98 research outputs found
3D plotted PCL scaffolds for stem cell based bone tissue engineering
The ability to control the architecture and strength of a bone tissue
engineering scaffold is critical to achieve a harmony between the scaffold and the
host tissue. Rapid prototyping (RP) technique is applied to tissue engineering to
satisfy this need and to create a scaffold directly from the scanned and digitized
image of the defect site. Design and construction of complex structures with
different shapes and sizes, at micro and macro scale, with fully interconnected pore
structure and appropriate mechanical properties are possible by using RP techniques.
In this study, RP was used for the production of poly(e-caprolactone) (PCL) scaffolds.
Scaffolds with four different architectures were produced by using different configurations
of the fibers (basic, basic-offset, crossed and crossed-offset) within the
architecture of the scaffold. The structure of the prepared scaffolds were examined by
scanning electron microscopy (SEM), porosity and its distribution were analyzed by
micro-computed tomography (m-CT), stiffness and modulus values were determined
by dynamic mechanical analysis (DMA). It was observed that the scaffolds had very
ordered structures with mean porosities about 60%, and having storage modulus
values about 1!107 Pa. These structures were then seeded with rat bone marrow
origin mesenchymal stem cells (MSCs) in order to investigate the effect of scaffold
structure on the cell behavior; the proliferation and differentiation of the cells on
the scaffolds were studied. It was observed that cell proliferation was higher on offset
scaffolds (262000 vs 235000 for basic, 287000 vs 222000 for crossed structure) and
stainings for actin filaments of the cells reveal successful attachment and spreading
at the surfaces of the fibers. Alkaline phosphatase (ALP) activity results were higher
for the samples with lower cell proliferation, as expected. Highest MSC differentiation
was observed for crossed scaffolds indicating the influence of scaffold structure on
cellular activities
Effect of scaffold architecture and BMP-2/BMP-7 delivery on in vitro bone regeneration
The aim of this study was to develop 3-D tissue engineered constructs that mimic the in vivo conditions through a self-contained growth factor delivery system. A set of nanoparticles providing the release of BMP-2 initially followed by the release of BMP-7 were incorporated in poly(Δ-caprolactone) scaffolds with different 3-D architectures produced by 3-D plotting and wet spinning. The release patterns were: each growth factor alone, simultaneous, and sequential. The orientation of the fibers did not have a significant effect on the kinetics of release of the model protein BSA; but affected proliferation of bone marrow mesenchymal stem cells. Cell proliferation on random scaffolds was significantly higher compared to the oriented ones. Delivery of BMP-2 alone suppressed MSC proliferation and increased the ALP activity to a higher level than that with BMP-7 delivery. Proliferation rate was suppressed the most by the sequential delivery of the two growth factors from the random scaffold on which the ALP activity was the highest. Results indicated the distinct effect of scaffold architecture and the mode of growth factor delivery on the proliferation and osteogenic differentiation of MSCs, enabling us to design multifunctional scaffolds capable of controlling bone healing.This project was conducted within the scope of the EU FP6 NoE Project Expertissues (NMP3-CT-2004-500283). We acknowledge the support to PY through the same project in the form of an integrated PhD grant. We also would like to acknowledge the support from Scientific and Technical Research Council of Turkey (TUBITAK) through project METUNANOBIOMAT (TBAG 105T508)
Evaluating oxygen tensions related to bone marrow and matrix for msc differentiation in 2d and 3d biomimetic lamellar scaffolds
The physiological O microenvironment of mesenchymal stem cells (MSCs) and osteoblasts and the dimensionality of a substrate are known to be important in regulating cell phenotype and function. By providing the physiologically normoxic environments of bone marrow (5%) and matrix (12%), we assessed their potential to maintain stemness, induce osteogenic differentiation, and enhance the material properties in the micropatterned collagen/silk fibroin scaffolds that were produced in 2D or 3D. Expression of osterix (OSX) and vascular endothelial growth factor A (VEGFA) was significantly enhanced in the 3D scaffold in all oxygen environments. At 21% O , OSX and VEGFA expressions in the 3D scaffold were respectively 13,200 and 270 times higher than those of the 2D scaffold. Markers for assessing stemness were significantly more pronounced on tissue culture polystyrene and 2D scaffold incubated at 5% O . At 21% O , we measured significant increases in ultimate tensile strength (p < 0.0001) and Youngâs modulus (p = 0.003) of the 3D scaffold compared to the 2D scaffold, whilst 5% O hindered the positive effect of cell seeding on tensile strength. In conclusion, we demonstrated that the 3D culture of MSCs in collagen/silk fibroin scaffolds provided biomimetic cues for bone progenitor cells toward differentiation and enhanced the tensile mechanical properties. 2 2 2 2
Chemical and topographical modification of PHBV surface to promote osteoblast alignment and confinement
Proper cell attachment and distribution, and thus stronger association in vivo between a bone implant and native tissue will improve the success of the implant. In this study, the aim was to achieve promotion of attachment and uniform distribution of rat mesenchymal stem cell-derived osteoblasts by introducing chemical and topographical cues on poly(3-hydroxybutyrate-co-3- hydroxyvalerate) (PHBV) film surfaces. As the chemical cues, either alkaline phosphatase was covalently immobilized on the film surface to induce deposition of calcium phosphate minerals or fibrinogen was adsorbed to improve cell adhesion. Microgrooves and micropits were introduced on the film surface by negative replication of micropatterned Si wafers. Both chemical cues improved cell attachment and even distribution on the PHBV films, but Fb was more effective especially when combined with the micropatterns. Cell alignment (<10° deviation angle) parallel to chemically modified microgrooves (1, 3, or 8 Όm groove width) and on 10 Όm-thick Fb lines printed on the unpatterned films was achieved. The cells on unpatterned and 5 Όm-deep micropitted films were distributed and oriented randomly. Results of this study proved that microtopographies on PHBV can improve osseointegration when combined with chemical cues, and that microgrooves and cell adhesive protein lines on PHBV can guide selective osteoblast adhesion and alignment. © 2007 Wiley Periodicals, Inc
Cornea engineering on polyester carriers
In this study, biodegradable polyester based carriers were designed for tissue engineering of the epithelial and the stromal layers of the cornea, and the final construct was tested in vitro. In the construction of the epithelial layer, micropatterned films were prepared from blends of biodegradable and biocompatible polyesters of natural (PHBV) and synthetic (P(L/DL)LA) origin, and these films were seeded with D407 (retinal pigment epithelial) cells. To improve cell adhesion and growth, the films were coated with fibronectin. To serve as the stromal layer of the cornea, highly porous foams of P(L/DL)LA-PHBV blends were seeded with 3T3 fibroblasts. Cell numbers on the polyester carriers were significantly higher than those on the tissue culture polystyrene control. The cells and the carriers were characterized scanning electron micrographs showed that the foam was highly porous and the pores were interconnected. 3T3 Fibroblasts were distributed quite homogeneously at the seeding site, but probably because of the high thickness of the carrier (âŒ6 mm); they could not sufficiently populate the core (central parts of the foam) during the test duration. The D407 cells formed multilayers on the micropatterned polyester film. Immunohistochemical studies showed that the cells retained their phenotype during culturing; D407 cells formed tight junctions characteristic of epithelial cells, and 3T3 cells deposited collagen type I into the foams. On the basis of these results, we concluded that the micropatterned films and the foams made of P(L/DL)LA-PHBV blends have a serious potential as tissue engineering carriers for the reconstruction of the epithelial and stromal layers of the cornea. © 2006 Wiley Periodicals, Inc
Small Bowel Obstruction due to Mesodiverticular Band of Meckel's Diverticulum: A Case Report
Meckel's diverticulum is the most common congenital anomaly of the small intestine. Common complications related to a Meckel's diverticulum include haemorrhage, intestinal obstruction, and inflammation. Small bowel obstruction due to mesodiverticular band of Meckel's diverticulum is a rare complication. Herein, we report the diagnosis and management of a small bowel obstruction occurring due to mesodiverticular band of a Meckel's diverticulum
Contact guidance enhances the quality of a tissue engineered corneal stroma
Corneal stroma is a very complex structure, composed of 200 lamellae of oriented collagen fibers. This highly complex nature of cornea is known to be important for its transparency and mechanical integrity. Thus, an artificial cornea design has to take into account this complex structure. In this study, behavior of human corneal keratocytes on collagen films patterned with parallel channels was investigated. Keratocytes proliferated well on films and reached confluency after 7 days in the incubation medium. Nearly all of the cells responded to the patterns and were aligned in contrast to the cells on unpatterned surfaces. Collagen type I and keratan sulfate secreted by keratocytes on patterned films appeared to be aligned in the direction of the patterns. The films showed an intermediate degradation over the course of a month. On the whole, transparency of the films increased with degradation and decreased by the presence of the cells. The decrease was, however, low and transparency level was maintained on the patterned films while on the unpatterned films a sharp decrease in transparency was followed by an improvement. This was due to the more organized distribution of cells and the oriented secretion of extracellular matrix molecules on patterned collagen films. Thus, these results suggest that application of contact guidance in cornea tissue engineering may facilitate the remodeling process, hence decrease the rehabilitation period. © 2007 Wiley Periodicals, Inc
Biodegradable nanomats produced by electrospinning : expanding multifunctionality and potential for tissue engineering
With increasing interest in nanotechnology, development of nanofibers (n-fibers) by using the
technique of electrospinning is gaining new momentum. Among important potential applications of
n-fiber-based structures, scaffolds for tissue-engineering represent an advancing front. Nanoscaffolds
(n-scaffolds) are closer to natural extracellular matrix (ECM) and its nanoscale fibrous structure.
Although the technique of electrospinning is relatively old, various improvements have been
made in the last decades to explore the spinning of submicron fibers from biodegradable polymers
and to develop also multifunctional drug-releasing and bioactive scaffolds. Various factors can
affect the properties of resulting nanostructures that can be classified into three main categories,
namely: (1) Substrate related, (2) Apparatus related, and (3) Environment related factors. Developed
n-scaffolds were tested for their cytocompatibility using different cell models and were seeded
with cells for to develop tissue engineering constructs. Most importantly, studies have looked at the
potential of using n-scaffolds for the development of blood vessels. There is a large area ahead
for further applications and development of the field. For instance, multifunctional scaffolds that
can be used as controlled delivery system do have a potential and have yet to be investigated for
engineering of various tissues. So far, in vivo data on n-scaffolds are scarce, but in future reports
are expected to emerge. With the convergence of the fields of nanotechnology, drug release and
tissue engineering, new solutions could be found for the current limitations of tissue engineering
scaffolds, which may enhance their functionality upon in vivo implantation. In this paper electrospinning
process, factors affecting it, used polymers, developed n-scaffolds and their characterization
are reviewed with focus on application in tissue engineering
Cellular GABAergic Neuroactive Steroid (3α,5α)-3-Hydroxy-Pregnan-20-One (3α,5α-THP) Immunostaining Levels Are Increased in the Ventral Tegmental Area of Human Alcohol Use Disorder Patients: A Postmortem Study
The GABAergic neuroactive steroid (3α,5α)-3-hydroxy-pregnan-20-one (3α,5α-THP, allopregnanolone) enhances GABAergic activity and produces subjective effects similar to ethanol. The effect of chronic alcohol exposure on 3α,5α-THP concentrations has been studied in mouse, rat, and monkey limbic brain areas. Chronic ethanol exposure produced divergent brain region and cell specific changes in 3α,5α-THP concentrations in animal studies. However, 3α,5α-THP levels in similar human brain regions have never been examined in individuals diagnosed with alcohol use disorder (AUD). Therefore, we used immunohistochemistry to examine 3α,5α-THP levels in the ventral tegmental area (VTA), substantia nigra pars medialis (SNM), and amygdala of human postmortem brains of patients diagnosed with AUD compared to social drinkers. The effects of sex and liver disease on 3α,5α-THP concentrations were examined in the aforementioned brain regions
Biodegradable Nanomats Produced by Electrospinning: Expanding Multifunctionality and Potential for Tissue Engineering
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