Innovative Anti-Ageing Cream with Hyaluronic Acid and Silk Proteins: Formulation, Safety and Skin Tolerance Assessment
Source abstract
Abstract
The increasing demand for advanced cosmetic formulations based on natural biopolymers has stimulated the design of multifunctional and sustainable skin care products. Hyaluronic acid (HA) and silk proteins are widely recognized for their hydrating, barrier-supportive, and biocompatible properties. This study aimed to develop a novel topical formulation, integrating low- and medium molecular weight hyaluronic acid (LMW-HA and MMW-HA), encapsulated sodium hyaluronate (NaHA), silk, and hydrolyzed silk as active components, aiming to enhance skin barrier function and biocompatibility. The formulation was subjected to comprehensive physicochemical characterization including evaluation of appearance, odor, color, pH, viscosity, and stability, all assessed over 30 days and microbiological stability testing under controlled storage conditions. Safety evaluation followed a dual-phase strategy: (i) in silico toxicological screening of individual ingredients, including sensitization, and mutagenicity predictions, and (ii) in vivo skin compatibility assessment in 25 human volunteers using a semi-occlusive patch test. The formulation demonstrated good physicochemical stability, as pH remained stable, and viscosity showed no significant changes, confirming structural integrity, indicating preserved structural and microbiological stability throughout the study period. The in silico assessment indicated no mutagenic and/or sensitizing alerts and favorable safety margins for all components, confirming the safety profile of each ingredient, supporting their suitability for dermocosmetic use, while in vivo evaluation revealed no significant adverse effects, with irritation scores indicating no skin reaction (erythema or edema) across the test population. These findings support the potential of this novel biopolymer-based formulation as a safe and well-tolerated dermocosmetic product, aligning with principles of sustainable development and biomimetic design.
Source: crossref · Quality: baseline
Research context
OpenAlex topics: Silk-based biomaterials and applications · Advancements in Transdermal Drug Delivery · Skin Protection and Aging
SciVX discovery tags are terminology-based. OpenAlex topics are source-provided classifications.
Affiliations
Department of Analytical Chemistry and Drug Analysis, Faculty of Pharmacy, George Emil Palade University of Medicine and Pharmacy, Sciences and Technology of Târgu Mureș, 540136 Târgu Mureș, Romania
Preclinic Department, Faculty of Medicine, “Lucian Blaga” University of Sibiu, 2A Lucian Blaga Str., 550169 Sibiu, Romania
References (100+)
Show source-provided references
1. (2025, February 17). Regulation (EC) No 1223/2009 of the European Parliament and of the Council of 30 November 2009 on Cosmetic Products. Available online: https://eur-lex.europa.eu/legal-content/EN/TXT/PDF/?uri=CELEX:32013R0655.
2. Loh, X.J. (2016). Natural Rheological Modifiers for Personal Care. Polymers for Personal Care Products and Cosmetics, Royal Society Of Chemistry.10.1039/9781782623984 ↗
3. Application of Fourier Transform Infrared Spectroscopy for the Characterization of Sustainable Cosmetics and Ingredients with Antioxidant Potential10.30638/eemj.2014.013 ↗
4. Determination of Synthetic and Natural Antioxidants in Cosmetic Preparations by Solid-Phase Extraction and Subsequent Gas and High Performance Liquid Chromatographic Analysis
5. Polymers for Personal Care and Cosmetics:Overview10.1021/bk-2013-1148.ch001 ↗
6. Alves, T.F.R., Morsink, M., Batain, F., Chaud, M.V., Almeida, T., Fernandes, D.A., da Silva, C.F., Souto, E.B., and Severino, P. (2020). Applications of Natural, Semi-Synthetic, and Synthetic Polymers in Cosmetic Formulations. Cosmetics, 7.10.3390/cosmetics7040075 ↗
7. Edible Polymers: An Insight into Its Application in Food, Biomedicine and Cosmetics10.1016/j.tifs.2020.05.025 ↗
8. Trends in Biopolymer Science Applied to Cosmetics10.1111/ics.12880 ↗
9. Morganti, P., Morganti, G., and Coltelli, M.B. (2023). Natural Polymers and Cosmeceuticals for a Healthy and Circular Life: The Examples of Chitin, Chitosan, and Lignin. Cosmetics, 10.10.3390/cosmetics10020042 ↗
10. Tafuro, G., Costantini, A., Baratto, G., Francescato, S., Busata, L., and Semenzato, A. (2021). Characterization of Polysaccharidic Associations for Cosmetic Use: Rheology and Texture Analysis. Cosmetics, 8.10.3390/cosmetics8030062 ↗
11. Gawade, R.P., Chinke, S.L., and Alegaonkar, P.S. (2020). Polymers in Cosmetics. Polymer Science and Innovative Applications: Materials, Techniques, and Future Developments, Elsevier.10.1016/B978-0-12-816808-0.00017-2 ↗
12. Biotechnology of Functional Proteins and Peptides for Hair Cosmetic Formulations10.1016/j.tibtech.2021.09.010 ↗
13. Zappelli, C., Barbulova, A., Apone, F., and Colucci, G. (2016). Effective Active Ingredients Obtained through Biotechnology. Cosmetics, 3.10.3390/cosmetics3040039 ↗
14. Biocosmetics: Technological Advances and Future Outlook10.1007/s11356-021-17567-3 ↗
15. Gomes, C., Silva, A.C., Marques, A.C., Lobo, J.S., and Amaral, M.H. (2020). Biotechnology Applied to Cosmetics and Aesthetic Medicines. Cosmetics, 7.10.3390/cosmetics7020033 ↗
16. The Colors of Biotechnology: General Overview and Developments of White, Green and Blue Areas 110.1093/femsle/fny239 ↗
17. Rainbow Code of Biotechnology
18. Innovative Approaches to an Eco-Friendly Cosmetic Industry: A Review of Sustainable Ingredients10.3390/cleantechnol6010011 ↗
19. Villa, C. (2018). Green Cosmetic Ingredients and Processes. Analysis of Cosmetic Products, Elsevier. [2nd ed.].10.1016/B978-0-444-63508-2.00013-8 ↗
20. Nhani, G.B.B., Di Filippo, L.D., de Paula, G.A., Mantovanelli, V.R., da Fonseca, P.P., Tashiro, F.M., Monteiro, D.C., Fonseca-Santos, B., Duarte, J.L., and Chorilli, M. (2024). High-Tech Sustainable Beauty: Exploring Nanotechnology for the Development of Cosmetics Using Plant and Animal By-Products. Cosmetics, 11.10.3390/cosmetics11040112 ↗
21. Aneklaphakij, C., Chamnanpuen, P., Bunsupa, S., and Satitpatipan, V. (2022). Recent Green Technologies in Natural Stilbenoids Production and Extraction: The Next Chapter in the Cosmetic Industry. Cosmetics, 9.10.3390/cosmetics9050091 ↗
22. Bouzroud, S., El Maaiden, E., Sobeh, M., Merghoub, N., Boukcim, H., Kouisni, L., and El Kharrassi, Y. (2023). Biotechnological Approaches to Producing Natural Antioxidants: Anti-Ageing and Skin Longevity Prospects. Int. J. Mol. Sci., 24.10.3390/ijms24021397 ↗
23. Siahaan, E.A., Pangestuti, R., Shin, K.H., and Kim, S.K. (2022). Potential Cosmetic Active Ingredients Derived from Marine By-Products. Mar. Drugs, 20.10.3390/md20120734 ↗
24. Costa, J.P., Custódio, L., and Reis, C.P. (2023). Exploring the Potential of Using Marine-Derived Ingredients: From the Extraction to Cutting-Edge Cosmetics. Mar. Drugs, 21.10.20944/preprints202310.1518.v1 ↗
25. Maldonado-Ruiz, K., Pedroza-Islas, R., and Pedraza-Segura, L. (2024). Blue Biotechnology: Marine Bacteria Bioproducts. Microorganisms, 12.10.3390/microorganisms12040697 ↗
26. Anestopoulos, I., Kiousi, D.E., Klavaris, A., Maijo, M., Serpico, A., Suarez, A., Sanchez, G., Salek, K., Chasapi, S.A., and Zompra, A.A. (2020). Marine--derived Surface Active Agents: Health—Promoting Properties and Blue Biotechnology--based Applications. Biomolecules, 10.10.3390/biom10060885 ↗
27. Sajna, K.V., Gottumukkala, L.D., Sukumaran, R.K., and Pandey, A. (2015). White Biotechnology in Cosmetics. Industrial Biorefineries and White Biotechnology, Elsevier.10.1016/B978-0-444-63453-5.00020-3 ↗
28. Preparation, Purification, and Characterization of Low-Molecular-Weight Hyaluronic Acid10.1007/s10529-020-03035-4 ↗
29. Comprehensive Review on Biotechnological Production of Hyaluronic Acid: Status, Innovation, Market and Applications10.1080/21655979.2022.2057760 ↗
30. Pérez-Morales, G., Poggi-Varaldo, H.M., Ponce-Noyola, T., Pérez-Valdespino, A., Curiel-Quesada, E., Galíndez-Mayer, J., Ruiz-Ordaz, N., and Sotelo-Navarro, P.X. (2024). A Review of the Production of Hyaluronic Acid in the Context of Its Integration into GBAER-Type Biorefineries. Fermentation, 10.10.3390/fermentation10060305 ↗
31. Microbial Production of Hyaluronic Acid: Current State, Challenges, and Perspectives10.1186/1475-2859-10-99 ↗
32. (2025, June 18). PrimalHyalTM 50_Givaudan. Available online: https://www.givaudan.com/fragrance-beauty/active-beauty/products/primalhyal-50.
33. Huang, L., Shi, J., Zhou, W., and Zhang, Q. (2023). Advances in Preparation and Properties of Regenerated Silk Fibroin. Int. J. Mol. Sci., 24.10.3390/ijms241713153 ↗
34. Sustainable Bombyx Mori’s Silk Fibroin for Biomedical Applications as a Molecular Biotechnology Challenge: A Review10.1016/j.ijbiomac.2024.130374 ↗
35. (2025, June 18). Silkgel_Givaudan. Available online: https://www.givaudan.com/fragrance-beauty/active-beauty/products/silkgel.
36. Silk-Based Biomaterials for Tissue Engineering10.1016/j.cis.2025.103413 ↗
37. Pérez-Rivero, C., and López-Gómez, J.P. (2023). Unlocking the Potential of Fermentation in Cosmetics: A Review. Fermentation, 9.10.3390/fermentation9050463 ↗
38. Recombinant Fibrous Protein Biomaterials Meet Skin Tissue Engineering10.3389/fbioe.2024.1411550 ↗
39. Hyaluronic Acid: A Key Molecule in Skin Aging10.4161/derm.21923 ↗
40. Galvez-Martin, P., Soto-Fernandez, C., Romero-Rueda, J., Cabañas, J., Torrent, A., Castells, G., and Martinez-Puig, D. (2023). A Novel Hyaluronic Acid Matrix Ingredient with Regenerative, Anti-Aging and Antioxidant Capacity. Int. J. Mol. Sci., 24.10.3390/ijms24054774 ↗
41. Potential of Silk Proteins in Cosmetics
42. Rieger, M. (2010). Introduction to Cosmetic Emulsions and Emulsification; Published on behalf of the International Federation of Societies of Cosmetic Chemists by Micelle, Micelle Press.
43. Formulation and Characterization of Some Oil in Water Cosmetic Emulsions Based on Collagen Hydrolysate and Vegetable Oils Mixtures10.1515/pac-2018-0911 ↗
44. Loretz, L.J., and Bailey, J.E. (2007). CTFA Preservative Challenge and Stability Testing Survey—Cosmetics & Toiletries, The Cosmetic Toiletry and Fragrance Association (CTFA).
45. Emollient Structure and Chemical Functionality Effects on the Biomechanical Function of Human Stratum Corneum10.1111/ics.12656 ↗
46. Tian, Y., Zhou, J., He, C., He, L., Li, X., and Sui, H. (2022). The Formation, Stabilization and Separation of Oil–Water Emulsions: A Review. Processes, 10.10.3390/pr10040738 ↗
47. Revitalizing the Skin: Exploring the Role of Barrier Repair Moisturizers10.1111/jocd.16171 ↗
48. Juncan, A.M., Morgovan, C., Rus, L.L., and Loghin, F. (2023). Development and Evaluation of a Novel Anti-Ageing Cream Based on Hyaluronic Acid and Other Innovative Cosmetic Actives. Polymers, 15.10.3390/polym15204134 ↗
49. Influence of Packaging and Stability Test Assessment of an Anti-Aging Cosmetic Cream10.37358/MP.18.3.5043 ↗
50. Alnuqaydan, A.M. (2024). The Dark Side of Beauty: An in-Depth Analysis of the Health Hazards and Toxicological Impact of Synthetic Cosmetics and Personal Care Products. Front. Public Health, 12.10.3389/fpubh.2024.1439027 ↗
51. Barthe, M., Bavoux, C., Finot, F., Mouche, I., Cuceu-Petrenci, C., Forreryd, A., Hansson, A.C., Johansson, H., Lemkine, G.F., and Thénot, J.P. (2021). Safety Testing of Cosmetic Products: Overview of Established Methods and New Approach Methodologies (Nams). Cosmetics, 8.10.3390/cosmetics8020050 ↗
52. Juncan, A.M., Rus, L.L., Morgovan, C., and Loghin, F. (2024). Evaluation of the Safety of Cosmetic Ingredients and Their Skin Compatibility through In Silico and In Vivo Assessments of a Newly Developed Eye Serum. Toxics, 12.10.3390/toxics12070451 ↗
53. Test Guidelines for Assessment of Skin Compatibility of Cosmetic Finished Products in Man10.1016/0278-6915(96)00029-4 ↗
54. (2025, February 16). Jojoba Esters (and) Helianthus Annuus (Sunflower) Seed Wax (and) Acacia Decurrens Flower Wax (and) Polyglycerin-3 ACTICIRE MB. Available online: https://www.ulprospector.com/documents/1520629.pdf?bs=3983&b=719008&st=1&sl=149104681&crit=a2V5d29yZDpbQUNUSUNJUkVd&k=ACTICIRE&r=eu&ind=personalcare.
55. (2025, February 16). Octyldodecyl Myristate MOD MB. Available online: https://www.ulprospector.com/documents/1395526.pdf?bs=3983&b=588277&st=1&sl=149105263&crit=a2V5d29yZDpbTU9EIE1CXQ%3d%3d&k=MOD|MB&r=eu&ind=personalcare.
56. How to Choose an Emollient? Pharmaceutical and Sensory Attributes for Product Selection10.1159/000543653 ↗
57. (2025, February 16). Caprylic/Capric Triglyceride LABRAFAC CC. Available online: https://www.ulprospector.com/documents/1395523.pdf?bs=3983&b=588276&st=1&sl=149105203&crit=a2V5d29yZDpbTGFicmFmYWMgQ0Nd&k=Labrafac|CC&r=eu&ind=personalcare.
58. Amended Safety Assessment of Triglycerides as Used in Cosmetics10.1177/10915818221123790 ↗
59. (2025, February 16). Hydrogenated Ethylhexyl Olivate (and) Hydrogenated Olive Oil Unsaponifiables SOFTOLIVE. Available online: https://www.ulprospector.com/documents/1597710.pdf?bs=830&b=724179&st=1&sl=149105673&crit=a2V5d29yZDpbU29mdG9saXZlXQ%3d%3d&k=Softolive&r=eu&ind=personalcare.
60. Safety Assessment of Plant-Derived Fatty Acid Oils10.1177/1091581817740569 ↗
61. (2025, February 16). Cetyl Alcohol (and) Glyceryl Stearate (and) PEG-75 Stearate (and) Ceteth-20 (and) Steareth-20 EMULIUM DELTA MB. Available online: https://www.ulprospector.com/documents/1520632.pdf?bs=3983&b=719010&st=1&sl=149105458&crit=a2V5d29yZDpbRW11bGl1bcKuIERlbHRhIE1CXQ%3d%3d&k=Emulium%c2%ae|Delta|MB&r=eu&ind=personalcare.
62. Moisture Retention of Glycerin Solutions with Various Concentrations: A Comparative Study10.1038/s41598-022-13452-2 ↗
63. Safety Assessment of Glycerin as Used in Cosmetics10.1177/1091581819883820 ↗
64. (2025, February 16). Carbomer Carbopol®®®®®® ETD 2050. Available online: https://www.ulprospector.com/documents/1172655.pdf?bs=77&b=3755&st=1&sl=149106076&crit=a2V5d29yZDpbQ2FyYm9wb2zCriBFVEQgMjA1MF0%3d&k=Carbopol%c2%ae|ETD|2050&r=eu&ind=personalcare.
65. Bergfeld, W.F., Donald, F.A.C.P., Belsito, V., Hill, R.A., Klaassen, C.D., Liebler, D.C., Marks, J.G., Shank, R.C., Slaga, T.J., and Snyder, P.W. (2019). Amended Safety Assessment of Acrylates Copolymers as Used in Cosmetics, CIR.
66. Allergic Contact Dermatitis from Carbomers: Two Case Report10.1111/cod.13616 ↗
67. The First Comprehensive Study Evaluating the Ecotoxicity and Biodegradability of Water-Soluble Polymers Used in Personal Care Products and Cosmetics10.1016/j.ecoenv.2021.113016 ↗
68. Simultaneous Analysis of Mono-, Di-, and Tri-Ethanolamine in Cosmetic Products Using Liquid Chromatography Coupled Tandem Mass Spectrometry10.1007/s12272-015-0677-5 ↗
69. Safety Assessment of Triethanolamine and Triethanolamine-Containing Ingredients as Used in Cosmetics10.1177/1091581813488804 ↗
70. Deciphering Trends in Replacing Preservatives in Cosmetics Intended for Infants and Sensitive Population10.1038/s41598-024-69624-9 ↗
71. (2025, February 19). Phenoxyethanol (and) Ethylhexylglycerin Euxyl PE 9010 Preservative. Available online: https://www.ulprospector.com/en/eu/PersonalCare/Detail/33934/1318586/euxyl-PE-9010?st=1&sl=359373241&crit=a2V5d29yZDpbRXV4eWwgUEUg.
72. Radrezza, S., Aiello, G., Baron, G., Aldini, G., Carini, M., and D’amato, A. (2021). Integratomics of Human Dermal Fibroblasts Treated with Low Molecular Weight Hyaluronic Acid. Molecules, 26.10.3390/molecules26165096 ↗
73. Human Skin Penetration of Hyaluronic Acid of Different Molecular Weights as Probed by Raman Spectroscopy10.1111/srt.12228 ↗
74. Juncan, A.M., Moisă, D.G., Santini, A., Morgovan, C., Rus, L.L., Vonica-Țincu, A.L., and Loghin, F. (2021). Advantages of Hyaluronic Acid and Its Combination with Other Bioactive Ingredients in Cosmeceuticals. Molecules, 26.10.3390/molecules26154429 ↗
75. (2025, February 19). Hydrolyzed Hyaluronic Acid PRIMALHYAL 300. Available online: https://www.ulprospector.com/documents/1597712.pdf?bs=830&b=724176&st=1&sl=149148732&crit=a2V5d29yZDpbUHJpbWFsSHlhbOKEoiAzMDBd&k=PrimalHyal%e2%84%a2|300&r=eu&ind=personalcare.
76. (2025, February 19). Mannitol Cellulose Calcium Sodium Borosilicate CI 77492 (US: Iron Oxides) Silica CI 77891 (US: Titanium Dioxide) CI 77480 (Gold) Tin Oxide Hydroxypropyl Methylcellulose HYALUSPHERE PF. Available online: https://www.ulprospector.com/documents/1597711.pdf?bs=830&b=724168&st=1&sl=149110383&crit=a2V5d29yZDpbSFlBTFVTUEhFUkVd&k=HYALUSPHERE&r=eu&ind=personalcare.
77. Nanda, S., Nanda, A., Lohan, S., Kaur, R., and Singh, B. (2016). Nanocosmetics: Performance enhancement and Safety Assurance. Nanobiomaterials in Galenic Formulations and Cosmetics Applications of Nanobiomaterials, William Andrew.10.1016/B978-0-323-42868-2.00003-6 ↗
78. (2025, August 05). Lipotec SNAP-8TM. Available online: https://www.ulprospector.com/en/eu/PersonalCare/Detail/2316/67495/SNAP-8-peptide-solution-C?st=1&sl=500143112&crit=U05BUC044oSi&ss=2.
79. Cosmeceutical Peptides in the Framework of Sustainable Wellness Economy10.3389/fchem.2020.572923 ↗
80. Nguyen, T.T.M., Yi, E.J., Jin, X., Zheng, Q., Park, S.J., Yi, G.S., Yang, S.J., and Yi, T.H. (2024). Sustainable Dynamic Wrinkle Efficacy: Non-Invasive Peptides as the Future of Botox Alternatives. Cosmetics, 11.10.3390/cosmetics11040118 ↗
81. Anti-Aging Peptides for Advanced Skincare: Focus on Nanodelivery Systems10.1016/j.jddst.2023.105087 ↗
82. (2025, August 05). Givaudan Flashwhite Unispheres®®. Available online: https://www.ulprospector.com/en/eu/PersonalCare/Detail/830/594872/Flashwhite-Unispheres?st=1&sl=500142730&crit=Rmxhc2h3aGl0ZSBVbmlzcGh.
83. Klimek-szczykutowicz, M., Szopa, A., and Ekiert, H. (2020). Citrus Limon (Lemon) Phenomenon—A Review of the Chemistry, Pharmacological Properties, Applications in the Modern Pharmaceutical, Food, and Cosmetics Industries, and Biotechnological Studies. Plants, 9.10.3390/plants9010119 ↗
84. Choi, H.Y., Lee, Y.J., Kim, C.M., and Lee, Y.M. (2024). Revolutionizing Cosmetic Ingredients: Harnessing the Power of Antioxidants, Probiotics, Plant Extracts, and Peptides in Personal and Skin Care Products. Cosmetics, 11.10.3390/cosmetics11050157 ↗
85. Cucumber (Cucumis sativus L.) with Heterologous Poly-γ-Glutamic Acid Has Skin Moisturizing, Whitening and Anti-Wrinkle Effects10.1016/j.ijbiomac.2024.130026 ↗
86. (2025, August 05). CosIng-Cosmetics Ingredients. Available online: https://ec.europa.eu/growth/tools-databases/cosing/details/35851.
87. Bergfeld, F.A.C.P., Donald, V., Belsito, D.E., Cohen, C.D., Klaassen, A.E., Rettie, D., Ross, T.J., Slaga, P.W., Snyder, D.V.M., and Tilton, S.C. (2023). Expert Panel for Cosmetic Ingredient Safety. Safety Assessment of Hyaluronates as Used in Cosmetics, Final Report: CIR.
88. Safety Assessment of Silk Protein Ingredients as Used in Cosmetics10.1177/1091581820966953 ↗
89. Silk Fibroin Biomaterials for Tissue Regenerations10.1016/j.addr.2012.09.043 ↗
90. Evaluation of Skin Care Concerns and Patient’s Perception of the Effect of NanoSilk Cream on Facial Skin10.1111/jocd.14198 ↗
91. Silk Fibroin for Cosmetic Dermatology10.1016/j.cej.2025.159986 ↗
92. Silva, A.S., Costa, E.C., Reis, S., Spencer, C., Calhelha, R.C., Miguel, S.P., Ribeiro, M.P., Barros, L., Vaz, J.A., and Coutinho, P. (2022). Silk Sericin: A Promising Sustainable Biomaterial for Biomedical and Pharmaceutical Applications. Polymers, 14.10.3390/polym14224931 ↗
93. (2025, June 15). Guidelines on Stability Testing of Cosmetic Products. Available online: https://cosmeticseurope.eu/resources/guidelines-on-stability-testing-of-cosmetics-ce-ctfa-2004/.
94. (2018). Cosmetics-Guidelines on the Stability Testing of Cosmetic Products (Standard No. ISO/TR:18811).
95. Packaging Evaluation and Safety Assessment of a Cosmetic Product10.37358/MP.18.4.5093 ↗
96. The Safety Assessment of Cosmetic Products
97. (2005). Sensory Analysis—Methodology—General Guidance (Standard No. ISO 6658:2005).
98. (2017). Cosmetics—Microbiology—Enumeration and Detection of Aerobic Mesophilic Bacteria (Standard No. ISO 21149:2017).
99. (2017). Cosmetics—Microbiology—Enumeration of Yeast and Mould (Standard No. ISO 16212:2017).
100. (2016). Cosmetics—Microbiology—Detection of Staphylococcus Aureus (Standard No. ISO 22718:2016).