Azizeh-Mitra Yousefi

Current strategies in multiphasic scaffold design for osteochondral tissue engineering: A review

The repair of osteochondral defects requires a tissue engineering approach that aims at mimicking the physiological properties and structure of two different tissues (cartilage and bone) using specifically designed scaffold-cell constructs. Biphasic and triphasic approaches utilize two or three different architectures, materials, or composites to produce a multilayered construct. This article gives an overview of some of the current strategies in multiphasic/gradient-based scaffold architectures and compositions for tissue engineering of osteochondral defects. In addition, the application of finite element analysis (FEA) in scaffold design and simulation of in vitro and in vivo cell growth outcomes has been briefly covered. FEA-based approaches can potentially be coupled with computer-assisted fabrication systems for controlled deposition and additive manufacturing of the simulated patterns. Finally, a summary of the existing challenges associated with the repair of osteochondral defects as well as some recommendations for future directions have been brought up in the concluding section of this article.

Effects of processing parameters in thermally induced phase separation technique on porous architecture of scaffolds for bone tissue engineering

Tissue engineering makes use of 3D scaffolds to sustain three-dimensional growth of cells and guide new tissue formation. To meet the multiple requirements for regeneration of biological tissues and organs, a wide range of scaffold fabrication techniques have been developed, aiming to produce porous constructs with the desired pore size range and pore morphology. Among different scaffold fabrication techniques, thermally induced phase separation (TIPS) method has been widely used in recent years because of its potential to produce highly porous scaffolds with interconnected pore morphology. The scaffold architecture can be closely controlled by adjusting the process parameters, including polymer type and concentration, solvent composition, quenching temperature and time, coarsening process, and incorporation of inorganic particles. The objective of this review is to provide information pertaining to the effect of these parameters on the architecture and properties of the scaffolds fabricated by the TIPS technique.

Prospect of Stem Cells in Bone Tissue Engineering: A Review

Mesenchymal stem cells (MSCs) have been the subject of many studies in recent years, ranging from basic science that looks into MSCs properties to studies that aim for developing bioengineered tissues and organs. Adult bone marrow-derived mesenchymal stem cells (BM-MSCs) have been the focus of most studies due to the inherent potential of these cells to differentiate into various cell types. Although, the discovery of induced pluripotent stem cells (iPSCs) represents a paradigm shift in our understanding of cellular differentiation. These cells are another attractive stem cell source because of their ability to be reprogramed, allowing the generation of multiple cell types from a single cell. This paper briefly covers various types of stem cell sources that have been used for tissue engineering applications, with a focus on bone regeneration. Then, an overview of some recent studies making use of MSC-seeded 3D scaffold systems for bone tissue engineering has been presented. The emphasis has been placed on the reported scaffold properties that tend to improve MSCs adhesion, proliferation, and osteogenic differentiation outcomes.

Three-dimensional porous scaffolds at the crossroads of tissue engineering and cell-based gene therapy

In the last 20 years, more than 1,500 gene therapy clinical trials have been approved worldwide targeting a variety of indications, from inherited monogenic diseases to acquired conditions such as cancer, cardiovascular and infectious diseases. However, concerns about the safety and efficacy of gene therapy pharmaceuticals justify the development of alternative strategies to ensure the clinical translation of this still promising field. In particular, ex vivo gene therapy strategies using autologous adult stem cells coupled to three‐dimensional (3D) porous scaffolds show great promises in preclinical studies. Developments in the fields of biomaterial sciences and tissue engineering have already helped understanding how we can harness to regenerative potential of many cell types to create artificial tissues and organs and vastly improve the engraftment of ex vivo manipulated adult stem cells. In this article, we will review the current state of the art in tissue engineering by exploring the various types of clinically available biomaterials and the methods used to process them into complex 3D scaffolds. We will then review how these technologies are applied in cell‐based gene therapy and identify novel avenues of research that may benefit patients in the near future. J. Cell. Biochem. 108: 537–546, 2009.

Design and Dynamic Culture of 3D-Scaffolds for Cartilage Tissue Engineering

Engineered scaffolds for tissue-engineering should be designed to match the stiffness and strength of healthy tissues while maintaining an interconnected pore network and a reasonable porosity. In this work, we have used 3D-plotting technique to produce poly-L-Lactide macroporous scaffolds with two different pore sizes. The ability of these macroporous scaffolds to support chondrocyte attachment and viability were compared under static and dynamic loading in vitro. Moreover, the 3D-plotting technique was combined with porogen-leaching, leading to macro/microporous scaffolds, so as to examine the effect of microporosity on the level of cell attachment and viability under similar loading condition. Canine chondrocytes’ cells were seeded onto the scaffolds with different topologies, and the constructs were cultured for up to 2 weeks under static conditions or in a bioreactor under dynamic compressive strain of 10% strain, at a frequency of 1 Hz. The attachment and cell growth of chondrocytes were examined by scanning electron microscopy and by 3-(4,5-dimethythiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay. A significant difference in cell attachment was observed in macroporous scaffolds with different pore sizes after 1, 7, and 14 days. Cell viability in the scaffolds was enhanced with decreasing pore size and increasing microporosity level throughout the culture period. Chondrocyte viability in the scaffolds cultured under dynamic loading was significantly higher (p<0.05) than the scaffolds cultured statically. Dynamic cell culture of the scaffolds improved cell viability and decreased the time of in vitro culture when compared to statically cultured constructs. Optimizing the culture conditions and scaffold properties could generate optimal tissue/constructs combination for cartilage repair.

Hierarchical scaffold design for mesenchymal stem cell-based gene therapy of hemophilia B.

Gene therapy for hemophilia B and other hereditary plasma protein deficiencies showed great promise in pre-clinical and early clinical trials. However, safety concerns about in vivo delivery of viral vectors and poor post-transplant survival of ex vivo modified cells remain key hurdles for clinical translation of gene therapy. We here describe a 3D scaffold system based on porous hydroxyapatite-PLGA composites coated with biomineralized collagen 1. When combined with autologous gene-engineered factor IX (hFIX) positive mesenchymal stem cells (MSCs) and implanted in hemophilic mice, these scaffolds supported long-term engraftment and systemic protein delivery by MSCs in vivo. Optimization of the scaffolds at the macro-, micro- and nanoscales provided efficient cell delivery capacity, MSC self-renewal and osteogenesis respectively, concurrent with sustained delivery of hFIX. In conclusion, the use of gene-enhanced MSC-seeded scaffolds may be of practical use for treatment of hemophilia B and other plasma protein deficiencies.

Solvent-free polymer/bioceramic scaffolds for bone tissue engineering: fabrication, analysis, and cell growth

This study examines the potential use of porous polycaprolactone (PCL) and polycaprolocatone/hydroxyapatite (PCL/HA) scaffolds fabricated through melt molding and porogen leaching for bone tissue engineering. While eliminating organic solvents is desirable, the process steps proposed in this study for uniformly dispersing HA particles (~5 μm in size) within the scaffold can also contribute to homogeneous properties for these porous composites. Poly(ethylene oxide) (PEO) was chosen as a porogen due to its similar density and melting point as PCL. Pore size of the scaffold was controlled by limiting the size of PCL and PEO particles used in fabrication. The percent of HA in the fabricated scaffolds was quantified by thermogravimetric analysis (TGA). Mechanical testing was used to compare the modulus of the scaffolds to that of bone, and the pore size distribution was examined with microcomputed tomography (μCT). Scanning electron microscopy (SEM) was used to examine the effect on scaffold morphology caused by the addition of HA particles. Both μCT and SEM results showed that HA could be incorporated into PCL scaffolds without negatively affecting scaffold morphology or pore formation. Energy-dispersive X-ray spectroscopy (EDS) and elemental mapping demonstrated a uniform distribution of HA within PCL/HA scaffolds. Murine calvaria-derived MC3T3-E1 cells were used to determine whether cells could attach on scaffolds and grow for up to 21 days. SEM images revealed an increase in cell attachment with the incorporation of HA into the scaffolds. Similarly, DNA content analysis showed a higher cell adhesion to PCL/HA scaffolds.

Physical and biological characteristics of nanohydroxyapatite and bioactive glasses used for bone tissue engineering

Abstract Critical-sized bone defects have, in many cases, posed challenges to the current gold standard treatments. Bioactive glasses are reported to be able to stimulate more bone regeneration than other bioactive ceramics; however, the difficulty in producing porous scaffolds made of bioactive glasses has limited their extensive use in bone regeneration. On the other hand, calcium phosphate ceramics such as synthetic hydroxyapatite and tricalcium phosphate are widely used in the clinic, but they stimulate less bone regeneration. This paper gives an overview of the recent developments in the field of bioactive nanoparticles, with a focus on nanohydroxyapatite and bioactive glasses for bone repair and regeneration. First, a brief overview of the chemical structure and common methods used to produce synthetic nanohydroxyapatite and bioactive glasses has been presented. The main body of the paper covers the physical and biological properties of these biomaterials, as well as their composites with biodegradable polymers used in bone regeneration. A summary of existing challenges and some recommendations for future directions have been brought in the concluding section of this paper.

Improving the homogeneity of tissue-mimicking cryogel phantoms for medical imaging

PURPOSE Tissue-mimicking phantoms can help in uncovering potential weaknesses in medical imaging systems. This work presents a new approach to developing phantoms for magnetic resonance elastography (MRE). Elastography requires sufficiently large and well-characterized phantoms to accurately validate motion estimation methods and to provide accurate stiffness measurements. Physically crosslinked polyvinyl alcohol hydrogels, prepared by the freeze-thaw technique, have been extensively used as MRE phantoms. However, the large cryogels developed by this technique usually exhibit variations in properties due to the low thermal conductivity of the polymeric solution. This leads to variations in freezing-thawing rates across the gels. Therefore, designing homogeneous large cryogels with tissue-mimicking mechanical properties poses a challenge to medical imaging researchers. METHODS Unlike conventional freeze-thaw techniques that use either sudden freezing or ramp freezing, the authors have developed a modified freeze-thaw process featuring a combination of multiple ramps and isotherms within a single freeze-thaw cycle. Aiming to develop brain-mimicking phantoms, they have blended three different water-soluble polymers (polyvinyl pyrrolidone, agarose, and polyacrylic acid) with polyvinyl alcohol and produced cryogels with a wide range of mechanical properties and swelling characteristics. The effect of the modified process on mechanical properties, swelling, and melting enthalpy of the produced gels has been investigated in this study. RESULTS It was demonstrated that imposing additional isotherms at the vicinity of phase change temperatures could effectively reduce the variations in properties within a typical large phantom (diameter vs height: 100 mm × 100 mm). While the conventional freeze-thaw process resulted in ∼16% variation in the enthalpy of fusion across the produced gels, the modified process reduced this variation to below 8%. The homogeneity in mechanical properties was also improved by over 50% compared to the conventional process. Upon comparing the mechanical properties of the gels with those of brain white matter, the authors have shown that a blend of polyvinyl alcohol and polyvinyl pyrrolidone can provide brain-mimicking properties, while leading to stable gels. CONCLUSIONS The modified freeze-thaw process enabled to minimize the temperature gradient within the large cryogel phantoms during the freeze-thaw cycle. The results of this study can help to fill the gaps in the scientific literature with regard to developing homogeneous phantoms for medical imaging. This work also provides a solid foundation for future studies in this field and could facilitate formulating new hydrogels to replicate the viscoelastic properties of soft tissues.

Post irradiation degradation of polypropylene radiation durability of polypropylene stabilized with phenolic stabilizer (II)

Abstract Post irradiation degradation of 60 Co gamma irradiated polypropylene stabilized with a phenolic type stabilizer has been followed up to two years after being irradiated. The effectiveness of the stabilizer at various concentrations on the polypropylene samples irradiated up to 150 kGy has been studied and the buildup of carbonyl group in the irradiated samples measured. The obtained results indicated that the oxidative reactions are retarded by addition of stabilizer. The variation of bend strength of the irradiated samples with and without stabilizer was also followed. This property was found to be greatly improved for the stabilized samples irradiated at low doses. However at high doses the effectiveness of the stabilizer declined. In order to correlate these results with the change in the molecular weight of polymer, trends of the change in rheological behaviour such as melt viscosity (μ a ) and flow behavior parameters ( K, n ) of both stabilized and unstabilized samples have been investigated. Results showed that at low doses, addition of the stabilizer greatly reduces the extent of chain scission, and therefore, the mechanical properties of the irradiated polymer are retained.