Effects of a Dietary Supplement on Collagen Production and Evaluation of Its Regenerative and Anti-Ageing Propertiess

Ageing of the skin is a multifactorial biological process characterized by progressive structural and functional decline of the dermis. Dermal fibroblasts are the principal cells responsible for the synthesis and maintenance of the extracellular matrix (ECM), including type I and type III collagen, which are essential for maintaining skin integrity, elasticity, and mechanical strength. During ageing, fibroblasts progressively lose proliferative capacity and enter a state of replicative senescence associated with irreversible cell-cycle arrest and altered cellular function. This transition is driven by mechanisms such as telomere shortening, oxidative stress, mitochondrial dysfunction, and persistent DNA damage responses, ultimately leading to reduced collagen synthesis and impaired tissue repair.

In parallel, ageing is accompanied by increased oxidative and glycative stress, which contribute to cumulative molecular damage. Oxidative modification of proteins results in the formation of protein carbonyls, widely recognized as stable markers of oxidative protein damage. Glycation reactions lead to the formation of Advanced Glycation End Products (AGEs), which accumulate in long-lived proteins such as collagen, promoting crosslinking, reduced tissue elasticity, and impaired extracellular matrix remodeling. These processes are further amplified by the accumulation of senescent fibroblasts, which secrete pro-inflammatory and matrix-remodeling factors, contributing to chronic low-grade inflammation and progressive tissue deterioration.

Targeting these interconnected mechanisms, including extracellular matrix metabolism, oxidative stress, glycation, and proteostasis, represents a promising strategy for supporting healthy ageing. Nutraceutical formulations containing natural bioactive compounds have emerged as potential modulators of these pathways, with evidence suggesting their ability to enhance antioxidant defenses, reduce protein damage, and support collagen biosynthesis.

The present study applies a translational framework to evaluate the biological effects of a collagen-supporting nutraceutical formulation. The investigation included an initial in vitro component using human fibroblasts to assess cellular responses related to ageing dynamics, followed by a randomized, placebo-controlled clinical intervention in healthy adults to determine systemic effects.

In the clinical study, participants received either the nutraceutical formulation or a matched placebo administered orally for six consecutive weeks. The formulation consisted of a combination of structural components of collagen, amino acids involved in collagen biosynthesis, antioxidants, enzymatic cofactors, and micronutrients supporting skin and cellular homeostasis. Specifically, each tablet contained hydrolyzed marine collagen peptides (200 mg) and hyaluronic acid (100 mg) to support extracellular matrix structure and hydration. It also included key amino acids required for collagen synthesis, namely glycine (120 mg), proline (60 mg), and hydroxyproline (60 mg), which contribute to collagen formation, stabilization, and maturation.

To address oxidative stress and molecular damage, the formulation incorporated antioxidant compounds including quercetin (50 mg), reduced glutathione (50 mg), and astaxanthin (0.1 mg), aiming to enhance cellular redox balance and protect proteins and membranes from oxidative modification. In addition, vitamin C (80 mg) and zinc (10 mg) were included as essential cofactors for enzymatic processes involved in collagen biosynthesis. The formulation was further supplemented with vitamin D3 (30 μg, 1200 IU) and biotin (100 μg) to support cellular homeostasis, skin health, and metabolic regulation. Excipients were included to ensure stability and appropriate tablet formulation.

Blood samples were collected at baseline and at the end of the intervention period to evaluate changes in circulating biomarkers associated with key biological processes relevant to ageing. The selected biomarkers included Pro-Collagen I Alpha as an indicator of collagen biosynthesis and extracellular matrix turnover, protein carbonyls as markers of oxidative protein damage, and Advanced Glycation End Products as markers of glycation burden. Together, these biomarkers provide complementary information on collagen metabolism, oxidative stress, and protein modification, allowing for an integrated assessment of biological ageing processes.

The study is designed to determine whether supplementation with the formulation can modulate these biomarkers in a manner consistent with improved extracellular matrix homeostasis and reduced molecular damage. By focusing on measurable systemic endpoints linked to fundamental mechanisms of ageing, the study aims to provide evidence for the potential role of targeted nutraceutical interventions in supporting skin health and broader healthy ageing trajectories.

Inclusion criteria

Adults aged 29 to 82 years Individuals with Fitzpatrick skin types I-VI Presence of mild to moderate fine lines and wrinkles Presence of mild to moderate photoaging consistent with chronological ageing Generally healthy individuals without known medical conditions that, in the opinion of the investigator, could interfere with study participation Ability to understand the study procedures and provide written informed consent

Exclusion criteria

• Body mass index (BMI) greater than 28 kg/m² Current diagnosis of cancer and/or undergoing chemotherapy or radiotherapy Use of dietary supplements during the study period Ongoing long-term treatment for autoimmune or chronic diseases Pregnancy, breastfeeding, or intention to become pregnant during the study period Severe photoaging beyond mild to moderate classification History of cosmetic procedures or dermatological conditions that, in the opinion of the investigator, could interfere with accurate skin assessment, including but not limited to rosacea, acne, or excessively oily or dry skin Known hypersensitivity or allergic reaction to any component of the study product