Analysis of metabolic changes in murine hair follicles treated with Procyanidin-B2 rich nutraceuticals by DI-MRMS

Significance of the Topic

Hair loss affects roughly half of adults worldwide and current pharmacological options have limited long-term efficacy and potential side effects.

Nutraceuticals enriched in Procyanidin B2 have emerged as promising alternatives due to antioxidant and anti-inflammatory properties and ability to induce anagen phase in vivo.

Understanding metabolic changes underlying hair follicle response is critical to optimize preventive and therapeutic strategies.

Objectives and Overview of the Study

This study aimed to profile intracellular metabolites in murine hair follicles following topical treatment with Procyanidin B2 rich Annurca apple extract using direct infusion magnetic resonance mass spectrometry.

The goal was to elucidate pathways modulated by treatment and link metabolic shifts to keratin biosynthesis and hair growth promotion.

Methodology and Instrumentation

Wild-type C57BL6 male mice received topical application of cosmetic foam containing Annurca apple extract or placebo twice weekly for four weeks.
Hair follicle cells were isolated by EDTA mediated detachment, methanol water extraction and centrifugation.
Metabolite analysis was performed by direct infusion electrospray on a solariX XR 7T MRMS in both positive and negative ion modes.
Data were acquired over m z 100–1500 with ultra-high mass accuracy (~0.17 ppm error) and processed in MetaboScape 4.0 using HMDB lists and mSigma filtering.
Statistical evaluation employed two way ANOVA with Bonferroni post hoc tests and PLS-DA for group separation.

Main Results and Discussion

Treatment induced distinct metabolic profiles separating AAE treated follicles from controls on PLS-DA.
Key modulated pathways include:

• Reduced pentose phosphate pathway activity with lower nucleotide and deoxynucleotide levels
• Decreased glutaminolysis and glutathione biosynthesis
• Accumulation of amino acids such as glutamine glycine and arginine derivatives
• Altered glycolytic and b oxidation intermediates

These changes suggest a redirection of precursors towards keratin production rather than energy generation or nucleotide synthesis and support the observed promotion of anagen phase.

Benefits and Practical Applications

• Fast high confidence metabolite profiling offers a tool for nutraceutical efficacy evaluation and quality control
• Insights into metabolic drivers of hair growth may guide design of targeted treatments with improved safety profiles
• Approach can be extended to other botanically derived hair growth promoters

Future Trends and Potential Applications

• Validation in larger cohorts and human studies to confirm translatability of murine findings
• Comparative analysis of different Procyanidin rich extracts to identify optimal formulations
• Integration with spatial imaging and multiomics to map metabolic fluxes within follicle microenvironments
• Development of personalized nutraceutical regimens based on metabolic phenotyping

Conclusion

DI MRMS enabled rapid and accurate detection of treatment related metabolic shifts in murine hair follicles.
Annurca apple extract redirects follicular metabolism from PPP and glutaminolysis towards a keratinogenic amino acid pool.
These findings support the nutraceutical’s role in promoting anagen entry and offer a framework for future hair growth research.

References

• 1 Sonthalia S Daulatabad D Tosti A 2016 Hair Restoration in Androgenetic Alopecia Beyond Minoxidil Finasteride Hair Transplantation 2 1 13