Edible duck feet gelatin (DFG)-based biocomposites with different glycerol (GLY) contents (15%, 25%, and 35% of dried DFG) were prepared. Physicochemical, mechanical, barrier, and heat seal properties of DFG films were characterized and compared as an alternative to bovine gelatin film. Increasing glycerol from 15 to 35% decreased the TS and YM and EB and HS increased, in value of 42.54-7.27 and 1240-157.10MPa and 22.82-50.33% and 42.06-347.15N/m respectively. The water vapor permeability (WVP) and oxygen permeability (OP) of films were increased from 4.78 to 5.6×10-11gm-1Pa-1s-1 and from 3.97 to 33.99cm3mμ/m2 d kPa respectively. GAB model estimations showed monolayer water content of films increased with the increase of plasticizer content. Moisture sorption isotherm modelling exhibited a type II BET classification. Fourier transform infrared (FTIR) spectra showed shifted peak at approximately 1024cm-1, which was related to glycerol. The results show that the properties of DFG film are suitable for use as an alternative material to bovine gelatin film.
Gelatin is a highly purified animal protein of pig, cow, and fish origins and is extensively used in food, pharmaceuticals, and personal care products. However, the acceptability of gelatin products greatly depends on the animal sources of the gelatin. Porcine and bovine gelatins have attractive features but limited acceptance because of religious prohibitions and potential zoonotic threats, whereas fish gelatin is welcomed in all religions and cultures. Thus, source authentication is a must for gelatin products but it is greatly challenging due to the breakdown of both protein and DNA biomarkers in processed gelatins. Therefore, several methods have been proposed for gelatin identification, but a comprehensive and systematic document that includes all of the techniques does not exist. This up-to-date review addresses this research gap and presents, in an accessible format, the major gelatin source authentication techniques, which are primarily nucleic acid and protein based. Instead of presenting these methods in paragraph form which needs much attention in reading, the major methods are schematically depicted, and their comparative features are tabulated. Future technologies are forecasted, and challenges are outlined. Overall, this review paper has the merit to serve as a reference guide for the production and application of gelatin in academia and industry and will act as a platform for the development of improved methods for gelatin authentication.
Gelatin is widely used in pharmaceuticals as a protective coating, such as soft and hard capsule shells. However, the animal source of gelatin is a sensitive issue because certain gelatins such as porcine and bovine gelatins are not welcome in Halal, Kosher and Hindus' consumer goods. Recently, we have documented DNA barcoding and multiplex PCR platforms for discriminating porcine, bovine and fish gelatins in various fish and confectionary products; but those assays were not self-authenticating and also not tested in highly refined pharmaceutical products. To address this knowledge gap, here we report a self-authenticating multiplex PCR-restriction fragment length polymorphism (RFLP) assay to identify animal sources of various gelatin in pharmaceutical capsules. Three different restriction enzymes, BsaAI, Hpy188I and BcoDI were used to yield distinctive RFLP patterns for gelatin-based bovine (26, 94 bp), fish (97, 198 bp) and porcine (17, 70 bp) DNA in control experiments. The specificity was cross-tested against 16 non-target species and the optimised assay was used to screen gelatin sources in 30 halal-branded pharmaceuticals capsule shells. Bovine and porcine DNA was found in 27 and 3 of the 30 different capsules products. The assay was suitable for detecting 0.1 to 0.01 ng total DNA extracted from pure and mixed gelatins. The study might be useful to authenticate and monitor halal, kosher, vegetarian and Hindu compliant pharmaceuticals, foods and cosmetics.
In this work, chitosan, gelatin and methylcellulose films incorporated with tannic acid (TA) were synthesised, characterised and applied for the first time to preserve cherry tomatoes (Solanum lycopersicum var. cerasiforme) and grapes (Vitis vinifera). The addition of TA at 15% (w/w) increased the transparency value of biopolymer films. The highest increment of transparency value was obtained for MC-TA film, increased from 0.572 to 4.73 A/mm. Based on antimicrobial study, the addition of TA improved the antibacterial properties of biopolymers against Escherichia coli (E. coli) and Staphylococcus aureus (S. aureus). The ability of films to preserve both fruits was evaluated in a 14-day preservation study. The application of biopolymer films treated with TA has decreased the weight loss and browning index of fruits, as compared to control films. A significant reduction in the weight loss of cherry tomatoes wrapped with chitosan (from 21.3 to 19.6%), gelatin (from 22.1 to 15.5%) and methylcellulose (26.2 to 20.5%) films were obtained following TA treatment. Overall, results obtained from this study highlight the effects of TA on physiochemical properties of biopolymer films and their ability to preserve fruits.
Composite films comprised of salmon (Salmo salar) skin gelatin and zein were prepared via crosslinking with glutaraldehyde. Response surface methodology (RSM) was used to optimize film composition to maximize tensile strength (TS) and elongation at break (EAB), and to minimize water solubility (WS) of the films. The significant (P gelatin-zein composite film was successfully crosslinked after the addition of glutaraldehyde, with the formation of crosslinked networks between proteins and a denser packed organization of proteins. Consequently, the resultant crosslinked composite film exhibited improvement on light transparency, water resistance and mechanical strength as a function of increasing humidity.
The issue of food authenticity has become a concern among religious adherents, particularly Muslims, due to the possible presence of nonhalal ingredients in foods as well as other commercial products. One of the nonhalal ingredients that commonly found in food and pharmaceutical products is gelatin which extracted from porcine source. Bovine and fish gelatin are also becoming the main commercial sources of gelatin. However, unclear information and labeling regarding the actual sources of gelatin in food and pharmaceutical products have become the main concern in halal authenticity issue since porcine consumption is prohibited for Muslims. Hence, numerous analytical methods involving chemical and chemometric analysis have been developed to identify the sources of gelatin. Chemical analysis techniques such as biochemical, chromatography, electrophoretic, and spectroscopic are usually combined with chemometric and mathematical methods such as principal component analysis, cluster, discriminant, and Fourier transform analysis for the gelatin classification. A sample result from Fourier transform infrared spectroscopy analysis, which combines Fourier transform and spectroscopic technique, is included in this paper. This paper presents an overview of chemical and chemometric methods involved in identification of different types of gelatin, which is important for halal authentication purposes.
Composite film dressings composed of pluronic F127 (PL)-pectin (PC) and pluronic (PL) F127-gelatin (GL) were investigated as potential drug delivery system for wound healing. Composite films were solvent cast by blending PL with PC or GL in different ratios using glycerol (2.5%) as plasticizer. Erythromycin (ER) (0.1%) was incorporated in films as model hydrophobic antibiotic. The optimized composite films were characterized for physical appearance, morphology, mechanical profile, and thermal behavior. In addition, drug release, antibacterial activity, and cytocompatibility of the films were investigated to assess their potential as drug delivery system. The composite films exhibited excellent wound dressing characters in terms of appearance, stability, and mechanical profile. Moreover, ER-loaded composite films released ER in controlled manner, exhibited antibacterial activity against Staphylococcus aureus, and were non-toxic to human skin fibroblast. These findings demonstrate that these composite films hold the potential to be formulated as antibacterial wound dressing.
This study aimed to explore a potential use of fish scale-derived gelatin nanofibrous scaffolds (GNS) in tissue engineering due to their biological and economical merits. Extraction of gelatin was achieved via decalcification, sonication and lyophilization of mixed fish scales. To fabricate nano-scale architecture of scaffolds analogous to natural extracellular matrix, gelatin was rendered into nanofibrous matrices through 6-h electrospinning, resulting in the average diameter of 48 ± 12 nm. In order to improve the water-resistant ability while retaining their biocompatibility, GNS were physically crosslinked with ultraviolet (UV) irradiation for 5 min (UGN5), 10 min (UGN10) and 20 min (UGN20). On average, the diameter of nanofibers increased by 3 folds after crosslinking, however, Fourier transform infrared spectroscopy analysis confirmed that no major alterations occurred in the functional groups of gelatin. A degradation assay showed that UGN5 and UGN10 scaffolds remained in minimum essential medium for 14 days, while UGN20 scaffolds degraded completely after 10 days. All UGN scaffolds promoted adhesion and proliferation of human keratinocytes, HaCaT, without causing an apparent cytotoxicity. UGN5 scaffolds were shown to stimulate a better growth of HaCaT cells compared to other scaffolds upon 1 day of incubation, whereas UGN20 had a long-term effect on cells exhibiting 25% higher cell proliferation than positive control after 7 days. In the wound scratch assay, UGN5 scaffolds induced a rapid cell migration closing up to 79% of an artificial wound within 24 h. The current findings provide a new insight of UGN scaffolds to serve as wound dressings in the future. In the wound scratch assay, UGN5 induced a rapid cell migration closing up to 79% of an artificial wound within 24 h.
Vaccine administration via the oral route is preferable to parenteral routes due to ease of administration. To date, most available oral vaccines comprises of live attenuated pathogens as oppose to peptide-based vaccines due to its low bioavailability within the gastrointestinal (GI) tract. Over the years, probiotic-based peptide delivery vehicles comprising of lactic acid bacteria such as Lactococcus lactis has emerged as an interesting alternative due to its generally recognized as safe (GRAS) status, a fully sequenced genome, transient gut colonization time, and is an efficient cellular factory for heterologous protein production. However, its survivability through the GI tract is low, thus better delivery approaches are being explored to improve its bioavailability. In this study, we employ the incorporation of a double coated mucoadhesive film consisting of sodium alginate and Lycoat RS 720 film as the inner coat. The formulated film exhibits good mechanical properties of tensile strength and percent elongation for manipulation and handling with an entrapment yield of 93.14±2.74%. The formulated mucoadhesive film is subsequently loaded into gelatin capsules with an outer enteric Eudragit L100-55 coating capable of a pH-dependent breakdown above pH 5.5 to protect against gastric digestion. The final product and unprotected controls were subjected to in vitro simulated gastrointestinal digestions to assess its survivability. The product demonstrated enhanced protection with an increase of 69.22±0.67% (gastric) and 40.61±8.23% (intestinal) survivability compared to unprotected controls after 6 hours of sequential digestion. This translates to a 3.5 fold increase in overall survivability. Owing to this, the proposed oral delivery system has shown promising potential as a live gastrointestinal vaccine delivery host. Further studies involving in vivo gastrointestinal survivability and mice immunization tests are currently being carried out to assess the efficacy of this novel oral delivery system in comparison to parenteral routes.
Gelatin and collagen are considered halal-critical ingredients as they are typically derived from either bovine or porcine animals. Current analytical methods for determining the sources of gelatin and collagen suffer from limitations in terms of robustness and false positives in peptide matching. Thus, the aim of this study was to investigate the utility of monitoring hydroxyproline, a signature amino acid for gelatin and collagen, for identifying potentially haram foodstuffs. To determine the hydroxyproline profiles among animal- and plant-based samples, one-way univariate analysis of variance followed by pair-wise comparison was used to establish statistical significance. Multivariate chemometric analysis through principal component analysis revealed a discrete distribution pattern among 59 samples due to hydroxyproline variability. Finally, inter- and intra-laboratory comparisons demonstrated the validity and robustness of hydroxyproline determination according to ISO 17025. Thus, this preliminary identification technique will aid the identification of potentially haram foodstuffs.
Gelatin hydrogels are attractive for wound applications owing to their well-defined structural, physical, and chemical properties as well as good cell adhesion and biocompatibility. This study aimed to develop gelatin hydrogels incorporated with bio-nanosilver functionalized with lactoferrin (Ag-LTF) as a dual-antimicrobial action dressing, to be used in treating infected wounds. The hydrogels were cross-linked using genipin prior to loading with Ag-LTF and characterized for their physical and swelling properties, rheology, polymer and actives interactions, and in vitro release of the actives. The hydrogel's anti-biofilm and antibacterial performances against S. aureus and P. aeruginosa as well as their cytotoxicity effects were assessed in vitro, including primary wound healing gene expression of human dermal fibroblasts (HDFs). The formulated hydrogels showed adequate release of AgNPs and LTF, with promising antimicrobial effects against both bacterial strains. The Ag-LTF-loaded hydrogel did not significantly interfere with the normal cellular functions as no alteration was detected for cell viability, migration rate, and expression of the target genes, suggesting the nontoxicity of Ag-LTF as well as the hydrogels. In conclusion, Ag-LTF-loaded genipin-cross-linked gelatin hydrogel was successfully synthesized as a new approach for fighting biofilms in infected wounds, which may be applied to accelerate healing of chronic wounds.
This study was aimed at engineering photocrosslinkable azithromycin (AZ)-laden gelatin methacryloyl fibers via electrospinning to serve as a localized and biodegradable drug delivery system for endodontic infection control. AZ at three distinct amounts was mixed with solubilized gelatin methacryloyl and the photoinitiator to obtain the following fibers: GelMA+5%AZ, GelMA+10%AZ, and GelMA+15%AZ. Fiber morphology, diameter, AZ incorporation, mechanical properties, degradation profile, and antimicrobial action against Aggregatibacter actinomycetemcomitans and Actinomyces naeslundii were also studied. In vitro compatibility with human-derived dental pulp stem cells and inflammatory response in vivo using a subcutaneous rat model were also determined. A bead-free fibrous microstructure with interconnected pores was observed for all groups. GelMA and GelMA+10%AZ had the highest fiber diameter means. The tensile strength of the GelMA-based fibers was reduced upon AZ addition. A similar pattern was observed for the degradation profile in vitro. GelMA+15%AZ fibers led to the highest bacterial inhibition. The presence of AZ, regardless of the concentration, did not pose significant toxicity. In vivo findings indicated higher blood vessel formation, mild inflammation, and mature and thick well-oriented collagen fibers interweaving with the engineered fibers. Altogether, AZ-laden photocrosslinkable GelMA fibers had adequate mechanical and degradation properties, with 15%AZ displaying significant antimicrobial activity without compromising biocompatibility.
To enhance the solubility of hydrophobic nutrients, the hydrophobicity of fish scale gelatin hydrolysate (FSGH) was increased with moderate acid or alkali hydrolysis. Acid-induced FSG hydrolysate (AcFSGH) at 3 h showed a superior curcumin loading efficiency (18.30 ± 0.38 μg/mL) among all FSGHs. Compared with FSG, the proportion of hydrophobic amino acids (from 41.1% to 46.4%) and the hydrophobic interaction (from 12.72 to 20.10 mg/mL) was significantly increased in the AcFSGH. Meanwhile, the transformation of the α-helix (from 12.8% to 4.9%) to the β-sheet (from 29.0% to 42.8%) was also observed in the AcFSGH. Based on the observation in the molecular weight and morphological analysis, AcFSGH acquired the best hydrophobic interaction with curcumin, presumably due to the formation of the flexible structure of the linear hydrolyzates. The above results call for an investigation of the role of FSG hydrolysate in the synthesis of nanoparticles loaded with bioactive lipophilic compounds.