Skin aging is characterized by wrinkles, hyperpigmentation, and decreased skin elasticity, which are partly caused by decreased collagen levels. One potential source of collagen to overcome skin aging is fish collagen which is rich in type I collagen. This study aims to develop hydrolyzed collagen (HC) from fish collagen using enzymatic hydrolysis with bromelain and thermal hydrolysis methods. The obtained HC was formulated into four microsphere formulas, namely native collagen (F1), unfractionated HC (F2), HC with a molecular weight of more than 30 kDa (F3), and HC with a molecular weight of less than 30 kDa (F4). The effectiveness of HC and microspheres was evaluated through in vitro and clinical tests. The enzymatic hydrolysis process produced a degree of hydrolysis (DH) of 60.9%; while thermal hydrolysis only reached 20%. HC characterization was carried out using SEM, electrophoresis, ATR-FTIR, and DSC, which showed significant differences between HC and native collagen. Microspheres F1, F2, F3, and F4 were successfully formulated with spherical shape, particle size between 2.23–5.00 µm, moisture content less than 5%, and yield more than 90%. The entrapment efficiency ranged from 38.86–61.80%, with drug loading values of 23.85% (F1); 22.29% (F2); 23.50% (F3); and 14.78% (F4). The characterization results showed that the type of collagen and HC affected drug loading and entrapment efficiency, but did not affect yield, morphology, and moisture content. In vitro testing showed that HC significantly increased NIH-3T3 fibroblast proliferation compared to control and native collagen, with a greater increase in HC with a molecular weight of less than 30 kDa. Clinical trials using a randomized, placebo-controlled, blind study method showed that HC microspheres significantly increased skin hydration, thickness, and density compared to other formulations. From this study, HC can be produced using bromelain. HC microspheres can be produced with a spherical shape and a size of less than 5 µm. The characteristics of HC are influenced by the molecular weight of collagen as an active ingredient. HC can increase the number of fibroblast cells and improve skin characteristics, namely reducing wrinkles, and increasing skin hydration, thickness and density.
Keywords: fish collagen, hydrolyzed collagen, microspheres, anti-aging
