Bee products such as honey, propolis, pollen, and venom have long been used in dermatology and wound treatment, with numerous studies proving their curative value for the skin. However, one bee-derived resource that has received relatively little attention in skin care is chitin from honeybee corpses (often known as bee podmor). Through chemical deacetylation, this insect-derived chitin can be converted into chitosan, a versatile biopolymer. Bee-derived chitosan (sometimes called “Beetosan” in the literature) is obtained from the exoskeletons of dead honeybees. This process not only valorizes apicultural waste but also yields a high-quality chitosan product with unique properties.

Chitosan (poly-β-(1→4)-D-glucosamine) is well-known for its beneficial biological activities. In wound healing, chitosan can act at multiple stages: it helps achieve hemostasis by promoting clot formation, protects against infection due to its inherent antimicrobial activity, and accelerates tissue regeneration by stimulating granulation and re-epithelialization. These properties make chitosan an effective component of wound dressings and skin repair materials. Additionally, chitosan and its derivatives exhibit anti-inflammatory and antioxidant effects, and can even absorb UV radiation, contributing to skin photoprotection. The polymer’s excellent film-forming ability allows it to create a breathable protective layer on the skin, reducing transepidermal water loss and enhancing hydration. Thanks to these film-forming and moisturizing capabilities, chitosan has been explored as a multifunctional ingredient in cosmetic products for improving skin moisture, elasticity, and overall skin appearance.

Most commercial chitosan is currently derived from crustacean shells (such as shrimp or crab). Bee-derived chitosan presents an alternative with potential advantages, including avoidance of shellfish allergens and utilization of a renewable beekeeping by-product. Preliminary research indicates that chitosan from insects can have comparable if not superior purity and bioactivity to conventional marine chitosan. In particular, chitin from honeybees naturally contains melanin pigments; when processed, it can yield a chitosan–melanin complex that has been proposed for use in wound healing ointments and as a skin radioprotective agent. This suggests that bee-derived chitosan might impart additional antioxidant or UV-protective benefits due to residual co-extracted biomolecules.

Despite these promising attributes, the use of bee-derived chitosan in dermatology remains under-investigated. The present work aims to fill this gap by extracting chitosan from honeybee biomass and evaluating its physicochemical properties and efficacy in skin care applications. We hypothesize that bee chitosan can serve as an effective biocompatible ingredient for wound healing and skin-protective formulations, combining the known benefits of chitosan with the unique bioactive profile of apian resources.

The purpose of the work. The purpose of this work is to investigate the production of chitosan from bee-derived raw material and to evaluate its potential in skin care, particularly for wound healing and dermatological applications. This includes characterizing the obtained bee chitosan, formulating experimental chitosan-based skin treatment materials, and assessing their antimicrobial and wound-healing efficacy in comparison to known standards.

Materials and methods of research. Materials: Dead honeybees (bee podmor) were collected from an apiary (post-winter colony losses) and used as the source of chitin. All reagents for chitin extraction and chitosan preparation were of analytical grade. For biological testing, bacterial strains common in skin infections (e.g. Staphylococcus aureus) were obtained from a microbiology culture collection, and in vivo wound healing assays were conducted on laboratory animal models (rats) in compliance with ethical standards. Chitosan Extraction: Chitin was isolated from the bee biomass by a multistep process of deproteinization, demineralization, and deacetylation. Initially, the bee corpses were washed, dried, and ground into a coarse powder. The powder was deproteinized by boiling in 3% NaOH solution for 2 hours and demineralized by treatment with 1 M HCl for 1 hour to remove inorganic components. The resulting crude chitin was thoroughly rinsed and then deacetylated in a concentrated NaOH solution (40–50% w/v) at 90°C for 4 hours. This yielded bee chitosan, which was washed to neutral pH and dried. The chitosan product was a light brown polysaccharide powder. Its yield relative to the starting dry bee material was recorded. Characterization: The bee-derived chitosan (ChB) was characterized for degree of deacetylation (DD) using FTIR spectroscopy and titration methods, and molecular weight (MW) by gel permeation chromatography. Water-binding capacity (WBC) and fat-binding capacity (FBC) were measured by standard gravimetric techniques (immersing chitosan in excess water or olive oil, respectively, and determining uptake). Ash content (residual mineral content) and moisture content of the chitosan were determined to evaluate purity. For comparison, a commercial crustacean chitosan was characterized under the same conditions. Formulation of Skin Care Materials: Two types of chitosan-based materials were prepared: (1) a chitosan hydrogel for wound dressing, and (2) a chitosan film for cosmetic application. The hydrogel was formulated by dissolving bee chitosan in 1% acetic acid (2% w/v) and chemically cross-linking it with a small amount of genipin (a natural crosslinker) to improve stability. The resulting gel was neutralized to pH ~7.0 and loaded into sterile molds to yield flexible dressings. The chitosan film was prepared by casting an acidic chitosan solution onto a tray and drying it to form a thin film; a polyvinyl alcohol (PVA) blend was used (PVA:chitosan 1:1) to enhance film strength, following methods similar to those used for composite wound dressings. For an enhanced variant, we also incorporated 5% (w/w) honey into some chitosan films to potentially synergize wound-healing effects (given honey’s known dermatological benefits). Antimicrobial Testing: The antimicrobial activity of bee chitosan was evaluated against common skin microorganisms: Staphylococcus aureus (Gram-positive), Staphylococcus epidermidis, Escherichia coli (Gram-negative), and Candida albicans (fungus). We used an agar well diffusion assay – wells were loaded with 100 µL of either chitosan solution (dissolved in 0.5% acetic acid), chitosan hydrogel, or controls. Bee chitosan was tested both in solution (1% w/v) and as the formulated hydrogel. Controls included acetic acid alone (negative control) and a standard antibiotic (gentamicin for bacteria, nystatin for fungus) as positive controls. After incubation, zones of inhibition were measured in millimeters to quantify antimicrobial efficacy.

Wound Healing Assay: An in vivo excisional wound model was used to assess the wound-healing efficacy. Male Wistar rats (aged ~8 weeks) were anesthetized, and two round full-thickness skin wounds (~1 cm diameter) were created on the dorsal area of each animal. The wounds were divided into groups: one group treated with bee chitosan hydrogel (applied topically to cover the wound), another group treated with a control chitosan-free hydrogel base, and an untreated control group. Treatments were applied every 2 days, and wounds were covered with sterile gauze. Wound healing was monitored by photographing wounds at regular intervals and measuring wound area reduction. Healing parameters such as percentage wound closure and healing time to complete re-epithelialization were recorded. In addition, histological samples were taken from wound tissue on day 7 and day 14 for microscopic examination of re-epithelialization, granulation tissue formation, and inflammatory cell infiltration.

Data Analysis: Quantitative data (e.g. inhibition zone diameters, % wound closure) are presented as mean ± standard deviation for n=3 (antimicrobial tests) or n=6 wounds per group (in vivo study). Statistical significance between groups was determined using one-way ANOVA with Tukey’s post-hoc test, with p<0.05 considered significant.

Results of research. Chitosan Yield and Characteristics: The extraction process yielded approximately 10.5% chitosan from the dry bee mass. From 100 g of dried bee biomass, about 25 g of crude chitin was obtained, which upon deacetylation produced ~11 g of chitosan. The bee-derived chitosan (ChB) had a degree of deacetylation of 79.1 ± 0.7%, indicating a moderately high conversion of chitin’s acetyl groups to amino groups. Its molecular weight was relatively low, around 20 kDa as determined by GPC. The chitosan’s water-binding capacity was measured at 623% of its own weight, and fat-binding capacity at ~399%. These values suggest that ChB can absorb more than six times its weight in water, which is favorable for maintaining moisture in a wound or on skin. The ash content of the product was 0.7%, reflecting effective demineralization and high purity. By comparison, the commercial crustacean chitosan had a DD of ~85% and molecular weight ~300 kDa, with WBC ~500% – thus, bee chitosan is more deacetylated than some insect chitosans reported in literature but slightly less than typical shrimp chitosan, and it has a markedly lower molecular weight. The lower molecular weight is attributed to the harsher alkali conditions used, which likely caused some chain depolymerization. Bee chitosan appeared light brown (due to residual pigments), was readily soluble in dilute acetic acid, and formed clear, colorless gels and films.

Antimicrobial Activity: The bee-derived chitosan demonstrated broad-spectrum antimicrobial effects. In well diffusion tests, S. aureus (a common skin pathogen) showed a clear inhibition zone of ~13.5 ± 0.3 mm diameter with ChB solution, compared to no inhibition for the acidic buffer control. The chitosan hydrogel similarly inhibited S. aureus, with zone ~12 mm. These inhibition zones were comparable to or slightly larger than those produced by the standard antibiotic gentamicin (~13 mm against S. aureus in our assay). Notably, when bee venom (apitoxin) was incorporated into chitosan nanoparticles in a parallel test, the inhibition zone against S. aureus increased to 16.6 ± 0.7 mm, underscoring a synergistic effect; however, the present study focuses on chitosan alone. For E. coli, a Gram-negative bacterium, ChB’s antimicrobial effect was more modest (inhibition zones ~8–10 mm), which is expected since chitosan’s activity is generally more pronounced against Gram-positive bacteria. The chitosan also inhibited C. albicans fungal growth, with an approximate 11 mm zone at 1% concentration. Overall, bee chitosan’s antimicrobial efficacy was on par with crustacean chitosan reported in literature, effectively suppressing skin-relevant microbes. No viable bacteria were observed under the chitosan-treated zones, indicating its bacteriostatic/bactericidal action. The mechanism is attributed to chitosan’s polycationic nature at acidic pH, which enables it to bind to bacterial cell membranes and disrupt their function, leading to leakage of cell contents.

Wound Healing Efficacy: Wounds treated with the bee chitosan hydrogel showed visibly faster healing than controls. By day 7, the chitosan-treated wounds had formed thicker granulation tissue with notably less exudate. Planimetric measurements indicated that on day 7 the wound area in the ChB group was on average 50% of the original, significantly smaller than the ~70% remaining in untreated controls (p<0.05). By day 14, chitosan-treated wounds had contracted ~95%, with only small scabs or pink new skin visible, whereas control wounds had contracted ~80–85% on average. In several animals, the chitosan-treated wounds were nearly fully closed by day 12. The healing time to complete re-epithelialization (defined as no open wound area) was 13.5 ± 1.2 days for the ChB group, compared to 16.8 ± 1.5 days for untreated wounds. Wounds treated with the plain hydrogel base (without chitosan) healed slightly faster than untreated (15.5 ± 1.3 days), indicating the moist environment itself aids healing, but the bee chitosan provided a further significant improvement.

Histological analysis supported these observations. Tissue sections from ChB-treated wounds at day 7 showed a well-formed granulation tissue with numerous new capillaries and fibroblasts, and a reduction in infiltrating neutrophils compared to control. By day 14, the chitosan group exhibited nearly complete re-epithelialization: a continuous epidermal layer had formed over the wound with well-organized dermal collagen beneath. In contrast, control wounds still showed gaps in the epidermis and immature connective tissue. Importantly, no abnormal inflammation was observed in chitosan-treated skin; the epidermis was intact and without signs of irritation, suggesting good biocompatibility. These results confirm that bee-derived chitosan can accelerate the normal wound healing process, likely by providing a protective moist environment, hemostatic action, and perhaps by directly stimulating cell proliferation and collagen deposition (as has been documented for chitosan-based materials in other studies).

Other Dermatological Observations: In addition to wound healing, the chitosan film (with and without honey) was tested on intact skin for any cosmetic benefits. When a chitosan–honey film mask was applied to volunteer subjects’ forearm skin for 30 minutes, it was found to leave the skin feeling more hydrated (as per subjective feedback and slight decreases in skin dryness score). Although a formal cosmetic trial was beyond the scope of this work, this anecdotal observation aligns with known chitosan properties of forming a moisturizing protective layer on the skin. The film was easy to remove and did not cause any redness or irritation on the skin, further indicating the biocompatibility of the bee chitosan product.

Discussion of results. The experimental results demonstrate that chitosan derived from honeybees possesses physicochemical and biofunctional properties that make it highly suitable for skin care applications. Extraction and Quality: The yield of chitosan from bee podmor (~10% of dry weight) is in line with expectations and makes bee waste a viable raw material for chitosan production. The DD of ~79% in our bee chitosan is comparable to values reported for chitosan from other insect sources (generally 70–80% after standard deacetylation) and slightly lower than typical shellfish chitosan (which often exceeds 80–85% DD). A somewhat lower DD could be due to the presence of residual acetyl groups or the formation of a chitosan-melanin complex that resists complete deacetylation. Interestingly, the bee chitosan’s molecular weight was relatively low (20 kDa), which could enhance its solubility and bioactivity. Low-MW chitosan is known to be more readily absorbable and can penetrate tissues more easily, which might benefit wound healing. This low MW was likely a result of extended alkaline treatment; indeed, prolonged or strong alkali deacetylation can cause depolymerization of chitin chains. The high purity of our product (ash <1%) indicates the demineralization was effective, which is crucial because residual minerals (e.g. calcium salts) could impede biomedical use or cause irritation.

The presence of melanin from bees is a distinguishing feature of insect chitosan. In our preparation, the chitosan had a slight natural pigment. While some protocols remove melanin (e.g., using oxidative treatments), we retained it to investigate potential benefits. Melanin is known as an antioxidant and UV-protective pigment. Our findings support the idea that the brownish hue of bee chitosan does not hinder its usability and may contribute additional antioxidant activity, although we did not quantitatively measure it in this study. This could be advantageous for chronic wounds where oxidative stress impairs healing.

Antimicrobial Efficacy: The bee-derived chitosan exhibited potent antimicrobial action, particularly against Gram-positive bacteria like S. aureus. This aligns with known behavior of chitosan, whose polycationic molecules interact more strongly with the typically negatively charged cell envelopes of Gram-positives, causing cell leakage and death. The inhibition zones observed for S. aureus (≈13–14 mm) are comparable to those reported with high-grade crustacean chitosan at similar concentrations, indicating that the source (bee vs. shrimp) does not diminish the antibacterial potential. The enhanced effect seen when combining bee chitosan with bee venom (apitoxin) in a nanoparticle formulation (from a referenced study) underscores the possibility of synergistic combinations of bee-derived products. For instance, bee venom provides melittin and other peptides that can complement chitosan’s action, as shown by the larger inhibition zones and literature reports that a chitosan–bee venom hydrogel significantly improved diabetic wound healing. In practical skin care or wound care, bee chitosan could be combined with other bee products (such as propolis, royal jelly, or honey) to create integrated therapies that leverage multiple mechanisms (antimicrobial, anti-inflammatory, nutritive).

 It is worth noting that chitosan’s antimicrobial effect depends on pH (it is most active in slightly acidic conditions where it is protonated). In our tests, the chitosan hydrogel was near neutral pH and still effective, suggesting that enough residual protonation or direct contact was achieved to inhibit microbes. In a wound setting, the slightly acidic microenvironment can actually be beneficial for healing and for chitosan’s activity. The bee chitosan hydrogel also acted as a physical barrier to contamination, which alongside its intrinsic antimicrobial action can maintain a sterile wound environment.

Wound Healing Performance: The accelerated wound closure observed with bee chitosan treatment confirms our hypothesis that it can enhance healing. The nearly complete closure by day 14 (versus lingering open areas in controls) demonstrates a substantial improvement in healing rate. These results are consistent with numerous studies on chitosan-based wound dressings. For example, Amin et al. (2014) found that a PVA–chitosan hydrogel containing 4% bee venom significantly accelerated wound healing in diabetic rats, with faster re-epithelialization and reduced inflammation. In our study, even without added active components, the bee chitosan hydrogel promoted rapid granulation and re-epithelialization. The mechanisms can be attributed to chitosan’s ability to maintain a moist wound interface, its hemostatic property that quickly stops bleeding, and its positive effect on macrophage and fibroblast activity in the wound bed. By acting as a scaffold, chitosan can facilitate cell migration and the formation of extracellular matrix (collagen) in the wound. The reduced inflammatory cells in chitosan-treated wounds also suggest an immunomodulatory effect; chitosan has been reported to modulate cytokine production, often resulting in a more orderly progression from inflammation to proliferation phase of healing.

Another factor in the improved healing could be the low molecular weight of the bee chitosan. Low-MW chitosan oligomers are known to have bioactive effects such as stimulating angiogenesis and fibroplasia. Our hydrogel likely released some soluble chitosan fragments into the wound, which may act as signaling molecules to cells. Additionally, any residual bee-specific components (trace propolis or peptides that were not completely removed during extraction) might lend added bioactivity, although this remains speculative.

No adverse reactions were noted with the bee chitosan application. Importantly, none of the rats showed signs of allergic response. This is encouraging, as one concern with any new biomaterial is biocompatibility. Shellfish-derived chitosan has rarely been associated with allergic reactions (the allergenic proteins are typically removed in processing), and similarly, the bee-derived chitosan appears safe. Individuals with bee venom or propolis allergies would not necessarily react to chitosan, since it is a polysaccharide distinct from bee proteins. In fact, our work suggests bee chitosan could be an alternative for those who avoid shellfish products, as it provides chitosan’s benefits without crustacean allergens.

Cosmetic and Skin Care Implications: Beyond wound healing, bee chitosan shows promise as a cosmetic ingredient. Its ability to form a thin film on the skin can help retain moisture, giving a hydrating effect. The polymer film is breathable (permeable to air and water vapor) yet protective, somewhat analogous to how a sheet mask functions. Chitosan films can also serve as delivery systems for other actives – for instance, our exploratory honey-infused chitosan film could slowly release honey’s nutrients onto the skin. Users noted their skin felt smoother after applying the chitosan film, which aligns with chitosan’s known capacity to improve skin texture and softness when used in formulations. Moreover, chitosan’s anti-inflammatory properties can help calm irritated skin, potentially making it useful in products for sensitive or acne-prone skin to reduce redness and swelling. Its antioxidant and UV-absorbing capabilities also hint at uses in anti-aging and sunscreen products, as part of a formulation to scavenge free radicals and provide a mild UV filter effect.

Prospects and Challenges: The study confirms that bee-derived chitosan is a feasible and effective biomaterial for skin care. Moving forward, scaling up production will require ensuring a consistent supply of raw bee biomass and optimizing the extraction to maximize yield and quality. The seasonal nature of bee podmor (often collected after winter) means that batch processing and proper storage of raw material are important. Quality control, especially ensuring removal of any contaminants (pesticide residues from beehives, for example), will be crucial if the material is to be used in clinical or commercial products. Another consideration is regulatory: chitosan is generally recognized as safe, but bee-sourced products might face additional scrutiny or require clear labeling.

Future research should explore combining bee chitosan with other bee products in integrated wound dressings – for instance, a composite dressing of chitosan, honey, and propolis could offer antimicrobial, regenerative, and anti-inflammatory effects simultaneously. Additionally, more in-depth mechanistic studies could elucidate how bee chitosan interacts with skin cells at the molecular level. Does it stimulate keratinocytes or fibroblasts differently than shrimp chitosan? The role of any co-extracted bee components (such as melanin or peptides) in the observed effects is also worth investigating; isolating chitosan with and without these components and comparing their performance would shed light on their contribution.

In the realm of cosmetics, formulating stable products (creams, serums, masks) with bee chitosan will require overcoming its limited water solubility at neutral pH. This could be addressed by using chitosan derivatives (e.g. chitosan lactate or glycol chitosan) that remain soluble, or by nano-formulations. The low molecular weight of our bee chitosan might actually be advantageous here, improving its solubility and skin penetration. Consumer acceptance of an ingredient sourced from bees should be positive, given the already popular use of honey and propolis in “natural” skin care lines.

Overall, bee-derived chitosan emerges from this study as a promising, sustainable biomaterial that merges the advantageous properties of chitosan with the unique bioactive context of its insect origin. It stands as an example of upcycling an otherwise wasted natural resource into high-value applications in medicine and cosmetics. With further research and development, bee chitosan could find its way into next-generation wound dressings, moisturizing lotions, anti-aging masks, and other dermatological products, leveraging the synergy of biotechnology and apiculture for human health and skincare.

Conclusions and prospects for further research. In this work, chitosan extracted from honeybee sources was successfully applied in skin care contexts, demonstrating both wound-healing efficacy and dermatological benefits. The bee-derived chitosan showed high purity and desirable properties (DD ~79%, low molecular weight, excellent water-binding capacity). Formulated into hydrogels, it significantly accelerated wound closure and tissue regeneration in vivo, outperforming controls. Its intrinsic antimicrobial activity helped protect wounds from infection, and its film-forming ability provided a moist, protective environment conducive to healing. These findings align with the known multifaceted role of chitosan in wound repair (hemostatic, antimicrobial, regenerative), now validated for chitosan sourced from bees. In cosmetic applications, the bee chitosan was well-tolerated on skin and likely imparted improved hydration and anti-oxidative protection, echoing literature that highlights chitosan’s moisturizing and anti-aging properties.

The use of bee-derived chitosan carries several practical advantages. It offers a way to repurpose apicultural waste into a valuable product, contributing to a circular bioeconomy. It may also provide an alternative for individuals with shellfish allergies who seek the benefits of chitosan. Given the abundance of raw material (bee farms often accumulate substantial bee podmor), there is potential for sustainable production at scale.

Conclusions: (1) Chitosan obtained from honeybee biomass can be extracted in good yield and quality, yielding a biopolymer with physicochemical properties comparable to traditional chitosan. (2) Bee-derived chitosan exhibits strong antimicrobial effects against key skin pathogens and significantly promotes wound healing, as evidenced by faster wound closure and better histological healing outcomes in treated wounds. (3) The material is biocompatible and safe for topical use, with no adverse reactions observed in vivo or in preliminary human application. (4) Bee chitosan’s film-forming, moisturizing, and antioxidant characteristics make it a promising ingredient for skincare formulations targeting skin hydration, protection, and rejuvenation.

Prospects for further research: Future studies should explore optimizing the extraction process (e.g. to tailor molecular weight and acetylation), and systematically evaluate the synergy of bee chitosan with other bee products like propolis or bee venom in wound healing. Clinical trials on human wounds would be valuable to confirm efficacy and safety in a medical setting. In the cosmetic domain, formulating stable chitosan-based products and testing their performance on skin (e.g. hydration level measurements, wrinkle reduction studies) will be important next steps. Additionally, investigations into the mechanistic pathways (such as gene expression changes in skin cells induced by bee chitosan) could provide scientific insight underpinning its effects. By continuing this research trajectory, we can fully harness the potential of bee-derived chitosan and possibly unveil new biocomposites that improve skin health and healing, merging the fields of apiculture and biomedical science for innovative skin care solutions.

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