Description

Hyaluronic Acid Peptide Powder

 

 

CAS Number	9004-61-9
Other Names	Hyaluronan, Hyaluronate, Hylan, Ácido Hialurónico, Glycoaminoglycan, and Glycoaminoglycane
IUPAC Name	(1→4)-(2-Acetamido-2-deoxy-D-gluco)-(1→3)-D-glucuronoglycan
Molecular Formula	(C₁₄H₂₁NO₁₁)ⁿ
Molecular Weight	20–200 kDa
Purity	≥99% Pure (LC-MS)
Liquid Availability	N/A
Powder Availability	15 grams, 150 capsules (100mg/capsule, 15grams bottle)
Storage Condition	Store in cool dry environment, away from direct sunlight.
Certificate Of Analysis	Due to this product’s nature, this chemical does not have a COA associated with it.
Terms	All products are for laboratory developmental research USE ONLY. Products are not for human consumption.

 

What is Hyaluronic Acid?

 

Hyaluronic acid (HA) is a naturally occurring glycosaminoglycan composed of repeating units of glucuronic acid and N-acetylglucosamine, widely distributed throughout connective, epithelial, and neural tissues. The compound functions synergistically with peptide-based molecules in maintaining skin hydration, elasticity, and structural integrity. Known for its exceptional water-binding capacity, hyaluronic acid plays a key role in tissue hydration, wound healing, and joint lubrication. In both biomedical and cosmetic applications, low–molecular weight and peptide-conjugated forms of hyaluronic acid are increasingly utilized to enhance bioavailability, stimulate collagen synthesis, and promote cellular regeneration.

 

Main Research

1) Application of hyaluronic acid-based hydrogel has the potential to treat diabetic wounds.

2) Hyaluronic acid-based nanomedicine shows enhanced potency against hepatocellular carcinoma.

 

Selected Data
1) This study completed by researchers Xu et al describes the synthesis, characterization, and biological evaluation of catechin-loaded HAMA-PBA hydrogels (HMPC hydrogels) designed for antioxidant and wound-healing applications, particularly in diabetic conditions.
Hyaluronic acid methacrylate (HAMA) was synthesized by dissolving hyaluronic acid (HA, 2 g) in deionized water (100 mL) and adding methacrylic anhydride (7 mL) dropwise while maintaining the pH between 8 and 10 with 1 M NaOH. The reaction proceeded for 24 hours in the dark. The resulting mixture was washed with ethanol, dialyzed in deionized water for five days, and then freeze-dried to obtain purified HAMA. To synthesize the boronic acid-functionalized derivative (HAMA-PBA), HAMA (1 g) was dissolved in 200 mL of deionized water and reacted with 3-aminomethyl phenylboronic acid (0.5 g) and a coupling reagent, at pH 6.5 for 72 hours at room temperature. The final product was dialyzed for five days and freeze-dried. The chemical structure of the synthesized polymers was confirmed through proton nuclear magnetic resonance (¹H NMR, D₂O) and Fourier-transform infrared (FTIR) spectroscopy [1].
Catechin, dissolved in ethanol, was added dropwise to a 2% HAMA-PBA aqueous solution and stirred at 1000 rpm at 25°C. Irgacure 2959 photoinitiator was then introduced, and the mixture was transferred into custom molds and polymerized under 365 nm ultraviolet light for 100 seconds. Hydrogel formulations are summarized in Table 1. To obtain extracts for biological tests, 100 mg of the hydrogel was soaked in 40 mL of phosphate-buffered saline for 24 hours to produce a 2.5 mg/mL extract solution.
Hydrogel microstructures were visualized via scanning electron microscopy. The water vapor transmission rate (WVTR) was determined as WVTR = (M₁ − M₂)/S, where M₁ − M₂ represents the change in water weight and S is the cup mouth area. Swelling behavior in PBS (pH 7.4) at 37°C was assessed using a standard method, with the swelling ratio (SR) calculated as SR = [(M₁ − M₂)/M₂] × 100%, where M₁ and M₂ are the weights of the swollen and dry hydrogel, respectively. Mechanical testing was performed using an Instron 5944 system under a strain rate of 1 mm/min, and Young’s modulus was calculated from the linear region between 5% and 20% strain [1].
Catechin release profiles were evaluated in PBS and 4 mg/mL PBS containing glucose to mimic diabetic wound conditions. Loaded hydrogels were incubated in 10 mL of buffer at 37°C and 100 rpm. At set intervals, 2 mL of medium was removed and replaced with a fresh buffer. Released catechin concentrations were measured by UV-Vis spectrophotometry at 230 nm.
Cytotoxicity, live/dead cell staining, cytoskeletal staining, and hemolysis tests were conducted according to detailed protocols in the supplementary information. The antioxidant potential was examined through both cell-free and cellular assays. The DPPH radical scavenging ability of the hydrogel extract at a dose of 2.5 mg/mL was measured by mixing with 200 μM DPPH methanol solution, incubating for 30 minutes at 37°C, and recording absorbance at 517 nm to calculate radical scavenging efficiency [1].
Intracellular reactive oxygen species (ROS) scavenging activity was evaluated in NIH 3T3 fibroblasts treated with 3 μg/mL hydrogen peroxide and 2.5 mg/mL hydrogel extract. Cells were stained with 2′,7′-dichlorofluorescin diacetate (DCFH-DA) and imaged using fluorescence microscopy and flow cytometry. Protective effects against oxidative damage were analyzed by acridine orange/ethidium bromide (AO/EB) staining. Additionally, antioxidant enzyme activities, superoxide dismutase (SOD), glutathione peroxidase (GPx), and malondialdehyde (MDA), were quantified using commercial assay kits after cell lysis [1].
In vitro scratch wound healing and angiogenesis assays were carried out as described in the supporting information, along with real-time quantitative PCR (RT-qPCR) to evaluate gene expression related to healing and angiogenesis
For in vivo testing, type 1 diabetes was induced in 4-6 week old female Sprague Dawley rats by intraperitoneal injection of 50 mg/kg streptozotocin. After one week, rats with blood glucose >16.7 mmol/L were confirmed diabetic and given full-thickness 1 cm wounds on their backs. Animals numbered three per group received treatments with either hydrogel or PBS, and wound healing progression was recorded. After 21 days, wound tissues were fixed in 4% formaldehyde for histological evaluation, including hematoxylin-eosin (H&E) and Masson’s trichrome staining. Immunohistochemical analysis was conducted for angiogenic and inflammatory markers CD31, VEGF, IL-6, and IL-10 [1].
2) The research team of Wang et al assessed the ability of hyaluronic acid to protect against hepatocellular carcinoma when used in nanomedicine. The compound PDC-DOX₂ was synthesized through an amide coupling reaction between the amino groups of DOX and the carboxyl groups of Pep-AA over three days. Specifically, Pep-AA and PyAOP were dissolved in anhydrous DMF with DIEA added as a base. The reaction mixture was maintained at 0 °C under nitrogen for five hours before the addition of DOX dissolved in DMF. The system was then stirred at room temperature for three days under nitrogen protection. After completion, the product was precipitated using cold diethyl ether and purified by reverse-phase high-performance liquid chromatography to yield PDC-DOX₂ [2].
To form HA-PDC-DOX₂ complexes, 0.1 mg/mL aqueous PDC-DOX₂ solution was mixed with varying concentrations of hyaluronic acid to achieve different weight ratios. The mixtures were vortexed for 30 seconds and incubated at room temperature for one hour before use. This process facilitated electrostatic and hydrophobic interactions between HA and PDC-DOX₂, forming stable nanoscale assemblies suitable for biological evaluation.
The structure and molecular weight of PDC-DOX₂ were confirmed using electrospray ionization mass spectrometry (ESI-MS) and Fourier transform infrared spectroscopy (FT-IR). The particle size distribution and zeta potential of PDC, PDC-DOX₂, and HA-PDC-DOX₂ were determined by dynamic light scattering (DLS) at 25 °C, using a Zetasizer Nano ZS90. The measurements were performed in triplicate at a concentration of 100 μM in water, and the effects of pH (1, 3, 5, 7) on stability were assessed by adjusting solutions with 1 M HCl or NaOH. Transmission electron microscopy (TEM) was used to examine nanoparticle morphology and confirm the nanoscale self-assembly of PDC-DOX₂ and HA-PDC-DOX₂ [2].
The self-assembly behavior of PDC-DOX₂ was further characterized by determining its critical aggregation concentration using pyrene as a hydrophobic fluorescence probe. Methanolic pyrene solution was dried overnight in dark conditions, and various concentrations of PDC-DOX₂ ranging from 1.75–400 μM were added. After ultrasonication for 10 minutes, fluorescence spectra were recorded and the intensity ratio of pyrene’s emission peaks was plotted against the logarithm of PDC-DOX₂ concentration to determine the CAC, indicating the concentration threshold for micelle formation [2].
To assess controlled drug release behavior, DOX release from both free DOX and PDC-DOX₂ nanomicelles was studied under physiological and mildly acidic conditions at 37 °C. Samples were placed in dialysis bags and immersed in PBS medium, with aliquots collected at scheduled intervals up to 72 h and replaced with fresh medium. The DOX concentration in each sample was determined using high-performance liquid chromatography Each test was performed in triplicate.
Human hepatocellular carcinoma (SMMC-7721) and murine hepatoma carcinoma (H22) cell lines were cultured under standard conditions in RPMI medium supplemented with 10% fetal bovine serum and antibiotics at 37 °C in a humidified atmosphere with 5% CO₂. Cytotoxicity was evaluated by the MTT assay. SMMC-7721 cells were seeded in 96-well plates and treated with varying concentrations of peptide, free DOX, PDC-DOX₂, and HA-PDC-DOX₂. After incubation, cells were treated with MTT reagent for 4 hours, and absorbance was measured at 490 nm. Cell viability was calculated relative to untreated controls [2].
The antitumor activity of PDC-DOX₂ and HA-PDC-DOX₂ was evaluated in C57BL/6 mice bearing H22 hepatoma xenografts. Tumors were established by subcutaneous injection of 2.5 × 10⁶ H22 cells into the flank region. When tumor volumes reached approximately 500 mm³, mice were randomly assigned to four treatment groups receiving PBS, free DOX at a dose of 5 mg/kg), PDC-DOX₂ at a dose equivalent to 5 mg/kg DOX, or HA-PDC-DOX₂ at a dose equivalent to 11 mg/kg PDC-DOX₂. Treatments were administered intravenously every three days, for a total of five doses. Tumor dimensions were measured every other day with calipers, and tumor volume (V) was calculated as V = (width² × length) × 0.5. After 18 days, mice were sacrificed, and tumor tissues were collected for histological evaluation via H&E staining [2].
Overall, these experimental methods established the synthesis, characterization, and biological evaluation of a hyaluronic acid-coated peptide–doxorubicin conjugate designed for targeted, pH-responsive drug delivery and enhanced anticancer efficacy.

 

Discussion
1) The study investigated the physicochemical properties, biocompatibility, antioxidant mechanisms, and therapeutic potential of a catechin-loaded HAMA-PBA hydrogel (HMPC hydrogel) for diabetic wound healing. The results demonstrated that this hydrogel possesses excellent structural, mechanical, and biological characteristics, as well as glucose-responsive drug release and antioxidative functions that collectively promote wound repair [1].
The chemical structure of HAMA-PBA was confirmed using ^1H NMR and FTIR spectroscopy, which showed approximately 58% methacrylation and 24% amidation of HA carboxyl groups by phenylboronic acid (PBA). FTIR spectra further verified the successful modification of hyaluronic acid (HA) and the formation of borate ester bonds between HAMA-PBA and catechin. Morphological analysis revealed a uniform, honeycomb-like porous architecture that enables efficient absorption of wound exudate and optimal water vapor transmission. The hydrogel exhibited a WVTR within the physiological range of healthy skin, between 240–1920 g/m², suggesting its ability to maintain a moist yet breathable wound environment. Swelling ratio (SR) studies showed rapid absorption during the first two hours, attributed to the hydrophilic nature of HA and its three-dimensional polymer network. However, increasing catechin content reduced swelling due to catechin’s hydrophobicity [1].
Mechanical testing revealed that the hydrogels displayed elastic behavior with Young’s moduli between 5 and 20 kPa, similar to normal skin tissue. Catechin release studies demonstrated glucose-responsive behavior. In phosphate-buffered saline (PBS), less than 20% of catechin was released within the first 12 hours, likely representing surface-bound catechin. In contrast, in glucose-containing PBS at a dose of 4 mg/mL, catechin release was at least twice as high, indicating that glucose molecules disrupted the borate bonds between catechin and the polymer. These results confirmed that the HMPC hydrogel combines desirable structural, mechanical, and glucose-responsive drug delivery characteristics suitable for diabetic wound applications.
MTT and live/dead cell assays demonstrated that HMPC hydrogels were non-toxic to fibroblasts, with cell viability exceeding 95% after 24 and 48 hours. Live/dead staining showed extensive green fluorescence, confirming high cell survival. Cytoskeletal staining with FITC-phalloidin revealed normal fibroblast morphology and adhesion comparable to control cells. Hemolysis tests showed less than 2% red blood cell lysis, indicating excellent hemocompatibility. Collectively, these results confirmed that the HMPC hydrogel was biocompatible, non-cytotoxic, and conducive to cell adhesion and proliferation, all essential for wound repair [1].
The HMPC hydrogel’s antioxidant properties were verified through both cell-free and cellular assays. DPPH radical scavenging tests showed that the HMP hydrogel without catechin had limited antioxidant activity demonstrated by 23.5% clearance, while catechin-loaded HMPC hydrogels achieved up to 92.1% scavenging in a catechin dose-dependent manner. Cellular ROS assays using DCFH-DA revealed that H₂O₂-treated cells displayed strong green fluorescence indicative of oxidative stress, which was markedly reduced in HMPC-treated cells. Flow cytometry confirmed a 40% reduction in ROS levels in the HMPC group compared with controls. AO/EB staining further showed that HMPC-treated cells remained predominantly viable under oxidative stress, demonstrating robust cytoprotection.
To investigate the antioxidative mechanism, enzymatic assays measured intracellular levels of superoxide dismutase (SOD), glutathione peroxidase (GPx), and malondialdehyde (MDA). H₂O₂ exposure reduced SOD activity to 39.8% and GPx by 65%, while increasing MDA content by 1.5-fold, all indicative of oxidative damage. Treatment with HMPC hydrogels restored SOD and GPx activity to near-normal levels and suppressed MDA accumulation. These findings confirm that HMPC hydrogels mitigate oxidative stress by preserving cellular antioxidant enzymes and reducing lipid peroxidation [1].
Scratch wound assays demonstrated that hydrogels with higher catechin concentrations (HMPC-2 and HMPC-3) significantly enhanced fibroblast migration, achieving wound closure rates of 89.6% and 95.2%, respectively, compared with minimal improvement in catechin-free HMP or low-dose HMPC-1 hydrogels. Angiogenesis assays using human umbilical vein endothelial cells (HUVECs) showed that HMPC-3 induced robust tube formation, with at least 15-fold and 20-fold increases in junction and branch formation, respectively, compared to the control. RT-qPCR analysis revealed that HMPC-3 treatment upregulated proangiogenic factors VEGF and CD31 by 2.1- and 2.3-fold, while downregulating inflammatory IL-6 by 80% and upregulating anti-inflammatory IL-10 by 2.5-fold. These results indicate that HMPC hydrogels promote angiogenesis and modulate inflammation through antioxidative mechanisms, thus supporting tissue regeneration [1].
In a diabetic rat model, wounds treated with HMPC-3 hydrogels showed a 97.05% closure rate after three weeks, compared with 89.06% in the HMP group and 78.25% in PBS controls. Histological analyses, including H&E and Masson’s staining, revealed enhanced re-epithelialization by 85.25% and collagen deposition by 86.34% in HMPC-3-treated wounds, compared with 58.96% and 62.52% in controls. These improvements were attributed to reduced ROS levels and enhanced fibroblast activity and angiogenesis.
Immunostaining confirmed elevated expression of angiogenic markers VEGF by 90.38% and CD31 by 91.03% in HMPC-3-treated tissues, along with increased IL-10 and decreased IL-6 levels, signifying effective inflammation suppression and vascular regeneration. Overall, the HMPC hydrogel exhibited excellent physicochemical properties, biocompatibility, antioxidant and anti-inflammatory effects, and glucose-responsive catechin release, collectively accelerating diabetic wound healing and tissue regeneration [1].
2) The study performed by Wang et al reports the successful synthesis, structural confirmation, and biological evaluation of a novel PDC-DOX₂ and its hyaluronic acid-coated form HA-PDC-DOX₂ for improved anticancer drug delivery. The section outlines the physicochemical characterization of the conjugates, their self-assembly behavior, pH-responsive release properties, and in vitro and in vivo antitumor activities [2].
To construct a peptide–drug conjugate containing two doxorubicin molecules, a modified peptide, K(adipic acid)₂IGLFRWR (Pep-AA), was designed with two carboxyl groups separated by a six-carbon linker to reduce steric hindrance and improve coupling efficiency. Electrospray ionization mass spectrometry (ESI-MS) confirmed the successful synthesis of Pep-AA, showing molecular ion peaks at m/z 1331.95 ([M+H]⁺) and 666 ([M+2H]²⁺), corresponding to its expected molecular weight. High-performance liquid chromatography (HPLC) analysis confirmed over 95% purity [2].
The assembly behavior of Pep-AA and PDC-DOX₂ was investigated using dynamic light scattering (DLS), zeta potential analysis, and transmission electron microscopy (TEM). At 100 μM concentration, PDC-DOX₂ formed stable nanoparticles that maintained a constant particle size over 24 hours, while Pep-AA alone showed progressive aggregation over time. TEM images revealed that Pep-AA self-assembled into fibrous structures, whereas PDC-DOX₂ formed uniform spherical micelles. This morphological shift was attributed to enhanced hydrophobic interactions arising from the two hydrophobic DOX molecules, which promoted spherical self-assembly.
Using pyrene fluorescence probing, the critical aggregation concentration (CAC) of PDC-DOX₂ was determined to be 19.95 μM, indicating that self-assembly occurred above this threshold concentration. Further studies revealed that pH strongly influenced PDC-DOX₂ aggregation due to electrostatic interactions. As pH increased from 1.0 to 7.0, the zeta potential became more positive, enhancing electrostatic repulsion and maintaining stable nanoparticles at neutral and mildly acidic conditions. Conversely, at lower pH, reduced charge led to aggregation, indicating that electrostatic forces governed nanomicelle formation. PDC-DOX₂ contained positively charged amino acids (two arginines and one lysine), which generated strong intermolecular repulsion essential for colloidal stability [2].
TEM confirmed that at mildly acidic pH, PDC-DOX₂ nanoparticles exhibited slight aggregation, while remaining stable under neutral conditions, further supporting the pH-dependent self-assembly mechanism. To enhance tumor targeting, negatively charged hyaluronic acid was electrostatically coated onto positively charged PDC-DOX₂ nanomicelles. HA, known for its biocompatibility and CD44 receptor-binding affinity, was added at different mass ratios.
The release of DOX from free DOX, PDC-DOX₂, and HA-PDC-DOX₂ was examined at physiological and mildly acidic conditions, simulating normal and tumor microenvironments, respectively. Free DOX rapidly diffused, reaching equilibrium within 24 hours. In contrast, PDC-DOX₂ and HA-PDC-DOX₂ exhibited sustained release over 72 hours. These findings indicate that PDC-DOX₂ nanomicelles are stable under physiological conditions but gradually release DOX in acidic tumor environments, improving targeted drug delivery and reducing systemic toxicity [2].
The cytotoxic effects of peptide, free DOX, PDC-DOX₂, and HA-PDC-DOX₂ were assessed using MTT assays in SMMC-7721 hepatocellular carcinoma cells. All DOX-containing samples showed dose-dependent inhibition of cell viability. The peptide alone exhibited no cytotoxicity, indicating high biocompatibility. Free DOX had the strongest cytotoxicity at 3.10 μg/mL, followed by PDC-DOX₂ at 7.45 μg/mL and HA-PDC-DOX₂ at 24.05 μg/mL. Although the conjugates were less potent in vitro, their design supports better tumor accumulation via the enhanced permeability and retention (EPR) effect and controlled pH-triggered release, potentially lowering systemic toxicity compared to free DOX [2].
The therapeutic performance of HA-PDC-DOX₂ was evaluated in H22 tumor-bearing C57BL/6 mice. Four groups received PBS, free DOX, PDC-DOX₂, or HA-PDC-DOX₂, each containing equivalent DOX doses of 5 mg/kg. Tumor growth and body weight were monitored every three days. Mice treated with HA-PDC-DOX₂ exhibited the most significant tumor growth inhibition, surpassing both free DOX and PDC-DOX₂ groups. This enhanced efficacy was attributed to HA-mediated targeting of CD44 receptors, facilitating tumor accumulation of nanoparticles. Body weight changes were minimal, indicating good tolerability and low systemic toxicity. Histological analysis of tumor tissues showed extensive nuclear fragmentation in treated groups, especially with HA-PDC-DOX₂, compared to intact nuclei in control tumors [2].
Collectively, these findings demonstrate that HA-PDC-DOX₂ exhibits stable self-assembly, pH-sensitive drug release, tumor-targeted accumulation, and strong antitumor efficacy with reduced toxicity, making it a promising nanoplatform for cancer therapy.

 

Disclaimer

**LAB USE ONLY**
*This information is for educational purposes only and does not constitute medical advice. THE PRODUCTS DESCRIBED HEREIN ARE FOR RESEARCH USE ONLY. All clinical research must be conducted with oversight from the appropriate Institutional Review Board (IRB). All preclinical research must be conducted with oversight from the appropriate Institutional Animal Care and Use Committee (IACUC) following the guidelines of the Animal Welfare Act (AWA).

 

Citations

[1] Xu Z, Liu G, Liu P, et al. Hyaluronic acid-based glucose-responsive antioxidant hydrogel platform for enhanced diabetic wound repair. Acta Biomater. 2022;147:147-157. doi:10.1016/j.actbio.2022.05.047

[2] Wang J, Qian Y, Xu L, et al. Hyaluronic acid-shelled, peptide drug conjugate-cored nanomedicine for the treatment of hepatocellular carcinoma. Mater Sci Eng C Mater Biol Appl. 2020;117:111261. doi:10.1016/j.msec.2020.111261

 

 

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