HYDROLYZED SPONGE

Dermal delivery is versatile in therapeutics as well as cosmetics in pursuit of enhancing safety/efficacy and alleviating pain/fear to alternate oral/injective administrations. Natural siliceous spicules offer a potential approach via simple topical medications to circumvent poor penetrations through the skin barrier by loading, carrying, and releasing the active ingredients in a highly efficient and controlled manner. The delivery of ingredients loaded on spicules is assessed to improve the dermal efficacy compared to simple topical treatments. First, needle-like spicules were isolated from natural freshwater sponges. Then, the active ingredient was loaded via liposome formations. Finally, the dermal efficiency was evaluated. Natural siliceous spicules were purified, sorted, and fully characterized to appear 250 μm of length and 12 μm of diameter on average. A model ingredient, pectolinarin, was efficiently loaded onto the internal space of spicules via lecithin-based liposome formations. The penetration was visualized using a porcine skin sample with a fluorescent dye and assessed quantitatively by a Franz diffusion cell system. Dermal absorption rate was measured 73.4%, while the percutaneous penetration rate was 2.2%. The release pattern turned out a simple diffusion analyzed by Fick's law and Higuchi model. The liposomes loaded onto spicules were further stabilized by a hydrophobic capsulation, which may benefit the overall efficacy of the ingredient. A novel dermal delivery system is beneficial to improve the topical efficacy of biologically active ingredients. The outcomes shed a light upon developing skin-protective/improving cosmetics and therapeutics with enhanced performance.

To study the effects and safety of Spongilla lacustris (SL) in 3% hydrogen peroxide (H2O) on rat skin. An experimental study was conducted on 3 groups of Wistar-Furth adult rats. The first group was applied with SL in H2O2 the second group with SL in 0.9% normal saline (NSS) and the control group with NSS. These agents were applied on and wiped off the rat skin weekly for four weeks, then the skin biopsies were done. The number of SL spicule and the depth of spicule penetration were examined by scanning electron microscope and by polarized light microscope respectively. Skin histopathology was determined by hematoxylin-eosin staining. The gross skin changes were observed. Under electron microscopic examination, SL was demonstrated as spicule which was sharp-edged, rod-shaped and smooth surface with approximate 150-300 microns in length and 10-20 microns in diameter. Spicule retention was found in the rat skin lasted until day 3 but was undetectable on day 7. The spicules could be detected deep into stratum basalis. Comparing among three groups, the thickness of epidermis in the second group was decreased with statistically significant difference (p = 0.044) by the end of week 7. The dermal thickness of all groups was increased by age. No any gross skin alteration of all groups was observed. The authors hypothesized that the spicule causes puncture that enhances H2O2 penetration into the skin. This solution was safe in the short-term usage. However, the long-term safety regarding granulomatous formation is still questionable.

Poor patient adherence with antiacne medications is a common clinical challenge. DMT310, a natural, topical product with a once-weekly application schedule, may alleviate this obstacle. Evaluate the safety, tolerability, and efficacy of DMT310 in treating moderate-to-severe acne. This 12-week, randomized, double-blind, placebo-controlled, multicenter clinical trial enrolled participants 12 years and older with moderate-to-severe acne. The intent-to-treat population included a total of 181 participants (DMT310, N = 91; placebo, N = 90). Participants who received DMT310 vs participants treated with placebo demonstrated a statistically significant greater reduction in the number of inflammatory and noninflammatory lesions at all time points: inflammatory lesion counts at week 12 (-15.64 vs -10.84, P < .001); noninflammatory lesion counts at week 12 (-18.26 vs -12.41, P < .001). DMT310-treated participants also had higher rates of Investigator's Global Assessment treatment success than participants in the placebo group at all time points: Investigator's Global Assessment at week 12 (44.40% vs 17.78%; P < .001). No serious treatment related adverse events occurred. DMT310 once-weekly topical treatment significantly reduced both inflammatory and noninflammatory lesions and yielded a greater proportion of Investigator's Global Assessment treatment success at all time points in participants with moderate-to-severe acne.

Wharton's Jelly-derived mesenchymal stem cell (WJ-MSC) secretome contains diverse bioactive factors with potential for skin regeneration, but its clinical efficacy is limited by poor transdermal delivery. In this study, we developed a dual-delivery system by nanoencapsulating WJ-MSC secretome and coating it onto marine sponge-derived spicules. Physicochemical characterization, in vitro assays (fibroblast and keratinocyte proliferation, keratinocyte migration, type I procollagen secretion, and antioxidant activity), and in vivo penetration studies were conducted. A single-arm clinical trial evaluated dermal absorption, pore characteristics, skin texture, wrinkles, and pigmentation following topical application. Transdermal penetration efficiency was significantly higher in the nano-coated spicule group than in the uncoated secretome control. In vitro, secretome treatment promoted fibroblast and keratinocyte activity, accelerated wound closure, and increased collagen synthesis. Clinically, a single application enhanced dermal absorption and significantly reduced pore number, while two weeks of treatment decreased wrinkles and pigmentation. Spicule-based nanoencapsulation effectively overcomes the skin barrier, enhances the regenerative activity of WJ-MSC secretome, and induces measurable clinical improvements in skin rejuvenation. This platform represents a promising cosmetic and therapeutic strategy in dermatology.

Silica spicules provide a natural transdermal conduit but require a linker that binds strongly under physiological conditions and releases payloads selectively in response to biological cues. Existing silane chemistries or polydopamine coatings lack enzyme responsiveness and show limited control over release. We created a 180-member peptide library with the motif L-X1-X2-[Y-F-Y]-A-L-G-P-H-C and screened for silica binding. Biophysical assays (circular dichroism, ζ-potential, quartz crystal microbalance, atomic force microscopy) and molecular dynamics identified high-affinity binders. The lead, P176, was tested for matrix metalloprotease (MMP)-responsive cleavage. Conjugation and release of Vitamin C and Stigmasterol were analyzed by HPLC and Franz diffusion cells. P176 showed high silica affinity (~55 µg mg -1 ), robust biophysical signals (Δf -35 to -38 Hz; rupture force ~154 pN; ζ shift -22 to-11.5 mV), and favorable adsorption energy (-48.5 kcal mol -1 , contact 4.5 nm 2 , 8.5 H-bonds). The MMP gate displayed efficient kinetics (Vmax 117.9 RFU·min -1 , Km 5.0 µM) with >90% cleavage at 60 min, reduced to 26% by inhibitor. Conjugation yields reached 87% (Vitamin C) and 77% (Stigmasterol). Franz diffusion showed MMP-dependent release (24 h: Vitamin C 90-96%, Stigmasterol 80-85%) with minimal basal leakage. Released Vitamin C enhanced collagen I to ~250% in fibroblasts, while Stigmasterol attenuated LPS-induced macrophage morphology; keratinocytes retained normal marker expression. This study demonstrates that a single amphipathic, sequence-programmed peptide can couple strong silica anchoring with protease-responsive release and broad payload compatibility, establishing a versatile platform for spicule-based transdermal and regenerative delivery.

The pathogenesis of atopic dermatitis (AD) is closely linked to both genetic and environmental factors, with patients often exhibiting a range of immunological abnormalities, including a pronounced Th2-type overreaction, which is a key feature of the disease. Cutibacterium acnes has been shown to induce a robust Th1 immune response through intraperitoneal injections, potentially preventing the development of AD. In this study, a novel nanoparticulate formulation of Cutibacterium acnes (NFCA) was developed with the formulation optimization for the dermal delivery. Sponge Haliclona sp. spicules (SHS) were isolated from the explants of sponge Haliclona sp . with our proprietary method. The NFCA was prepared by high-speed grinding followed by film extrusion. The skin penetration of the model drugs in NFCA with SHS were visualized using confocal microscopy. The therapeutic effects of NFCA coupled with SHSs against AD in mice were assessed by using pathohistological examination and cytokine ELISA assay. The NFCA particle size was 254.1±39.4 nm, with a PDI of 0.29±0.08 and a Zeta potential of -7.9±0.6 mV. SHS significantly enhanced total skin absorption of FD10K (39.6±6.7%, p =0.00076) as well as deposition in the viable epidermis (3.2±1.6%, p =0.08) and deep skin (dermis & receptor) (36.0±5.9%, p =1.82E-5) compared to the control. In vitro cytotoxicity tests showed that NFCA had low toxicity to HaCaT cells (IC50=63.8 mg/mL). The study confirmed that NFCA can activate immune signaling pathways, promoting the high expression of IL-6 and IL-8 in keratinocytes, enhancing TNF-α and IL-1β expression in macrophages, and inducing Th1 and Th17-type immune responses. Furthermore, we demonstrated that the dermal delivery of NFCA using SHS in vivo significantly reduced epidermal thickness, serum IgE levels, and tissue IL-4 levels, thereby accelerating skin repair and mitigating Th2 polarization. SHS were employed to effectively deliver NFCA to the deeper skin layers to exert its immune functions. Moreover, the combination of SHS and NFCA can significantly cure mice with atopic dermatitis.

Transdermal drug delivery systems hold promise, but their effectiveness is often constrained by the skin's barrier. Microneedles (MNs) improve drug permeability by creating micro-channels in the skin, yet they continue to face challenges such as infection risks and safety concerns. To overcome these challenges, a novel antimicrobial sponge MNs (ASMNs@PVP-INS) modified with polyvinylpyrrolidone (PVP) for insulin (INS) delivery was designed. Mechanical testing demonstrated that these MNs possess excellent mechanical strength, capable of withstanding at least 0.11 N per needle without rupture. In vitro drug penetration tests revealed that the MNs consistently released over 75 % of INS within a 6 h. In an animal model, ASMNs@PVP-INS reduced initial blood glucose levels from 22.4 to 5.72 mmol/L, effectively maintaining glucose control for more than 6 h without inducing hypoglycemia. Additionally, agar diffusion assays indicated that INS loading did not compromise the antimicrobial properties of antimicrobial sponge MNs (ASMNs). Skin irritation tests showed that ASMNs@PVP-INS exhibited mild irritation (PII < 0.6), with skin damage fully recovering within 8 h. Safety assessments indicated no significant toxicity to mice, with biochemical markers remaining within normal ranges, thereby confirming their good biocompatibility. In conclusion, ASMNs@PVP-INS hold promise as a novel drug delivery vehicle.

We report on the fabrication of mesoporous silicon dioxide coated Haliclona sp. spicules (mSHS) to enhance the delivery of the insoluble photosensitizer protoporphyrin IX (PpIX) into deep skin layers and mediate photodynamic therapy for metastatic melanoma in mice. The mSHS are dispersed sharp edged and rod-like micro-particles with a length of approximate 143.6 ± 6.4 μm and a specific surface area of 14.9 ± 3.4 m 2 /g. The mSHS can be topically applied to the skin, adapting to any desired skin area and lesion site. The insoluble PpIX were incorporated into the mesoporous silica coating layers of mSHS (mSHS@PpIX) with the maximum PpIX loading capacity of 120.3 ± 3.8 μg/mg. The mSHS@PpIX significantly enhanced the deposition of PpIX in the viable epidermis (5.1 ± 0.4 μg/cm 2 ) and in the dermis (0.5 ± 0.2 μg/cm 2 ), which was 154 ± 11-fold and 22 ± tenfold higher than those achieved by SHS, respectively. Topical delivery of PpIX using mSHS (mSHS@PpIX) completely eradicated the primary melanoma in mice in 10 days without recurrence or metastasis over 60 days. These results demonstrate that mSHS can be a promising topical drug delivery platform for the treatment of diverse cutaneous diseases, such as metastatic melanoma.

Acyclovir is an acyclic purine nucleoside analog that is highly effective in inhibiting the herpes simplex virus. However, topical acyclovir has poor efficacy because of its low skin permeability. This study aimed to develop an acyclovir gel plaster containing sponge spicules (AGP-SS) to achieve synergistic improvements in skin absorption and deposition of acyclovir. The process of preparing the gel plaster was optimized by orthogonal experiments, while the composition of the formulation was optimized using the Plackett-Burman and Box-Behnken experimental designs. The selected formula was tested for physical properties, in vitro release, stability, ex vivo permeation, skin irritation, and pharmacokinetics. The optimized formulation exhibited good physical characteristics. In vitro release and ex vivo permeation studies showed that acyclovir release from AGP-SS was dominated by diffusion with significantly higher skin permeation (20.00 ± 1.07 μg/cm 2 ) than that of the controls (p < 0.05). Dermatopharmacokinetic analyses revealed that the maximum concentration (78.74 ± 11.12 μg/g), area under the curve (1091.81 ± 29.05 μg/g/h) and relative bioavailability (197.12) of AGP-SS were higher than those of the controls. Therefore, gel plaster containing sponge spicules show potential for development as transdermal delivery systems to achieve higher skin absorption and deposition of acyclovir, especially in deep skin layers.

This study reports that the use of low-frequency sonophoresis (LFS) in combination with sponge Haliclona sp. spicules (SHS), referred to as cSoSp (combined Sonophoresis and Spicules), can enhance the transdermal drug delivery in a synergistic manner. The topical application of cSoSp in vitro significantly enhanced the skin absorption of Fluorescent-Dextrans (4000 Da, FD-4K), a model drug of low-molecular-weight heparin (LMWH). The utilization of cSoSp dramatically increased the transdermal flux of FD-4K (188.6 ± 93.7 ng cm -2 h -1 ) compared to LFS (5.8 ± 3.1 ng cm -2 h -1 ) and SHS (3.2 ± 1.2 ng cm -2 h -1 ) among others. The mechanism of action of cSoSp could be attributed to the synergism between plenty of long-lasting nano-channels created by SHS and the disorders of SC lipids made by shock waves of LFS, which improves the homogeneity of the cavitation effects. Furthermore, LMWH (3000 Da) was transdermally delivered by using cSoSp to treat both superficial venous thrombosis (SVT) and deep venous thrombosis (DVT) in the marginal ear vein of rabbits with a good therapeutic effect. Furthermore, skin irritation and toxicity studies using guinea pigs indicated that cSoSp was nonirritating without any morphological changes in the keratinocytes. cSoSp offers a promising strategy to enhance the transdermal delivery of hydrophilic macromolecules such as heparin.

We report the development of sponge Haliclona sp. spicules, referred to as SHS, and its topical application in skin delivery of hydrophilic biomacromolecules, a series of fluorescein isothiocyanate-dextrans (FDs). SHS are silicious oxeas which are sharp-edged and rod-shaped (∼120 μm in length and ∼7 μm in diameter). SHS can physically disrupt skin in a dose-dependent manner and retain within the skin over at least 72 h, which allows sustained skin penetration of hydrophilic biomacromolecules. The magnitude of enhancement of FD delivery into skin induced by SHS treatment was dependent on its molecular weight. Specifically, SHS topical application enhanced FD-10 (MW: 10 kDa) penetration into porcine skin in vitro by 33.09 ± 7.16-fold compared to control group (p < 0.01). SHS dramatically increased the accumulation of FD-10 into and across the dermis by 62.32 ± 13.48-fold compared to the control group (p < 0.01). In vivo experiments performed using BALB/c mice also confirmed the effectiveness of SHS topical application; the skin absorption of FD-10 with SHS topical application was 72.14 ± 48.75-fold (p < 0.05) and 15.39 ± 9.91-fold (p < 0.05) higher than those from the PBS and Dermaroller microneedling, respectively. Further, skin irritation study and transepidermal water loss (TEWL) measurement using guinea pig skin in vivo indicated that skin disruption induced by SHS treatment is self-limited and can be recovered with time and efficiently. SHS can offer a safe, effective, and sustained skin delivery of hydrophilic biomacromolecules and presents a promising platform technology for a wide range of cosmetic and medical applications.

Chemotherapy-induced hair loss is a distressing side effect of cancer treatment, and medical interventions are often needed to address this problem. The objectives of this study were to evaluate the bioactivity of goat placenta (GP) extract on both normal and chemotherapy-induced hair cells and to develop PEGylated liposomes (PL) and microspicule (MS) formulations for promoting hair growth in patients with chemotherapy-induced hair loss. The bioactivities of GP extract on human follicle dermal papilla (HFDP) cells and cells damaged by chemotherapy were assessed. GP extract was incorporated into PLs and MS gel (PL-MS) and then investigated in vitro skin permeation and in vivo studies on the scalps of patients with chemotherapy-induced hair loss. GP extract stimulated HFDP cell proliferation in both normal and cisplatin-damaged cells. PL nanovesicles and MS gel worked synergistically to deliver macromolecular proteins into the skin and hair follicles. The application of GP extract-loaded PL-MS to the scalps of chemotherapy-treated patients for 12 weeks significantly enhanced the hair growth rate, without causing skin irritation. In conclusion, GP extract promoted the proliferation of hair cells damaged by chemotherapy, when this extract, combined with PL-MS, effectively delivered bioactive macromolecules across the skin and hair follicles, resulting in successful regrowth of hair post-chemotherapy.

Boesenbergia rotunda (L.) Mansf contained a potent anti-obesity agent. The objectives of this study were to investigate the anti-adipogenesis and lipolysis effects of panduratin A from B. rotunda extract and develop extract-loaded lipolytic body microspicule (MS) serum. Panduratin A that was separated from the ethanolic extract of B. rotunda in fraction 3 (BP-3) were studied the bioactivity of 3T3-L1 preadipocyte cells. The extract-loaded MS serum was formulated and evaluated for safety and efficacy. The BP-3 extract containing panduratin A at 0.29 g per g of the extract was not toxic to the cells at concentrations lower than 10 µg/mL, and the antiadipogenesis and lipolysis effects of the BP-3 extract were strong at 10 µg/mL. To deliver bioactive panduratin A into and through the skin, MS serum was successfully formulated. Application of BP-3 extract-loaded MS serum to the human thigh for 14 d reduced the thigh circumference and increased skin hydration and firmness. Although the skin erythema was increased, no severe redness or pain was found. In conclusion, BP-3 extract acts as a potent bioactive compound to inhibit adipocyte cells, and the antiadipogenesis and lipolysis effects of BP-3 extract in MS serum might play an important role as a potential lipolytic body product for reducing human subcutaneous fat mass.

Deer antler velvet (DAV) extract is important in stimulating the growth of skin and hair cells. The aim of this study was to develop DAV extract-loaded niosomes (NIs) and microspicules (MS) serum for effective hair growth. Bioactivities of DAV extract on human dermal papilla cells were studied. To develop protein delivery, NIs were prepared to entrap DAV extract and then loaded into MS serum (NI serum MS). Physicochemical properties and in vitro skin permeations were evaluated. The formulations were applied on human scalp and relative efficacies were evaluated. DAV extract at 2,000 µg/ml significantly enhanced cell proliferation and aggregation. DAV extract-loaded NI exhibited nanovesicle, narrow size distribution and negative surface charge. The vesicles were able to load into MS serum and showed significantly highest macromolecular protein permeation through the skin, with deposition into the deepest skin layer compared with other formulations. Applying the serum on human scalp for 14 and 30 days significantly enhanced hair elongation and melanin content, with increased skin hydration and decreased the erythema index, thereby promoting hair growth without skin irritation. NI serum MS containing DAV extract played an important role to deliver biomacromolecular protein through the skin and hair follicles, leading to effective hair growth.

Micro-needle patches have been recently used to increase skin permeability, which improves drug delivery, and for cosmetic purposes. However, these patches may often have limited efficacy due to insufficient skin penetration and reduced compliance caused by discomfort. We evaluated the efficacy and the safety of soluble micro-spicule containing epidermal growth factor (MS-EGF) for the treatment of periocular wrinkles. Twenty healthy volunteers aged 33 to 54 years were enrolled in a randomized, controlled, split-face study. For 4 weeks, a periocular wrinkle was treated daily with either a soluble MS-EGF cream or a cream containing EGF alone. All subjects underwent 8 weeks of follow-up. Efficacy was assessed using an ultrasonic measurement of dermal depth and density, digital skin image analysis, 5-point photonumeric scale for periocular wrinkles and subjective satisfaction. MS-EGF group showed statistically significant increase of dermal depth and density compared to EGF alone group after 4 and 8 weeks. In addition, there was a marked improvement shown in clinical and 3-dimensional skin image in MS-EGF group. The treatments were well-tolerated; no significant side-effect was noted. The MS-EGF formulation may represent an effective and biocompatible advance in the treatment of periocular wrinkles.

We report the development of sponge Haliclona sp. spicules, referred to as SHS, and its topical application in skin delivery of hydrophilic biomacromolecules, a series of fluorescein isothiocyanate-dextrans (FDs). SHS are silicious oxeas which are sharp-edged and rod-shaped (∼120 μm in length and ∼7 μm in diameter). SHS can physically disrupt skin in a dose-dependent manner and retain within the skin over at least 72 h, which allows sustained skin penetration of hydrophilic biomacromolecules. The magnitude of enhancement of FD delivery into skin induced by SHS treatment was dependent on its molecular weight. Specifically, SHS topical application enhanced FD-10 (MW: 10 kDa) penetration into porcine skin in vitro by 33.09 ± 7.16-fold compared to control group (p < 0.01). SHS dramatically increased the accumulation of FD-10 into and across the dermis by 62.32 ± 13.48-fold compared to the control group (p < 0.01). In vivo experiments performed using BALB/c mice also confirmed the effectiveness of SHS topical application; the skin absorption of FD-10 with SHS topical application was 72.14 ± 48.75-fold (p < 0.05) and 15.39 ± 9.91-fold (p < 0.05) higher than those from the PBS and Dermaroller microneedling, respectively. Further, skin irritation study and transepidermal water loss (TEWL) measurement using guinea pig skin in vivo indicated that skin disruption induced by SHS treatment is self-limited and can be recovered with time and efficiently. SHS can offer a safe, effective, and sustained skin delivery of hydrophilic biomacromolecules and presents a promising platform technology for a wide range of cosmetic and medical applications.

Micro-needle patches have been recently used to increase skin permeability, which improves drug delivery, and for cosmetic purposes. However, these patches may often have limited efficacy due to insufficient skin penetration and reduced compliance caused by discomfort. We evaluated the efficacy and the safety of soluble micro-spicule containing epidermal growth factor (MS-EGF) for the treatment of periocular wrinkles. Twenty healthy volunteers aged 33 to 54 years were enrolled in a randomized, controlled, split-face study. For 4 weeks, a periocular wrinkle was treated daily with either a soluble MS-EGF cream or a cream containing EGF alone. All subjects underwent 8 weeks of follow-up. Efficacy was assessed using an ultrasonic measurement of dermal depth and density, digital skin image analysis, 5-point photonumeric scale for periocular wrinkles and subjective satisfaction. MS-EGF group showed statistically significant increase of dermal depth and density compared to EGF alone group after 4 and 8 weeks. In addition, there was a marked improvement shown in clinical and 3-dimensional skin image in MS-EGF group. The treatments were well-tolerated; no significant side-effect was noted. The MS-EGF formulation may represent an effective and biocompatible advance in the treatment of periocular wrinkles.

We demonstrated that the topical combined use of sponge Haliclona sp. spicules (SHS) and flexible liposomes (FL), referred to as SFLS (SHS-Flexible Liposomes combined System), can result in synergy to improve the skin absorption and deposition of hyaluronic acid (HA), especially in deep skin layers, both in vitro and in vivo. SHS treatment can result in skin micro-channels which are continuous, deep enough (48.6 ± 13.5 μm) and available in large quantities (850 ± 125 micro-channels per mm2). These micro-channels gradually closed up in 120 h and also allowed the intact vesicles of flexible liposomes and vesicle-bound or vesicle-encapsulated HA to penetrate into the skin-deep layers under the driving force of transdermal osmotic gradients. Specifically, SFLS topical application enhanced the penetration of FITC-HA (MW: 250 kDa) into porcine skin in vitro up to 23.2 ± 3.7%, which is 19.4 ± 3.1-fold (p < 0.001) that of a Phosphate Buffered Saline (PBS) group, 3.4 ± 0.5-fold (p < 0.01) that of an SHS group and 3.6 ± 0.6-fold (p < 0.01) that from the combined use of a Dermaroller and flexible liposomes. Moreover, SFLS can lead to significantly enhanced skin deposition of HA in all skin layers, especially in deep skin layers: up to 86.8 ± 4.1% of HA absorbed by skin was accumulated in deep skin layers. The effectiveness of SFLS topical application was also confirmed in vivo by using BALB/c mice. In addition, a skin irritation and toxicity study showed that the SFLS treatment may cause very minimal redness and the skin can recover in a short time. In sum, the combined use of SHS and FL (SFLS) offers a promising strategy to safely and effectively improve the skin delivery of hydrophilic biomacromolecules such as HA.

Wharton's Jelly-derived mesenchymal stem cell (WJ-MSC) secretome contains diverse bioactive factors with potential for skin regeneration, but its clinical efficacy is limited by poor transdermal delivery. In this study, we developed a dual-delivery system by nanoencapsulating WJ-MSC secretome and coating it onto marine sponge-derived spicules. Physicochemical characterization, in vitro assays (fibroblast and keratinocyte proliferation, keratinocyte migration, type I procollagen secretion, and antioxidant activity), and in vivo penetration studies were conducted. A single-arm clinical trial evaluated dermal absorption, pore characteristics, skin texture, wrinkles, and pigmentation following topical application. Transdermal penetration efficiency was significantly higher in the nano-coated spicule group than in the uncoated secretome control. In vitro, secretome treatment promoted fibroblast and keratinocyte activity, accelerated wound closure, and increased collagen synthesis. Clinically, a single application enhanced dermal absorption and significantly reduced pore number, while two weeks of treatment decreased wrinkles and pigmentation. Spicule-based nanoencapsulation effectively overcomes the skin barrier, enhances the regenerative activity of WJ-MSC secretome, and induces measurable clinical improvements in skin rejuvenation. This platform represents a promising cosmetic and therapeutic strategy in dermatology.

The influence of physiological factors on the solubility of drug compounds has been thoroughly investigated in humans. However, as these factors vary between species and since many in vivo studies are carried out in rats or mice, it has been difficult to establish sufficient in vitro in vivo relations. The aim of this study was to develop a physiologically relevant in vitro dissolution model simulating the gastrointestinal (GI) fluids of fasted rats and compare it to previously published in vitro and in vivo data. To develop the in vitro model, the pH was measured in situ in six segments of the GI tract of anesthetised rats, then the fluids from the stomach, the proximal and the distal small intestine were collected and characterized with regard to osmolality, and bile acid and phospholipid concentration. The pH and osmolality were found to increase throughout the GI tract. The bile acids and phospholipids were present in high concentrations in the proximal small intestine, and the bile acid concentration doubled in the distal part, where the phospholipid concentration decreased. Matrix-assisted laser desorption ionisation mass spectrometry imaging was applied on a cross section of the small intestine, to study which bile acids and phospholipid classes were present in the small intestine of rats. Both cholic acid, taurocholic acid and glycocholic acid were detected, and phosphatidylcholine (34:2) was found to be mainly present in the intestinal wall or mucus, whereas lysophosphatidylcholine (16:0) was also detected in the lumen. Based on these observations, biorelevant media were developed to simulate fluids in the stomach and the proximal part of the small intestine in fasted rats. The media were implemented in a two-step in vitro dissolution model, which was found to better predict the in vivo performance of furosemide, when compared to previously published in vitro and in vivo data.

We demonstrated for the first time that a marine-derived antimicrobial peptide (AMP), Sph12-38, exhibit high antimicrobial activity against P. acnes with a minimum bactericidal concentration (MBC) value of 7 μM. Meanwhile, Sph12-38 has no significant cytotoxicity to human keratinocytes (HKs) at its high concentration (33.5 μM). The topical application of sponge Haliclona sp. spicules (SHS) dramatically enhanced the skin penetration of Sph12-38 up to 40.9 ± 5.9% (p < 0.01), which was 6.1 ± 0.9-fold higher than that of Sph12-38 alone. Further, SHS resulted in the accumulation of most Sph12-38 in viable epidermis and dermis. Further, the combined use of Sph12-38 and SHS resulted in a cure rate of 100% for rabbit ear acne treatment in vivo for two weeks, while the one induced by other groups was 40%, 0% and 0% for SHS alone, Sph12-38 alone and control group, respectively. The strategy of combined using AMP and SHS can also be applied in a rational designed topical delivery system for the management of other deep infection of the skin. The effectiveness of SHS by itself on the treatment of acne was also demonstrated by clinical trials. After 14 days of treatment by 1% SHS gel. The number of skin lesions decreased by 51.4%.

Collagen microparticles were prepared using marine sponge collagen. For this purpose a previous method by Rössler et al. (J. Microencapsul. 12 (1995) 49) of emulsification and cross-linking of native calf collagen was modified. The modified method for sponge collagen microparticles (SCMPs) achieved a yield of 10%. Scanning electromicroscopic photographs showed spherical particles with a diameter of 120-300 nm and photon correlation spectroscopic measurements indicated particle size range from 126 (+/-2.9) to 2179 (+/-342) nm. This broad size distribution was caused by some agglomerates that could not be destroyed by ultrasonication. The surface charge was measured as a function of pH. At pH 2.8 the particles were nearly uncharged, at pH 9.0 the particles showed a strong negative charge of about -60 mV. The preformed SCMPs were loaded by adsorption of all-trans retinol. A loading of up to 8% was obtained. Retinol-loaded SCMPs were incorporated into hydrogels and drug stability was investigated. The in vitro penetration of retinol into hairless mice skin in this formulation was compared to retinol formulations without microparticles. The SCMPs had no influence on the chemical stability of retinol in the hydrogel. The dermal penetration of retinol into the skin increased significantly by approximately two-fold.

Transdermal administration of estradiol offers advantages over oral estrogens for hormone replacement therapy regarding side effects by bypassing the hepatic presystemic metabolism. The objective of this study was to develop nanoparticles of Chondrosia reniformis sponge collagen as penetration enhancers for the transdermal drug delivery of 17beta-estradiol-hemihydrate in hormone replacement therapy. Collagen nanoparticles were prepared by controlled alkaline hydrolysis and characterized using atomic force microscopy and photon correlation spectroscopy. Estradiol-hemihydrate was loaded to the nanoparticles by adsorption to their surface, whereupon a drug loading up to 13.1% of sponge collagen particle mass was found. After incorporation of drug-loaded nanoparticles in a hydrogel, the estradiol transdermal delivery from the gel was compared with that from a commercial gel that did not contain nanoparticles. Saliva samples in postmenopausal patients showed significantly higher estradiol levels after application of the gel with nanoparticles. The area under the curve (AUC) for estradiol time-concentration curves over 24 hours was 2.3- to 3.4-fold higher and estradiol levels 24 hours after administration of estradiol were at least twofold higher with the nanoparticle gel. The hydrogel with estradiol-loaded collagen nanoparticles enabled a prolonged estradiol release compared to a commercial gel and yielded a considerably enhanced estradiol absorption. Consequently, sponge collagen nanoparticles represent promising carriers for transdermal drug delivery.

In vitro anti-tumour and anti-radical activities of the acetone extract of the freshwater sponge Ochridaspongia rotunda were the subject of this study. The extract was found to be highly cytotoxic to human lung tumour cell line A-549 reaching IC 50 value of 5.01 ± 0.21 μg/mL. Indeed, it displayed only 2-fold less anti-tumour activity than doxorubicin (IC 50 value 2.42 ± 0.13 μg/mL) used as a positive control. The same extract was also found to be almost 37-fold more selective against A-549 vs. MRC-5 (normal) lung cells, in difference to weak selectivity of doxorubicin (less than 3-fold). Its profound anti-DPPH radical activity comparable to that of quercetin (IC 50 values 3.68 ± 0.19 and 3.14 ± 0.09 μg/mL, respectively) coupled with no signs of genotoxicity in the comet assay (MRC-5 cell line, vs. doxorubicin) has actually implicated the importance of this animal bioresource in searching for pharmaceutically useful bioactive compounds of natural origin.

We demonstrated that the topical combined use of sponge Haliclona sp. spicules (SHS) and flexible liposomes (FL), referred to as SFLS (SHS-Flexible Liposomes combined System), can result in synergy to improve the skin absorption and deposition of hyaluronic acid (HA), especially in deep skin layers, both in vitro and in vivo. SHS treatment can result in skin micro-channels which are continuous, deep enough (48.6 ± 13.5 μm) and available in large quantities (850 ± 125 micro-channels per mm2). These micro-channels gradually closed up in 120 h and also allowed the intact vesicles of flexible liposomes and vesicle-bound or vesicle-encapsulated HA to penetrate into the skin-deep layers under the driving force of transdermal osmotic gradients. Specifically, SFLS topical application enhanced the penetration of FITC-HA (MW: 250 kDa) into porcine skin in vitro up to 23.2 ± 3.7%, which is 19.4 ± 3.1-fold (p < 0.001) that of a Phosphate Buffered Saline (PBS) group, 3.4 ± 0.5-fold (p < 0.01) that of an SHS group and 3.6 ± 0.6-fold (p < 0.01) that from the combined use of a Dermaroller and flexible liposomes. Moreover, SFLS can lead to significantly enhanced skin deposition of HA in all skin layers, especially in deep skin layers: up to 86.8 ± 4.1% of HA absorbed by skin was accumulated in deep skin layers. The effectiveness of SFLS topical application was also confirmed in vivo by using BALB/c mice. In addition, a skin irritation and toxicity study showed that the SFLS treatment may cause very minimal redness and the skin can recover in a short time. In sum, the combined use of SHS and FL (SFLS) offers a promising strategy to safely and effectively improve the skin delivery of hydrophilic biomacromolecules such as HA.

Sponges, which are parasitic on plants widely found in lakes and oceans, represent a vast resource that has yet to be effectively utilised. Sponge spicules (SS), which contain high amounts of silica dioxide, form after long-term biomineralisation. In this study, SS attached to plant bodies were subjected to acid and heat treatments, followed by grinding, to obtain 10-40-nm siliceous sponge spicules (SSS). SSS and polylactic acid (PLA) were then combined to create 50-450-nm PLA/SSS composite nanofibers. The morphology and bioactivity of the electrospun PLA/SSS nanofibers were examined; the tensile, thermal, and water-resistant properties of the fibers were also evaluated. Our results showed a dramatic enhancement in the thermal and tensile properties of PLA with increasing SSS content; specifically, a 3 wt% increase in SSS content resulted in a 47 °C increase in the initial decomposition temperature and a 73.3-MPa increase in Young's modulus. The water resistance of PLA/SSS increased with SSS content, as indicated by the increase in the water contact angle compared with PLA nanofibers. PLA/SSS nanofibers also exhibited slightly enhanced human foreskin fibroblast cell proliferation, good cytocompatibility, and an antibacterial effect. The enhanced antibacterial and biodegradable properties of PLA/SSS are expected to be useful in biomedical material applications.

The topical application of exosomes secreted by mesenchymal stem cells (MSC-Exos) on the skin is a very new and interesting topic in the medical field. In this study, we aimed to investigate whether marine sponge Haliclona sp. spicules (SHSs) could effectively enhance the skin delivery of human umbilical cord-derived MSC-Exos (hucMSC-Exos), and further evaluate the topical application of hucMSC-Exos combined with SHSs in rejuvenating photoaged mouse skin. SHSs were isolated from the explants of sponge Haliclona sp. with our proprietary method, and hucMSC-Exos were prepared from the conditioned medium of hucMSCs using ultracentrifugation. The effects of SHSs on the skin penetration of fluorescently labeled hucMSC-Exos were determined using confocal microscopy in vitro (porcine skin) and in vivo (mouse skin). The therapeutic effects of hucMSC-Exos coupled with SHSs against UV-induced photoaging in mice were assessed by using microwrinkles analysis, pathohistological examination and real-time RT-PCR. We also tested the skin irritation caused by the combination of hucMSC-Exos and SHSs in guinea pigs. In vitro results showed that hucMSC-Exos could not readily penetrate through porcine skin by themselves. However, SHSs increased the skin absorption of exosomes by a factor of 5.87 through creating microchannels. Similar penetration enhancement of hucMSC-Exos was observed after SHSs treatment in mice. The combined use of hucMSC-Exos and SHSs showed significant anti-photoaging effects in mice, including reducing microwrinkles, alleviating histopathological changes, and promoting the expression of extracellular matrix constituents, whereas hucMSC-Exos alone produced considerably weaker effects. Skin irritation test showed that the combination of hucMSC-Exos and SHSs caused slight irritation, and the skin recovered shortly. SHSs provide a safe and effective way to enhance the skin delivery of MSC-Exos. Moreover, the combination of MSC-Exos and SHSs may be of much use in the treatment of photoaging.

The present study aimed to understand the killing effects of seaweed derived metabolite Callophycin A (Cal A). In vitro studies confirmed that the beneficial effects of Cal A on the viability of C. albicans. To enhance the biological activity, we used to demonstrated that chitosan and spicules as a drug carrier. The Callophycin A loading was confirmed by spectral variation of FT-IR and morphological variation by SEM. Moreover, around 65% and 38% of Cal A was successfully loaded in chitosan and spicules respectively. Further, VVC induced animal model experiments confirmed that the candidicidal activity of 1% clotrimazole, Cal A, Cal@Chi and Cal@Spi. After 6 days of treatment Cal@Chi produces a significant reduction in the fungal burden of vaginal lavage. The histo-morphological alterations also evidenced that the protective role of Cal@Chi in VVC model. The present investigations are known to be the first and foremost study to discriminate the potentiality of Cal A composites. Cal A loaded chitosan nanoparticles could be used as an alternative strategy for the development of the novel marine natural product based topical applications.

Dermal delivery is versatile in therapeutics as well as cosmetics in pursuit of enhancing safety/efficacy and alleviating pain/fear to alternate oral/injective administrations. Natural siliceous spicules offer a potential approach via simple topical medications to circumvent poor penetrations through the skin barrier by loading, carrying, and releasing the active ingredients in a highly efficient and controlled manner. The delivery of ingredients loaded on spicules is assessed to improve the dermal efficacy compared to simple topical treatments. First, needle-like spicules were isolated from natural freshwater sponges. Then, the active ingredient was loaded via liposome formations. Finally, the dermal efficiency was evaluated. Natural siliceous spicules were purified, sorted, and fully characterized to appear 250 μm of length and 12 μm of diameter on average. A model ingredient, pectolinarin, was efficiently loaded onto the internal space of spicules via lecithin-based liposome formations. The penetration was visualized using a porcine skin sample with a fluorescent dye and assessed quantitatively by a Franz diffusion cell system. Dermal absorption rate was measured 73.4%, while the percutaneous penetration rate was 2.2%. The release pattern turned out a simple diffusion analyzed by Fick's law and Higuchi model. The liposomes loaded onto spicules were further stabilized by a hydrophobic capsulation, which may benefit the overall efficacy of the ingredient. A novel dermal delivery system is beneficial to improve the topical efficacy of biologically active ingredients. The outcomes shed a light upon developing skin-protective/improving cosmetics and therapeutics with enhanced performance.

Poor patient adherence with antiacne medications is a common clinical challenge. DMT310, a natural, topical product with a once-weekly application schedule, may alleviate this obstacle. Evaluate the safety, tolerability, and efficacy of DMT310 in treating moderate-to-severe acne. This 12-week, randomized, double-blind, placebo-controlled, multicenter clinical trial enrolled participants 12 years and older with moderate-to-severe acne. The intent-to-treat population included a total of 181 participants (DMT310, N = 91; placebo, N = 90). Participants who received DMT310 vs participants treated with placebo demonstrated a statistically significant greater reduction in the number of inflammatory and noninflammatory lesions at all time points: inflammatory lesion counts at week 12 (-15.64 vs -10.84, P < .001); noninflammatory lesion counts at week 12 (-18.26 vs -12.41, P < .001). DMT310-treated participants also had higher rates of Investigator's Global Assessment treatment success than participants in the placebo group at all time points: Investigator's Global Assessment at week 12 (44.40% vs 17.78%; P < .001). No serious treatment related adverse events occurred. DMT310 once-weekly topical treatment significantly reduced both inflammatory and noninflammatory lesions and yielded a greater proportion of Investigator's Global Assessment treatment success at all time points in participants with moderate-to-severe acne.

Marine-sponge-derived spicule microparticles (SPMs) possess unique structural and compositional features suitable for bone tissue engineering. However, significant challenges remain in establishing their osteogenic mechanism and practical application in animal models. This study explores the biomimetic potential of SPM in orchestrating biomineralization behavior and modulating the Yes-associated protein 1/transcriptional coactivator with PDZ-binding motif (YAP/TAZ) pathway both in vitro and in vivo. Characterization of SPM revealed a structure comprising amorphous silica oxide mixed with collagen and trace amounts of calcium and phosphate ions, which have the potential to facilitate biomineralization. Structural analysis indicated dynamic biomineralization from SPM to hydroxyapatite, contributing to both in vitro and in vivo osteoconductions. In vitro assessment demonstrated dose-dependent increases in osteogenic gene expression and bone morphogenetic protein-2 protein in response to SPM. In addition, focal adhesion mediated by silica diatoms induced cell spreading on the surface of SPM, leading to cell alignment in the direction of SPM. Mechanical signals from SPM subsequently increased the expression of YAP/TAZ, thereby inducing osteogenic mechanotransduction. The osteogenic activity of SPM-reinforced injectable hydrogel was evaluated in a mouse calvaria defect model, demonstrating rapid vascularized bone regeneration. These findings suggest that biomimetic SPM holds significant promise for regenerating bone tissue.

We demonstrated for the first time that a marine-derived antimicrobial peptide (AMP), Sph12-38, exhibit high antimicrobial activity against P. acnes with a minimum bactericidal concentration (MBC) value of 7 μM. Meanwhile, Sph12-38 has no significant cytotoxicity to human keratinocytes (HKs) at its high concentration (33.5 μM). The topical application of sponge Haliclona sp. spicules (SHS) dramatically enhanced the skin penetration of Sph12-38 up to 40.9 ± 5.9% (p < 0.01), which was 6.1 ± 0.9-fold higher than that of Sph12-38 alone. Further, SHS resulted in the accumulation of most Sph12-38 in viable epidermis and dermis. Further, the combined use of Sph12-38 and SHS resulted in a cure rate of 100% for rabbit ear acne treatment in vivo for two weeks, while the one induced by other groups was 40%, 0% and 0% for SHS alone, Sph12-38 alone and control group, respectively. The strategy of combined using AMP and SHS can also be applied in a rational designed topical delivery system for the management of other deep infection of the skin. The effectiveness of SHS by itself on the treatment of acne was also demonstrated by clinical trials. After 14 days of treatment by 1% SHS gel. The number of skin lesions decreased by 51.4%.

Psoriasis is an autoimmune skin disease where dendritic cells play a crucial role. In psoriasis topical treatment, pathologically-induced thickening of the stratum corneum and epidermal hyperplasia severely hinder the penetration of drugs into the deep skin, thereby reducing therapeutic efficacy. Here, we developed the curcumin-POPG phospholipid complex coated Sponge Haliclona sp. Spicules (CUR-Ph-SHS) with the supercritical antisolvent-fluidized bed coating (SAS-FB) technology. This innovative formulation was designed to enhance skin permeation and suppress dendritic cell activation in the psoriasis mouse model. Through the process of SAS-FB, the phospholipid complex formed a uniform amorphous coating on CUR-Ph-SHS, which increased the dissolution of the active ingredient and self-assembled into nanoparticles after release. Moreover, the nanoparticles released from CUR-Ph-SHS could be efficiently internalized by dendritic cells, demonstrating superior anti-maturation activity in vitro compared to raw curcumin. Owing to its dual-tip structure and excellent wettability, CUR-Ph-SHS could effectively penetrate the stratum corneum, releasing self-assembled phospholipid complex nanoparticles into the deep skin and draining lymph nodes of psoriasis mice. In addition, the in-vivo anti-psoriasis experiment revealed that CUR-Ph-SHS relieved the psoriasis symptoms and inhibited the expression of inflammatory cytokines in the skin lesions. Meanwhile, CUR-Ph-SHS successfully inhibited the activation of dendritic cells while inducing the proliferation of regulatory T cells. Overall, these findings highlight CUR-Ph-SHS as novel coated microneedles for remodeling skin immune homeostasis in psoriasis topical therapy, suggesting potential applications in treating other skin inflammatory diseases.

Diagnostic and therapeutic drug monitoring are necessary means of precision medicine. Our previous study demonstrated that the sponge Haliclona sp. spicules (SHS) can be employed as a new class of microneedles, capable of forming a substantial number of microchannels within the skin. In this study, dermal interstitial fluid (ISF) was obtained via percutaneous extraction using sponge Haliclona sp. spicules (SHS) to explore the correlation between the concentration of target compounds in the extract and their content in the skin or blood. Methodologically, we compared SHS with dermaroller, ultrasound, and reverse iontophoresis for interstitial fluid (ISF) extraction efficacy in vitro, while optimizing extraction parameters (dosage of SHS, extraction solvent, extraction time) via orthogonal experimental design. Target compounds spanning diverse physicochemical properties (fluorescein sodium, rhodamine B, salicylic acid, FD 1 k, urea, glucose) were evaluated. We also detected the concentration of target compounds and their levels in the blood in vivo. Key results showed that robust linear correlations were observed between the detected concentration of target compounds through SHS and their levels in various skin layers, as well as in the blood. Additionally, we developed a mathematical model predicting extraction efficiency log kp = 2.9711 + 0.00235MV - 0.059526 log P + 6.7506 log MW-0.6. These findings provide a new non-invasive percutaneous sampling strategy.

Topical anesthesia offers a painless alternative to injections but is limited by the low skin permeability of local anesthetics through the stratum corneum, leading to a slow onset. In this study, we introduce a novel spicule-assisted topical delivery system using Sponge Haliclona sp. spicules (SHS) combined with Carbomer gel to enhance the transdermal absorption of lidocaine hydrochloride (LH). SHS act as dispersed microneedles, creating numerous microchannels that significantly improve skin permeability. In vitro, SHS increased total skin absorption of LH nearly tenfold (78.45 ± 6.96%) and accelerated drug penetration into deeper skin layers (97.5% in dermis and receptor compartments). In vivo, SHS-LH treatment achieved a maximum anesthetic effect within 10 min, markedly faster and stronger than conventional topical application. A pilot clinical trial confirmed that the SHS-Carbomer gel system halved the onset time of lidocaine and significantly prolonged its anesthetic effect. These findings demonstrate that the SHS-based dispersed microneedle system provides a rapid, safe, and needle-free alternative for local anesthesia, offering a substantial improvement over conventional topical formulations.

Diabetes mellitus is a chronic metabolic disorder that requires long-term management, as persistent hyperglycemia may lead to severe complications, thereby making real-time glucose monitoring and precise control essential. Traditional subcutaneous insulin injections are inconvenient, exhibit poor patient compliance and dosing accuracy, and do not provide real-time feedback, thereby hindering personalized therapy. Microneedles (MNs) offers a minimally invasive, user-friendly, and self-administered alternative; however, existing MN systems still exhibit low drug-loading capacity, insufficient controlled-release performance, a lack of glucose-monitoring capability, and inherent infection risks, which limit their application in long-term diabetes management. To address these limitations, we developed a novel antimicrobial sponge-hydrogel bilayer MNs system (ASMNs@PVA/SA-INS) that integrates insulin delivery, glucose sensing, and antimicrobial protection. The system comprises a polyvinyl alcohol/sodium alginate (PVA/SA) hydrogel layer for controlled insulin release and a sponge MN base functionalized silver nanoparticles to provide antibacterial activity. Experiments results demonstrated that over 83 % of insulin was released within 24 h, effectively controlling blood glucose for more than 8 h. Additionally, the system responded to glucose level changes within 10 min through a visible color change, indicating real-time sensing capability. This platform offers a promising strategy for intelligent and individualized diabetes management.

Sponge spicules have emerged as promising biomaterial scaffolds due to their biocompatibility and unique structural properties; however, achieving stable and bioactive functionalization remains a key challenge. The tripeptide GHK is known to promote collagen synthesis and wound repair, yet its therapeutic efficacy is often limited by rapid diffusion and instability. Here, we report ALTUM, a thiol-functionalized sponge spicule composite in which GHK is covalently conjugated via disulfide linkage to enable controlled and redox-responsive peptide delivery. ALTUM exhibited sustained GHK retention under physiological and storage conditions, while exposure to reduced glutathione (GSH) selectively accelerated peptide release through disulfide bond cleavage. This dual release behavior-long-term stability combined with reduction-triggered activation-distinguishes ALTUM from conventional delivery systems. The composite also demonstrated structural stability under thermal, cyclic, and photostability conditions. In an artificial human skin model, ALTUM enhanced dermal penetration of GHK and significantly increased collagen deposition in the dermal layer, demonstrating its capacity to promote collagen production within deeper skin tissue, compared to simple spicule-peptide mixtures. ALTUM was fabricated at an optimized spicule-to-peptide ratio of 3% ( w / w ), preserving the needle-shaped spicule morphology after surface modification. In vitro, ALTUM exhibited a sustained release profile, with GHK release markedly accelerated in the presence of 10 mM glutathione (GSH) compared with non-reductive conditions, reaching approximately 60% cumulative release over 35 days. In the bioprinted artificial human skin model, ALTUM delivered 9.72 ng/cm 2 of GHK, more than five-fold higher than the physical mixture of spicules and free GHK (1.9 ng/cm 2 ), and significantly increased type I collagen expression in human dermal fibroblasts. Mechanistically, ALTUM-mediated delivery was associated with increased TGF-β expression and engagement of the SMAD signaling pathway, as indicated by increased phosphorylation of SMAD2/3, consistent with involvement of the TGF-β-SMAD axis in the observed collagen induction. Collectively, these findings establish ALTUM as a structurally stable, redox-responsive dermal delivery platform that enhances collagen synthesis and skin regeneration.

Marine collagens are environmentally friendly and biologically compatible collagens derived from aquatic organisms, including invertebrates like sponges and jellyfish. Starting from marine collagens as raw materials, sophisticated purification procedures lead to low-molecular-weight, bioactive fragments that are valuable functional ingredients for nutritional supplements, cosmeceuticals, and medical devices in regenerative medicine. From the mid-2010s to the late 2010s, several high-quality studies highlighted the skin bioregenerative properties of marine collagen-derived oligopeptides. The purpose of this review is to discuss these properties and their rationale. This review only analyzes studies focused on skin regeneration published in indexed journals with significant impact factors (the rule was rigid for human studies), supplemented by a few contributions from Google Scholar for methodologically sound in vitro and animal studies. Activation of skin fibroblasts with high systemic bioavailability after oral intake supports the bioregenerative properties of hydrolyzed marine collagen. Marine collagen hydrolysates do not cause irritation or inflammation and have a negligible impact on pro-inflammatory mediators in animal studies. Preclinical research indicates that 50 µg/mL of hydrolyzed marine collagen peptides accelerates cell migration almost as effectively as 10 µg/mL of recombinant human epidermal growth factor, although the predictive value of in vitro studies for humans remains uncertain. Accelerated wound healing with collagen neosynthesis is associated with increased expression in immunohistochemistry of platelet-endothelial cell adhesion molecule-1, basic fibroblast growth factor, and transforming growth factor ß-1. Furthermore, enzyme-treated hydrolysates produced under acidic conditions exhibit antioxidant effects without affecting pro-inflammatory cytokines, while enzyme-treated hydrolysates under alkaline conditions have the opposite effect. An increasing number of preclinical and human studies highlight the skin and overall bioregenerative properties of hydrolyzed marine collagens. Their high systemic intestinal absorption after oral intake distinguishes them from hydrolysates from other sources. Long-term safety should be a primary concern of future research.

Mounting evidence showing that local nitric oxide (NO) delivery may significantly improve the wound healing process has stimulated the development of wound dressings capable of releasing NO topically. Herein, we describe the preparation of a self-expandable NO-releasing hydrolyzed collagen sponge (CS), charged with the endogenously found NO donor, S-nitrosoglutathione (GSNO). We show that cold pressed and GSNO-charged CS (CS/GSNO) undergo self-expansion to its original 3D shape upon water absorption to a swelling degree of 2,300 wt%, triggering the release of free NO. Topical application of compressed CS/GSNO on wounds in an animal model showed that exudate absorption by CS/GSNO leads to the release of higher NO doses during the inflammatory phase and progressively lower NO doses at later stages of the healing process. Moreover, treated animals showed significant increase in the mRNA expression levels of monocyte chemoattractant protein-1 (MCP-1), murine macrophage marker (F4/80), transforming growth factor beta (TGF-β), stromal cell-derived factor 1 (SDF-1), insulin-like growth factor-1 (IGF-1), nitric oxide synthase(iNOS), and matrix metalloproteinase(MMP-9). Cluster differentiation 31 (CD31), vascular endothelial growth factor (VEGF), and F4/80 were measured on Days 7 and 12 by immunohistochemistry in the cicatricial tissue. These results indicate that the topical delivery of NO enhances the migration and infiltration of leucocytes, macrophages, and keratinocytes to the wounded tissue, as well as the neovascularization and collagen deposition, which are correlated with an accelerated wound closure. Thus, self-expandable CS/GSNO may represent a novel biocompatible and active wound dress for the topical delivery of NO on wounds.

This study demonstrated both adipose-derived stem cells (ASCs) in vitro and in vivo combined with three-dimensional (3D) porous sponge matrices on implant wound healing. Sponge matrices were created from hyaluronic acid (HA), collagen (Col), and gelatin (Gel), constructing two types: HA-L (low content) and HA-H (high content), to be cross-linked with 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC). Fourier transform infrared spectroscopy method verified carboxyl groups of HA and amino groups of Col and Gel reacting between the raw materials and scaffolds to identify the successive cross-linking. The swelling ratios of two types of sponge matrices were analyzed by water absorption capabilities, and the results displayed both over 30-fold dry scaffold weight enhancements. In biodegradation tests, matrices were hydrolyzed over time by three cutaneous enzymes, hyaluronidase, lysozyme, and collagenase I. ASCs from rats were cultured within the HA-H scaffold, demonstrating higher antioxidative abilities and secretions on related genes and proteins compared to the other two groups. The ASC HA-H matrix promoted cell proliferation to stimulate capillary angiogenesis inducer secretions, including vascular endothelial growth factor (VEGF) and transforming growth factor- β (TGF- β ). In vivo histological examinations showed ASCs from implanted HA-H implant transported into the subcutis, and rat skin cells also infiltrated into the original matrix zone to increase the extracellular matrix (ECM) reconstructions. Our experimental data revealed that the ASC HA-H sponge implant was effective in improving wound repair.

This two-arm, parallel-group, double-blind, randomized clinical trial design evaluated the risk of postoperative bleeding in anticoagulated patients undergoing dental extraction treated with topical TXA in comparison to collagen-gelatin sponge. Forty patients were randomly included in one of the study groups: (1) topical use of 4.8% TXA solution; and (2) resorbable hydrolyzed collagen-gelatin sponge applied to the surgical alveolus. Primary outcomes were postoperative bleeding episodes and secondary outcomes were thromboembolic events and postoperative INR values. The relative risk (RR), the absolute risk reduction (RAR) and the number needed to treat (NNT) were used as effect estimates and calculated from the counting of bleeding episodes observed during the first postoperative week. The bleeding rate under the TXA treatment was 22.2%, while in the collagen-gelatin sponge group it was 45.7%, resulting in a RR of 0.49 (95% CI 0.24-099; p = 0.046), RAR 23.5% and NNT 4.3. TXA was more effective in reducing bleeding in surgical sites located in the mandible (RR = 0.10; 95% CI 0.01-0.71; p = 0.021) and the posterior region (RR = 0.39; 95% CI 0.18-0.84; p = 0.016). Within the limitations of the study it seems that topical TXA is more effective in controlling bleeding after tooth extractions in anticoagulated patients than collagen-gelatin sponge. CLINICAL TRIAL REGISTRATION: RBR-83qw93.

Marine animal by-products of the food industry are a great source of valuable biomolecules. Skins and bones are rich in collagen, a protein with various applications in food, cosmetic, healthcare, and medical industries in its native form or partially hydrolyzed (gelatin). Salmon gelatin is a candidate of interest due to its high biomass production available through salmon consumption, its biodegradability, and its high biocompatibility. However, its low mechanical and thermal properties can be an obstacle for various applications requiring cohesive material. Thus, gelatin modification by cross-linking is necessary. Enzymatic cross-linking by microbial transglutaminase (MTG) is preferred to chemical cross-linking to avoid the formation of potentially cytotoxic residues. In this work, the potential of salmon skin gelatin was investigated, in a comparative study with porcine gelatin, and an enzymatic versus chemical cross-linking analysis. For this purpose, the two cross-linking methods were applied to produce three-dimensional, porous, and mechanically reinforced hydrogels and sponges with different MTG ratios (2%, 5%, and 10% w/w gelatin). Their biochemical, rheological, and structural properties were characterized, as well as the stability of the material, including the degree of syneresis and the water-binding capacity. The results showed that gelatin enzymatically cross-linked produced material with high cross-linking densities over 70% of free amines. The MTG addition seemed to play a crucial role, as shown by the increase in mechanical and thermal resistances with the production of a cohesive material stable above 40 °C for at least 7 days and comparable to porcine and chemically cross-linked gelatins. Two prototypes were obtained with similar thermal resistances but different microstructures and viscoelastic properties, due to different formation dynamics of the covalent network. Considering these results, the enzymatically cross-linked salmon gelatin is a relevant candidate as a biopolymer for the production of matrix for a wide range of biotechnological applications such as food packaging, cosmetic patch, wound healing dressing, or tissue substitute.