2 |
2 | Detailed explanation of sample size calculation | NA | NA | NA | NA | NA | NA | NA | NA | NA | NA | NA |
3 | Detailed explanation of the sampling technique | NA | NA | NA | 2 | 2 | NA | 2 | 2 | 2 | 2 | 2 |
4 | Details of the comparison group | 2 | 1 | 1 | 1 | 2 | 1 | 1 | 2 | 2 | 2 | 1 |
5 | Detailed explanation of the methodology | 2 | 2 | 2 | 2 | 2 | 2 | 2 | 2 | 2 | 2 | 2 |
6 | Operator details | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 |
7 | Randomization | NA | NA | NA | NA | NA | NA | NA | NA | NA | NA | NA |
8 | Method of measurement of outcome | 2 | 2 | 2 | 2 | 2 | 2 | 2 | 2 | 2 | 2 | 2 |
9 | Outcome assessor details | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 |
10 | Blinding | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 |
11 | Statistical analysis | 0 | 0 | 0 | 2 | 2 | 2 | 0 | 2 | 2 | 2 | 2 |
12 | Presentation of results | 2 | 2 | 2 | 2 | 2 | 2 | 0 | 2 | 2 | 2 | 2 |
Total score | 55.55% | 44.44% | 50% | 65% | 70% | 61.11% | 40% | 70% | 70% | 70% | 65% | |
Risk of bias | Medium | High | Medium | Medium | Medium | Medium | High | Medium | Medium | Medium | Medium |
Results
Study selection
Study characteristics
Author (Year) | Type(s) of Evaluation | Method(s) | Main Outcome(s) | References |
---|---|---|---|---|
Yang et al. (2020) | Foam cell formation | Oil Red O staining | The knockdown of limp2 reduces the rate of foam cell formation and enhances cholesterol export. The interaction of LIMP2 and caveolin-1 (CAV1) in the lysosome of macrophages may play a key role in this regard. | [41] |
Cathepsin L activity | Magic red cathepsin L assay | |||
RNA sequencing | RT-PCR | |||
Protein detection | Western blot | |||
Protein detection | Co-immunoprecipitation | |||
Gupta et al. (2019) | Binding and uptake of oxLDL | Fluorescence intensity microscopy | TRPV4 plays a key part in foam cell formation and inflammatory genes upregulation, which is subsequent to LDL oxidation. This process was also induced by P. gingivalis LPS. | [44] |
Foam cell formation | Oil Red O Staining | |||
Expression levels of TRPV4, actin, or CD36 | Immunoblot and immunofluorescence assay | |||
Kim et al. (2018) | Oxidation extent of HDL or LDL | TBARS assay | HDL incubated with P. gingivalis showed significantly higher oxidation levels and TNF- α production. P. gingivalis induces HDL oxidation, by proinflammatory response in interaction with macrophages. | [48] |
TNF-α | ELISA | |||
The activity of MMPs, and Gelatinase | Electrophoresis, Gelatin zymography | |||
Foam cells | Oil Red O staining | |||
Liang et al. (2016) | Foam cell formation | Oil Red O staining | P. gingivalis can induce foam cell formation through the upregulation of CD36 expression in macrophages. CD36 expression in the presence of P. gingivalis is mediated by NF-κB, ERK1/2, and p65. | [46] |
NF-κB activity | RT-PCR | |||
CD36 protein levels | Western blot | |||
NF-κB activity | Luciferase reporter assay | |||
The interaction of NF-κB and CD36 promoters | EMSA | |||
The interaction of NF-κB and CD36 promoters | Chromatin immunoprecipitation assay | |||
CD36 protein levels | Flow cytometry | |||
Shaik-Dasthagirisheb et al. (2016) | Foam cell formation | Oil Red O Staining | Both P. gingivalis and C. pneumonia can induce foam cell formation in macrophages. | [43] |
Lipid peroxidation | TBARS assay for level of oxidized LDL | P. gingivalis enhances LDL oxidation while no statistical difference was reported between the species. | ||
Inflammatory cytokines secretion | ELISA | Both P. gingivalis and C. pneumonia enhance TNF-α and IL-6 secretion from LDL-treated macrophages. | ||
Gene expression | PCR | Despite the differences between P. gingivalis and C. pneumonia, they indicate a similar pattern in activation and down-regulation of genes in macrophages. | ||
Li et al. (2013) | Foam cell formation | Oil Red O staining | P. gingivalis LPS can promote foam cell formation in ox-LDL-treated macrophages. P. gingivalis LPS could enhance CD36 mRNA expression which acts as a mediator receptor for lipid uptake and decrease the cholesterol efflux by down-regulation of ABCA1. | [47] |
Cholesterol efflux | Cholesterol efflux assay | |||
Expression of ABCA1, CD36 | RT-PCR | |||
HO-shRNA level | Western blot | |||
Shaik-Dasthagirisaheb et al. (2013) | Foam cell formation | Oil Red O staining | The sole addition of P. gingivalis to macrophages could enhance foam cell formation; however, the sole addition of LDL did not demonstrate the same effect. Moreover, heat-killed P. gingivalis had a similar effect on foam cell formation compared to alive P. gingivalis, regardless of the presence or the absence of LDL. | [42] |
MyD88 and lps2 gene’s role in foam cell formation | Oil Red O staining | In both concurrent and uncoupled methods, MyD88 gene knockout demonstrated substantial reductions in a number of foam cells compared to the naïve types. However, in the presence of LDL lps2-knockout mice formed foam cells similar to naïve types. | ||
Effect of P. gingivalis dose on Foam cell formation | Oil Red O staining | Enhanced concentrations of P. gingivalis (MOI of 1, 10, and 100), regardless of the concurrent or uncoupled LDL treatment, elicited a greater percentage of foam cells | ||
Effect of LDL on the production of inflammatory cytokines | ELISA | The elevated levels of LDL significantly decrease the pro-inflammatory cytokine production by macrophages cultured with P. gingivalis. | ||
Giacona et al. (2004) | Foam cell formation | Oil Red O staining | The results indicate the higher effect of naïve P.g compared to fimbria-deficient P. gingivalis to induce foam cell formation. | [37] |
Recovery of viable P. gingivalis from antibiotic-treated macrophages | Antibiotic protection assay | Recovery of naïve P. gingivalis species was significantly higher than the fimbria-deficient ones. | ||
Uptake of P. gingivalis by macrophages | Transmission electron microscopy | The naïve P. gingivalis types are more capable in adhering and entering the macrophage cells than the fimbria-deficient ones. | ||
Miyakawa et al. (2004) | Foam cell formation by aggregated LDL | Oil Red O staining | P. gingivalis and its OMVs induce dose-dependent LDL aggregation and eventually foam cell formation, which is in part performed by the proteolysis of apo B-100 protein that is involved in the transportation of LDL. | [36] |
LDL aggregation | Transmission electron microscope | |||
SDS–PAGE and western blotting | ||||
LDL modification | Relative electrophoresis mobility (REM) shift assays | |||
Kuramitso et al. (2003) | Foam cell formation | Oil Red O staining | P. gingivalis promotes foam cell formation which the most important element in this regard seems to be the P. gingivalis LPS. Moreover, P. gingivalis can induce MCP-1 secretion in endothelial cells. | [45] |
MCP-1 | ELISA | |||
Qi et al. (2003) | Effect of P. g foam cell formation | Oil Red O staining | P. gingivalis LPS alone cannot induce foam cell formation by itself. The presence of P. gingivalis and its’ OMVs can modify LDL and induce foam cell formation. | [35] |
Effect of OMV on foam cell formation | ||||
Effect of LPS on foam cell formation | ||||
Effect of LDL-uptake on foam cell formation | Fluorescence imaging of LDL binding to macrophages | |||
LDL modification by P. gingivalis during foam cell formation | Agarose gel electrophoresis | |||
LDL peroxidation induced by P. gingivalis | TBARS assay |
Results of individual studies
Authors (Year) | Key molecular element | Mechanism of Action | References |
---|---|---|---|
Yang et al. (2020) | LIMP2 | P. gingivalis induces foam cell formation via NF-κB and JNK pathways, which enhance the expression of LIMP2, caveolin-1 (CAV-1), and their interactions. | [41] |
Gupta et al. (2019) | TRPV4 | TRPV4 can regulate oxLDL uptake in macrophages and this mechanosensitive channel is sensitive to the extracellular matrix stiffness induced by P. gingivalis LPS. | [44] |
Kim et al. (2018) | HDL | P. gingivalis can induce HDL oxidation, which prevents its athero-protective effects and promotes athero-inductive effects by eliciting pro-inflammatory cytokines secretion. | [48] |
Liang et al. (2016) | CD36, NF-κB, ERK1/2, and p65 | The P. gingivalis infection can cause CD36 upregulation through the pathways mediated by NF-κB, ERK1/2, and p65. | [46] |
Shaik-Dasthagirisaheb et al. (2016) | Modification of genes subsequent in macrophage-infected P. gingivalis | P. gingivalis can up-regulate and down-regulate the genes involved in lipid uptake and efflux, respectively. P. gingivalis can also enhance the expression of genes associated with inflammatory biomarkers, cell adhesion, and ECM modification. | [43] |
Li et al. (2013) | P. gingivalis LPS, CD36, ABCA-1, calpain, HO-1 | P. gingivalis LPS induces foam cell formation through HO-1 expression, which results in the activation of the cJun/AP-1 pathway that can promote upregulation of CD36 and downregulation of ABCA-1via upregulation of calpain activity. | [47] |
Shaik-Dasthagirisaheb et al. (2013) | P. gingivalis LPS, Myeloid differentiation factor 88 (MyD88) | P. gingivalis LPS can induce foam cell formation, regardless of the presence or the absence of LDL. Moreover, the knockout of the MyD88 gene can markedly reduce foam cell formation. | [42] |
Miyakawa et al. (2004) | OMV | P. gingivalis and its OMVs could induce LDL aggregation in a dose-dependent manner by proteolysis of apo B-100 protein and modification of LDL to induce higher mobility of the final LDLs. | [36] |
Giacona et al. (2004) | P. gingivalis fimbria | The major fimbria of P. gingivalis plays a key role in inducing foam cell formation and P. gingivalis invasion into the macrophage cells. Moreover, the major fimbria enhances the recovery of P. gingivalis in the presence of antibiotics. | [37] |
Kuramitso et al. (2003) | P. gingivalis fimbria, P. gingivalis LPS, MCP-1 | The induction of MCP-1 secretion from the endothelial cells, caused by P. gingivalis, can attract more monocytes to the site and accelerate the process of foam cell formation and eventually, atherosclerosis. | [45] |
Qi et al. (2003) | P. gingivalis LPS, OMV | Induction of cholesterol binding and intake by macrophages | [35] |