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result(s) for
"Chlup, Hynek"
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Designing of PLA scaffolds for bone tissue replacement fabricated by ordinary commercial 3D printer
2017
Background
The primary objective of Tissue engineering is a regeneration or replacement of tissues or organs damaged by disease, injury, or congenital anomalies. At present, Tissue engineering repairs damaged tissues and organs with artificial supporting structures called scaffolds. These are used for attachment and subsequent growth of appropriate cells. During the cell growth gradual biodegradation of the scaffold occurs and the final product is a new tissue with the desired shape and properties.
In recent years, research workplaces are focused on developing scaffold by bio-fabrication techniques to achieve fast, precise and cheap automatic manufacturing of these structures. Most promising techniques seem to be Rapid prototyping due to its high level of precision and controlling. However, this technique is still to solve various issues before it is easily used for scaffold fabrication.
In this article we tested printing of clinically applicable scaffolds with use of commercially available devices and materials. Research presented in this article is in general focused on “scaffolding” on a field of bone tissue replacement.
Results
Commercially available 3D printer and Polylactic acid were used to create originally designed and possibly suitable scaffold structures for bone tissue engineering. We tested printing of scaffolds with different geometrical structures. Based on the osteosarcoma cells proliferation experiment and mechanical testing of designed scaffold samples, it will be stated that it is likely not necessary to keep the recommended porosity of the scaffold for bone tissue replacement at about 90%, and it will also be clarified why this fact eliminates mechanical properties issue. Moreover, it is demonstrated that the size of an individual pore could be double the size of the recommended range between 0.2–0.35 mm without affecting the cell proliferation.
Conclusion
Rapid prototyping technique based on Fused deposition modelling was used for the fabrication of designed scaffold structures. All the experiments were performed in order to show how to possibly solve certain limitations and issues that are currently reported by research workplaces on the field of scaffold bio-fabrication. These results should provide new valuable knowledge for further research.
Journal Article
Properties of Bovine Collagen as Influenced by High-Pressure Processing
2023
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 Ton 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.
Journal Article
In Vivo Evaluation of Short-Term Performance of New Three-Layer Collagen-Based Vascular Graft Designed for Low-Flow Peripheral Vascular Reconstructions
2018
Aim. The aim of this study was to evaluate short-term patency of the new prosthetic graft and its structural changes after explantation. Methods. The study team developed a three-layer conduit composed of a scaffold made from polyester coated with collagen from the inner and outer side with an internal diameter of 6 mm. The conduit was implanted as a bilateral bypass to the carotid artery in 7 sheep and stenosis was created in selected animals. After a period of 161 days, the explants were evaluated as gross and microscopic specimens. Results. The initial flow rate (median ± IQR) in grafts with and without artificial stenosis was 120±79 ml/min and 255±255 ml/min, respectively. Graft occlusion occurred after 99 days in one of 13 conduits (patency rate: 92%). Wall-adherent thrombi occurred only in sharp curvatures in two grafts. Microscopic evaluation showed good engraftment and preserved structure in seven conduits; inflammatory changes with foci of bleeding, necrosis, and disintegration in four conduits; and narrowing of the graft due to thickening of the wall with multifocal separation of the outer layer in two conduits. Conclusions. This study demonstrates good short-term patency rates of a newly designed three-layer vascular graft even in low-flow conditions in a sheep model.
Journal Article
Online elasticity estimation and material sorting using standard robot grippers
by
Chlup, Hynek
,
Stoudek, Pavel
,
Patni, Shubhan P.
in
CAE) and Design
,
Computer-Aided Engineering (CAD
,
Engineering
2024
Stiffness or elasticity estimation of everyday objects using robot grippers is highly desired for object recognition or classification in application areas like food handling and single-stream object sorting. However, standard robot grippers are not designed for material recognition. We experimentally evaluated the accuracy with which material properties can be estimated through object compression by two standard parallel jaw grippers and a force/torque sensor mounted at the robot wrist, with a professional biaxial compression device used as reference. Gripper effort versus position curves were obtained and transformed into stress/strain curves. The modulus of elasticity was estimated at different strain points and the effect of multiple compression cycles (precycling), compression speed, and the gripper surface area on estimation was studied. Viscoelasticity was estimated using the energy absorbed in a compression/decompression cycle, the Kelvin-Voigt, and Hunt-Crossley models. We found that (1) slower compression speeds improved elasticity estimation, while precycling or surface area did not; (2) the robot grippers, even after calibration, were found to have a limited capability of delivering accurate estimates of absolute values of Young’s modulus and viscoelasticity; (3) relative ordering of material characteristics was largely consistent across different grippers; (4) despite the nonlinear characteristics of deformable objects, fitting linear stress/strain approximations led to more stable results than local estimates of Young’s modulus; and (5) the Hunt-Crossley model worked best to estimate viscoelasticity, from a single object compression. A two-dimensional space formed by elasticity and viscoelasticity estimates obtained from a single grasp is advantageous for the discrimination of the object material properties. We demonstrated the applicability of our findings in a mock single-stream recycling scenario, where plastic, paper, and metal objects were correctly separated from a single grasp, even when compressed at different locations on the object. The data and code are publicly available.
Journal Article
Effects of Extrusion and Irradiation on the Mechanical Properties of a Water–Collagen Solution
2022
This article describes 1D extension tests on bovine collagen samples (8% collagen in water). At such a high collagen concentration, the mechanical properties of semi-solid samples can be approximated by hyperelastic models (two-parametric HGO and Misof models were used), or simply by Hooke’s law and the modulus of elasticity E. The experiments confirm a significant increase in the E-modulus of the samples irradiated with high-energy electrons. The modulus E ~ 9 kPa of non-irradiated samples increases monotonically up to E ~ 250 kPa for samples absorbing an e-beam dose of ~3300 Gy. This amplification is attributed to the formation of cross-links by irradiation. However, E-modulus can be increased not only by irradiation but also by exposure to a high strain rate. For example, soft isotropic collagen extruded through a 200 mm long capillary increases the modulus of elasticity from 9 kPa to 30 kPa, and the increase is almost isotropic. This stiffening occurs when the corrugated collagen fibers are straightened and are aligned in the flow direction. It seems that the permanent structural changes caused by extrusion mitigate the effects of the ex post applied irradiation. Irradiation of extruded samples by 3300 Gy increases the modulus of E-elasticity only three times (from 30 kPa to approximately 90 kPa). Extruded and ex post irradiated samples show slight anisotropy (the stiffness in the longitudinal direction is on an average greater than the transverse stiffness).
Journal Article
Evaluation of the Immunogenicity of a Vascular Graft Covered with Collagen Derived from the European Carp (Cyprinus carpio) and Bovine Collagen
2019
Aim. To assess the systemic and local immunological response to subcutaneous implants of a vascular graft covered with collagen extracted from the European carp (freshwater fish) or with collagen of bovine origin. Methods. Pieces of a vascular graft covered by pure bovine (Bos taurus, BOV, n=14) or carp (Cyprinus carpio, CYP, n=14) collagen 5 mm in size were implanted subcutaneously in the dorsum of a Balb/cOla mice. A sham operation group of 12 animals served as the control. At 7 and 14 days after the operation, one-half of each group was terminated and blood for serum, spleen, and implant with surrounding tissue were collected. Mean cytokine (TNF-α, IL-10, IL-4, IL-1β, IL-13, and IFN-γ) levels in serum were determined using ELISA. Spleen cell cultures were used for in vitro testing of lymphocyte proliferation and cytokine secretion. Local expressions of IL-6, IL-10, TNF-α, TGF-β, CCL-2, and CCL-3 were determined using PCR. Results. We found no significant difference among control, BOV, and CYP groups in mean cytokine serum levels at seven days. At day 14, the BOV group had higher levels of TNF-α (P=.018) and both the BOV and CYP groups had lower levels of IL-4 (P=.011 and P=.047, respectively) compared with the control group. Both tested implants showed only a minimal effect on the production of selected cytokines. Cell proliferation in the CYP group stimulated by CYP gel at 14 days was significantly lower than by BOV gel in BOV group (P=.0031) or by CYP gel in the control group (P=.041). The difference between the groups in the local RNA expression of all the tested mediators both at 7 and at 14 days was not significant apart from a lower level of TNF-α in the BOV group compared to CYP at 14 days (P=.013). Conclusions. Implants covered with carp collagen induce an immunological response that is comparable to that of bovine collagen covered implants in a mouse model.
Journal Article
Composite Polylactide/Polycaprolactone Foams with Hierarchical Porous Structure for Pre-Vascularized Tissue Engineering
by
Bačáková, Lucie
,
Chlup, Hynek
,
Svobodová, Lucie
in
Adipose Tissue - cytology
,
Biocompatible Materials - chemistry
,
Biodegradation
2025
Modern tissue engineering requires not only degradable materials promoting cell growth and differentiation, but also vascularization of the engineered tissue. Porous polylactide/polycaprolactone (PLA/PCL, ratio 3/5) foam scaffolds were prepared by a combined porogen leaching and freeze-drying technique using NaCl (crystal size 250–500 µm) and a water-soluble cellulose derivative (KlucelTM E; 10–100% w/w relative to the total PLA/PCL concentration) as porogens. Scanning electron microscopy, micro-CT, and Brunauer–Emmett–Teller analysis showed that all scaffolds contained a trimodal range of pore sizes, i.e., macropores (average diameter 298–539 μm), micropores (100 nm to 10 μm), and nanopores (mostly around 3.0 nm). All scaffolds had an open porosity of about 90%, and the pores were interconnected. The size of the macropores and the nanoporosity were higher in the scaffolds prepared with Klucel. Nanoporosity increased water uptake by the scaffolds, while macroporosity promoted cell ingrowth, which was most evident in scaffolds prepared with 25% Klucel. Human adipose-derived stem cells co-cultured with endothelial cells formed pre-vascular structures in the scaffolds, which was further enhanced in a dynamic cell culture system. The scaffolds are promising for the engineering of pre-vascularized soft tissues (relatively pliable 10% Klucel scaffolds) and hard tissues (mechanically stronger 25% and 50% Klucel scaffolds).
Journal Article
Rifampin-Releasing Triple-Layer Cross-Linked Fresh Water Fish Collagen Sponges as Wound Dressings
by
Chlup, Hynek
,
Hartinger, Jan Miroslav
,
Popková, Michaela
in
Animals
,
Anti-Bacterial Agents - pharmacology
,
Antibiotics
2020
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.
Journal Article
Age estimation based on a combined arteriosclerotic index
2012
This study introduces a new quantity, the combined arteriosclerotic index (CAI), which is defined as the ratio between the diameter and the longitudinal prestrain of an artery. The longitudinal prestrain has been adopted as the ratio between the in situ length and the excised length of the abdominal aorta, and is a measure of arterial elasticity. During ageing, arteriosclerosis is manifested by the loss of pretension and by enlargement of the diameter of the artery. CAI combines these two effects. A sample of 61 female and 194 male autopsy measurements of human abdominal aortas shows that CAI correlates significantly with chronological age (
R
= 0.916/0.921; female/male). The sample had the following parameters: age 53 ± 19/48 ± 16 years; diameter of the abdominal aorta 12.4 ± 2.2/13.4 ± 2.1 mm; and longitudinal prestrain 1.13 ± 0.10/1.15 ± 0.10 (mean ± sample standard deviation; female/male). The resulting CAI was 11.2 ± 2.7/11.9 ± 2.6 mm. The classical linear regression model was employed for age estimation by CAI. The model gave a residual standard deviation of 7.6/6.3 years and a 95% prediction interval range of ± 15.4/12.5 years (female/male). A two-sample
t
-test confirmed that there are significant differences between the female and male population during ageing, reflected by CAI, unlike longitudinal prestrain. It was concluded that CAI is a suitable predictor of age at time of death and is easily obtainable in the autopsy room.
Journal Article
Age-related distribution of longitudinal pre-strain in abdominal aorta with emphasis on forensic application
2012
It is a well-known fact that the length of an artery in situ and the length of an excised artery differs. Retraction of blood vessels is usually observed. This pre-tension plays crucial role in arterial biomechanics. It augments an artery wall load-bearing capacity. This paper presents the longitudinal pre-strain of the human aorta as an index of human age. The length of abdominal aortas was measured during autopsies before and after segment resection. The longitudinal pre-strain was calculated in 130 donors; 100 male and 30 female bodies. The pre-strain was defined as the ratio between in situ length and the length after the excision. The mean pre-strain was found to be 1.18
±
0.10 for male and 1.14
±
0.10 for female sample (mean
±
standard deviation). The age in the male group was 41.6
±
15.9 years; and 47.7
±
17.7 years in the female group. Statistical analysis revealed the correlation coefficient between age and pre-strain
r
=
−0.821 and
r
=
−0.839 in male and female group, respectively. The analysis also confirmed close correlation between aortic circumference and age; and between circumference and pre-strain. Linear and power law regression equations were employed and prediction intervals were computed. The power law estimates the age more accurately than linear one model. Nevertheless, especially for small values of the pre-strain (aged individuals) the linear model can be advantageous.
Journal Article