Introduction

Trehalose is commonly used as a moisturizer in skincare products, but its effects do not arise solely from its multiple hydroxyl groups attracting water.

Trehalose consists of two D-glucopyranose units. The anomeric carbon C1 of each glucose unit is linked through an oxygen atom to form an α,α-1,1-O-glycosidic bond, namely a C1—O—C1 linkage. Because both anomeric positions are occupied by the glycosidic bond, trehalose has no free reducing end and is therefore a nonreducing disaccharide. The remaining eight hydroxyl groups in the molecule remain free and can participate in hydration and hydrogen-bonding interactions.

In aqueous environments, trehalose participates in sugar—water networks through multiple hydrogen bonds, affecting the arrangement and motion of local water molecules. As water content gradually decreases, trehalose can also interact with proteins, the polar groups of phospholipids, and residual water, and it can form a highly viscous amorphous matrix under low-moisture conditions.

The effects of trehalose can be summarized as follows: the α,α-1,1-O-glycosidic bond and eight free hydroxyl groups confer nonreducing properties and multivalent hydration capacity; the dynamic sugar—water hydrogen-bonding network slows the reorientation of hydration water; and as water content decreases, trehalose can also participate in interfacial hydrogen bonding and residual-water retention, which may help maintain the relative stability of the hydration environment at the skin surface and in the outer stratum corneum.

1 Molecular Structure of Trehalose

1.1 Two Glucose Units Are Linked Through Their Anomeric Positions

The molecular formula of trehalose is C₁₂H₂₂O₁₁, and its relative molecular mass is 342.30. Its systematic structural name is:

α-D-glucopyranosyl α-D-glucopyranoside

Its structure can be abbreviated as:

α-D-Glcp-(1↔1)-α-D-Glcp

Here, Glcp represents a glucopyranose residue. Both glucose residues in trehalose are in the D-pyranose form, and both anomeric carbon atoms, C1, have the α configuration. They are linked through a bridging oxygen atom to form a C1—O—C1-type O-glycosidic bond; therefore, this linkage is also referred to as an α,α-1,1-O-glycosidic bond.[1] Trehalose consists of two glucose units linked through an α,α-1,1-glycosidic bond and is a nonreducing disaccharide.

1.2 The α,α-1,1 Structure Makes Trehalose a Nonreducing Sugar

Free glucose can interconvert between a cyclic hemiacetal structure and an open-chain aldehyde structure. The carbonyl group in the open-chain structure can participate in reduction reactions, nonenzymatic glycation, and browning reactions.

Maltose, for example, contains a glycosidic bond linking C1 of the first glucose unit to C4 of the second glucose unit. C1 of the second glucose unit retains a free anomeric hydroxyl group and can interconvert between the cyclic hemiacetal structure and the open-chain form. Maltose is therefore a reducing sugar. In trehalose, both C1 anomeric positions participate in glycosidic-bond formation. There is no free anomeric hydroxyl group or hemiacetal end, and a free aldehyde group cannot form through the normal ring-opening equilibrium. Trehalose is therefore a nonreducing disaccharide.

This structure provides three properties relevant to skincare formulations:

The nonreducing property does not mean that trehalose remains unchanged under all conditions. Strong acids, high temperatures, or trehalase can hydrolyze the glycosidic bond. Under common skincare-product storage conditions, however, trehalose is generally less chemically reactive than sugars with a free reducing end.

1.3 The Characteristics of Trehalose Are Not Determined Solely by the Number of Hydroxyl Groups

Trehalose contains eight free hydroxyl groups, but the number of hydroxyl groups in a single molecule does not directly determine its actual moisturizing capacity.

The relative molecular mass of trehalose is approximately 342.30, compared with approximately 180.16 for glucose and 92.09 for glycerol. At the same mass concentration, glycerol contains substantially more molecules than trehalose. Therefore, the fact that trehalose contains multiple hydroxyl groups alone does not demonstrate that its immediate hygroscopic capacity is stronger than that of glycerol or ordinary monosaccharides.

The structural characteristics of trehalose arise from the following combination:

Nonreducing property ＋ multivalent hydration ＋ stable glycosidic linkage ＋ ability to form an amorphous phase under low-moisture conditions

This combination of properties enables trehalose not only to participate in hygroscopic moisturization but also to provide a degree of protection to biomolecules and membrane structures as water content decreases.

2 How Trehalose Regulates the State of Water

2.1 Hydroxyl Groups Form a Dynamic Sugar—Water Hydrogen-Bonding Network

After trehalose dissolves, the hydroxyl groups in the molecule can form hydrogen bonds with water. A water molecule can interact with one or more hydroxyl groups on trehalose and may also bridge two different trehalose molecules.

The interaction can be represented as:

Trehalose—OH···OH₂···OH—Trehalose

The dotted lines represent hydrogen bonds.

These hydrogen bonds are continuously formed, broken, and rearranged. Trehalose does not permanently immobilize water. Instead, it increases the probability that water molecules remain within the hydration environment of the sugar molecules, making the local state of water different from that of pure water.

2.2 Trehalose Prolongs the Time Required for Surrounding Water Molecules to Reorient and Reorganize Their Hydrogen Bonds

When evaluating the hydration effects of sugars, two concepts must be distinguished:

① Hydration number: the number of water molecules around a sugar molecule that are influenced by it.

② Hydration dynamics: the rates at which these water molecules reorient, rotate, and diffuse.

A 2022 broadband dielectric spectroscopy study showed that, compared with maltose, trehalose strengthens hydrogen-bonding interactions among water molecules and forms a relatively uniform aqueous environment, while the difference in hydration number between the two sugars is relatively small.[2]

A 2025 comparative study of trehalose, maltose, and sucrose further found that the three sugars did not show a clear trehalose-specific difference in hydration number or in the cooperativity of water-molecule reorientation. However, the reorientation time of water molecules surrounding trehalose was longer.[3]

This process can be represented as:

Water molecules enter the trehalose hydration environment → hydrogen bonds are continuously exchanged → reorientation time is prolonged → local hydration dynamics change at the molecular scale

2.3 Trehalose Moisturization Involves Three Levels

3 How Trehalose Protects Biological Structures as Water Content Decreases

3.1 Water Loss from the Stratum Corneum Is Accompanied by Structural Changes

Water in the stratum corneum participates in regulating keratin flexibility, the dissolved state of natural moisturizing factors, and the headgroup spacing and phase state of intercellular lipids.

As water content decreases, the following changes may occur in the stratum corneum:

① The keratin hydration layer becomes thinner, and interactions among proteins become stronger.

② The flexibility of corneocytes decreases, and mechanical brittleness increases.

③ The dissolution and distribution of natural moisturizing factors change.

④ The headgroup spacing, lamellar structure, and phase state of intercellular lipids may change.

⑤ The skin surface may become tight, rough, and flaky.

Studies using protein and phospholipid-membrane models suggest that trehalose can participate in hydrogen-bonding networks among water, proteins, and lipids during water loss, thereby reducing some of the structural disturbances caused by rapid changes in the hydration environment. The actual extent of this effect in the human stratum corneum remains to be verified.

3.2 Water-Replacement Mechanism

Under fully hydrated conditions, water molecules are arranged around carbonyl, hydroxyl, amino, and charged groups on protein surfaces. Water molecules also form hydrogen bonds with phosphate and carbonyl groups in phospholipid headgroups.

When interfacial water decreases, the spacing between phospholipid headgroups may decrease, and new interactions may form within proteins or between different proteins.

Under low-moisture conditions, the hydroxyl groups of trehalose can form hydrogen bonds with polar groups in proteins or phospholipids, partially assuming the role previously played by interfacial water:

Protein or phospholipid polar group ←hydrogen bond→ Trehalose

Molecular dynamics studies have shown that trehalose can affect the area occupied by lipid molecules and the fluidity of the membrane core in dehydrated POPC bilayers, providing partial support for the water-replacement hypothesis. However, dehydrated membranes containing trehalose do not fully return to the structure of fully hydrated membranes.[4] Real-time drying experiments have shown that, during the initial stage of drying, trehalose primarily affects the phase behavior of phospholipid membranes by slowing water loss and increasing residual water content.[5]

Water replacement is a form of partial interfacial compensation. Trehalose cannot restore all the physical and biological functions of water, nor can it make a fully dehydrated membrane identical to a fully hydrated membrane.

3.3 Residual-Water Retention Mechanism

In addition to directly participating in polar interfacial interactions, trehalose can retain a small amount of interfacial water near biomolecules. In this case, trehalose does not directly replace all the water but instead works together with residual water to maintain the hydration state of polar groups.

This structure can be represented as:

Protein surface ←hydrogen bond→ Residual water ←hydrogen bond→ Trehalose

Molecular simulations have shown that trehalose can form localized clusters on protein surfaces, allowing water replacement and residual-water retention to occur simultaneously.[6]

The respective roles of the two mechanisms can be summarized as follows:

① Trehalose directly forms hydrogen bonds with polar groups, reducing interfacial changes before and after dehydration.

② Trehalose retains a small amount of water around biomolecules, maintaining the necessary local hydration.

3.4 Glass Transition and Restriction of Molecular Motion

When a trehalose system loses water to a sufficiently low moisture content, or is rapidly cooled under appropriate conditions, the sugar molecules may not have enough time to form regular crystals and may instead form a highly viscous amorphous structure.

The glass-transition temperature is denoted as Tg, meaning glass transition temperature. Below Tg, an amorphous system is in a glassy state with relatively slow molecular motion. Above Tg, the system enters a rubbery state with greater molecular mobility.

Decrease in water content → increase in trehalose concentration and system viscosity → reduction in molecular motion → formation of a highly viscous amorphous or glassy matrix

Water is an important plasticizer in amorphous trehalose systems:

Increase in water content → increase in molecular mobility → decrease in Tg → reduction in glass-state stability

Once trehalose enters a glassy state, it can restrict changes in protein conformation, membrane-lipid rearrangement, molecular diffusion, and certain degradation reactions. Water-diffusion behavior in trehalose—water systems also changes markedly near the glass transition.[7,8]

It should be noted that trehalose may also undergo a transition from the dihydrate to an anhydrous crystalline form. Amorphization, vitrification, and crystallization should therefore be evaluated separately.[9]

3.5 Three Mechanisms Jointly Participate in Low-Moisture Protection

The low-moisture protection of proteins and membrane structures by trehalose can generally be explained by three complementary mechanisms:

Interfacial hydrogen-bond replacement ＋ residual-water retention ＋ restriction of molecular motion by an amorphous matrix

The principal effects at different levels of water content are shown below:

Ordinary serums, emulsions, and creams have relatively high water contents. Trehalose therefore exists mainly in a dissolved state and primarily contributes to hygroscopic hydration and regulation of water dynamics.

After a product is applied, local trehalose concentration and system viscosity may increase as water evaporates. However, whether a stable glassy state forms depends on the trehalose concentration, environmental humidity, film thickness, and the presence of other components such as glycerol, salts, and polymers.

The vitrification mechanism is more directly relevant to lyophilized powders, low-water-activity preparations, and solid-state systems containing protein-based active ingredients.

4 Trehalose and the Skin Barrier

4.1 Its Fundamental Effects Arise Primarily from Stratum Corneum Hydration

Topically applied trehalose is initially distributed in the aqueous film formed by the product, on the skin surface, and in any superficial stratum corneum layers that it may reach. In these regions, trehalose can form dynamic hydrogen bonds with water and hydrophilic groups and may buffer rapid changes in the local hydration environment during water evaporation.

Its fundamental effects include:

① Participating in the local hydration network at the skin surface and in the superficial stratum corneum.

② Delaying rapid changes in the state of water in the surface film and outer stratum corneum.

③ Reducing the increase in tightness and brittleness caused by gradual water loss.

④ Improving sugar—water interactions in combined moisturizing systems.

Whether trehalose can penetrate the stratum corneum at a sufficient concentration to reach the viable epidermis requires skin-delivery and penetration studies using the specific formulation. Under conventional topical-use conditions, its fundamental moisturizing effect still mainly arises from hydration of the skin surface and stratum corneum.

4.2 Effects of High-Concentration Trehalose on IL-33- and NRF2-Related Signaling

A study published in the Journal of Investigative Dermatology in 2025 used primary human keratinocytes and an in vitro three-dimensional living skin equivalent model and investigated the effects of 60 mg/mL trehalose. This concentration corresponds to approximately 6% weight/volume.[10]

The study used interleukin-4 (IL-4) and interleukin-13 (IL-13) to establish a barrier-impairment model associated with T helper 2-type inflammation.

The following signaling changes were observed after trehalose treatment:

Trehalose → enhancement of MEK5—ERK5 signaling and attenuation of MEK1/2—ERK signaling → reduced IL-33 expression and nuclear accumulation

Here, MEK refers to mitogen-activated protein kinase kinase, ERK refers to extracellular signal-regulated kinase, and IL-33 refers to interleukin-33.

Another pathway was:

Trehalose → enhancement of JNK—NRF2—HMOX1 signaling → attenuation of IL-4/IL-13-associated oxidative stress

Here:

① JNK refers to c-Jun N-terminal kinase.

② NRF2 refers to nuclear factor erythroid 2-related factor 2.

③ HMOX1 refers to heme oxygenase 1.

Trehalose also reduced the activation of signal transducer and activator of transcription 3 and 6, namely STAT3 and STAT6, and restored the expression of some epidermal differentiation markers and antimicrobial proteins suppressed by IL-4 and IL-13.

4.3 Scope of Evidence from Skin-Barrier Signaling Research

Research on NRF2- and IL-33-related signaling indicates that high-concentration trehalose, when in direct contact with keratinocytes, can regulate signaling related to oxidative stress, cytokines, and epidermal differentiation.

The scope of evidence from this study is shown below:

This study supports the potential of trehalose to regulate specific barrier-stress signaling pathways. However, it does not directly demonstrate that low-concentration skincare products produce anti-inflammatory or barrier-repair effects of the same magnitude.

4.4 Claudin-1 Has Not Been Confirmed as a Principal Target in the Above Study

Claudin-1 is an important transmembrane protein in epidermal tight junctions and primarily contributes to the paracellular barrier between cells in the granular layer.

The above study observed restored expression of certain epidermal differentiation markers and antimicrobial proteins. However, a complete chain of evidence demonstrating that ordinary trehalose directly upregulates Claudin-1 and enhances tight-junction function has not yet been established.

Confirmation of an effect of trehalose on Claudin-1 would require simultaneous evaluation of:

① CLDN1 gene expression.

② Claudin-1 protein content.

③ Localization of Claudin-1 at the cell membrane.

④ Transepithelial electrical resistance or paracellular permeability.

⑤ Skin changes under actual topical-use conditions with the finished product.

5 Differences Between the Effects of Trehalose, Glycerol, and Hyaluronic Acid

5.1 Glycerol Primarily Increases Stratum Corneum Water Content

Glycerol is a relatively small molecule that can enter the stratum corneum and increase its water content. Clinical research has shown that glycerol primarily increases skin hydration, whereas petrolatum mainly improves barrier function by reducing transepidermal water loss.[11]

5.2 Hyaluronic Acid Primarily Provides Surface Hydration and Film Formation

Hyaluronic acid forms a hydrated network through its carboxyl groups, hydroxyl groups, and polymer chains. High-molecular-weight hyaluronic acid mainly provides surface film formation and lubrication, whereas lower-molecular-weight materials may differ in distribution and skin feel.

Human studies have shown that hyaluronic acids of different molecular weights may differ in the extent to which they improve skin water content. However, an increase in water content is not necessarily accompanied by a decrease in transepidermal water loss.[12]

5.3 Trehalose Primarily Contributes to Hydration Stability and Drying Protection

The distinguishing effects of trehalose include:

① Providing multivalent hydration through its nonreducing disaccharide structure.

② Slowing the reorientation of hydration water.

③ Buffering interfacial changes as water content decreases.

④ Restricting the structural motion of proteins and membrane lipids in low-moisture systems.

⑤ Regulating skin-barrier stress signaling in high-concentration in vitro models.

6 Role of Trehalose in Combined Moisturizing Systems

A complete moisturizing system must simultaneously address water supplementation, hydration stability, surface film formation, lipid structure, and evaporation control.

The respective roles of different ingredients can be represented as:

Small-molecule humectants such as glycerol increase stratum corneum water content → trehalose regulates the hydration environment and buffers drying-related changes → hyaluronic acid forms a hydrated surface film → barrier lipids and occlusive agents reduce further water loss

The specific functions are as follows:

① Humectants such as glycerol: increase water and hydrophilic solutes in the stratum corneum.

② Trehalose: participates in the sugar—water hydrogen-bonding network and regulates local water dynamics.

③ Hydrophilic polymers such as hyaluronic acid: form a hydrated surface film and improve lubrication and skin feel.

④ Ceramides, cholesterol, and fatty acids: supplement the intercellular lipids of the stratum corneum.

⑤ Petrolatum, hydrocarbons, oils, and film-forming agents: reduce the rate at which water evaporates into the external environment.

Trehalose occupies a position between hygroscopic moisturization and drying protection and cannot independently replace barrier lipids or occlusive agents.

7 Use Concentration and Efficacy Evaluation of Trehalose

7.1 The Effective Concentration Depends on the Evaluation Objective

There is no single minimum effective concentration of trehalose that applies to all formulations. “Effectiveness” may refer to increasing water content, prolonging moisturization, reducing dryness and tightness, stabilizing proteins, or regulating cellular signaling.

Historical industry-use data summarized in the CIR safety assessment reported a maximum trehalose concentration of approximately 2% in leave-on products. This value reflects market-use conditions and safety-assessment data from a particular period. It is not a legally prescribed maximum permitted concentration or a universal threshold for moisturizing efficacy.[13]

7.2 Concentration-Gradient Screening Can Be Used for Basic Moisturization

For ordinary serums, emulsions, and creams, 0.5%, 1.0%, and 2.0% may be used as internal formulation-screening concentrations to establish an initial concentration—effect relationship.

These three concentrations are starting points for research and development testing and do not mean that all formulations will produce the same results at the same concentrations.

Testing should include a compositionally identical blank base without trehalose and should evaluate:

① Immediate stratum corneum water content: to determine short-term hydration changes after application.

② Water retention at 2, 4, and 8 hours: to observe the duration of the moisturizing effect; specific time points may be adjusted according to product positioning.

③ Transepidermal water loss: to determine whether the product also improves control of water evaporation.

④ Performance under low-relative-humidity conditions: to assess hydration retention in dry environments.

⑤ Tackiness and tightness after the applied film has dried: to evaluate the effect of trehalose concentration on sensory properties.

⑥ Low-temperature, freeze—thaw, and long-term storage stability: to observe changes in viscosity, appearance, and system structure.

⑦ Trehalose crystallization: to determine whether crystallization occurs at high concentrations, low temperatures, or after water evaporation.

7.3 Concentrations Used for Cellular Signaling Must Be Validated Separately

The NRF2- and IL-33-related study used 60 mg/mL trehalose, corresponding to approximately 6% weight/volume. This concentration is substantially higher than the levels used in many ordinary moisturizing products.

When a product is intended to be evaluated for barrier-repair or soothing effects, the following factors must be confirmed using the final formulation and actual use concentration:

① The effective concentration and stability of trehalose in the formulation.

② The actual distribution of trehalose at the skin surface and within the stratum corneum.

③ Whether the formulation can deliver a sufficient concentration to the relevant cellular regions.

④ Whether the final formulation can reproduce changes in signaling markers such as NRF2 and IL-33.

⑤ Whether human studies can confirm improvements in barrier function and subjective symptoms.

Concentrations used for basic moisturization and those used for in vitro cellular-signaling studies belong to different evaluation systems and cannot substitute for one another.

8 Classification and Research Applications of Representative Chemicals Related to Trehalose Structure, Water Dynamics, Low-Moisture Protection, and Skin Barrier Research

Table 1. Core Trehalose Materials and Structural Reference Sugars

Table 2. Products Related to Hygroscopic Moisturization, Natural Moisturizing Factors, and Osmoprotection

Table 3. Stratum Corneum Barrier Lipids, Emollients, and Occlusive Materials

Table 4. Model Materials for Protein Stabilization and Phospholipid Membrane Protection

Note: The products listed above are representative Aladdin products related to scientific research and formulation studies. Additional product specifications, grades, and COA information can be searched on the Aladdin website using the product name, CAS number, or catalog number.

References

[1] Elbein A D, Pan Y T, Pastuszak I, Carroll D. New insights on trehalose: a multifunctional molecule. Glycobiology, 2003, 13(4): 17R–27R. DOI: 10.1093/glycob/cwg047.

[2] Hu J, Liao Z, Yano Y, Yamahara H, Tabata H. Broadband dielectric spectroscopic analysis toward characterization of the hydration state and bioprotective superiority of trehalose. The Journal of Physical Chemistry B, 2022, 126(3): 708–715. DOI: 10.1021/acs.jpcb.1c09941.

[3] Yamamoto M, Yasuhara A, Kondo N, Shiraga K. Slower reorientation of hydration water surrounding trehalose compared to maltose and sucrose. The Journal of Chemical Physics, 2025, 163(3): 035101. DOI: 10.1063/5.0257433.

[4] Golovina E A, Golovin A V, Hoekstra F A, Faller R. Water replacement hypothesis in atomic details: effect of trehalose on the structure of single dehydrated POPC bilayers. Langmuir, 2010, 26(13): 11118–11126. DOI: 10.1021/la100891x.

[5] Wolkers W F, Oldenhof H, Glasmacher B. Dehydrating phospholipid vesicles measured in real-time using attenuated total reflection Fourier transform infrared spectroscopy. Cryobiology, 2010, 61(1): 108–114. DOI: 10.1016/j.cryobiol.2010.06.001.

[6] Fedorov M V, Goodman J M, Nerukh D, Schumm S. Self-assembly of trehalose molecules on a lysozyme surface: the broken glass hypothesis. Physical Chemistry Chemical Physics, 2011, 13(6): 2294–2299. DOI: 10.1039/C0CP01705A.

[7] Simperler A, Kornherr A, Chopra R, Jones W, Motherwell W D S, Zifferer G. The glass transition temperatures of amorphous trehalose-water mixtures and the mobility of water: an experimental and in silico study. Carbohydrate Research, 2007, 342(11): 1470–1479. DOI: 10.1016/j.carres.2007.04.011.

[8] Malferrari M, Francia F, Venturoli G. Retardation of protein dynamics by trehalose in dehydrated systems of photosynthetic reaction centers: insights from electron transfer and thermal denaturation kinetics. The Journal of Physical Chemistry B, 2015, 119(43): 13600–13618. DOI: 10.1021/acs.jpcb.5b02986.

[9] Kilburn D, Townrow S, Meunier V, Richardson R, Alam A, Ubbink J. Structural evolution of the dihydrate to anhydrate crystalline transition of α,α-trehalose. The Journal of Physical Chemistry B, 2009, 113(4): 1004–1009. DOI: 10.1021/jp807704n.

[10] Dai X, Mizukami Y, Watanabe K, Tsuda T, Shidahara M, Yoshida S, Yatsuzuka K, Shiraishi K, Mori H, Murakami M, Kawakami R, Imamura T, Fujisawa Y, Muto J. Trehalose prevents IL-4/IL-13-induced skin barrier impairment by suppressing IL-33 expression and increasing NRF2 activation in human keratinocytes in vitro. Journal of Investigative Dermatology, 2025, 145(6): 1422–1432.e10. DOI: 10.1016/j.jid.2024.08.038.

[11] Vaillant L, Georgescou G, Rivollier C, Delarue A. Combined effects of glycerol and petrolatum in an emollient cream: a randomized, double-blind, crossover study in healthy volunteers with dry skin. Journal of Cosmetic Dermatology, 2020, 19(6): 1399–1403. DOI: 10.1111/jocd.13163.

[12] Muhammad P, Novianto E, Setyorini M, Legiawati L, Yusharyahya S N, Menaldi S L, Budianti W K. Effectiveness of topical hyaluronic acid of different molecular weights in xerosis cutis treatment in elderly: a double-blind, randomized controlled trial. Archives of Dermatological Research, 2024, 316: 329. DOI: 10.1007/s00403-024-03003-2.

[13] Fiume M M, Bergfeld W F, Belsito D V, Hill R A, Klaassen C D, Liebler D C, Marks J G Jr, Shank R C, Slaga T J, Snyder P W, Gill L J, Heldreth B. Safety assessment of monosaccharides, disaccharides, and related ingredients as used in cosmetics. International Journal of Toxicology, 2019, 38(Suppl 1): 5S–38S. DOI: 10.1177/1091581818814189.

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