Collagen nanofibrous materials have become integral to tissue engineering due to their exceptional properties and biocompatibility. Dehydrothermal crosslinking (DHT) enhances stability and maintains structural integrity without the formation of toxic residues. The study involved the crosslinking of electrospun collagen, applying DHT with access to air and under vacuum conditions. Various DHT exposure times of up to 72 h were applied to examine the time dependance of the DHT process. The DHT crosslinked collagen was subsequently chemically crosslinked using carbodiimides. The material crosslinked in this way evinced elevated Young's modulus values and ultimate tensile strength values, a lower swelling rate and lower shrinkage ratio during crosslinking, and a higher degree of resistance to degradation than the material crosslinked solely with DHT or carbodiimides. It was shown that the crosslinking mechanism using DHT occupies different binding sites than those using chemical crosslinking. Access to air for 12 h or less did not exert a significant impact on the material properties compared to DHT under vacuum conditions. However, concerning longer exposure times, it was determined that access to air results in the deterioration of the properties of the material and that reactions take place that occupy the free bonding sites, which subsequently reduces the effectiveness of chemical crosslinking using carbodiimides.

• Keywords
• Collagen, EDC/NHS, chemical crosslinking, crosslinking, degradation, dehydrothermal crosslinking, swelling, uniaxial tensile tests,
• Publication type
• Journal Article MeSH

Bone defects resulting from trauma, surgery, and congenital, infectious, or oncological diseases are a functional and aesthetic burden for patients. Bone regeneration is a demanding procedure, involving a spectrum of molecular processes and requiring the use of various scaffolds and substances, often yielding an unsatisfactory result. Recently, the new collagen sponge and its structural derivatives manufactured from European carp (Cyprinus carpio) were introduced and patented. Due to its fish origin, the novel scaffold poses no risk of allergic reactions or transfer of zoonoses and additionally shows superior biocompatibility, mechanical stability, adjustable degradation rate, and porosity. In this review, we focus on the basic principles of bone regeneration and describe the characteristics of an "ideal" bone scaffold focusing on guided bone regeneration. Moreover, we suggest several possible applications of this novel material in bone regeneration processes, thus opening new horizons for further research.

• Keywords
• GBR membrane, bioactive scaffold, bone regeneration, carp collagen, tissue engineering,
• Publication type
• Journal Article MeSH
• Review MeSH

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.

• Keywords
• XRD, blood plasma, collagen, mass loss, mechanical properties, micro-CT, porosity, scaffold, simulated body fluid, structural parameters,
• Publication type
• Journal Article MeSH

The study presents a novel vancomycin-releasing collagen wound dressing derived from Cyprinus carpio collagen type I cross-linked with carbodiimide which retarded the degradation rate and increased the stability of the sponge. Following lyophilization, the dressings were subjected to gamma sterilization. The structure was evaluated via scanning electron microscopy images, micro-computed tomography, and infrared spectrometry. The structural stability and vancomycin release properties were evaluated in phosphate buffered saline. Microbiological testing and a rat model of a wound infected with methicillin-resistant Staphylococcus aureus (MRSA) were then employed to test the efficacy of the treatment of the infected wound. Following an initial mass loss due to the release of vancomycin, the sponges remained stable. After 7 days of exposure in phosphate buffered saline (37°C), 60% of the material remained with a preserved collagen secondary structure together with a high degree of open porosity (over 80%). The analysis of the release of vancomycin revealed homogeneous distribution of the antibiotic both across and between the sponges. The release of vancomycin was retarded as proved by in vitro testing and further confirmed by the animal model from which measurable concentrations were observed in blood samples 24 hours after the subcutaneous implantation of the sponge, which was more than observed following intraperitoneal administration. The sponge was also highly effective in terms of reducing the number of colony-forming units in biopsies extracted from the infected wounds 4 days following the inoculation of the wounds with the MRSA solution. The presented sponges have ideal properties to serve as wound dressing for prevention of surgical site infection or treatment of already infected wounds.

• MeSH
• Anti-Bacterial Agents pharmacokinetics   MeSH
• Wound Healing drug effects   MeSH
• Carps MeSH
• Carbodiimides pharmacokinetics   MeSH
• Collagen pharmacokinetics   MeSH
• Rats MeSH
• Methicillin-Resistant Staphylococcus aureus drug effects   MeSH
• Bandages MeSH
• Vancomycin pharmacokinetics   MeSH
• Animals MeSH
• Check Tag
• Rats MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Names of Substances
• Anti-Bacterial Agents MeSH
• Carbodiimides MeSH
• Collagen MeSH
• Vancomycin MeSH

OBJECTIVES: Surgical wounds resulting from biofilm-producing microorganisms represent a major healthcare problem that requires new and innovative treatment methods. Rifampin is one of a small number of antibiotics that is able to penetrate such biofilms, and its local administration has the potential to serve as an ideal surgical site infection protection and/or treatment agent. This paper presents two types (homogeneous and sandwich structured) of rifampin-releasing carbodiimide-cross-linked fresh water fish collagen wound dressings. METHODS: The dressings were prepared by means of the double-lyophilization method and sterilized via gamma irradiation so as to allow for testing in a form that is able to serve for direct clinical use. The mechanical properties were studied via the uniaxial tensile testing method. The in vivo rifampin-release properties were tested by means of a series of incubations in phosphate-buffered saline. The microbiological activity was tested against methicillin-resistant staphylococcus aureus (MRSA) employing disc diffusion tests, and the in vivo pharmacokinetics was tested using a rat model. A histological examination was conducted for the study of the biocompatibility of the dressings. RESULTS: The sandwich-structured dressing demonstrated better mechanical properties due to its exhibiting ability to bear a higher load than the homogeneous sponges, a property that was further improved via the addition of rifampin. The sponges retarded the release of rifampin in vitro, which translated into at least 22 hours of rifampin release in the rat model. This was significantly longer than was achieved via the administration of a subcutaneous rifampin solution. Microbiological activity was proven by the results of the disc diffusion tests. Both sponges exhibited excellent biocompatibility as the cells penetrated into the scaffold, and virtually no signs of local irritation were observed. CONCLUSIONS: We present a novel rifampin-releasing sandwich-structured fresh water fish collagen wound dressing that has the potential to serve as an ideal surgical site infection protection and/or treatment agent.

• MeSH
• Anti-Bacterial Agents pharmacology   MeSH
• Biofilms drug effects   MeSH
• Wound Healing drug effects   MeSH
• Surgical Wound Infection drug therapy   MeSH
• Collagen pharmacology   MeSH
• Rats MeSH
• Methicillin-Resistant Staphylococcus aureus drug effects   MeSH
• Bandages MeSH
• Rats, Wistar MeSH
• Rifampin pharmacology   MeSH
• Fishes metabolism   MeSH
• Fresh Water MeSH
• Animals MeSH
• Check Tag
• Rats MeSH
• Male MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Names of Substances
• Anti-Bacterial Agents MeSH
• Collagen MeSH
• Rifampin MeSH

One of the main aims of bone tissue engineering, regenerative medicine and cell therapy is development of an optimal artificial environment (scaffold) that can trigger a favorable response within the host tissue, it is well colonized by resident cells of organism and ideally, it can be in vitro pre-colonized by cells of interest to intensify the process of tissue regeneration. The aim of this study was to develop an effective tool for regenerative medicine, which combines the optimal bone-like scaffold and colonization technique suitable for cell application. Accordingly, this study includes material (physical, chemical and structural) and in vitro biological evaluation of scaffolds prior to in vivo study. Thus, porosity, permeability or elasticity of two types of bone-like scaffolds differing in the ratio of collagen type I and natural calcium phosphate nanoparticles (bCaP) were determined, then analyzes of scaffold interaction with mesenchymal stem cells (MSCs) were performed. Simultaneously, dynamic seeding using a perfusion bioreactor followed by static cultivation was compared with standard static cultivation for the whole period of cultivation. In summary, cell colonization ability was estimated by determination of cell distribution within the scaffold (number, depth and homogeneity), matrix metalloproteinase activity and gene expression analysis of signaling molecules and differentiation markers. Results showed, the used dynamic colonization technique together with the newly-developed collagen-based scaffold with high content of bCaP to be an effective combined tool for producing bone grafts for bone implantology and regenerative medicine.

• Keywords
• Bone tissue engineering, Collagen scaffolds, Dynamic seeding, Mesenchymal stem cells, Static cultivation,
• MeSH
• Cell Differentiation MeSH
• Calcium Phosphates metabolism   MeSH
• Collagen chemistry   MeSH
• Bone and Bones chemistry   MeSH
• Cells, Cultured MeSH
• Mesenchymal Stem Cells metabolism   MeSH
• Nanoparticles MeSH
• Osteogenesis drug effects   MeSH
• Swine MeSH
• Regenerative Medicine MeSH
• Tissue Engineering methods   MeSH
• Tissue Scaffolds chemistry   MeSH
• Mesenchymal Stem Cell Transplantation methods   MeSH
• Animals MeSH
• Check Tag
• Female MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Names of Substances
• calcium phosphate MeSH Browser
• Calcium Phosphates MeSH
• Collagen MeSH

BACKGROUND: Collagen-based scaffolds provide a promising option for the treatment of bone defects. One of the key parameters of such scaffolds consists of porosity, including pore size. However, to date, no agreement has been found with respect to the methodology for pore size evaluation. Since the determination of the exact pore size value is not possible, the comparison of the various methods applied is complicated. Hence, this study focuses on the comparison of two widely-used methods for the characterization of porosity-scanning electron microscopy (SEM) and micro-computed tomography (micro-CT). METHODS: 7 types of collagen-based composite scaffold models were prepared by means of lyophilization and collagen cross-linking. Micro-CT analysis was performed in 3D and in 2D (pore size parameters were: major diameter, mean thickness, biggest inner circle diameter and area-equivalent circle diameter). Afterwards, pore sizes were analyzed in the same specimens by an image analysis of SEM microphotographs. The results were statistically evaluated. The comparison of the various approaches to the evaluation of pore size was based on coefficients of variance and the semi-quantitative assessment of selected qualities (e.g. the potential for direct 3D analysis, whole specimen analysis, non-destructivity). RESULTS: The pore size values differed significantly with respect to the parameters applied. Median values of pore size values were ranging from 20 to 490 µm. The SEM values were approximately 3 times higher than micro-CT 3D values for each specimen. The Mean thickness was the most advantageous micro-CT 2D approach. Coefficient of variance revealed no differences among pore size parameters (except major diameter). The semi-quantitative comparison approach presented pore size parameters in descending order with regard to the advantages thereof as follows: (1) micro-CT 3D, (2) mean thickness and SEM, (3) biggest inner circle diameter, major diameter and area equivalent circle diameter. CONCLUSION: The results indicated that micro-CT 3D evaluation provides the most beneficial overall approach. Micro-CT 2D analysis (mean thickness) is advantageous in terms of its time efficacy. SEM is still considered as gold standard for its widespread use and high resolution. However, exact comparison of pore size analysis in scaffold materials remains a challenge.

• Keywords
• Bone regeneration, Micro-CT, Pore size, Porosity, SEM, Scaffold,
• MeSH
• Collagen chemistry   MeSH
• Porosity MeSH
• X-Ray Microtomography MeSH
• Tissue Engineering * MeSH
• Tissue Scaffolds * MeSH
• Publication type
• Journal Article MeSH
• Names of Substances
• Collagen MeSH

Collagen composite scaffolds have been used for a number of studies in tissue engineering. The hydration of such highly porous and hydrophilic structures may influence mechanical behaviour and porosity due to swelling. The differences in physical properties following hydration would represent a significant limiting factor for the seeding, growth and differentiation of cells in vitro and the overall applicability of such hydrophilic materials in vivo. Scaffolds based on collagen matrix, poly(DL-lactide) nanofibers, calcium phosphate particles and sodium hyaluronate with 8 different material compositions were characterised in the dry and hydrated states using X-ray microcomputed tomography, compression tests, hydraulic permeability measurement, degradation tests and infrared spectrometry. Hydration, simulating the conditions of cell seeding and cultivation up to 48 h and 576 h, was found to exert a minor effect on the morphological parameters and permeability. Conversely, hydration had a major statistically significant effect on the mechanical behaviour of all the tested scaffolds. The elastic modulus and compressive strength of all the scaffolds decreased by ~95%. The quantitative results provided confirm the importance of analysing scaffolds in the hydrated rather than the dry state since the former more precisely simulates the real environment for which such materials are designed.

• MeSH
• Biocompatible Materials chemistry   MeSH
• Calcium Phosphates chemistry   MeSH
• Collagen chemistry   MeSH
• Hyaluronic Acid chemistry   MeSH
• Mechanical Phenomena MeSH
• Elastic Modulus MeSH
• Compressive Strength MeSH
• Polyesters chemistry   MeSH
• Porosity MeSH
• X-Ray Microtomography MeSH
• Materials Testing MeSH
• Tissue Engineering methods   MeSH
• Tissue Scaffolds chemistry   MeSH
• Water chemistry   MeSH
• Desiccation * MeSH
• Publication type
• Journal Article MeSH
• Names of Substances
• Biocompatible Materials MeSH
• calcium phosphate MeSH Browser
• Calcium Phosphates MeSH
• Collagen MeSH
• Hyaluronic Acid MeSH
• poly(lactide) MeSH Browser
• Polyesters MeSH
• Water MeSH