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1.
Electron J Biotechnol ; 49: 5-13, Jan. 2021. ilus, tab, graf
Article in English | LILACS | ID: biblio-1291618

ABSTRACT

BACKGROUND: Opsonization, is the molecular mechanism by which target molecules promote interactions with phagocyte cell surface receptors to remove unwanted cells by induced phagocytosis. We designed an in vitro system to demonstrate that this procedure could be driven to eliminate adipocytes, using peptides mimicking regions of the complement protein C3b to promote opsonization and enhance phagocytosis. Two cell lines were used: (1) THP-1 monocytes differentiated to macrophages, expressing the C3b opsonin receptor CR1 in charge of the removal of unwanted coated complexes; (2) 3T3-L1 fibroblasts differentiated to adipocytes, expressing AQP7, to evaluate the potential of peptides to stimulate opsonization. (3) A co-culture of the two cell lines to demonstrate that phagocytosis could be driven to cell withdrawal with high efficiency and specificity. RESULTS: An array of peptides were designed and chemically synthesized p3691 and p3931 joined bound to the CR1 receptor activating phagocytosis (p < 0.033) while p3727 joined the AQP7 protein (p < 0.001) suggesting that opsonization of adipocytes could occur. In the co-culture system p3980 and p3981 increased lipid uptake to 91.2% and 89.0%, respectively, as an indicator of potential adipocyte phagocytosis. CONCLUSIONS: This in vitro model could help understand the receptor­ligand interaction in the withdrawal of unwanted macromolecules in vivo. The adipocyte-phagocytosis discussed may help to control obesity, since peptides of C3b stimulated the CR1 receptor, promoting opsonisation and phagocytosis of lipidcontaining structures, and recognition of AQP7 in the differentiated adipocytes, favored the phagocytic activity of macrophages, robustly supported by the co-culture strategy.


Subject(s)
Phagocytosis , Complement System Proteins , Adipocytes , In Vitro Techniques , Opsonin Proteins , Coculture Techniques , Foam Cells , Macrophages , Microscopy, Fluorescence
2.
Mater Sci Eng C Mater Biol Appl ; 61: 712-9, 2016 Apr 01.
Article in English | MEDLINE | ID: mdl-26838901

ABSTRACT

We coated transcutaneous implants made of titanium alloy Ti6Al4V with copolymer dimethyl (2-methacryloyloxy-ethyl) phosphonate and 4-vinylpyridine and investigated the tissue reaction with respect to its biocompatible and antimicrobial properties in vivo. We distinguished between clinically observable superficial inflammations and histologically detectable deep infections. The vinylpyridine moieties were transferred into cationic pyridinium groups by reaction with hexyl bromide. Thus polymers with both antimicrobial capacity and good biocompatibility were obtained. In a short-term study, we implanted specially designed bare or coated implants in hairless but immunocompetent mice and analyzed the tissue reaction histologically. No difference was found between bare and coated implants in the initial healing phase of up to 14 days; however, after 21 days the scar tissue formation was higher in the bare implant group. The degree of epithelial downgrowth was comparable in both groups at any time point. In a long-term study of up to 168 days, we analyzed resistance to infection. In the bare implant group, 7 of the 12 implantation sites became infected deep whereas in the coated implant group only two deep infections were observed. The other implantation sites showed only superficial signs of inflammation. These results generally accord with previous in-vitro studies.


Subject(s)
Anti-Infective Agents/chemistry , Coated Materials, Biocompatible/chemistry , Polymers/chemistry , Titanium/chemistry , Alloys , Animals , Anti-Infective Agents/therapeutic use , Bacterial Infections/prevention & control , Cicatrix/metabolism , Cicatrix/pathology , Coated Materials, Biocompatible/pharmacology , Mice , Mice, Hairless , Prostheses and Implants , Re-Epithelialization/drug effects , Skin/pathology
3.
Spine (Phila Pa 1976) ; 39(20): E1220-7, 2014 Sep 15.
Article in English | MEDLINE | ID: mdl-25010099

ABSTRACT

STUDY DESIGN: An experimental study using a sheep cervical spine interbody fusion model. OBJECTIVE: First, to compare anterior cervical discectomy and fusion of an experimental bioabsorbable cage consisting of a magnesium alloy and a polymer (poly-ε-caprolactone, PCL) with an autologous tricortical iliac crest bone graft. Second, to determine the degradation kinetics of the cage, assess the 2 fusion devices for biomechanical stability, and determine their histological characteristics. SUMMARY OF BACKGROUND DATA: Bioabsorbable cages are not routinely used in spine surgery at present, due to some undesirable effects such as cracks and foreign body reactions. This study involved the manufacture of a bioabsorbable cage from a magnesium alloy and the polymer PCL, which was then used as a device for anterior cervical discectomy and fusion in a sheep cervical spine fusion model. METHODS: Twenty-four sheep had anterior cervical discectomy and fusion of C3-C4 and C5-C6 with an experimental bioabsorbable cage consisting of the magnesium alloy AZ31, which was infiltrated and covered with PCL at 1 level and with an autologous tricortical iliac crest bone graft at a second level. The sheep were divided into 4 groups. After 3, 6, 12, or 24 weeks postimplantation, the animals were killed and the cervical spines were harvested. The intervertebral spaces with the cage were investigated using µ-computed tomographic images to calculate degradation kinetics. Stiffness of all monosegments was determined through biomechanical testing. Histological analysis was performed to evaluate fusion status and to detect any foreign body reactions. The results from both implants were compared. RESULTS: The magnesium-PCL cage showed nonlinear degradation over time. Both implants demonstrated time-dependent increases in stability, with a significantly greater stiffness of the bone graft after 24 weeks in all directions of motion. Histologically, the cage showed no signs of fusion with progressive encapsulation over time. CONCLUSION: In comparison with the bone graft, the bioabsorbable cage showed inferior stiffness and fusion properties. Thus, further component modifications are necessary. LEVEL OF EVIDENCE: N/A.


Subject(s)
Absorbable Implants , Bone Transplantation , Cervical Vertebrae/surgery , Materials Testing , Spinal Fusion/instrumentation , Animals , Biomechanical Phenomena , Diskectomy/instrumentation , Diskectomy/methods , Magnesium , Polymers , Sheep , Spinal Fusion/methods
4.
J Biomater Appl ; 28(5): 667-75, 2014 Jan.
Article in English | MEDLINE | ID: mdl-23292720

ABSTRACT

Biodegradable magnesium-based implants are currently being developed for use in orthopedic applications. The aim of this study was to investigate the acute, subacute, and chronic local effects on bone tissue as well as the systemic reactions to a magnesium-based (MgYREZr-alloy) screw containing rare earth elements. The upper part of the screw was implanted into the marrow cavity of the left femora of 15 adult rabbits (New Zealand White), and animals were euthanized 1 week, 12 weeks, and 52 weeks postoperatively. Blood samples were analyzed at set times, and radiographic examinations were performed to evaluate gas formation. There were no significant increased changes in blood values compared to normal levels. Histological examination revealed moderate bone formation with direct implant contact without a fibrous capsule. Histopathological evaluation of lung, liver, intestine, kidneys, pancreas, and spleen tissue samples showed no abnormalities. In summary, our data indicate that these magnesium-based screws containing rare earth elements have good biocompatibility and osteoconductivity without acute, subacute, or chronic toxicity.


Subject(s)
Alloys , Biocompatible Materials , Bone Screws , Models, Animal , Animals , Female , Rabbits , Spectrometry, X-Ray Emission
5.
Acta Biomater ; 10(1): 34-9, 2014 Jan.
Article in English | MEDLINE | ID: mdl-24021229

ABSTRACT

An appropriate pH level is an important prerequisite for the physiologal functioning of cells and tissues. Changes in the extracellular pH often lead to specific cellular reactions and an altered metabolism of cells and tissues influences the extracellular pH range. Thus a method to monitor the extracellular pH is a valuable tool to track specific tissue reactions. In this article we describe a method for the determination of the pH range adjacent to degradable biomaterials using wireless in vivo imaging. Using hairless but immunocompetent mice the fluorophor 5-(6)-carboxy SNARF-1 and the in vivo fluorescence and multispectral acquisition and analysis system Maestro it is possible to track shifts in pH in small living animals over a longer period of time. This method is especially suitable for studies which focus on the interaction of degrading biomaterials with their adjacent tissues.


Subject(s)
Biochemistry/methods , Biocompatible Materials/chemistry , Animals , Benzopyrans/metabolism , Buffers , Fluorescence , Foreign-Body Reaction/pathology , Hydrogen-Ion Concentration , Imaging, Three-Dimensional , Limit of Detection , Mice , Naphthols/metabolism , Rhodamines/metabolism , Solutions , Staining and Labeling
6.
Springerplus ; 2: 418, 2013.
Article in English | MEDLINE | ID: mdl-24024102

ABSTRACT

BACKGROUND: Bioabsorbable implants are not widely used in spine surgery. This study investigated the clinical and radiological findings after anterior cervical discectomy and fusion (ACDF) in an ovine animal model with an experimental bioabsorbable cage consisting of magnesium and polymer (poly-ϵ-caprolactone, PCL) in comparison to a tricortical bone graft as the gold standard procedure. MATERIALS AND METHODS: 24 full-grown sheep had ACDF of C3/4 and C5/6 with an experimental bioabsorbable implant (magnesium and PCL) in one level and an autologous tricortical bone graft in the second level. The sheep were divided into 4 groups (6 sheep each). After 3, 6, 12, or 24 weeks postoperatively, the cervical spines were harvested and conventional x-rays of each operated segment were conducted. The progress of interbody fusion was classified according to a three-point scoring system. RESULTS: There were no operation related complications except for one intraoperative fracture of the anterior superior iliac spine and two cases of screw loosening and sinking, respectively. In particular, no vascular, neurologic, wound healing or infectious problems were observed. According to the time of follow-up, both interbody fusion devices showed similar behaviour with increasing intervertebral osseointegration and complete arthrodesis in 10 of 12 (83.3%) motion segments after 24 weeks. CONCLUSIONS: The bioabsorbable magnesium-PCL cage used in this experimental animal study showed clinically no signs of incompatibility such as infectious or wound healing problems. The radiographic results regarding the osseointegration are comparable between the cage and the bone graft group.

7.
Biomed Tech (Berl) ; 58(4): 359-67, 2013 Aug.
Article in English | MEDLINE | ID: mdl-23729531

ABSTRACT

Novel surface coatings of percutaneous implants need to be tested in biocompatibility studies. The use of animal models for testing usually involves numerous lethal biopsies for the analysis of the implant-tissue interface. In this study, optical coherence tomography (OCT) was used to monitor the reaction of the skin to a percutaneous implant in an animal model of hairless but immunocompetent mice. In vivo optical biopsies with OCT were taken at days 7 and 21 after implantation and post mortem on the day of noticeable inflammation. A Fourier-domain OCT was programmed for spoke pattern scanning schemes centered at the implant midpoint to reduce motion artifacts during in vivo imaging. Image segmentation allowed the automatic detection and morphometric analysis of the skin contour and the subcutaneous implant anchor. On the basis of the segmentation, the overall refractive index of the tissue within one OCT data set was estimated as a free parameter of a fitting algorithm, which corrects for the curved distortion of the planar implant base in the OCT images. OCT in combination with the spoke scanning scheme and image processing provided time-resolved three-dimensional optical biopsies around the implants to assess tissue morphology.


Subject(s)
Dermatitis/etiology , Dermatitis/pathology , Dermatologic Surgical Procedures/methods , Equipment Failure Analysis/methods , Prostheses and Implants/adverse effects , Skin/pathology , Tomography, Optical Coherence/methods , Animals , Mice , Mice, Nude
8.
Acta Biomater ; 9(10): 8509-17, 2013 Nov.
Article in English | MEDLINE | ID: mdl-23416472

ABSTRACT

Biodegradable magnesium-based alloys are very promising materials for temporary implants. However, the clinical use of magnesium-based alloys is often limited by rapid corrosion and by insufficient mechanical stability. Here we investigated RS66, a magnesium-based alloy with extraordinary physicochemical properties of high tensile strength combined with a high ductility and a homogeneous grain size of ~1 µm which was obtained by rapid solidification processing and reciprocal extrusion. Using a series of in vitro and in vivo experiments, we analyzed the biodegradation behavior and the biocompatibility of this alloy. In vitro, RS66 had no cytotoxic effects in physiological concentrations on the viability and the proliferation of primary human osteoblasts. In vivo, RS66 cylinders were implanted into femur condyles, under the skin and in the muscle of adult rabbits and were monitored for 1, 2, 3, 4 and 8 weeks. After explantation, the RS66 cylinders were first analyzed by microtomography to determine the remaining RS66 alloy and calculate the corrosion rates. Then, the implantation sites were examined histologically for healing processes and foreign body reactions. We found that RS66 was corroded fastest subcutaneously followed by intramuscular and bony implantation of the samples. No clinical harm with transient gas cavities during the first 6 weeks in subcutaneous and intramuscular implantation sites was observed. No gas cavities were formed around the implantation site in bone. The corrosion rates in the different anatomical locations correlated well with the local blood flow prior to implantation. A normal foreign body reaction occurred in all tissues. Interestingly, no enhanced bone formation could be observed around the corroding samples in the condyles. These data show that RS66 is biocompatible, and due to its interesting physicochemical properties, this magnesium alloy is a promising material for biodegradable implants.


Subject(s)
Absorbable Implants , Alloys/pharmacology , Biocompatible Materials/pharmacology , Magnesium/pharmacology , Materials Testing , Alloys/toxicity , Animals , Biocompatible Materials/toxicity , Bone and Bones/drug effects , Cell Death/drug effects , Cell Proliferation/drug effects , Cell Survival/drug effects , Cells, Cultured , Corrosion , Female , Humans , Implants, Experimental , Magnesium/toxicity , Neutrophils/cytology , Neutrophils/drug effects , Organ Size/drug effects , Osteoblasts/cytology , Osteoblasts/drug effects , Osteogenesis/drug effects , Prosthesis Implantation , Rabbits , Regional Blood Flow/drug effects , Staining and Labeling , Tomography
9.
Acta Biomater ; 9(10): 8714-21, 2013 Nov.
Article in English | MEDLINE | ID: mdl-23069319

ABSTRACT

Magnesium materials are of increasing interest in the development of biodegradable implants as they exhibit properties that make them promising candidates. However, the formation of gas cavities after implantation of magnesium alloys has been widely reported in the literature. The composition of the gas and the concentration of its components in these cavities are not known as only a few studies using non-specific techniques were done about 60 years ago. Currently many researchers assume that these cavities contain primarily hydrogen because it is a product of magnesium corrosion in aqueous media. In order to clearly answer this question we implanted rare earth-containing magnesium alloy disks in mice and determined the concentration of hydrogen gas for up to 10 days using an amperometric hydrogen sensor and mass spectrometric measurements. We were able to directly monitor the hydrogen concentration over a period of 10 days and show that the gas cavities contained only a low concentration of hydrogen gas, even shortly after formation of the cavities. This means that hydrogen must be exchanged very quickly after implantation. To confirm these results hydrogen gas was directly injected subcutaneously. Most of the hydrogen gas was found to exchange within 1h after injection. Overall, our results disprove the common misbelief that these cavities mainly contain hydrogen and show how quickly this gas is exchanged with the surrounding tissue.


Subject(s)
Hydrogen/analysis , Magnesium/pharmacology , Prostheses and Implants , Alloys , Animals , Corrosion , Electrochemical Techniques , Female , Immunocompetence , Mass Spectrometry , Mice , Mice, Hairless , Photoelectron Spectroscopy , Skin/cytology , Skin/drug effects
10.
Biomaterials ; 33(29): 6926-32, 2012 Oct.
Article in English | MEDLINE | ID: mdl-22818983

ABSTRACT

Quantification of apoptotic tissues during inflammatory processes induced by biomaterials is challenging in vivo. Here we present a non-invasive method using a fluorescence imaging system which facilitates intermittent snap shots of the current state of local apoptotic tissue. For this purpose, apoptotic cells around two different subcutaneously implanted materials (titanium discs and copper-coated titanium discs) in hairless but immunocompetent mice were quantified after 4, 8 and 23 days of implantation. For validation, the results of fluorescence signals were compared to the histology of the inflammatory tissue using apoptotic-specific TUNEL-, macrophage-specific F4/80-, neutrophile-specific NIMP-R14- and chloroacetate esterase-staining. We could demonstrate that the fluorescence signals were well suited to quantify the extent of apoptosis in vivo and this is a good indication for the biocompatibility of biomaterials. This study shows that non-invasive monitoring of tissue processes following the implantation of biomaterials is possible in vivo and may help to reduce the number of animals in studies addressing biocompatibility.


Subject(s)
Apoptosis , Biocompatible Materials/chemistry , Coated Materials, Biocompatible/chemistry , Animals , Carboxylic Ester Hydrolases/pharmacology , Copper/chemistry , Female , Foreign-Body Reaction , Immunohistochemistry/methods , In Situ Nick-End Labeling , Inflammation , Macrophages/metabolism , Mice , Microscopy, Fluorescence/methods , Neutrophils/metabolism , Time Factors , Titanium/chemistry
11.
Adv Biochem Eng Biotechnol ; 126: 285-96, 2012.
Article in English | MEDLINE | ID: mdl-21989489

ABSTRACT

Biohybrid has been used as a fashionable term in scientific publications during the past years to describe a functional unit consisting of a bioactive and a structural component. The bioactive part of the biohybrid could consist of cells or bioactive molecules, while the structural part is of biological or non-biological origin. Biohybrids are currently used as implants and transplants in regenerative medicine or in vitro applications such as assays, biosensors or bioreactors. However, a clear definition of a biohybrid has not been given yet. This chapter reviews the current applications of biohybrids and identifies the challenges of biohybrids in in vivo applications. A classification of biohybrids according to their functional use and application is provided.


Subject(s)
Biocompatible Materials/chemical synthesis , Biological Products/chemistry , Prostheses and Implants , Tissue Engineering/methods , Prosthesis Design , Surface Properties
12.
Med Image Comput Comput Assist Interv ; 14(Pt 3): 454-62, 2011.
Article in English | MEDLINE | ID: mdl-22003731

ABSTRACT

Optical Coherence Tomography (OCT) is a noninvasive imaging technique which is used here for in vivo biocompatibility studies of percutaneous implants. A prerequisite for a morphometric analysis of the OCT images is the correction of optical distortions caused by the index of refraction in the tissue. We propose a fully automatic approach for 3D segmentation of percutaneous implants using Markov random fields. Refraction correction is done by using the subcutaneous implant base as a prior for model based estimation of the refractive index using a generalized Hough transform. Experiments show the competitiveness of our algorithm towards manual segmentations done by experts.


Subject(s)
Image Interpretation, Computer-Assisted/methods , Imaging, Three-Dimensional/methods , Tomography, Optical Coherence/methods , Algorithms , Animals , Biocompatible Materials/chemistry , Diagnostic Imaging/methods , Humans , Markov Chains , Mice , Prostheses and Implants , Software
13.
J Appl Biomater Biomech ; 9(1): 26-33, 2011.
Article in English | MEDLINE | ID: mdl-21607935

ABSTRACT

PURPOSE: In modern orthopedics aseptic loosening caused by the formation of micro-wear particles remains a problem for endoprosthetic joint replacements as revision surgery is necessary with corresponding costs and exertions by patients. This study is devoted to the question of how the osseous ingrowth of implants can be supported. It was investigated whether the developed copolymer, p-VBP-co-GMA, coated on the surface of the implants, supports bone healing. In addition, it was analyzed whether covalent linkage of Bone Morphogenetic Protein 2 (BMP-2) to the copolymer layer enhances bone formation. METHODS: Eight adult New Zealand White Rabbits were implanted with four different foils (control, copolymer, copolymer + BMP-2, control + BMP-2) each. The histomorphometric analysis of all samples was made 28 days after implantation. RESULTS: The copolymer had a positive effect on bone remodeling compared to the control group. We observed that the copolymer group had a significantly increased bone volume per tissue volume ratio and bone density to the control group. In contrast, this in-vivo study showed that the immobilization of BMP-2 onto the copolymer layer did not enhance bone healing. The bone volume per tissue volume ratio was decreased as well as the bone density compared to control + BMP-2 group. CONCLUSION: The analysis showed that the bone remodeling process in the copolymer + BMP-2-group is in an early phase comparable to the control group. These results suggest that the coating with the developed copolymer has major potential for medical use as it enhances bone mass around the implant.


Subject(s)
Bone Regeneration/drug effects , Coated Materials, Biocompatible/pharmacology , Femur/physiology , Femur/surgery , Polymers/pharmacology , Prostheses and Implants , Titanium/chemistry , Animals , Bone Morphogenetic Protein 2/therapeutic use , Femur/drug effects , Rabbits , Treatment Outcome
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