Levobupivacaine-Loaded Liposome Associated with Thermogel for Prolonged Analgesia.

Pain is a phenomenon present in the majority of the population, affecting, among others, the elderly, overweight people, and especially recently operated patients, analgesia being necessary. In the specific case of relief of postoperative pain, different kinds of anesthetics are being used, among them bupivacaine, a widely used drug which promotes long-lasting analgesic effects. However, cardiotoxicity and neurotoxicity are related to its repetitive use. To overcome these shortcomings, Novabupi® (a racemic mixture) was developed and is marketed as an injectable solution. This formulation contains an enantiomeric excess of the levogyre isomer, which has reduced toxicity effects. Seeking to rationalize its use by extending the duration of effect and reducing the number of applications, the objectives of this work were to develop and evaluate liposomes containing Novabupi (LBPV), followed by incorporation into thermogel. Liposomes were prepared using the lipid hydration method, followed by size reduction using sonication, and the developed formulations were characterized by hydrodynamic diameter, polydispersity index (PDI), surface zeta potential, and encapsulation efficiency. The selected optimal liposomal formulation was successfully incorporated into a thermogel without loss of thermoresponsive properties, being suitable for administration as a subcutaneous injection. In the ex vivo permeation studies with fresh rodent skin, the thermogel with liposomes loaded with 0.5% LBPV (T-gel formulation 3) showed higher permeation rates compared to the starting formulation, thermogel with 0.5% LBPV (T-Gel 1), which will probably translate into better therapeutic benefits for treatment of postoperative analgesia, especially with regard to the number of doses applied.

Antibacterial and Antibiofilm Potential of Bacterial Cellulose Hydrogel Containing Vancomycin against Multidrug-Resistant Staphylococcus aureus and Staphylococcus epidermidis.

The present study aimed to evaluate the in vitro antibacterial and antibiofilm activity of bacterial cellulose hydrogel produced by Zoogloea sp. (HYDROGEL) containing vancomycin (VAN) against bacterial strains that cause wound infections, such as multidrug-resistant (MDR) Staphylococcus aureus and Staphylococcus epidermidis. Initially, HYDROGEL was obtained from sugar cane molasses, and scanning electron microscopy (SEM) was performed to determine morphological characteristics. Then, VAN was incorporated into HYDROGEL (VAN-HYDROGEL). The antibacterial activity of VAN, HYDROGEL, and VAN-HYDROGEL was assessed using the broth microdilution method to determine the minimum inhibitory concentration (MIC) and the minimum bactericidal concentration (MBC) against methicillin-sensitive S. aureus (MSSA) ATCC 25923, methicillin-resistant S. aureus (MRSA) ATCC 33591, S. epidermidis INCQS 00016 (ATCC 12228), five clinical isolates of MRSA, and nine clinical isolates of methicillin-resistant S. epidermidis, following the Clinical and Laboratory Standards Institute (CLSI) guidelines. Additionally, the antibacterial activity of VAN, HYDROGEL, and VAN-HYDROGEL was studied using the time-kill assay. Subsequently, the antibiofilm activity of VAN, HYDROGEL, and VAN-HYDROGEL was evaluated using crystal violet and Congo red methods, as well as SEM analysis. VAN and VAN-HYDROGEL showed bacteriostatic and bactericidal activity against MRSA and methicillin-resistant S. epidermidis strains. HYDROGEL did not show any antibacterial activity. Analysis of the time-kill assay indicated that HYDROGEL maintained the antibacterial efficacy of VAN, highlighting its efficiency as a promising carrier. Regarding antibiofilm activity, VAN and HYDROGEL inhibited biofilm formation but did not demonstrate biofilm eradication activity against methicillin-resistant S. aureus and S. epidermidis strains. However, it was observed that the biofilm eradication potential of VAN was enhanced after incorporation into HYDROGEL, a result also proven through images obtained by SEM. From the methods carried out in this study, it was possible to observe that HYDROGEL preserved the antibacterial activity of vancomycin, aside from exhibiting antibiofilm activity and enhancing the antibiofilm effect of VAN. In conclusion, this study demonstrated the potential of HYDROGEL as a candidate and/or vehicle for antibiotics against MDR bacteria that cause wound infections.

Effect of citral nanoemulsion on the inactivation of Listeria monocytogenes and sensory properties of fresh-cut melon and papaya during storage.

This study evaluated the survival of Listeria monocytogenes on fresh-cut melon and papaya treated with citral nanoemulsion (CN) during 7 days of storage at 4, 8, 12, and 16 °C. CN was prepared by catastrophic phase inversion, and fresh-cut melon and papaya were artificially inoculated, resulting in 5 log cfu/g of L. monocytogenes. Then, they were treated with 0.30 (CN-0.3) and 0.15 (CN-0.15) µL/mL of CN. CN presented droplet size below 200 nm, monodisperse distribution, and negative surface charge. CN-0.3 reduced the L. monocytogenes counts more efficiently, with counts below the detection limit (1 log cfu/g) in both fruits after 48 h at 4 °C, and 72 h at 8 °C and 12 °C. At 16 °C, L. monocytogenes counts were below the detection limit for CN-0.3 after 120 h in papaya, but it survived the other treatments for 7 days. Both CN-0.3 and CN-0.15 decreased the indigenous microbiota. Scanning electron microscopy (SEM) showed bubbles in L. monocytogenes membrane and cell disruption in fruits treated with CN-0.3. Finally, CN-0.3 treated melon and papaya showed greater brightness, herbal flavor and aroma, firmness, and juiciness, as well as lower sugar and organic acid profile changes than the control samples during storage. Results indicate citral nanoemulsion's efficiency in controlling L. monocytogenes growth on fresh-cut melon and papaya stored at refrigerated temperatures without negatively influencing the sensory parameters.

Fucoidan-Coated Liposomes: A Target System to Deliver the Antimicrobial Drug Usnic Acid to Macrophages Infected with Mycobacterium tuberculosis.

The present study describes the use of fucoidan, a negative sulfated polysaccharide, as a coating material for the development of liposomes targeted to macrophages infected with Mycobacterium tuberculosis. First, fucoidan was chemically modified to obtain a hydrophobized-fucoidan derivative (cholesteryl-fucoidan) using a two-step microwave-assisted (µW) method. The total reaction time was decreased from 14 hours to 1 hour while maintaining the overall yield. Cholesterylfucoidan was then used to prepare surface-modified liposomes containing usnic acid (UA-LipoFuc), an antimicrobial lichen derivative. UA-LipoFuc was evaluated for mean particle size, polydispersity index (PDI), surface charge (ζ), and UA encapsulation efficiency. In addition, a cytotoxicity study, competition assay and an evaluation of antimycobacterial activity against macrophages infected with M. tuberculosis (H37Ra) were performed. When the amount of fucoidan was increased (from 5 to 20 mg), vesicle size increased (from 168 ± 2.82 nm to 1.18 ± 0.01 µm). Changes in from +20 ± 0.41 mV for uncoated liposomes to -5.41 ± 0.23 mV for UA-LipoFuc suggested that the fucoidan was placed on the surface of the liposomes. UA-LipoFuc exhibited a lower IC50 (8.26 ± 1.11 µM) than uncoated liposomes (18.37 ± 3.34 µM), probably due to its higher uptake. UA-LipoFuc5 was internalized through the C-type carbohydrate recognition domain of the cell membrane. Finally, usnic acid, both in its free form and encapsulated in fucoidan-coated liposomes (UA-LipoFuc5), was effective against infected macrophages. Hence, this preliminary investigation suggests that encapsulated usnic acid will aid in further studies related to infected macrophages and may be a potential option for tuberculosis treatment.

Microencapsulation of Lactobacillus acidophilus La-05 and incorporation in vegan milks: Physicochemical characteristics and survival during storage, exposure to stress conditions, and simulated gastrointestinal digestion.

The effect of microencapsulation of L. acidophilus La-05 (8 log CFU/mL) by external ionic gelation technique in alginate (30 g/L; AM) and alginate coated with a low molecular weight chitosan solution (5 g/L; AC5M) on the survival of the freeze-dried probiotic culture during storage (7 °C; 0, 7, 15, 30, 60, 90 and 120 days), and exposure to temperature (72, 85 and 90 °C), pH (2, 4, and 6), and NaCl (10, 15 and 20 g/L) were studied. Furthermore, vegan milks (soybean and rice milks) added with microencapsulated probiotic cultures were evaluated for the physicochemical characteristics and survival of the probiotic culture during refrigerated storage (7 °C; 7 days) and in vitro digestion. Free cells were used as control. AM and AC5M showed similar microencapsulation yield (>90%) with uniform and spherical microparticles dispersed without agglomeration. Scanning electron microscopy showed that chitosan was able to cover the porous structure of the alginate particles, resulting in a more stabilized microparticle. The microencapsulation provided higher probiotic protection to storage, thermal treatment, NaCl and pH (decreases of ~1 log CFU/mL) compared to the free cells (decreases of ~3, 4, 2 and 3 log CFU/mL, respectively), and increased probiotic survival during refrigerated storage and in vitro digestion of vegan milks compared to free cells (decreases of ~1 and 4 log CFU/mL, respectively). Only microencapsulated probiotic cultures (AM and AC5M) maintained suitable probiotic counts (>6 log CFU/mL) during storage, exposure to stress conditions and simulated gastrointestinal digestion. Chitosan coating increased the probiotic survival in the vegan milks during refrigerated storage. Microencapsulation by external ionic gelation in alginate proved to be a suitable microencapsulation technique to improve the probiotic survival to storage, stress conditions (temperature, pH and NaCl) and simulated gastrointestinal conditions. This was the first study that evaluated the addition of probiotic cultures to rice and soybean milks, proving that the vegan milks could be considered suitable carriers for microencapsulated probiotic cultures.