Risk-of-bias assessment of in vitro studies evaluating anti-infective drugs in vascular or endothelial cell models.
| Ref # | Study type | Drug/Class | Model/Population | Tools used | Key bias domains (summary) | Overall risk | Ageing effects (biomarkers) |
|---|---|---|---|---|---|---|---|
| [39] | In vitro | Ciprofloxacin | VSMCs | Narrative | Single cell line of human microvascular endothelial cells; high dose; endpoints limited; no replication | High | ↑ ROS, ↑ p16 expression, ↑ SA-β-gal activity (senescence) |
| [127] | In vitro | Itraconazole | Endothelial cells | Narrative | VEGFR2 assay; high dose; limited replication | Some concerns | Inhibited angiogenesis; impaired VEGFR2 signaling; vascular aging surrogate |
| [128] | In vitro | Itraconazole | HUVECs | Narrative | Glycosylation/Trafficking assays; selective endpoints | Some concerns | ↓ VEGFR2 glycosylation; impaired endothelial proliferation |
| [129] | In vitro | Itraconazole | Angiogenesis assays | Narrative | Tube formation only; no replication | Some concerns | ↓ Endothelial tube formation (angiogenesis proxy of ageing) |
| [131] | In vitro | Amphotericin B | EA.hy926 endothelial | Narrative | Supratherapeutic dose; reproducibility unclear | High | ↑ ROS, ↓ cell viability, mitochondrial dysfunction |
| [132] | In vitro | Ciclopirox | HPV+ cancer cells | Narrative | Senescence/Apoptosis endpoints only | High | ↑ SA-β-gal, ↑ p21 expression, induced senescence + apoptosis |
| [133] | In vitro | Thiabendazole | Endothelial networks | Narrative | Repurposing assay; no long-term validation | Some concerns | Vascular disruption; mimics endothelial dysfunction (an aging hallmark) |
| [31] | In vitro | Antibiotics (mitochondrial) | C. elegans fibroblasts | Narrative | Lifespan assays; mechanism not confirmed | Some concerns | ↑ Lifespan extension, ↓ mitochondrial respiration stress |
| [52] | In vitro | Bactericidal antibiotics | Mammalian cells | Narrative | ROS induction at high dose | High | ↑ ROS, mitochondrial dysfunction, DNA damage |
| [60] | In vitro | Senolytic (mitochondria) | Fibroblast cultures | Narrative | ROS dependency assays; no replication | Some concerns | ↓ Senescence phenotype when mitochondria were disrupted |
| [86] | In vitro | Aminoglycosides | Ribosomal assays | Narrative | Error clustering only | Some concerns | ↑ Translation errors; proteostasis impairment |
| [88] | In vitro | Aminoglycosides | Mammalian cells | Narrative | Aggregation assays only | Some concerns | ↑ Protein aggregation; impaired proteostasis; senescence link |
| [90] | In vitro | Azithromycin | CF epithelial cells | Narrative | Autophagy blockade assays | High | ↓ Autophagy, ↑ infection susceptibility, and senescence risk |
| [103] | In vitro | Antifungal RNA | Vulvovaginal candidiasis model | Narrative | LncRNA assays only | High | ↓ Fungal proliferation; indirect impact on senescence genes |
| [104] | In vitro | Antiviral RNA | COVID-19 assays | Narrative | Mimic/Inhibitor studies only | Some concerns | Modulated miRNA; impact on senescence/innate immune pathways |
| [107] | In vitro | Diet-microbiota | Host tissue cultures | Narrative | Epigenetic endpoints only | Some concerns | Altered DNA methylation; histone modification; epigenetic ageing link |
| [108] | In vitro | Microbiota-HDAC | Hepatocyte cultures | Narrative | Histone acetylation only | Some concerns | Global histone acetylation changes; ↑ epigenetic ageing risk |
| [114] | In vitro | HAART drugs | Human endothelial | Narrative | ROS/Endothelial stress only | High | ↑ ROS, ↑ endothelial senescence markers, ↓ NO bioavailability |
| [135] | In vitro | Niclosamide | Ageing models (cellular) | Narrative | mTORC1 suppression assays only | Some concerns | ↓ mTORC1 hyperactivation; improved frailty markers |
Narrative appraisal was applied, focusing on dose relevance, replication, and mechanistic endpoints. Ageing biomarker effects (e.g., SA-β-gal, p16/p21, mitochondrial dysfunction, ROS) are summarized (n = 19). ↑: increased/upregulated/elevated, ↓: decreased/downregulated/reduced. HAART: highly active antiretroviral therapy; HDAC: histone deacetylase; NO: nitric oxide; ROS: reactive oxygen species; VSMCs: vascular smooth muscle cells; mTORC1: mechanistic target of rapamycin complex 1; HUVECs: human umbilical vein endothelial cells.
The authors thank Lusaka Apex Medical University, School of Health Sciences, and Faculty of Medicine, for their academic and institutional support towards the completion of this manuscript.
CWS: Conceptualization, Writing—original draft, Writing—review & editing. JM: Data curation, Resources, Writing—review & editing. SKD: Methodology, Writing—review & editing, Supervision. AC: Data curation, Visualization, Writing—review & editing. LS: Validation, Resources, Writing—review & editing. KM: Investigation, Formal analysis, Writing—review & editing. All authors read and approved the submitted version.
The authors declare that they have no conflicts of interest.
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The dataset used during the study is available from the corresponding author upon request.
No funding was received for this manuscript.
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