Bone Broth Protein

Polymyxins are last-line antibiotics against multidrug-resistant Klebsiella pneumoniae but using polymyxins alone may not be effective due to emerging resistance. A previous study found that combining polymyxin B with chloramphenicol effectively kills MDR K. pneumoniae, although the bone marrow toxicity of chloramphenicol is concerning. The aim of this study is to assess the antibacterial efficacy and cytotoxicity of polymyxin B when combined with chloramphenicol and its derivatives, namely thiamphenicol and florfenicol (reported to have lesser toxicity compared to chloramphenicol). The antibacterial activity was evaluated with antimicrobial susceptibility testing using broth microdilution and time-kill assays, while the cytotoxic effect on normal bone marrow cell line, HS-5 was evaluated using the MTT assay. All bacterial isolates tested were found to be susceptible to polymyxin B, but resistant to chloramphenicol, thiamphenicol, and florfenicol when used alone. The use of polymyxin B alone showed bacterial regrowth for all isolates at 24 h. The combination of polymyxin B and florfenicol demonstrated additive and synergistic effects against all isolates (≥ 2 log10 cfu ml-1 reduction) at 4 and 24 h, respectively, while the combination of polymyxin B and thiamphenicol resulted in synergistic killing at 24 h against ATCC BAA-2146. Furthermore, the combination of polymyxin B with florfenicol had the lowest cytotoxic effect on the HS-5 cells compared to polymyxin B combination with chloramphenicol and thiamphenicol. Overall, the combination of polymyxin B with florfenicol enhanced bacterial killing against MDR K. pneumoniae and exerted minimal cytotoxic effect on HS-5 cell line. © 2023. The Author(s), under exclusive licence to the Japan Antibiotics Research Association.

Bone broth readily undergoes oil-water separation due to its complex emulsified structure, which limits its industrial utilization. This study evaluated the influence of xanthan gum (XG) and guar gum (GG) blends on the stability of bovine bone broth. The 6:4 XG:GG ratio exhibited the strongest synergistic effect, achieving a droplet size of 0.99 μm and a zeta potential of -25.1 mV. The system showed pseudoplastic behavior with G' consistently exceeding G″. Spectroscopic results indicated an 18.4% increase in β-sheet content and the highest interfacial protein adsorption (37 mg/m2), suggesting the formation of a compact interfacial film. Molecular docking revealed that XG interacts with collagen through electrostatic bonding, while GG contributes hydrophobic interactions. After 60 days at 25 °C, POV and TBARS decreased to 45% and 40% of the control, confirming improved oxidative and storage stability. These findings provide a basis for designing stable high-fat emulsified foods.

This study developed a bio-enhanced acidolysis process using Acetobacter fermentation to efficiently release and transform calcium from animal-derived byproducts, including eggshell, oyster shell, beef bone, and chicken bone powders. Shell-derived broths released markedly more calcium than bone-derived broths, around a 5.6-fold difference. Released calcium may be present in multiple forms: free ionic Ca2+, protein calcium, peptide calcium, and chelated complexes with organic and amino acids. FTIR, XRD, and BET analyses indicated that biologically produced acetic acid selectively dissolved disordered or amorphous regions, increasing residual crystallinity and particle size. Furthermore, the microbial system integrates organic acid generation with acid-driven mineral dissolution and collagen matrix loosening, forming a synergistic mechanism for calcium solubilization. Overall, this Acetobacter mediated bio-acidification process provides a green, economical, and multifunctional pathway for converting low-value animal byproducts into bioaccessible calcium ingredients, supporting sustainable development in functional food bioprocessing.

Umami peptides are valuable taste substances due to their exceptional taste and beneficial properties. In this study, purification of fermented goose bone broth was performed using continuous chromatography and sensory analysis, and after identification through nano-LC-MS/MS, four umami peptides were screened out by umami activity prediction and molecular docking, which are VGYDAE, GATGRDGAR, GETGEAGER, and GETGEAGERG derived from collagen. Sensory analysis indicated that they were also umami-enhancing, with thresholds ranging from 0.41 to 1.15 mmol/L, among which GER9 was the best. Combining the results of docking and molecular dynamics simulation, it was known that hydrogen bond and electrostatic interactions were vital in driving the umami formation. Moreover, Glu, Ser, and Asp of umami receptor T1R1/T1R3 were the key residues for the binding between four umami peptides and T1R1/T1R3. These findings provide novel insights into the high-value utilization of goose bones and offer profound theoretical guidance for understanding the umami mechanism.

Membrane exposure is a widely reported and relatively common complication in Guided Bone Regeneration (GBR) procedures. The introduction of micro-porous dPTFE barriers, which are impervious to bacterial cells, could reduce the technique sensitivity to membrane exposure, even if there are no studies investigating the potential passage of bacterial metabolites through the barrier. Aim of this study was the in vitro evaluation of the permeability of three different GBR membranes (dPTFE, native and cross-linked collagen membranes) to Porphyromonas gingivalis; in those cases, where bacterial penetration could not be observed, another purpose was the analysis of the viability and differentiation capability of an osteosarcoma (U2OS) cell line in presence of bacteria eluate obtained through membrane percolation. A system leading to the percolation of P. gingivalis broth culture through the experimental membranes was arranged to assess the permeability to bacteria after 24 and 72 h of incubation. The obtained solution was then added to U2OS cell cultures which underwent, after 10 days of incubation, MTT and red alizarin essays. The dPTFE membrane showed resistance to bacterial penetration, while both types of collagen membranes were crossed by P. gingivalis after 24 h. The bacteria eluate filtered through dPTFE membrane didn't show any toxicity on U2OS cells. Results of this study demonstrate that dPTFE membranes can contrast the penetration of both P. gingivalis and its metabolites toxic for osteoblast-like cells. The toxicity analysis was not possible for the collagen membranes, since permeability to bacterial cells was observed within the first period of incubation. © 2022. The Author(s), under exclusive licence to The Society of The Nippon Dental University.

Introduction: Recent evidence suggests that the consumption of essential amino acids (AA) and/or those abundantly present in collagen may have the capacity to influence the synthesis of new collagen in ligaments and tendons, when tissue perfusion is optimized (e.g., during exercise). However, little is currently known about the bioavailability of these AAs in blood after the consumption of various collagen and diary protein sources: such information is needed to develop potentially useful dietary and supplement intake strategies. Objectives: The aim of the current study was to characterize blood AA concentrations in response to consumption of collagen and dairy protein sources; specifically, maximum concentrations, the timing of maximum concentration, and total (area under the curve) exposure above baseline. Methods: A 20 g serve of various dairy and collagen proteins, and a 300 mL serve of bone broth were consumed by healthy, recreationally active males after an overnight fast. Blood samples were drawn every 20 min for a total of 180 min, for analysis of plasma AA concentrations. Total AA, essential AA and collagen specific AAs were analyzed for maximum concentration, timing of peak, and area under the curve. Results: In general, protein intake was associated with a similar increase in total and collagen specific AAs, except for collagen proteins being a superior source of glycine (683 ± 166 μmol/L) compared to 260 ± 65 μmol/L for dairy proteins (P < 0.0001), whilst dairy proteins were a superior source of leucine (267 ± 77 μmol/L) compared to 189 ± μmol/L for collagen proteins (P < 0.04). Although there were several differences in the bioavailability of hydrolysed compared to non-hydrolysed proteins, this only reached statistical significance within the dairy proteins, but not for collagen proteins. Conclusions: The intake of collagen proteins result in higher plasma peaks of glycine, whilst the intake of dairy proteins result in higher plasma peaks of leucine. This information may support further investigations, and identification of key AAs that may support exercise in the synthesis of collagen.

Bone-derived protein stocks are used in food industry to enhance taste of soups, sauces, and a range of other products. Both during commercial manufacturing and when used for culinary purposes, the stocks may be exposed to high temperatures for an extended time period. The present study investigated the effect of retention at 90°C for 0, 3, 6, 9, 24, 48, 72, and 168 hr on the functional attributes of concentrated bone-derived beef stocks (57% Dry matter (DM)). Visual inspection and rheological analyses showed that during increasing heat exposure, the gel strength as well as viscosity of the concentrated stocks decreased incrementally and significantly (P > 0.001). Nuclear magnetic resonance (NMR) relaxation measurements conducted on the beef stocks also revealed strong effects of heat exposure on the transverse (T2 ) relaxation time, which increased incrementally and significantly (P > 0.001) with longer heat exposure. Thus, the present study demonstrated that heat-induced changes in rheological properties of bone-derived beef stocks can be ascribed to changes in intrinsic water-protein interactions and water attributes as a result of heat-induced protein modifications. In conclusion, the study proves that NMR relaxometry is a valuable tool for monitoring changes in intrinsic water mobility that are manifested in modified functional attributes of concentrated beef stocks. © 2019 John Wiley & Sons, Ltd.

Intake of dietary sources of collagen may support the synthesis of collagen in varying tissues, with the availability of key amino acids being a likely contributor to its effectiveness. This study analyzed commonly consumed preparations of bone broth (BB) to assess the amount and consistency of its amino acid content. Commercial and laboratory-prepared samples, made with standardized and variable (nonstandardized) protocols, were analyzed for key amino acids (glycine, lysine, proline, leucine, hydroxyproline, and hydroxylysine). The main finding of this study was that amino acid concentrations in BB made to a standardized recipe were significantly lower for hydroxyproline, glycine, and proline (p = .003) and hydroxylysine, leucine, and lysine (p = .004) than those provided by a potentially therapeutic dose (20 g) of reference collagen supplements (p > .05). There was a large variability in the amino acid content of BB made to nonstandardized recipes, with the highest levels of all amino acids found within the café-prepared varieties. For standardized preparations, commercial BBs were lower in all amino acids than the self-prepared varieties. There were no differences (p > .05) in the amino acid content of different batches of BB when prepared according to a standardized recipe. If the intake of collagen precursors is proven to support the synthesis of new collagen in vivo, it is unlikely that BB can provide a consistently reliable source of key amino acids. Research on the provision of key amino acids from dietary sources should continue to focus on the standard sources currently being researched.

Nanostructured calcium phosphate porous microspheres are of great potential in drug delivery and bone regeneration due to their large specific surface area, biocompatibility, and similarity to inorganic component of osseous tissue. In this work, strontium (Sr)-doped amorphous calcium phosphate porous microspheres (SrAPMs) were synthesized through a microwave-hydrothermal method using fructose 1,6-bisphosphate trisodium salt as the source of phosphate ions. The SrAPMs showed a mesoporous structure and a relatively high specific area. Compared with the hydroxyapatite nanorods prepared by using Na2HPO4·12H2O as the phosphorus source, the SrAPMs with a higher specific surface area were more effective in drug loading using vancomycin as the antiobiotics of choice and consequently having a higher antibacterial efficiency both on agar plates and in broths. Furthermore, to assess the potential application of SrAPMs in bone defect repair, a novel biomimetic bone tissue-engineering scaffold consisting of collagen (Coll) and SrAPMs was constructed using a freeze-drying fabrication process. Incorporation of the SrAPMs not only improved the mechanical properties, but also enhanced the osteogenesis of rat bone marrow mesenchymal stem cells. The in vivo experiments demonstrated that the SrAPMs/Coll scaffolds remarkably enhanced new bone formation compared with the Coll and APMs/Coll scaffolds in a rat critical-sized calvarial defect model at 8 weeks postimplantation. In summary, SrAPMs developed in this work are promising as antibiotic carriers and may encourage bone formation when combined with collagen.

BACKGROUND: The present study aimed to determine if salty and sweet taste preferences in children are related to each other, to markers of growth, and to genetic differences. METHODS: We conducted a 2-day, single-blind experimental study using the Monell two-series, forced-choice, paired-comparison tracking method to determine taste preferences. The volunteer sample consisted of a racially/ethnically diverse group of children, 5-10 years of age (n = 108), and their mothers (n = 83). After excluding those mothers who did not meet eligibility and children who did not understand or comply with study procedures, the final sample was 101 children and 76 adults. The main outcome measures were most preferred concentration of salt in broth and crackers; most preferred concentration of sucrose in water and jelly; reported dietary intake of salty and sweet foods; levels of a bone growth marker; anthropometric measurements such as height, weight, and percent body fat; and TAS1R3 (sweet taste receptor) genotype. RESULTS: Children preferred higher concentrations of salt in broth and sucrose in water than did adults, and for both groups, salty and sweet taste preferences were significantly and positively correlated. In children, preference measures were related to reported intake of sodium but not of added sugars. Children who were tall for their age preferred sweeter solutions than did those that were shorter and percent body fat was correlated with salt preference. In mothers but not in children, sweet preference correlated with TAS1R3 genotype. CONCLUSIONS AND RELEVANCE: For children, sweet and salty taste preferences were positively correlated and related to some aspects of real-world food intake. Complying with recommendations to reduce added sugars and salt may be more difficult for some children, which emphasizes the need for new strategies to improve children's diets.

Development of a material for simultaneous sustained and localized delivery of antibiotics and induction of spontaneous regeneration of hard tissues affected by osteomyelitis stands for an important clinical need. In this work, a comparative analysis of the bacterial and osteoblastic cell response to two different nanoparticulate carriers of clindamycin, an antibiotic commonly prescribed in the treatment of bone infection, one composed of calcium phosphate and the other comprising poly-(D,L-lactide-co-glycolide)-coated calcium phosphate, was carried out. Three different non-cytotoxic phases of calcium phosphate, exhibiting dissolution and drug release profiles in the range of one week to two months to one year, respectively, were included in the analysis: monetite, amorphous calcium phosphate and hydroxyapatite. Spherical morphologies and narrow size distribution of both types of nanopowders were confirmed in transmission and scanning electron microscopic analyses. The antibiotic-containing powders exhibited sustained drug release contingent upon the degradation rate of the carrier. Assessment of the antibacterial performance of the antibiotic-encapsulated powders against Staphylococcus aureus, the most common pathogen isolated from infected bone, yielded satisfactory results both in broths and on blood agar plates for all the analyzed powders. In contrast, no cytotoxic behavior was detected upon the incubation of the antibiotic powders with the osteoblastic MC3T3-E1 cell line for up to three weeks. The cells were shown to engage in a close contact with the antibiotic-containing particles, irrespective of their internal or surface phase composition, polymeric or mineral. At the same time, both types of particles upregulated the expression of osteogenic markers osteocalcin, osteopontin, Runx2 and protocollagen type I, suggesting their ability to promote osteogenesis and enhance remineralization of the infected site in addition to eliminating the bacterial source of infection.

A staphylococal surface protein capable of binding several extracellular matrix glycoproteins was purified as a result of our attempts to identify a receptor(s) for bone sialoprotein (BSP) on Staphylococcus aureus cells. Proteins from different staphylococcal strains were solubilized in sodium lauryl sulfate, separated by polyacrylamide gel electrophoresis, blotted onto Immobilon P membranes, and probed with 125I-BSP. Several bacterial proteins bound the radiolabeled ligand, and various strains expressed different repertoirs of BSP-binding proteins. Major BSP-binding proteins with apparent M(r)s of 72,000 or 60,000 were present on most strains, and these proteins were further studied. The 72- and 60-kDa proteins were preferentially expressed when bacteria were cultured in Luria broth compared with when they were cultured on tryptic soy broth, and the abundance of the proteins could be correlated to an increased 125I-BSP binding. Both the 72-kDa and the 60-kDa proteins were solubilized by extraction of cells with 1 M LiCl and were purified by cation-exchange chromatography. Amino acid composition analysis of the purified 72-kDa protein indicated a high content of lysine (11.9%) and hydrophobic amino acids (28.0% combined). In Western ligand blotting (immunoblotting) experiments, the 72-kDa protein bound not only BSP but also radiolabeled fibronectin, fibrinogen, vitronectin, thrombospondin, and, to some extent, collagen. Addition of the purified 60-kDa protein to S. aureus cells did not inhibit binding of the different ligands but in some cases resulted in an augmentation of the binding of 125I-ligand. Purified 60-kDa protein could hemagglutinate sheep erythrocytes at a concentration of 61 micrograms/ml. The agglutination reaction was inhibited by high concentrations of fucose, mannose, or melibiose. These data suggest that the purified proteins may serve as bacterial receptors with broad specificity for matrix glycoproteins and that the proteins may act as carbohydrate-binding proteins.

Bone broth is a traditional nutrient revered by different people from ancient times to the modern era as a remedy for various illnesses. This review investigates the nutritional components of bone broth, focusing primarily on the most abundant amino acids and minerals saturated in bone broth and their impact on health, particularly in the context of intestinal barrier integrity, intestinal permeability, inflammation, and their application in inflammatory bowel disease. Through comprehensive reviews of animal and human studies, this research highlights that bone broth includes amino acids (glutamine, glycine, proline, histidine, arginine), minerals (Ca, P, K, Mg, Zn) that are beneficial and not just a traditional remedy, resolving questions that have been posed for generations. The benefits documented for components in bone broth support the enhancement of gut health, alleviate inflammation in the intestinal barrier, improve intestinal barrier function in health and disease states, particularly in inflammatory bowel disease, as well as enhancing nutrient absorption. Bone broth offers a nutrient-dense option for enhancing overall health and may offer an alternative to dietary supplements with claims for enhanced gut health. We aim to foster interest in and provide evidence to substantiate claims for bone broth as a potential remedy, particularly for maintaining remission in conditions like IBD and possibly functional diarrhea and to encourage further research. © 2025. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.