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Biomedical subjects

John A Jansen

Publications and source records attributed to John A Jansen.

At least 19 recordsLinked to original sources

Histological assessment of titanium and polypropylene fiber mesh implantation with and without fibrin tissue glue.

Polypropylene (PP) and titanium (Ti) meshes are well-known surgical implants that provoke a relative low foreign body reaction. Firm stabilization of the implant is important to prevent migration and subsequent failure of the operation. Fibrin tissue glues are commercially available adhesives and are widely accepted and applied in the medical field for hemorrhage, surgical bleeding, support of wound healing, wound and tissue gluing, sealing, and closure but also as antiadhesive agent in certain applications. The objective of this study was to evaluate the additional histological effect of fibrin glue application combined with two different types of meshes. Six pieces of mesh of each were subcutaneously implanted for 3, 6, and 12 weeks, with and without fibrin glue. After excision, processing, and staining, light microscopic analysis was performed on the sections, using subjective histological description and histomorphometry. Capsule quality, capsule thickness, interstitial quality, and total score were evaluated. To compare the samples with glue and without glue, analysis of variance (ANOVA) tests were carried out. No complications were observed. In general, the glue remnants remained visible at 3 and 6 weeks of implantation, accompanied by an inflammatory reaction and macrophage activity. At 12 weeks, all samples showed good tissue integration without evidence of glue. Evidently, the samples with glue demonstrated a prolonged inflammatory response and were surrounded by fibrous tissue capsules that were significantly thicker compared with the samples without glue (p < 0.05).

Analysis of Variance↗

Nanobiomaterial applications in orthopedics.

Advancements in nanobiotechnology are revolutionizing our capability to understand biological intricacies and resolve biological and medical problems by developing subtle biomimetic techniques. Nanocomposites and nanostructured materials are believed to play a pivotal role in orthopedic research since bone itself is a typical example of a nanocomposite. This article reviews current strategies using nanobiomaterials to improve current orthopedic materials and examines their applications in bone tissue engineering. Preliminary investigations support the potential of nanobiomaterials in orthopedic applications; however, significant advancements are necessary to achieve clinical use. Overall, current trends in nanobiotechnology foreshadow a bright future through the use of nanobiomaterials in the orthopedic domain.

Biocompatible Materials↗

Enrichment of osteogenic cell populations from rat bone marrow stroma.

The presence of multiple cell types in bone marrow and their varying proportions from isolation to isolation may count for the considerable variation in the outcome of different experiments. The presence of these multiple subpopulations suggests a need for a method that can purify the osteogenic component, i.e. osteoprogenitors, from other components. The availabilities of monoclonal antibodies recognizing subpopulations of osteoblasts are providing means for antibody-based methods. The cell surface antigens STRO-1, ALP and HOP-26 were used for cell sorting experiments with fluorescence activated cell sorting (FACS). These cell populations were analyzed on differential gene expression, cell proliferation and differentiation into the osteoblastic lineage. The oligo-microarray results showed that only the ALP positive cell population expressed genes of the extracellular matrix; like different collagens, ECM-1 and matrix protease MMP-14. The real-time polymerase chain reaction (QPCR) results showed that STRO-1 and ALP positive cells had an upregulation in expression of lipoprotein lipase, osteocalcin, and collagen type I. Integrin beta-3 was only upregulated for ALP positive cells, while for these cells downregulation occurred for the genes myosine, alkaline phosphatase and integrin beta-1. HOP-26 positive cells showed an upregulation in collagen type I compared to control group. The DNA analysis revealed that the cells of the control group and the HOP-26 positive cells showed a 5 times higher cell growth compared to the STRO-1 and ALP positive cells. The alkaline phosphatase activity showed no activity for the control group. The STRO-1 and ALP positive cells had a higher activity compared to the HOP-26 positive. The calcium measurements revealed only for the control group calcium at day 24. Based on the results of our study, we conclude that the FACS method had no negative effect on the proliferating as well as differentiating response of the cells. Further, we conclude that by using an antibody-based cell selection method, different cell populations with different mRNA expression profiles and different osteogenic characteristics can be obtained.

Animals↗

The performance of human dental pulp stem cells on different three-dimensional scaffold materials.

The aim of this study was to investigate the in vitro and in vivo behavior of human dental pulp stem cells (DPSCs) isolated from impacted third molars, when seeded onto different 3-dimensional (3-D) scaffold materials: i.e. a spongeous collagen, a porous ceramic, and a fibrous titanium mesh. Scaffolds were loaded with DPSC, and subsequently divided into two groups. The first group was cultured in osteogenic differentiation medium in vitro for 4 weeks. The second group of samples was implanted subcutaneously in nude mice for 6 or 12 weeks. Samples cultured in vitro were analyzed by scanning electron microscopy and RT-PCR for dentin sialophosphoprotein (DSPP) expression. In vivo samples were evaluated by histology, RT-PCR and immunohistochemistry. The results indicated that in vitro, cells developed abundant deposition of mineralized extracellular matrix (ECM) with expression of DSPP in all 3-D materials. The simultaneous implantation experiment showed formation of tissue that was DSPP positive in all three scaffolds materials. However, the aspect of the formed tissues in all scaffolds resembled more connective tissue than a dentin-like tissue. Limited calcification of the ECM was only seen in the ceramic scaffold. In both experiments, no other differences could be attributed to the different materials used. In conclusion, the in vivo behavior of DPSC and their relations with 3-D scaffold materials should be further studied before clinical use can be considered.

Animals↗

Mechanical evaluation of implanted calcium phosphate cement incorporated with PLGA microparticles.

In this study, the mechanical properties of an implanted calcium phosphate (CaP) cement incorporated with 20wt% poly (dl-lactic-co-glycolic acid) (PLGA) microparticles were investigated in a rat cranial defect. After 2, 4 and 8 weeks of implantation, implants were evaluated mechanically (push-out test) and morphologically (Scanning Electron Microscopy (SEM) and histology). The results of the push-out test showed that after 2 weeks the shear strength of the implants was 0.44+/-0.44MPa (average+/-sd), which increased to 1.34+/-1.05MPa at 4 weeks and finally resulted in 2.60+/-2.78MPa at 8 weeks. SEM examination showed a fracture plane at the bone-cement interface at 2 weeks, while the 4- and 8-week specimens created a fracture plane into the CaP/PLGA composites, indicating an increased strength of the bone-cement interface. Histological evaluation revealed that the two weeks implantation period resulted in minimal bone ingrowth, while at 4 weeks of implantation the peripheral PLGA microparticles were degraded and replaced by deposition of newly formed bone. Finally, after 8 weeks of implantation the degradation of the PLGA microparticles was almost completed, which was observed by the bone ingrowth throughout the CaP/PLGA composites. On basis of our results, we conclude that the shear strength of the bone-cement interface increased over time due to bone ingrowth into the CaP/PLGA composites. Although the bone-cement contact could be optimized with an injectable CaP cement to enhance bone ingrowth, still the mechanical properties of the composites after 8 weeks of implantation are insufficient for load-bearing purposes.

Absorbable Implants↗

Inhibition of cardiomyocyte automaticity by electrotonic application of inward rectifier current from Kir2.1 expressing cells.

A biological pacemaker might be created by generation of a cellular construct consisting of cardiac cells that display spontaneous membrane depolarization, and that are electrotonically coupled to surrounding myocardial cells by means of gap junctions. Depending on the frequency of the spontaneously beating cells, frequency regulation might be required. We hypothesized that application of Kir2.1 expressing non-cardiac cells, which provide I (K1) to spontaneously active neonatal cardiomyocytes (NCMs) by electrotonic coupling in such a cellular construct, would generate an opportunity for pacemaker frequency control. Non-cardiac Kir2.1 expressing cells were co-cultured with spontaneously active rat NCMs. Electrotonic coupling between the two cell types resulted in hyperpolarization of the cardiomyocyte membrane potential and silencing of spontaneous activity. Either blocking of gap-junctional communication by halothane or inhibition of I (K1) by BaCl(2) restored the original membrane potential and spontaneous activity of the NCMs. Our results demonstrate the power of electrotonic coupling for the application of specific ion currents into an engineered cellular construct such as a biological pacemaker.

Action Potentials↗

In vitro and in vivo reactivity of porous, electrosprayed calcium phosphate coatings.

The dissolution and/or precipitation behaviour of porous calcium phosphate (CaP) coatings, deposited using electrostatic spray deposition (ESD), was investigated (a) in vitro after soaking in simulated body fluid (SBF) for several time periods (2, 4, 8, and 12 weeks), and (b) in vivo after subcutaneous implantation of CaP-coated implants in the back of goats for identical time periods. Physical and chemical properties of coatings were characterized before and after in vitro/vivo testing by means of scanning electron microscopy, X-ray diffraction, Fourier-transform infrared spectroscopy, and energy dispersive spectroscopy. Moreover, part of the explants was prepared for light microscopical evaluation of the tissue response. In vitro, all apatitic ESD-coatings induced the formation of homogeneous and adherent CaP precipitation layers. Amorphous CaP, however, displayed a delayed precipitation of poorly adherent CaP layers, whereas heterogeneous calcification was observed on top of beta-TCP-coated substrates, indicating that beta-TCP and amorphous CaP coatings exhibit a poor ability of inducing calcification in SBF as compared to crystalline apatitic coatings. In vivo, no adverse tissue reactions (toxic effects/inflammatory cells) were observed using light microscopy, and all coatings became surrounded by a dense, fibrous tissue capsule after implantation. All ESD-coatings degraded gradually at a dissolution rate depending on the chemical phase (order of relative solubility: amorphous CaP approximately carbonate apatite>beta-TCP>carbonated hydroxyapatite), thereby enabling synthesis of CaP coatings with a tailored degradation rate.

Animals↗

In vitro generated extracellular matrix and fluid shear stress synergistically enhance 3D osteoblastic differentiation.

This study instituted a unique approach to bone tissue engineering by combining effects of mechanical stimulation in the form of fluid shear stresses and the presence of bone-like extracellular matrix (ECM) on osteodifferentiation. Rat marrow stromal cells (MSCs) harvested from bone marrow were cultured on titanium (Ti) fiber mesh discs for 12 days in a flow perfusion system to generate constructs containing bone-like ECM. To observe osteodifferentiation and bone-like matrix deposition, these decellularized constructs and plain Ti fiber meshes were seeded with MSCs (Ti/ECM and Ti, respectively) and cultured in the presence of fluid shear stresses either with or without the osteogenic culture supplement dexamethasone. The calcium content, alkaline phosphatase activity, and osteopontin secretion were monitored as indicators of MSC differentiation. Ti/ECM constructs demonstrated a 75-fold increase in calcium content compared with their Ti counterparts after 16 days of culture. After 16 days, the presence of dexamethasone enhanced the effects of fluid shear stress and the bone-like ECM by increasing mineralization 50-fold for Ti/ECM constructs; even in the absence of dexamethasone, the Ti/ECM constructs exhibited approximately a 40-fold increase in mineralization compared with their Ti counterparts. Additionally, denatured Ti/ECM* constructs demonstrated a 60-fold decrease in calcium content compared with Ti/ECM constructs after 4 days of culture. These results indicate that the inherent osteoinductive potential of bone-like ECM along with fluid shear stresses synergistically enhance the osteodifferentiation of MSCs with profound implications on bone-tissue-engineering applications.

Alkaline Phosphatase↗

Bone-like tissue formation using an equine COLLOSS E-filled titanium scaffolding material.

COLLOSS, a bovine extracellular matrix product containing native BMPs has already shown osteoinductive properties. To overcome problems with risk of transmissable spongiform encephalopathy (TSE) infection, an equine derived version was investigated in this study, named COLLOSS E. Disc- and tube-shaped implants were made from titanium fibre mesh. The central space of tubes was filled and the discs were impregnated with the COLLOSS E material to assess osteo-induction. These implants and non-loaded controls were implanted subcutaneously into the back of Wistar rats. After implantation periods of 2, 8, and 12 weeks, the implants were retrieved and sections were made. Histology showed a thin fibrous capsule surrounding the titanium mesh and a very mild tissue reaction. The disc implants, loaded or non-loaded, showed no bone formation at all. After 2 weeks of implantation 3 out of 5 of the loaded tubes showed bone formation with a mean of 0.3 mm2 areas of new formed bone, after 8 weeks 3 out of 6 and 0.7 mm2, and after 12 weeks this increased to 6 out of 6 and 1.0 mm2. In the non-loaded tubes only connective tissue in growth was seen. In conclusion, it was demonstrated that COLLOSS E material, loaded in a titanium fibre mesh tube shows osteoinductive properties. The effect of COLLOSS E has to be investigated further in orthotopic sites, which resemble more the final clinical application for bone reconstruction.

Animals↗

Experimental results of the tracheoesophageal tissue connector for improved fixation of shunt valves in laryngectomized patients.

BACKGROUND: After total laryngectomy and voice rehabilitation using a tracheoesophageal shunt valve, patients often have valve-related complications such as leakage. To solve these problems, a tracheoesophageal tissue connector (TE-TC) was devised to serve as an interface between the patient's tissue (trachea and esophagus) and the shunt valve. METHODS: The TE-TC is a permucosal connection constructed from a titanium ring (filled with a silicon rubber plug) combined with polypropylene or titanium mesh. After implantation in adult goats for 12 weeks the implants were submitted to histologic investigation. RESULTS: Firm implant fixation was achieved. In nearly all (18/19), no signs of infection of the implant were seen; 11 of 19 animals died before the end of the experiment owing to complications not related to the implant. CONCLUSIONS: The TE-TC is a new device with potential in the solution for fixation-related problems in tracheoesophageal voice rehabilitation.

Animals↗

Evaluation of bone regeneration by DNA release from composites of oligo(poly(ethylene glycol) fumarate) and cationized gelatin microspheres in a critical-sized calvarial defect.

This research examines the bone formation response to release of plasmid DNA encoding human Bone Morphogenetic Protein-2 from hydrogel composites consisting of cationized gelatin microspheres (CGMS) embedded within a crosslinked oligo(poly(ethylene glycol) fumarate) (OPF) hydrogel network in a critical-sized rat cranial defect model after 30 days. Four composite groups were investigated: (1) composites with 10 microg DNA loaded into the CGMS phase, (2) composites with 10 microg DNA loaded into the OPF phase, (3) composites with 100 microg DNA loaded into the OPF phase, and (4) composites without DNA (material control). Light microscopy revealed no enhancement in bone formation for groups releasing plasmid DNA, relative to the material control group. Limited formation of new bone was observed from the defect margins and within the defect for some samples. The hydrogels swelled appreciably and fragmentation of the implants was noted to varying degrees among samples within groups, with a presence of inflammatory cells related to the degree of fragmentation. The lack of enhancement in bone formation indicates that the release of plasmid DNA from the composites was not sufficient to elicit a bone regeneration response.

Animals↗

Modulation of cell differentiation in bone tissue engineering constructs cultured in a bioreactor.

In summary, many factors can influence the osteoblastic differentiation of marrow stromal cells when cultivated on three-dimensional tissue engineering scaffolds. In creating ideal bone tissue engineering constructs consisting of a combination of a scaffold, cells, and bioactive factors; a flow perfusion bioreactor is a much more suitable culture environment than static culture in well plates. The bioreactor eliminates mass transport limitations to the scaffold interior and provides mechanical stimulation to the seeded cells through fluid shear. Scaffold properties such as pore size impact cell differentiation, especially in flow perfusion culture. In addition, the bone-like extracellular matrix created by the in vitro culture of marrow stromal cells on porous scaffolds creates an osteoinductive environment for the differentiation of other marrow stromal cell populations. Therefore, bone tissue engineering constructs created by in vitro culture have excellent potential for bone regeneration applications in the clinic. However, more work is required to optimize this tissue engineering strategy. A biodegradable material with mechanical integrity similar to native bone and degradation properties similar to the rate of bone formation would be a more ideal scaffold material. It is also yet unclear what the optimum scaffold pore size and amount of in vitro generated extracellular matrix are to maximize bone formation. Finally, better characterization of the flow patterns within the flow perfusion bioreactor is needed to better understand the relationship between fluid shear and cell differentiation for creation of the ideal scaffold/culture combination.

Biocompatible Materials↗

Transforming growth factor-beta1 release from a porous electrostatic spray deposition-derived calcium phosphate coating.

This study evaluated the utilization of a porous coating, derived with electrostatic spray deposition (ESD), as a carrier material for transforming growth factor-beta1 (TGF-beta1). A porous beta-tricalcium phosphate coating was deposited with ESD, and 10 ng of (125) I-labeled TGF-beta1 was loaded on the substrates. A burst release during the first hour of incubation of >90% was observed, in either culture medium or phosphate-buffered saline (PBS). Ninety-nine percent of the growth factor was released after 10 days of incubation. All samples were able to inhibit epithelial cell growth, indicating that the growth factor had remained bioactive after release. Thereafter, osteoblast-like cells were seeded upon substrates with or without 10 ng of TGF-beta1. While proliferation of osteoblast-like cells was increased on TGF-beta1-loaded substrates, differentiation was inhibited or delayed. In conclusion, a porous ESD-derived calcium phosphate coating can be used as a carrier material for TGF-beta1, when a burst release is desired.

Animals↗

The effect of platelet-rich plasma on the bone healing around calcium phosphate-coated and non-coated oral implants in trabecular bone.

The effect of local application of autologous platelet-rich plasma (PRP) on bone healing in combination with the use of titanium implants with 2 different surface configurations was investigated. PRP fractions were obtained from venous blood sample of 6 goats and applied via gel preparation and subsequent installation in the implant site or via dipping of the implant in PRP liquid before insertion. Thirty-six implants (18 non-coated and 18 calcium phosphate (CaP) coated) were placed into the goat femoral condyles (trabecular bone). The animals were sacrificed at 6 weeks after implantation, and implants with surrounding tissue were processed for light microscopical evaluation. In addition to subjective description of the histological findings, histomorphometrical variables were also evaluated (the bone-implant contact and the bone mass adjacent to the implant). Significantly more interfacial bone-to-implant contact was observed for all 3 groups of CaP-coated implants and the titanium / liquid group (non-coated implant with PRP liquid) than for the other 2 non-coated titanium groups (with PRP gel or without PRP). The evaluation of the bone mass close to implant surface indicated that all the groups induced a significant increase of the bone mass except the PRP gel groups. On the basis of the observations, it was concluded that magnetron-sputtered CaP coatings can improve the integration of oral implants in trabecular bone. The additional use of PRP did not offer any significant effect on the bone response to the CaP-coated implants, whereas PRP in a liquid form showed a significant effect on bone apposition to roughened titanium implants during the early post-implantation healing phase.

Animals↗

Porous poly(DL-lactic-co-glycolic acid)/calcium phosphate cement composite for reconstruction of bone defects.

Calcium phosphate (Ca-P) cements are injectable, self-setting ceramic pastes generally known for their favorable bone response. Ingrowth of bone and subsequent degradation rates can be enhanced by the inclusion of macropores. Initial porosity can be induced by CO(2) foaming during setting of the cement, whereas secondary porosity can develop after hydrolysis of incorporated poly(DL-lactic- co-glycolic acid) (PLGA) microparticles. In this study, we focused on the biological response to porous PLGA/Ca-P cement composites. Pre-set composite discs of four formulations (4 wt% or 15 wt% PLGA microparticles and low or high CO(2) induced porosity) were implanted subcutaneously and in cranial defects in rats for 12 weeks. Histological analysis of the explanted composites revealed that bone and fibrous tissue ingrowth was facilitated by addition of PLGA microparticles (number average diameter of 66 +/- 25 microm). No adverse tissue reaction was observed in any of the composites. Significant increases in composite density due to bone ingrowth in cranial implants were found in all formulations. The results suggest that the PLGA pores are suitable for bone ingrowth and may be sufficient to enable complete tissue ingrowth without initial CO(2) induced porosity. Finally, bone-like mineralization in subcutaneous implants suggests that, under appropriate conditions and architecture, porous PLGA/Ca-P cement composites can exhibit osteoinductive properties. These PLGA/Ca-P composites are a promising scaffolding material for bone regeneration and bone tissue engineering.

Absorbable Implants↗

The effects of implant surface roughness and surgical technique on implant fixation in an in vitro model.

OBJECTIVES: The aim of the present study was to determine the relationship between implant surface parameters, surgical approach and initial implant fixation. MATERIAL AND METHODS: Sixty tapered, conical, screw-shaped implants with machined or etched surface topography were implanted into the explanted femoral condyle of goats. The implant sites were prepared either by a conventional technique, by undersized preparation, or by the osteotome technique. Peak insertion & removal torque, bone-to-implant contacts (BIC) and morphological bone appearance were assessed by scanning electron microscope (SEM) and micro-computer tomography. (micro-CT). RESULTS: Insertion and removal torque values were significantly higher for etched implants inserted with the undersized technique (115.2 +/- 31.1, 102.9 +/- 36.4 N cm) respectively. Also, the average BIC value was higher for the etched implants placed with the undersized technique (87.5 +/- 5.6), which was statistically significant compared with machined and etched implants inserted by conventional technique. CONCLUSION: In conclusion, this study shows that the surgical technique has a decisive effect on implant fixation (represented in this study by installation torque value/removal torque value and histomorphometric evaluation) in trabecular bone. Nevertheless, additional in vivo studies have to be done to prove the importance of surgical protocol for the final implant-bone response.

Animals↗

The bone regenerative effect of platelet-rich plasma in combination with an osteoconductive material in rat cranial defects.

The effect of platelet-rich plasma (PRP) on bone regeneration, in combination with an osteoconductive material, was evaluated in a rat model. Cranial defects, 6.2 mm in diameter, were filled with HA/beta-TCP particles, HA/beta-TCP particles combined with PRP and HA/beta-TCP particles combined with PRP gel, where some were left empty as a control. After 4 weeks of implantation histological, histomorphometrical and micro-computed tomography analyses revealed no difference in new bone formation among the groups. Further, no additional effect of PRP gel in comparison with PRP liquid was detected, except for the increased handling capacity of the graft. These findings suggest that PRP had no positive effect on bone formation in addition to an osteoconductive material after an implantation period of 4 weeks. Also, no negative effect was seen, and neither PRP nor HA/beta-TCP hampered bone ingrowth into the defects.

Animals↗