Characterization of gelatin-based wound dressing biomaterials containing increasing coconut oil concentrations.

This study determined the influence and ideal ratios of various coconut oil (CO) amounts in gelatin (G) based-films as wound dressings since there are limited comparative studies to evaluate the sole effect of increasing CO on protein-based biomaterials. Homogenous films at G:CO ratio of 4:0,4:2,4:3,4:4 (w:w) corresponding to CO-0, CO-2, CO-3, CO-4, respectively, were obtained using solution casting. SEM showed CO caused rougher surfaces decreasing mechanical strength. However, no pores were observed in CO-4 due to bigger clusters of oil improving stretchability compared to CO-3; and durability since aging of CO-4 was >10% lower than CO-0 in aqueous media. FTIR showed triglycerides' band only in CO films with increasing amplitude. Moreover, amide-I of CO-2 was involved in more hydrogen bonding, therefore, CO-2 had the highest melt-like transition temperatures (Tmax) at ∼163 °C while others' were at ∼133 °C; and had more ideal mechanical properties among CO films. XTT showed that increased CO improved 3T3 cell viability as CO-0 significantly decreased viability at 10,50,75,100 µg/mL (p < 0.05), whereas CO-2 and CO-3 within 5-75 µg/mL and CO-4 within 5-100 µg/mL range increased viability ≥100% suggesting proliferation. All CO samples at 25 µg/mL stimulated 3T3 cell migration in Scratch Assay indicating wound healing. CO amounts mainly improved thermal and healing properties of gelatin-based biomaterial. CO-2 was more thermally stable and CO-4 had better influence on cell viability and wound healing than CO-0. Therefore, increased CO ratios, specifically 4:2 and 4:4, G:CO (w:w), in gelatin-based films can be ideal candidates for wound dressing materials.

Synthesis characterisation and neuroprotectivity of Silybum marianum extract loaded chitosan nanoparticles.

AIM: Silybum marianum extract (SME) possesses neuroprotective potency through its high antioxidant content. We attempted to increase the effectiveness of SME by encapsulating them in chitosan. Neuroprotective potency of SME and SME-loaded chitosan nanoparticles (SME-CNPs) were shown in SH-SY5Y cell line against H2O2-induced oxidative stress. METHODS: We produced CNPs and SME-CNPs by ionic gelation method and properly determined their physical characteristics. Encapsulation efficiency, loading capacity, and in vitro release tests were performed for SME-CNPs. The neurotoxicity and neuroprotective efficiency in SH-SY5Y cell line against H2O2 was also investigated. RESULTS: The size of SME-CNPs was 168.2 ± 11.12 nm with zeta potential 10.6 ± 1.0 mV. The encapsulation efficiency and loading capacity were successfully achieved at 96.6% and 1.89% respectively. SME and SME-CNPs improved cell viability higher than 80%, and SME-CNPs exhibited significant neuroprotective effects against H2O2 damage. CONCLUSIONS: It was concluded that SME and SME-CNPs highly prevent damage caused by H2O2 and reduce cell damage in vitro by their neuroprotective effects.

Production and characterization of a coconut oil incorporated gelatin-based film and its potential biomedical application.

The influence of coconut oil (CO) on a gelatin-based film was investigated when used as a potential wound dressing material. There is limited study on CO in protein-based wound dressing materials. Therefore, in this study a self-supporting, continuous and homogenous CO incorporated gelatin-based film was formulated and obtained by solution casting method. The influence of CO on physicochemical and thermal properties of gelatin-based film was also determined. Moreover, the effect CO in gelatin films on cell viability and cell migration was analysed with a preliminary cell culture study. Homogenous dispersion of 10% (w/w) CO was obtained in films when 3% (v/w) Tween 80, a surfactant, was incorporated to 20% (w/w) plasticized gelatin film forming solution. Effect of CO on gelatin-based film was observed via phase separation by scanning electron microscopy analysis. Water uptake of gelatin film with no CO, GE film; and 10% (w/w) CO incorporated GE film, GE-CO, were 320% and 210%, respectively, after 3 h in water. Fourier transform infrared spectroscopy analysis showed triglyceride component of CO and increased hydrogen bonding between NH groups of gelatin in GE-CO films. Differential scanning calorimetry results suggested a more ordered structure of GE-CO film due to an increase in melt-like transition temperature and melting enthalpy of GE-CO film. CO content also increased cell viability, assessed by XTT assay since cell viability was approximately 100% when L929 cell culture was incubated with GE-CO of 5-100 µg ml-1. Moreover, GE-CO samples within 5-25 µg ml-1concentration range, increased proliferation of L929 cells since cell viability was significantly higher than the 100% viable cell culture control (P< 0.05) which is also an indication of efficient healing. However, GE decreased viability of L929 cells significantly at 100-10 µg ml-1concentration range (P< 0.05) and were toxic at concentrations of 100, 75 and 50 µg ml-1which decreased ∼50% of the viability of the cells. Scratch Assay to assessin vitrowound healing showed cell migration towards scratch after 24 h as an indication of wound healing only in GE-CO samples. This study showed that, CO could efficiently be added to gelatin-based films for preparation of a primary wound dressing biomaterial which is also demonstrated to have a promising wound healing effect for minor wounds.

Conjugation, characterization and toxicity of lipophosphoglycan-polyacrylic acid conjugate for vaccination against leishmaniasis.

Research on the conjugates of synthetic polyelectrolytes with antigenic molecules, such as proteins, peptides, or carbohydrates, is an attractive area due to their highly immunogenic character in comparison to classical adjuvants. For example, polyacrylic acid (PAA) is a weak polyelectrolyte and has been used in several biomedical applications such as immunological studies, drug delivery, and enzyme immobilization. However, to our knowledge, there are no studies that document immune-stimulant properties of PAA in Leishmania infection. Therefore, we aimed to develop a potential vaccine candidate against leishmaniasis by covalently conjugating PAA with an immunologically vital molecule of lipophosphoglycan (LPG) found in Leishmania parasites. In the study, LPG and PAA were conjugated by a multi-step procedure, and final products were analyzed with GPC and MALDI-TOF MS techniques. In cytotoxicity experiments, LPG-PAA conjugates did not indicate toxic effects on L929 and J774 murine macrophage cells. We assume that LPG-PAA conjugate can be a potential vaccine candidate, and will be immunologically characterized in further studies to prove its potential.

Utility of the microculture method in non-invasive samples obtained from an experimental murine model with asymptomatic leishmaniasis.

The sensitivity of diagnostic methods for visceral leishmaniasis (VL) decreases because of the low number of parasites and antibody amounts in asymptomatic healthy donors who are not suitable for invasive sample acquisition procedures. Therefore, new studies are urgently needed to improve the sensitivity and specificity of the diagnostic approaches in non-invasive samples. In this study, the sensitivity of the microculture method (MCM) was compared with polymerase chain reaction (PCR), enzyme-linked immunosorbent assay (ELISA), and immunofluorescent antibody test (IFAT) methods in an experimental murine model with asymptomatic leishmaniasis. Results showed that the percent of positive samples in ELISA, IFAT, and peripheral blood (PB) -PCR tests were 17.64%, 8.82%, and 5.88%, respectively, whereas 100% positive results were obtained with MCM and MCM-PCR methods. Thus, this study, for the first time, showed that MCM is more sensitive, specific, and economic than other methods, and the sensitivity of PCR that was performed to samples obtained from MCM was higher than sensitivity of the PCR method sampled by PB.