Physicochemical characterization of surfactant incorporating vesicles that incorporate colloidal magnetite.

Drug administration through the transdermal route has optimized for the comfort of patients and easy application. However, the main limitation of transdermal drug delivery is the impermeability of the human skin. Recent advances on improvement of drug transport through the skin include elastic liposomes as a penetration enhancer. Entrapment of ferrofluids in the core of liposomes produces magnetoliposomes, which can be driven by a high-gradient magnetic field. The association of both strategies could enhance the penetration of elastic liposomes. This work relies on the preparation and characterization of elastic-magnetic liposomes designed to permeate through the skin. The incorporation of colloidal magnetite and the elastic component, octaethylene glycol laurate (PEG-8-L), in the structure of liposomes were evaluated. The capability of the elastic magnetoliposomes for permeation through nanopores of two stacked polycarbonate membranes was compared to conventional and elastic liposomes. Magnetite incorporation was dependent on vesicle diameter and size distribution as well as PEG-8-L incorporation into liposomes, demonstrating the capability of the fluid bilayer to accommodate the surfactant without disruption. On the contrary, PEG-8-L incorporation into magnetoliposomes promoted a decrease of average diameter and a lower PEG-8-L incorporation percentage as a result of reduction on the fluidity of the bilayer imparted by iron incorporation into the lipid structure. Elastic liposomes demonstrated an enhancement of the deformation capability, as compared with conventional liposomes. Conventional and elastic magnetoliposomes presented a reduced capability for deformation and permeation.

Hyaluronic acid behavior in oral administration and perspectives for nanotechnology-based formulations: A review.

Hyaluronic acid (HA) is a ubiquitous polysaccharide with diverse biological functions. Is known that in the intestinal epithelium, the exogenous HA of molar mass ≥105 Da orally administered antagonizes TLR4 overexpression resulting from dysbiosis and promotes immunomodulation in multifactorial crosstalk, thus helping to treat or to prevent injuries. As macromolecules mediate cell signaling, the three-dimensional structure of HA plays a vital role in those functions. Introducing HA in terms of its molecular structure, its spatial architecture as dependent on pH, concentration and molar mass, occurrence, biological functions and turnover in the tissues, this review addresses the HA in the gastrointestinal system, the molecular dynamics of intestinal uptake and signaling, immunomodulation at intestinal and systemic levels and HA fate to other tissues. Finally, at the light of these behaviors, a nanotechnological approach is presented as progress in the field of the oral HA administration and discussed with perspectives for future developments.

Adsorption of antiphospholipid antibodies on affinity magnetoliposomes.

Phosphatidylcholine-based magnetoliposomes containing specific ligands for biological molecules, so-called affinity magnetoliposomes (AML), may prove to be useful as adsorbents in applications such as diagnosis or anchoring and delivery of drugs at specific sites in the human body. In the present study, the performance of affinity magnetoliposomes to adsorb anticardiolipin antibodies (aCL) from a previously characterized pool of patients with autoimmune diseases is described. The magnetic vesicles were prepared by enrobing nanometer-sized colloidal magnetite particles with a phospholipid bilayer composed of dimyristoylphosphatidylcholine (DMPC) and the affinity lipid ligand cardiolipin (CL). Adsorption of antibodies onto the affinity magnetoliposomes assayed using a high-gradient magnetophoresis (HGM) system, in which the magnetoliposomes were first magnetically captured on stainless steel fibers, and which were subsequently overflowed either with a pool of sera from autoimmune patients or sera of healthy individuals as a control. The spectrophotometric assay showed stronger changes in absorbance spectra when the affinity magnetoliposomes containing cardiolipin were added to sera of autoimmune patients than when they were added to sera of healthy individuals. The breakthrough curves obtained from a frontal analyses of the adsorption in the magnetophoresis system showed a 10% difference for total adsorbed IgG when sera of autoimmune and healthy individuals were assayed on magnetoliposomes containing cardiolipin.

Antimicrobial activity of polymyxin-loaded solid lipid nanoparticles (PLX-SLN): Characterization of physicochemical properties and in vitro efficacy.

Antimicrobial resistance is a current public health concern, limiting the available therapeutic options used for the treatment of common bacterial infections. The development of new drug entities via biotechnological processes is however expensive and time-consuming. Therefore, old antimicrobial agents have been recovered for clinical use. An example of these drugs is polymyxin, which is known for its serious adverse side effects, such as nephrotoxicity, neurotoxicity and promotion of skin pigmentation. To overcome these limitations, the use of biodegradable nanoparticles has been proposed to allow site-specific targeting, increasing the drug's bioavailability and decreasing its side effects. The aim of this work was the development of an optimized pharmaceutical formulation composed of solid lipid nanoparticles (SLN) loading polymyxin B sulphate (PLX) for the treatment of bacterial infections. The PLX-loaded SLN were produced by a double emulsion method (w/o/w), obtaining particles with a mean size of approximately 200nm, polydispersity of 0.3 and zeta potential of -30mV. The encapsulation efficiency reached values above 90% for all developed formulations. SLN remained stable for a period of 6months of storage at room temperature. The occlusive properties of the SLN was shown to be dependent on the type of lipid, while the antimicrobial properties of PLX-loaded SLN were effective against resistant strains of Pseudomonas aeruginosa. Results from the differential scanning calorimetry (DSC), wide angle X-ray diffraction (WAXD) and small angle X-ray scattering (SAXS) analyses confirmed the crystallinity of the inner SLN matrices, suggesting the capacity of these particles to modify the release profile of the loaded drug.

Randomized controlled trial comparing hyaluronic acid, platelet-rich plasma and the combination of both in the treatment of mild and moderate osteoarthritis of the knee.

Objective: This study aims at evaluating the clinical effects of Platelet Rich Plasma (PRP) and Hyaluronic Acid (HA) as individual treatments for mild to moderate Osteoarthritis (OA) and it also examines the potential synergistic effects of PRP in combination with HA. Research continues to emerge examining the potential therapeutic efficacy of HA and PRP as autologous injectable treatments for joint arthritis. However, there is a paucity of research investigating the effects of combining HA and PRP on pain and functional status in patients with OA. Design: In this multi-center, randomized, controlled, double blind, prospective trial, 105 patients with mild to moderate knee osteoarthritis, who met the study criteria, were randomly allocated to one of three interventions: HA (n=36), PRP (n=36), or HA+PRP (n=33). Each patient received 3 intra-articular knee injections of their assigned substance, with 2 week intervals between each injection. Clinical outcomes were evaluated using the Western Ontario and McMaster Universities Arthritis Index (WOMAC) and Visual Analogue Scale (VAS) questionnaire at baseline and after 1,3,6 and 12 months. Results: The study showed that the PRP group have significant reduction in VAS scores at 1 (p= 0.003), 3 (p= 0.0001), 6 (p= 0.0001) and 12 (p= 0.000) months when compared to HA. In addition, the PRP group illustrated greater improvement in WOMAC physical activity scale at 12 months (p= 0.008) when compared to the HA group. Combining HA and PRP resulted in a significant decreases in pain (p=0.0001) and functional limitation (p=0.0001) when compared to HA alone at 1 year post treatment; and significantly increased physical function at 1 (p=0.0004) and 3 (p=.011) months when compared to PRP alone. Conclusion: The findings of the study support the use of autologous PRP as an effective treatment of mild to moderate knee osteoarthritis. It also shows that the combination of HA and PRP resulted to better outcomes than HA alone up to 1 year and PRP alone up to 3 months. Furthermore, the results suggest that combination of PRP and HA could potentially provide better functional outcomes in the first 30 days after treatment with both PRP and HA alone.

Physico-chemical characterization of benzocaine-beta-cyclodextrin inclusion complexes.

Local anesthetics are able to induce pain relief by binding to the sodium channel of excitable membranes, blocking the influx of sodium ions and the propagation of the nervous impulse. Benzocaine (BZC) is a local anesthetic whose low water-solubility limits its application to topical formulations. The present work focuses on the characterization of inclusion complexes of BZC in beta-cyclodextrin (beta-CD). Differential scanning calorimetry and electron microscopy gave evidences of the formation and the morphology of the complex. Fluorescence spectroscopy showed a BZC/beta-CD 1:1 stoichiometry. Phase-solubility diagrams allowed the determination of the association constants between BZC and beta-CD (549 M(-1)) and revealed that a three-fold increase in BZC solubility can be reached upon complexation with beta-CD. The details of BZC/beta-CD molecular interaction were analyzed by 1H 2D NMR allowing the proposition of an inclusion model for BZC into beta-CD where the aromatic ring of the anesthetic is located near the head of the beta-CD cavity. Moreover, in preliminary toxicity studies, the complex seems to be less toxic than BZC alone, since it induced a decrease in the in vitro oxidation of human hemoglobin. These results suggest that the BZC/beta-CD complex represents an effective novel formulation to enhance BZC solubility in water, turning it promising for use outside its traditional application, i.e., in infiltrative anesthesia.

Didanosine-loaded chitosan microspheres optimized by surface-response methodology: a modified "Maximum Likelihood Classification" approach formulation for reverse transcriptase inhibitors.

Didanosine-loaded chitosan microspheres were developed applying a surface-response methodology and using a modified Maximum Likelihood Classification. The operational conditions were optimized with the aim of maintaining the active form of didanosine (ddI), which is sensitive to acid pH, and to develop a modified and mucoadhesive formulation. The loading of the drug within the chitosan microspheres was carried out by ionotropic gelation technique with sodium tripolyphosphate (TPP) as cross-linking agent and magnesium hydroxide (Mg(OH)2) to assure the stability of ddI. The optimization conditions were set using a surface-response methodology and applying the "Maximum Likelihood Classification", where the initial chitosan concentration, TPP and ddI concentration were set as the independent variables. The maximum ddI-loaded in microspheres (i.e. 1433 mg of ddI/g chitosan), was obtained with 2% (w/v) chitosan and 10% TPP. The microspheres depicted an average diameter of 11.42 µm and ddI was gradually released during 2 h in simulated enteric fluid.

Development and characterization of a cationic lipid nanocarrier as non-viral vector for gene therapy.

The aim of the present work was to produce a cationic solid lipid nanoparticle (SLN) as non-viral vector for protein delivery. Cationic SLN were produced by double emulsion method, composed of softisan(®) 100, cetyltrimethylammonium bromide (CTAB), Tween(®) 80, Span(®) 80, glycerol and lipoid(®) S75 loading insulin as model protein. The formulation was characterized in terms of mean hydrodynamic diameter (z-ave), polydispersity index (PI), zeta potential (ZP), stability during storage time, stability after lyophilization, effect of toxicity and transfection ability in HeLa cells, in vitro release profile and morphology. SLN were stable for 30days and showed minimal changes in their physicochemical properties after lyophilization. The particles exhibited a relatively slow release, spherical morphology and were able to transfect HeLa cells, but toxicity remained an obstacle. Results suggest that SLN are nevertheless promising for delivery of proteins or nucleic acids for gene therapy.

Sodium alginate-cross-linked polymyxin B sulphate-loaded solid lipid nanoparticles: Antibiotic resistance tests and HaCat and NIH/3T3 cell viability studies.

Polymyxins are a group of antibiotics with a common structure of a cyclic peptide with a long hydrophobic tail. Polymyxin B sulphate (PLX) has cationic charge, which is an obstacle for the efficient loading into Solid Lipid Nanoparticles (SLN). In the present paper, we describe an innovative method to load PLX into SLN to achieve the sustained release of the drug. PLX was firstly cross-linked with sodium alginate (SA) at different ratios (1:1, 1:2 and 1:3 SA/PLX), and loaded into SLN produced by high pressure homogenization (HPH). Optimized SLN were produced applying 500bar pressure and 5 homogenization cycles. The best results were obtained with SA/PLX (1:1), recording 99.08±1.2% for the association efficiency of the drug with SA, 0.99±10g for the loading capacity and 212.07±5.84% degree of swelling. The rheological profile of aqueous SA solution followed the typical behaviour of concentrated polymeric solutions, whereas aqueous SA/PLX solution exhibited a gel-like dynamic behaviour. Micrographs show that SA/PLX depicted a porous and discontinuous amorphous phase in different ratios. The encapsulation efficiency of SA/PLX (1:1) in SLN, the mean particle diameter, polydispersity index and zeta potential were, respectively, 82.7±5.5%; 439.5±20.42nm, 0.241±0.050 and -34.8±0.55mV. The effect of SLN on cell viability was checked in HaCat and NIH/3T3 cell lines, and the minimal inhibitory concentrations (MIC) were determined in Pseudomonas aeruginosa strains. SA/PLX-loaded SLN were shown to be less toxic than free PLX. Minimal inhibitory concentrations (MIC) showed the presence of the cross-linker polymer-drug complex, and SLN were shown to enhance MIC in the evaluated strains.

In vivo absorption of didanosine formulated in pellets composed of chitosan microspheres.

The in vivo absorption of didanosine was studied, focusing on the performance of a novel pharmaceutical formulation for didanosine, composed of chitosan granules containing didanosine incorporated in chitosan microspheres. This novel formulation is aimed at oral administration in AIDS therapy. The experimental results in male adult dogs showed controlled delivery of didanosine along 36 h, with a 2-fold increase in the absorption time of didanosine compared to the commercial granules, gastro-resistant didanosine and tablets. The higher absorption is due to adhesion to the intestinal membrane, improving absorption through increase of residence time, permeation and release. Furthermore, the novel formulation facilitates handling and deglutition, especially in the elderly and children, as well as enhances the taste and reduces the frequency of doses and collateral effects associated with a high concentration of the buffer agents usually used in other formulations.

Relevant aspects of centrifugation step in the preparation of platelet-rich plasma.

Introduction. Platelet-Rich Plasma (PRP) is rich in growth factors, playing important role in tissue healing. The wide variation of reported protocols for preparation of PRP leads to variable compositions, which induce different biological responses and prevent results comparison. This study aims to highlight relevant aspects of the centrifugation step to obtain reproducible results and overall quality. Material and Methods. Samples of blood were collected from 20 healthy donors that have signed free informed consent. Two centrifugation steps (spins) were analyzed for the influence of centrifugal acceleration, time, processed volume, and platelet gradient. The Pure Platelet-Rich Plasma (P-PRP) was characterized as platelet concentration, integrity, and viability (sP-selectin measurement). Results. Lower centrifugal accelerations favour platelet separation. The processing of 3.5 mL of blood at 100 ×g for 10 min (1st spin), 400 ×g for 10 min (2nd spin), withdrawing 2/3 of remnant plasma, promoted high platelet recovery (70-80%) and concentration (5x) maintaining platelet integrity and viability. The recovery of platelets was reduced for a larger WB volume (8.5 mL) processed. Conclusion. Centrifugal acceleration, time, WB processed volume, and minimization of the platelet gradient before sampling are relevant aspects to ensure reproducible compositions within the autologous nature of PRP.

Solid lipid nanoparticles for hydrophilic biotech drugs: optimization and cell viability studies (Caco-2 & HEPG-2 cell lines).

Insulin was used as model protein to developed innovative Solid Lipid Nanoparticles (SLNs) for the delivery of hydrophilic biotech drugs, with potential use in medicinal chemistry. SLNs were prepared by double emulsion with the purpose of promoting stability and enhancing the protein bioavailability. Softisan(®)100 was selected as solid lipid matrix. The surfactants (Tween(®)80, Span(®)80 and Lipoid(®)S75) and insulin were chosen applying a 2(2) factorial design with triplicate of central point, evaluating the influence of dependents variables as polydispersity index (PI), mean particle size (z-AVE), zeta potential (ZP) and encapsulation efficiency (EE) by factorial design using the ANOVA test. Therefore, thermodynamic stability, polymorphism and matrix crystallinity were checked by Differential Scanning Calorimetry (DSC) and Wide Angle X-ray Diffraction (WAXD), whereas the effect of toxicity of SLNs was check in HepG2 and Caco-2 cells. Results showed a mean particle size (z-AVE) width between 294.6 nm and 627.0 nm, a PI in the range of 0.425-0.750, ZP about -3 mV, and the EE between 38.39% and 81.20%. After tempering the bulk lipid (mimicking the end process of production), the lipid showed amorphous characteristics, with a melting point of ca. 30 °C. The toxicity of SLNs was evaluated in two distinct cell lines (HEPG-2 and Caco-2), showing to be dependent on the concentration of particles in HEPG-2 cells, while no toxicity in was reported in Caco-2 cells. SLNs were stable for 24 h in in vitro human serum albumin (HSA) solution. The resulting SLNs fabricated by double emulsion may provide a promising approach for administration of protein therapeutics and antigens.

Encapsulation of antioxidants in gastrointestinal-resistant nanoparticulate carriers.

Reactive oxygen species (ROS) are known to cause several human pathologies. For this reason, antioxidants have gained utmost importance because of their potential as prophylactic and therapeutic agents in many diseases. Examples of their application include their use in diabetic patients, as aging drugs, in cancer diseases, Parkinson's, Alzheimer's, autoimmune disorders, and also in inflammation. Antioxidants have limited absorption profiles, therefore low bioavailability and low concentrations at the target site. Efforts have been done towards loading antioxidant molecules in advanced nanoparticulate carriers, e.g., liposomes, polymeric nanoparticles, solid lipid nanoparticles, self-emulsifying drug delivery system. Examples of -successful achievements include the encapsulation of drugs and other active ingredients, e.g., coenzyme Q10, vitamin E and vitamin A, resveratrol and polyphenols, curcumin, lycopene, silymarin, and superoxide dismutase. This review focuses on the comprehensive analysis of using nanoparticulate carriers for loading these molecules for oral administration.

Cashew apple juice as microbial cultivation medium for non-immunogenic hyaluronic acid production.

In this work, natural cashew apple juice was used as cultivation medium as an alternative to substitute brain heart infusion medium. The effect of aeration and juice supplementation with yeast extract on the production of hyaluronic acid in batch fermentation was also investigated. Similar levels of cell mass were obtained in inoculum using cashew apple juice supplemented with yeast extract or the conventional brain heart infusion medium. Fermentation in Erlenmeyer flasks produced low biomass and hyaluronic acid concentrations. The hyaluronic acid concentration and viscosity increased from 0.15 g/L and 3.87 cP (no aeration or medium supplementation) to 1.76 g/L and 107 cP, when aeration (2 vvm) and 60 g/L of yeast extract were used. The results suggest the production of low-molecular weight hyaluronic acid oligomers instead of the high molecular weight polymer.

Prediction and modulation of platelet recovery by discontinuous centrifugation of whole blood for the preparation of pure platelet-rich plasma.

The aim of this study was to describe the behavior of the separation of red blood cells (RBCs) by discontinuous centrifugation (DC) of whole blood to modulate and control the platelet recovery in the preparation of pure platelet-rich plasma (P-PRP). P-PRP is a platelet-rich plasma (PRP) in which the white blood cell layer is not included. To achieve this goal, an analytical model was derived that takes into account the packing of RBCs and predicts the behavior of platelet and plasma recovery efficiencies (PtPlRE) based on the volume of whole blood, the hematocrit, and the volume of supernatant, as a function of the operating variables, centrifugal acceleration, and time. The model was derived from the basic equation of DC, which originates from the equilibrium balance of forces on a particle, and included the addition of one factor that corrected the terminal velocity of RBCs and was also correlated to the PtPlRE in the supernatant. This factor was the ratio between the fractional volume concentrations of plasma and RBCs in the centrifugation pellet after centrifugation. The model was validated and the variability of the data was determined using experimental data from 10 healthy donors in the age range of 25-35 years. The predicted behavior for the packing of RBCs and the PtPlRE was consistent with the behavior seen in the experimental data. Thus, the PtPlRE could be modulated and controlled through centrifugal acceleration, time, and hematocrit. Use of this model based on a physical description of events is the first step of a reliable standardization of PRP preparations.

Optimizing SLN and NLC by 2(2) full factorial design: effect of homogenization technique.

Solid lipid nanoparticles (SLN) and nanostructured lipid carrier (NLC) have been employed in pharmaceutics and biomedical formulations. The present study focuses on the optimization of the production process of SLN and NLC by High Shear Homogenization (HSH) and High Pressure Homogenization (HPH). To build up the surface response charts, a 2(2) full factorial design based on 2 independent variables was used to obtain an optimized formulation. The effects of the production process on the mean particle size, polydispersity index (PI) and zeta potential (ZP) were investigated. Optimized SLN were produced applying 20,000 rpm HSH and 500 bar HPH pressure and NLC process 15,000 rpm HSH and 700 bar HPH pressure, respectively. This factorial design study has proven to be a useful tool in optimizing SLN (~100 nm) and NLC (~300 nm) formulations. The present results highlight the benefit of applying statistical designs in the preparation of lipid nanoparticles.

Current State-of-Art and New Trends on Lipid Nanoparticles (SLN and NLC) for Oral Drug Delivery.

Lipids and lipid nanoparticles are extensively employed as oral-delivery systems for drugs and other active ingredients. These have been exploited for many features in the field of pharmaceutical technology. Lipids usually enhance drug absorption in the gastrointestinal tract (GIT), and when formulated as nanoparticles, these molecules improve mucosal adhesion due to small particle size and increasing their GIT residence time. In addition, lipid nanoparticles may also protect the loaded drugs from chemical and enzymatic degradation and gradually release drug molecules from the lipid matrix into blood, resulting in improved therapeutic profiles compared to free drug. Therefore, due to their physiological and biodegradable properties, lipid molecules may decrease adverse side effects and chronic toxicity of the drug-delivery systems when compared to other of polymeric nature. This paper highlights the importance of lipid nanoparticles to modify the release profile and the pharmacokinetic parameters of drugs when administrated through oral route.

Analysis of in vivo absorption of didanosine tablets in male adult dogs by HPLC.

Didanosine is an effective antiviral drug in untreated and antiretroviral therapy-experienced patients with Human Immunodeficiency Virus (HIV). An automated system using on-line solid extraction and High Performance Liquid Chromatography (HPLC) with ultraviolet (UV) detection was developed and validated for pharmacokinetic analysis of didanosine in dog plasma. Modifications were introduced on a previous methodology for simultaneous analysis of antiretroviral drugs in human plasma. Extraction was carried out on C18 cartridges, with high extraction yield as stationary phase, whereas mobile phase consisted of a mixture of 0.02 M potassium phosphate buffer, acetonitrile (KH2PO4: acetonitrile: 96:4, v/v) and 0.5% (w/v) of heptane sulphonic acid. The pH was adjusted to 6.5 with triethylamine. All samples and standard solutions were chromatographed at 28 °C. For an isocratic run, the flux was 1.0 mL/min, detection was at 250 nm and injected volume was 20 µL. The method was selective and linear for concentrations between 50 and 5000 ng/mL. Drug stability data ranged from 96% to 98%, and limit of quantification was 25 ng/mL. Extraction yield was up to 95%. Drug stability in dog plasma was kept frozen at -20 °C for one month after three freeze-thaw cycles, and for 24 h after processing in the auto sampler. Assay was successfully applied to measure didanosine concentrations in plasma dogs.

Metabolic effects of the initial glucose concentration on microbial production of hyaluronic acid.

The objective of the present work was to evaluate the metabolic effects induced by the initial glucose concentration (IGC) on the cultivation of Streptococcus zooepidemicus for the production of hyaluronic acid (HA). These effects were monitored along non-controlled pH cultivations, carried out in 250-mL Erlenmeyer flasks (natural aeration) and in a 3-L bioreactor (forced aeration) as well. Effects of the IGC were observed with focus on the main metabolites, cell growth, production, and average molecular weight of HA. The absence of glucose resulted in a mixed acid metabolism independent of the oxygen supply, while, for IGCs ranging from 5 to 90 g L(-1), the homolactic metabolism was prevalent. The IGC had no influence on the amounts of either biomass or HA produced in the cultivations carried out in flasks; however, cultivations in 3-L bioreactor were found to be strongly dependent on it. The highest concentration of HA (1.21 g L(-1)) was obtained from 25 g L(-1) IGC, the only cultivation where the conversion of glucose to HA was higher than the one of glucose to biomass. Average molecular weight of HA increased concomitant with the IGC, independently of aeration; nevertheless, it decreased along cultivation under forced aeration, due to the shear imparted by stirring.

Cashew apple juice as microbial cultivation medium for non-immunogenic hyaluronic acid production

In this work, natural cashew apple juice was used as cultivation medium as an alternative to substitute brain heart infusion medium. The effect of aeration and juice supplementation with yeast extract on the production of hyaluronic acid in batch fermentation was also investigated. Similar levels of cell mass were obtained in inoculum using cashew apple juice supplemented with yeast extract or the conventional brain heart infusion medium. Fermentation in Erlenmeyer flasks produced low biomass and hyaluronic acid concentrations. The hyaluronic acid concentration and viscosity increased from 0.15 g/L and 3.87 cP (no aeration or medium supplementation) to 1.76 g/L and 107 cP, when aeration (2 vvm) and 60 g/L of yeast extract were used. The results suggest the production of low-molecular weight hyaluronic acid oligomers instead of the high molecular weight polymer.