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Magnesium is an excellent material in terms of biocompatibility and its corrosion products can serve as an active source for new bone formation. However, localized corrosion and H2 generation limit the potential of Mg-based implants. Utilizing low-alloyed Mg-Zn wires can strongly reduce problems with large H2 bubbles and improve the mechanical properties considerably while maintaining excellent long-term biocompatibility. Acidic pickling and a polymer coating can be effectively used to lower the rate of in vivo degradation. In this work, microstructural, mechanical, and in vitro characterization of 250 µm and 300 µm extruded wires made from ultra-pure Mg, commercially pure Mg, Mg-0.15Zn, Mg-0.4Zn and Mg-1Zn was performed. Additionally, Mg-0.4Zn wires together with a variant coated with a copolymer of l-lactide and ε-caprolactone were tested in vivo on artificially damaged Wistar rat femurs. Based on the observed Mg-induced osteogenesis, polymer-coated Mg wires with a small addition of Zn are a perspective material for bone-support applications, such as cerclage and fixation wires.

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Various studies have correlated the mechanical properties of the aortic wall with its biochemical parameters and inner structure. Very few studies have addressed correlations with the cohesive properties, which are crucial for understanding fracture phenomena such as aortic dissection, i.e. a life-threatening process. Aimed at filling this gap, we conducted a comprehensive biochemical and histological analysis of human aortas (the ascending and descending thoracic and infrarenal abdominal aorta) from 34 cadavers obtained post-mortem during regular autopsies. The pentosidine, hydroxyproline and calcium contents, calcium/phosphorus molar ratio, degree of atherosclerosis, area fraction of elastin, collagen type I and III, alpha smooth muscle actin, vasa vasorum, vasa vasorum density, aortic wall thickness, thicknesses of the adventitia, media and intima were determined and correlated with the delamination forces in the longitudinal and circumferential directions of the vessel as determined from identical cadavers. The majority of the parameters determined did not indicate significant correlation with age, except for the calcium content and collagen maturation (enzymatic crosslinking). The main results concern differences between enzymatic and non-enzymatic crosslinking and those caused by the presence of atherosclerosis. The enzymatic crosslinking of collagen increased with age and was accompanied by a decrease in the delamination strength, while non-enzymatic crosslinking tended to decrease with age and was accompanied by an increase in the delamination strength. As the rate of calcification increased, the presence of atherosclerosis led to the formation of calcium phosphate plaques with higher solubility than the tissue without or with only mild signs of atherosclerosis. STATEMENT OF SIGNIFICANCE: This study presents a detailed biochemical and histological analysis of human aortic samples (ascending thoracic aorta, descending thoracic aorta and infrarenal abdominal aorta) taken from 34 cadavers. The contribution of this scientific study lies in the detailed biochemical comparison of the enzymatic and non-enzymatic glycosylation-derived crosslinks of vascular tissues and their influence on the delamination strength of the human aorta since, to the best of our knowledge, no such comprehensive studies exist in the literature. A further benefit concerns the notification of the limitations of the various analytical methods applied; an important factor that must be taken into account in such studies., Competing Interests: Declaration of interests The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.)

The aim of our study was to describe the impact of collagen in the gel and dry state to various doses of electron beam radiation (1, 10 and 25 kGy) which are using for food processing and sterilization. The changes in the chemical compositions (water, amino acids, lipids, glycosaminoglycans) were analyzed and the changes in the structure (triple-helix or β-sheet, the integrity of the collagen) were assessed. Subsequently, the impact of the applied doses on the mechanical properties, stability in the enzymatic environment, swelling and morphology were determined. The irradiated gels evinced enhanced degrees of cross-linking with only partial degradation. Nevertheless, an increase was observed in their stability manifested via a higher degree of resistance to the enzymatic environment, a reduction in swelling and, in terms of the mechanical behaviour, an approximation to the non-linear behavior of native tissues. In contrast, irradiation in the dry state exerted a somewhat negative impact on the observed properties and was manifested mainly via the scission of the collagen molecule and via a lower degree of stability in the aqueous and enzymatic environments. Neither the chemical composition nor the morphology was affected by irradiation., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier B.V. All rights reserved.)

The aim of this study was to investigate batch-to-batch inconsistencies in the processing of pig and fish collagen isolates processed using two protocols that differed in terms of the acetic acid concentrations applied and the pre- and post-extraction steps, and which were previously tested in our laboratory with the intention of preserving the biological structures and functions of the collagen isolates for biomedical purposes. Both the major and minor components such as the amino acids, lipids, water, glycosaminoglycan and ash contents and elemental content, as well as the structure and morphology of the raw sources and the resulting batches of isolates were subsequently examined in detail applying standardized analytical methods including high perfomance liquid chromatography, ultraviolet-visible and infrared spectrometry, polyacrylamide gel electrophoresis, energy dispersive spectroscopy and scanning electron microscopy. All the fish isolates provided severalfold higher yields (8-45 wt%) than did the pig isolates (3-9 wt%). In addition, the variability of the fish isolate yields (the coefficient of variation for processing A: 16.4-32.9 % and B: 6.8-17.4 %) was significantly lower (p ≤ 0.05, n = 5) than that of the pig isolates (A: 27.7-69.8 %; B: 35.3-87.9 %). In general, the fish skin batches had significantly higher protein contents (˃60 wt%) and lower lipid contents (<10 wt%) than the pig skin batches (<55 wt% protein and up to 66 wt% lipid). In addition, the fish skin batches did not differ significantly in terms of their composition applying the same processing method, whereas the pig skin batches exhibited considerable variations in terms of their compositions, particularly regarding the protein and lipid contents. It can be stated that, concerning the fish isolates, processing B was, in most cases, slightly more efficient and reproducible than processing A. However, concerning the pig isolates, although processing A appeared to be more efficient than processing B in terms of the yield, it resulted in the production of isolates that contained a certain level of contaminants. The study provides a comprehensive discussion on the suitability of the processing protocol in terms of producing batches of reproducible quality according to the specific type of biomaterial processed from different animal species., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

The physical properties and structure of collagen treated with high-pressure technologies have not yet been investigated in detail. The main goal of this work was to determine whether this modern gentle technology significantly changes the properties of collagen. High pressure in the range of 0-400 MPa was used, and the rheological, mechanical, thermal, and structural properties of collagen were measured. The rheological properties measured in the area of linear viscoelasticity do not statistically significantly change due to the influence of pressure or the duration of pressure exposure. In addition, the mechanical properties measured by compression between two plates are not statistically significantly influenced by pressure value or pressure hold time. The thermal properties T on and ∆H measured by differential calorimetry depend on pressure value and pressure hold time. Results from amino acids and FTIR analyses show that exposure of collagenous gels to high pressure (400 MPa), regardless of applied time (5 and 10 min), caused only minor changes in the primary and secondary structure and preserved collagenous polymeric integrity. SEM analysis did not show changes in collagen fibril ordering orientation over longer distances after applying 400 MPa of pressure for 10 min., Competing Interests: The authors declare no conflict of interest.

This study aims to point out the main drawback with respect to the design of simulated body environments. Three media commonly used for the simulation of the identical body environment were selected, i.e., Kokubo's simulated body fluid that simulates the inorganic component of human blood plasma, human blood plasma, and phosphate buffer saline. A comparison was performed of the effects of the media on collagen scaffolds. The mechanical and structural effects of the media were determined via the application of compression mechanical tests, the determination of mass loss, and image and micro-CT analyses. The adsorption of various components from the media was characterized employing energy-dispersive spectrometry. The phase composition of the materials before and after exposure was determined using X-ray diffraction. Infrared spectroscopy was employed for the interpretation of changes in the collagen secondary structure. Major differences in terms of the mechanical properties and mass loss were observed between the three media. Conversely, only minor structural changes were detected. Since no general recommendation exists for selecting the simulated body environment, it is necessary to avoid the simplification of the results and, ideally, to utilize alternative methods to describe the various aspects of degradation processes that occur in the media.

The aim of the study was to develop an orthopedic implant coating in the form of vancomycin-loaded collagen/hydroxyapatite layers (COLHA+V) that combine the ability to prevent bone infection with the ability to promote enhanced osseointegration. The ability to prevent bone infection was investigated employing a rat model that simulated the clinically relevant implant-related introduction of bacterial contamination to the bone during a surgical procedure using a clinical isolate of Staphylococcus epidermidis . The ability to enhance osseointegration was investigated employing a model of a minipig with terminated growth. Six weeks following implantation, the infected rat femurs treated with the implants without vancomycin (COLHA+ S. epidermidis ) exhibited the obvious destruction of cortical bone as evinced via a cortical bone porosity of up to 20% greater than that of the infected rat femurs treated with the implants containing vancomycin (COLHA+V+ S. epidermidis ) (3%) and the non-infected rat femurs (COLHA+V) (2%). The alteration of the bone structure of the infected COLHA+ S. epidermidis group was further demonstrated by a 3% decrease in the average Ca/P molar ratio of the bone mineral. Finally, the determination of the concentration of vancomycin released into the blood stream indicated a negligible systemic load. Six months following implantation in the pigs, the quantified ratio of new bone indicated an improvement in osseointegration, with a two-fold bone ingrowth on the COLHA (47%) and COLHA+V (52%) compared to the control implants without a COLHA layer (27%). Therefore, it can be concluded that COLHA+V layers are able to significantly prevent the destruction of bone structure related to bacterial infection with a minimal systemic load and, simultaneously, enhance the rate of osseointegration.

The aim of this study was to develop an osteo-inductive resorbable layer allowing the controlled elution of antibiotics to be used as a bone/implant bioactive interface particularly in the case of prosthetic joint infections, or as a preventative procedure with respect to primary joint replacement at a potentially infected site. An evaluation was performed of the vancomycin release kinetics, antimicrobial efficiency and cytocompatibility of collagen/hydroxyapatite layers containing vancomycin prepared employing different hydroxyapatite concentrations. Collagen layers with various levels of porosity and structure were prepared using three different methods: by means of the lyophilisation and electrospinning of dispersions with 0, 5 and 15wt% of hydroxyapatite and 10wt% of vancomycin, and by means of the electrospinning of dispersions with 0, 5 and 15wt% of hydroxyapatite followed by impregnation with 10wt% of vancomycin. The maximum concentration of the released active form of vancomycin characterised by means of HPLC was achieved via the vancomycin impregnation of the electrospun layers, whereas the lowest concentration was determined for those layers electrospun directly from a collagen solution containing vancomycin. Agar diffusion testing revealed that the electrospun impregnated layers exhibited the highest level of activity. It was determined that modification using hydroxyapatite exerts no strong effect on vancomycin evolution. All the tested samples exhibited sufficient cytocompatibility with no indication of cytotoxic effects using human osteoblastic cells in direct contact with the layers or in 24-hour infusions thereof. The results herein suggest that nano-structured collagen-hydroxyapatite layers impregnated with vancomycin following cross-linking provide suitable candidates for use as local drug delivery carriers., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Infections of the musculoskeletal system present a serious problem with regard to the field of orthopedic and trauma medicine. The aim of the experiment described in this study was to develop a resorbable nanostructured composite layer with the controlled elution of antibiotics. The layer is composed of collagen, hydroxyapatite nanoparticles, and vancomycin hydrochloride (10 wt%). The stability of the collagen was enhanced by means of cross-linking. Four cross-linking agents were studied, namely an ethanol solution, a phosphate buffer solution of N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride/N-hydroxysuccinimide, genipin, and nordihydroguaiaretic acid. High performance liquid chromatography was used so as to characterize the in vitro release rates of the vancomycin and its crystalline degradation antibiotically inactive products over a 21-day period. The maximum concentration of the released active form of vancomycin (approximately 265 mg/L) exceeded the minimum inhibitory concentration up to an order of 17 times without triggering the burst releasing effect. At the end of the experiment, the minimum inhibitory concentration was exceeded by up to 6 times (approximately 100 mg/L). It was determined that the modification of collagen with hydroxyapatite nanoparticles does not negatively influence the sustainable release of vancomycin. The balance of vancomycin and its degradation products was observed after 14 days of incubation., (Copyright © 2016. Published by Elsevier Inc.)

Nanocomposite scaffolds which aimed to imitate a bone extracellular matrix were prepared for bone surgery applications. The scaffolds consisted of polylactide electrospun nano/sub-micron fibres, a natural collagen matrix supplemented with sodium hyaluronate and natural calcium phosphate nano-particles (bioapatite). The mechanical properties of the scaffolds were improved by means of three different cross-linking agents: N-(3-dimethylamino propyl)-N'-ethylcarbodiimide hydrochloride and N-hydroxysuccinimide in an ethanol solution (EDC/NHS/EtOH), EDC/NHS in a phosphate buffer saline solution (EDC/NHS/PBS) and genipin. The effect of the various cross-linking conditions on the pore size, structure and mechanical properties of the scaffolds were subsequently studied. In addition, the mass loss, the swelling ratio and the pH of the scaffolds were determined following their immersion in a cell culture medium. Furthermore, the metabolic activity of human mesenchymal stem cells (hMSCs) cultivated in scaffold infusions for 2 and 7 days was assessed. Finally, studies were conducted of cell adhesion, proliferation and penetration into the scaffolds. With regard to the structural stability of the tested scaffolds, it was determined that EDC/NHS/PBS and genipin formed the most effectively cross-linked materials. Moreover, it was discovered that the genipin cross-linked scaffold also provided the best conditions for hMSC cultivation. In addition, the infusions from all the scaffolds were found to be non-cytotoxic. Thus, the genipin and EDC/NHS/PBS cross-linked scaffolds can be considered to be promising biomaterials for further in vivo testing and bone surgery applications.