Senescence in Skin Aging - Quantifying methods and anti-aging treatments
Typ
Examensarbete för masterexamen
Master's Thesis
Master's Thesis
Program
Biotechnology (MPBIO), MSc
Publicerad
2024
Författare
Öberg, Ylva
Modellbyggare
Tidskriftstitel
ISSN
Volymtitel
Utgivare
Sammanfattning
Aging is characterized by gradual and continuous deterioration of physiological functions over time. Aging research explores the underlying cellular mechanisms and genetic pathways involved in the decline, with the final goal of therapeutic and cosmetic interventions for age-related alterations and pathologies. Senescence has been identified as a central hallmark of aging. With age, our immune system and DNA damage repair mechanisms become less efficient, resulting in an accumulation of senescent cells. They cease to proliferate and grow, irreversibly exit the cell cycle, thus causing a loss of genome integrity. The development of anti-aging interventions relies substantially on research into methods to measure and explore the underlying mechanisms of senescence.
To understand the process of skin aging, this study is looking at cellular senescence, with special emphasis on replicative-induced senescence in Human Dermal Fibroblasts. In the quest for multiple tools to objectively measure cellular senescence in this cell model, an in-vitro cell-based model was developed based on established quantifying methods: NAD+/NADH-Glokit, Quantitative Real-Time PCR (qPCR), Flow Cytometry, SA-β-Galactosidase staining and Immunofluorescence for C12FDG. The assays included in the proposed model were validated by demonstrating a statistically significant difference between high and low-passage HDFs. The model is a good starting point to objectively quantify cellular senescence in skin aging research. However, the model at this stage does not describe the senescent phenotype in its full complexity; more biomarkers need to be taken into account. Research is ongoing and further research is needed to expand the tools in the toolbox.
Furthermore, this study applied the NAD+/NADH ratio of the model to test the effects certain flavonoid treatments (Quercetin, Apigenin and Kaempferol) have on skin senescence modulation, as compared to the chemical 78c treatment proven to reverse age-associated metabolic decline. Only treatment with low concentrations of Apigenin successfully increased the NAD+/NADH ratio in high-passage cells. However, further research is needed to confirm the results of the limited sample size in this study and to draw any steadfast conclusions.
Beskrivning
Ämne/nyckelord
skin aging , replicative-induced senescence , aging research , assay , human dermal fibroblasts , skin senescence modulation , NAD+/NADH-Glo™assay , RT-qPCR , SA-β-Galactosidase staining , C12FDG , flow cytometry , 78c , Quercetin , Apigenin , Kaempferol