Design Of Tissue Engineering Scaffolds Research Articles

Numerical simulations of bioextruded polymer scaffolds for tissue engineering applications

Scaffolds provide a temporary mechanical and vascular support for tissue regeneration while shaping the in-growth tissues. These scaffolds must be biocompatible, biodegradable, enclose appropriate porosity, pore structure and pore distribution, and have optimal structural and vascular performance, with both surface and structural compatibility. Surface compatibility means a chemical, biological and physical suitability to the host tissue. Structural compatibility corresponds to an optimal adaptation to the mechanical behaviour of the host tissue. Recent advances in the design of tissue engineering scaffolds are increasingly relying on computer-aided design modelling and numerical simulations. The design of optimized scaffolds based on fundamental knowledge of their macro microstructure is a relevant topic of research. This research work presents a comparison between experimental compressive data and numerical simulations of bioextruded polymer scaffolds with different pore sizes for the elastic and plastic domain. Constitutive behaviour models of cellular structures are used in numerical simulations to compare numerical data with the experimental compressive data. Vascular simulation is also used in the design process of the extrusion-based scaffolds in order to define an optimized scaffold design. © 2013 Society of Chemical Industry

Functionally heterogeneous porous scaffold design for tissue engineering

Porous scaffolds with interconnected and continuous pores have recently been considered as one of the most successful tissue engineering strategies. In the literature, it has been concluded that properly interconnected and continuous pores with their spatial distribution could contribute to perform diverse mechanical, biological and chemical functions of a scaffold. Thus, there has been a need for reproducible and fabricatable scaffold design with controllable and functional gradient porosity. Improvements in Additive Manufacturing (AM) processes for tissue engineering and their design methodologies have enabled the development of controlled and interconnected scaffold structures. However homogeneous scaffolds with uniform porosity do not capture the intricate spatial internal micro architecture of the replaced tissue and thus are not capable of capturing the design. In this work, a novel heterogeneous scaffold modeling is proposed for layered-based additive manufacturing processes. First, layers are generated along the optimum build direction considering the heterogeneous micro structure of tissue. Each layer is divided into functional regions based on the spatial homogeneity factor. An area weight based method is developed to generate the spatial porosity function that determines the deposition pattern for the desired gradient porosity. To design a multi-functional scaffold, an optimum deposition angle is determined at each layer by minimizing the heterogeneity along the deposition path. The proposed methodology is implemented and illustrative examples are also provided. The effective porosity is compared between the proposed design and the conventional uniform porous scaffold design. Sample designed structures have also been fabricated with a novel micro-nozzle biomaterial deposition system. The result has shown that the proposed methodology generates scaffolds with functionally gradient porosity.

The effects of pore size in bilayered poly(lactide-co-glycolide) scaffolds on restoring osteochondral defects in rabbits

Bilayered porous scaffolds have recently attracted interest because of their considerable promise for repairing osteochondral defects. However, determination of optimal pore size in bilayered porous scaffolds remains an important issue. This study investigated the in vivo effects of pore size in bilayered scaffolds using a rabbit model of osteochondral defects. We fabricated five types of integrated bilayered poly(lactide-co-glycolide) (PLGA) scaffolds with different pore sizes in the chondral and osseous layers (50-100 µm, 100-200 µm, 200-300 µm, and 300-450 µm). A subset of bilayered scaffolds seeded with or without allogenic bone marrow mesenchymal stem cells (BMSCs) was implanted in rabbit osteochondral defects. All of the cell/scaffold composite constructs supported the simultaneous regeneration of articular cartilage and subchondral bone, but the best results were observed in cell-seeded PLGA scaffolds with 100-200 µm pores in the chondral layer and 300-450 µm pores in the osseous layer. Our study supports the concept that the effects of pore size on osteochondral repair should be taken into consideration during scaffold design for tissue engineering.

Characterization of mechanical and biochemical properties of developing embryonic tendon

Tendons have uniquely high tensile strength, critical to their function to transfer force from muscle to bone. When injured, their innate healing response results in aberrant matrix organization and functional properties. Efforts to regenerate tendon are challenged by limited understanding of its normal development. Consequently, there are few known markers to assess tendon formation and parameters to design tissue engineering scaffolds. We profiled mechanical and biological properties of embryonic tendon and demonstrated functional properties of developing tendon are not wholly reflected by protein expression and tissue morphology. Using force volume-atomic force microscopy, we found that nano- and microscale tendon elastic moduli increase nonlinearly and become increasingly spatially heterogeneous during embryonic development. When we analyzed potential biochemical contributors to modulus, we found statistically significant but weak correlation between elastic modulus and collagen content, and no correlation with DNA or glycosaminoglycan content, indicating there are additional contributors to mechanical properties. To investigate collagen cross-linking as a potential contributor, we inhibited lysyl oxidase-mediated collagen cross-linking, which significantly reduced tendon elastic modulus without affecting collagen morphology or DNA, glycosaminoglycan, and collagen content. This suggests that lysyl oxidase-mediated cross-linking plays a significant role in the development of embryonic tendon functional properties and demonstrates that changes in cross-links alter mechanical properties without affecting matrix content and organization. Taken together, these data demonstrate the importance of functional markers to assess tendon development and provide a profile of tenogenic mechanical properties that may be implemented in tissue engineering scaffold design to mechanoregulate new tendon regeneration.

Decellularized musculofascial extracellular matrix for tissue engineering

Ideal scaffolds that represent native extracellular matrix (ECM) properties of musculofascial tissues have great importance in musculofascial tissue engineering. However, detailed characterization of musculofascial tissues' ECM (particularly, of fascia) from large animals is still lacking. In this study, we developed a decellularization protocol for processing pig composite musculofascial tissues. Decellularized muscle (D-muscle) and decellularized fascia (D-fascia), which are two important components of decellularized musculofascial extracellular matrix (DMM), were comprehensively characterized. D-muscle and D-fascia retained intact three-dimensional architecture, strong mechanical properties, and bioactivity of compositions such as collagen, laminin, glycosaminoglycan, and vascular endothelial growth factor. D-muscle and D-fascia provided a compatible niche for human adipose-derived stem cell integration and proliferation. Heterotopic and orthotopic implantation of D-muscle and D-fascia in a rodent model further proved their biocompatibility and myogenic properties during the remodeling process. The differing characteristics of D-muscle from D-fascia (e.g. D-muscle's strong pro-angiogenic and pro-myogenic properties vs. D-fascia's strong mechanical properties) indicate different clinical application opportunities of D-muscle vs. D-fascia scaffolds. DMM comprising muscle and fascia ECM as a whole unit can thus provide not only a clinically translatable platform for musculofascial tissue repair and regeneration but also a useful standard for scaffold design in musculofascial tissue engineering.

New paradigms in hierarchical porous scaffold design for tissue engineering

This paper presented an effective method for the three-dimensional (3D) hierarchical porous scaffold design for tissue engineering. To achieve such a hierarchical porous structure with accurately controlled internal pore architectures, the recursive intersection Boolean operation (RIBO) was proposed in order to satisfy computational efficiency and biological function requirements of a porous scaffold. After generating the distance field (DF) for the given anatomic model and required pore architectures, the recursive DF modifications enable us to design hierarchical porous scaffolds with complex combinations of pore morphologies. A variety of experimental results showed that the proposed hierarchical porous scaffold design method has the potential benefits for accurately controlling both the porosity and the pore architecture gradients while preserving the advantages of triply periodic minimal surface pore geometries.

Scaffolds and stem cells: delivery of cell transplants for retinal degenerations

Retinal degenerations and optic neuropathies often lead to death of photoreceptors or retinal ganglion cells, respectively. Stem cell therapies are showing promise for these diseases in preclinical models and are beginning to transition into human trials, but cell delivery and integration remain major challenges. Focusing on photoreceptor- and progenitor-directed approaches, in this article, the authors review how advances in tissue engineering and cell scaffold design are enhancing cell therapies for retinal degeneration.

The incorporation and controlled release of platelet‐rich plasma‐derived biomolecules from polymeric tissue engineering scaffolds

AbstractPlatelet‐rich plasma (PRP) has been gaining popularity in recent years as a cost‐effective material capable of stimulating healing in a number of different clinical applications. As the clinical role ofPRPhas been growing so too has its prevalence in the fields of tissue engineering and regenerative medicine, particularly in the field of extracellular matrix (ECM) analogue scaffold fabrication. As polymeric scaffold fabrication techniques strive to create structures that ever more closely replicate the nativeECM's form and function, the need for increased scaffold bioactivity becomes more pronounced.PRP, which has been shown to contain over 300 bioactive molecules, has the potential to deliver a combination of growth factors and cytokines capable of stimulating cellular activity through enhanced chemotaxis, proliferation andECMproduction. The ability to incorporate such a potent bioactive milieu into a polymeric tissue engineering scaffold, which lacks intrinsic cell signaling molecules, may help to promote scaffold integration with native tissues and increase the overall patency of polymericECManalogue structures. This mini‐review briefly discusses the physiological basis ofPRPand its current clinical use, as well as the potential role thatPRPmay play in the future of polymeric tissue engineering scaffold design. Copyright © 2012 Society of Chemical Industry

In vitro and in vivo biocompatibility study on laser 3D microstructurable polymers

Films and microstructured scaffolds have been fabricated using direct laser writing out of different polymers: hybrid organic-inorganic ORMOCORE b59, acrylate-based AKRE23, novel organic-inorganic Zr containing hybrid SZ2080, and biodegradable PEG-DA-258. Adult myogenic stem cells were grown on these surfaces in vitro. Their adhesion, growth, and viability test results suggest good potential applicability of the materials in biomedical practice. Pieces of these polymers were implanted in rat’s paravertebral back tissue. Histological examination of the implants and surrounding tissue ex vivo after 3 weeks of implantation was conducted and results show the materials to be at least as biocompatible as surgical clips or sutures. The applied direct laser writing technique seems to offer good future prospects in a polymeric 3D scaffold design for artificial tissue engineering with autologous stem cells.

Heterogeneous porous scaffold design for tissue engineering using triply periodic minimal surfaces

Recently, a paradigm shift is taking place in tissue engineering scaffold design from homogeneous porous scaffolds to functionally graded scaffolds that have heterogeneous internal structures with controlled porosity levels and architectures. This paper presents a new heterogeneous modeling methodology for designing tissue engineering scaffolds with precisely controlled porosity and internal architectures using triply periodic minimal surfaces. The internal architectures and porosity at the spatial locations of the scaffolds are determined based on a given distribution of architectures and porosity levels specified at a few selected points on the model. After generating the hexahedral elements for a 3D anatomical shape using the distance field algorithm, the unit cell libraries composed of triply periodic minimal surfaces are mapped into the subdivided hexahedral elements using the shape function widely used in the finite element method. By simply allocating parameter values related to the porosity and architecture type to the corner nodes in each hexahedral element, we can easily and precisely control the pore size, porosity, and architecture type at each region of the scaffold while preserving perfectly interconnected pore networks across the entire scaffold.

Homogeneous Chitosan/Poly(L-Lactide) Composite Scaffolds Prepared by Emulsion Freeze-Drying

The combination between chitosan (CS)-based hydrophilic extracellular matrix polysaccharide and polylactide (PLA)-based hydrophobic biodegradable aliphatic polyester is a challenge in the biomaterials field. This study investigated the formation of homogeneous chitosan/poly(L-lactide) (CS/PLLA) porous composite scaffold using a novel emulsion freeze-drying technique. An oil-in-water (O/W) emulsification system was used in the presence of surfactant Tween-80, in which CS solution was used as the water phase and PLLA solution was used as the oil phase. The composite scaffolds showed well interconnected pore structures and homogenous distribution of CS and PLLA when the PLLA volume fraction was not higher than 50%. Once the PLLA content increased to 75%, SEM micrographs demonstrated that the two components present phase separation region. FT-IR analysis revealed that there are strong hydrogen bond interactions between CS and PLLA components. The porosity of the CS/PLLA composites was in the range of 85–90% and showed a slight decrease with increasing PLLA dose. The mechanical properties of the composites lay between that of the pure CS and the PLLA scaffold. The compressive strength increased from 0.17 to 0.21 MPa, while the compressive modulus increased from 2.37 to 3.38 MPa as the PLLA contents increased from 25 to 75%. In vitro cytotoxicity was evaluated by MTT assay. The results indicated that MC3T3-E1 cell viability and proliferation in the CS/PLLA scaffold were comparable to that in the CS scaffold, and much higher than that in the PLLA scaffold. The successful hydrophilic polysaccharide and hydrophobic polyester system offers a new delivery method of growth factors and a novel scaffold design for tissue engineering.

Topographic characterization and protein quantification of esophageal basement membrane for scaffold design reference in tissue engineering

Basement membrane (BM) has been investigated widely and applied in the field of bio-synthesized scaffold design in tissue engineering. However, this respect of investigation has seen scarcely in the field of esophageal tissue engineering. This study reports BM's basic topographic characters and quantification of structural and major proteins involving collagen IV, laminin, entactin, proteoglycans of porcine esophagus. Several methods were adopted to strip epithelium from mucosa tissue. For example, ethylene diamine tetraacetic acid, sonication, dithiothreitol, cold sodium chloride (NaCl), mechanical force of glass coverslip, and cold trypsin and so forth. Assessed experimental results under different conditions, the optimal condition to isolate epithelium from BM was established. After the reaction of cold trypsin at 4°C for 15 h, the BM was isolated from epithelium and exposed integratedly. Hematoxylin and eosin staining and immunohistochemical staining verified the effectiveness of epithelium removal and the integrity of BM. The topographic features of the exposed BM were observed under transmission electron microscopy and scanning electron microscopy. It displayed a rugged surface and 3-D topography consisting of pores and fibers with sub-100 nm range via ImageJ software. The major proteins existing in BM were quantitatively analyzed by enzyme-linked immune sorbent assay. All these results will provide references for the design of synthesized scaffolds and protein modification in esophageal tissue-engineering research.

A Hydrogel-Mineral Composite Scaffold for Osteochondral Interface Tissue Engineering

Osteoarthritis is the leading cause of physical disability among Americans, and tissue engineered cartilage grafts have emerged as a promising treatment option for this debilitating condition. Currently, the formation of a stable interface between the cartilage graft and subchondral bone remains a significant challenge. This study evaluates the potential of a hybrid scaffold of hydroxyapatite (HA) and alginate hydrogel for the regeneration of the osteochondral interface. Specifically, the effects of HA on the response of chondrocytes were determined, focusing on changes in matrix production and mineralization, as well as scaffold mechanical properties over time. Additionally, the optimal chondrocyte population for interface tissue engineering was evaluated. It was observed that the HA phase of the composite scaffold promoted the formation of a proteoglycan- and type II collagen-rich matrix when seeded with deep zone chondrocytes. More importantly, the elevated biosynthesis translated into significant increases in both compressive and shear moduli relative to the mineral-free control. Presence of HA also promoted chondrocyte hypertrophy and type X collagen deposition. These results demonstrate that the hydrogel-calcium phosphate composite supported the formation of a calcified cartilage-like matrix and is a promising scaffold design for osteochondral interface tissue engineering.

Evaluation of Cell‐Material Interactions on Newly Designed, Printable Polymers for Tissue Engineering Applications

AbstractThe generation of advanced scaffolds for tissue engineering is increasingly implemented by rapid prototyping techniques and therefore designed materials. Two novel polymeric networks based on newly synthesized carboxyfunctionalized polyacrylates were created to fit the requirements of 3D printing processes regarding viscosity and their surface tension. The polyacrylates presented carboxylic groups either without any spacer, or, in the case of mono‐(2‐acryloxyethyl)‐succinate, with a substantial spacing from the polymer backbone. The material properties were characterized with rheometry, FTIR, XPS, and contact angle measurements. Polymer surfaces were prepared and seeded with primary human dermal microvascular endothelial cells (HDMEC). Cell–material interaction with the different polymer surfaces were investigated by light microscopy, water soluble tetrazolium‐1 (WST‐1) viability assay, fluorescein diacetate/propidium iodide (FDA/PI) staining, and acetylated low‐density lipoprotein (acLDL) assay. Thus, the cell confluence, viability, and functionality of the HDMEC were evaluated. The non‐spaced methacrylic acid derivative was proved in cell culture regarding its biocompatibility. Along with its appropriate physical properties, the novel network‐generating polymeric blends constitute promising candidates for future scaffold design in tissue engineering.

Micromechanical regulation in cardiac myocytes and fibroblasts: implications for tissue remodeling

Cells of the myocardium are at home in one of the most mechanically dynamic environments in the body. At the cellular level, pulsatile stimuli of chamber filling and emptying are experienced as cyclic strains (relative deformation) and stresses (force per unit area). The intrinsic characteristics of tension-generating myocytes and fibroblasts thus have a continuous mechanical interplay with their extrinsic surroundings. This review explores the ways that the micromechanics at the scale of single cardiac myocytes and fibroblasts have been measured, modeled, and recapitulated in vitro in the context of adaptation. Both types of cardiac cells respond to externally applied strain, and many of the intracellular mechanosensing pathways have been identified with the careful manipulation of experimental variables. In addition to strain, the extent of loading in myocytes and fibroblasts is also regulated by cues from the microenvironment such as substrate surface chemistry, stiffness, and topography. Combinations of these structural cues in three dimensions are needed to mimic the micromechanical complexity derived from the extracellular matrix of the developing, healthy, or pathophysiologic heart. An understanding of cardiac cell micromechanics can therefore inform the design and composition of tissue engineering scaffolds or stem cell niches for future applications in regenerative medicine.

Computer-aided porous scaffold design for tissue engineering using triply periodic minimal surfaces

Recently, computer aided geometric design of triply periodic minimal surfaces (TPMS) has received considerable attention in the area of computer aided nano design on account of its ability to efficiently construct a large number of complex surfaces. In this paper, a TPMS is described with periodic surfaces composed of simple trigonometric functions, thus enabling easy generation of TPMS for use in various mechanical, chemical, and physical applications. We first describe a TPMS with mesh surface using conventional marching cube algorithm. We then propose various related algorithms for generating complete solid model for various applications, ranging from thickened solid, through voxel solid, to complexshaped solid. The validity of this new technique is demonstrated for a variety of TPMSs, including the P, G, D, I-WP, F-RD, L, and I2-Y** surfaces. Finally, a new control approach for pore size distribution in tissue scaffold design is presented based on the pore-making element composed of TPMS and conformal refinement of all-hexahedral mesh in order to show the practical applicability of the newly suggested modeling approach.

Methylprednisolone release from agar-Carbomer-based hydrogel: a promising tool for local drug delivery

AbstractA number of studies and works in drug delivery literature are focused on the understanding and modelling of transport phenomena, the pivotal point of a good scaffold design for tissue engineering. Accurate knowledge of the diffusion coefficient of an active drug plays a key role in the analysis, prediction of their kinetics and formulation of efficient drug delivery systems. In this work, the kinetics of the release of methylprednisolone from agar-Carbomer hydrogel were studied taking into consideration the different drug concentrations and clearances typically achieved in in vitro or in vivo tests. Starting from the experiments it is possible to model the transport phenomenon and to calculate the diffusion coefficient through the hydrogel matrix.

Fabrication of Biodegradable Polyester Nanocomposites by Electrospinning for Tissue Engineering

Recently, nanocomposites have emerged as an efficient strategy to upgrade the structural and functional properties of synthetic polymers. Polyesters have attracted wide attention because of their biodegradability and biocompatibility. A logic consequence has been the introduction of natural extracellular matrix (ECM) molecules, organic or inorganic nanostructures to biodegradable polymers to produce nanocomposites with enhanced properties. Consequently, the improvement of the interfacial adhesion between biodegradable polymers and natural ECM molecules or nanostructures has become the key technique in the fabrication of nanocomposites. Electrospinning has been employed extensively in the design and development of tissue engineering scaffolds to generate nanofibrous substrates of synthetic biodegradable polymers and to simulate the cellular microenvironment. In this paper, several types of biodegradable polyester nanocomposites were prepared by electrospinning, with the aim of being used as tissue engineering scaffolds. The combination of biodegradable nanofibrous polymers and natural ECM molecules or nanostructures opens new paradigms for tissue engineering applications.

Aligned natural–synthetic polyblend nanofibers for peripheral nerve regeneration

Peripheral nerve regeneration remains a significant clinical challenge to researchers. Progress in the design of tissue engineering scaffolds provides an alternative approach for neural regeneration. In this study aligned silk fibroin (SF) blended poly( l-lactic acid-co-ε-caprolactone) (P(LLA-CL)) nanofibrous scaffolds were fabricated by electrospinning methods and then reeled into aligned nerve guidance conduits (NGC) to promote nerve regeneration. The aligned SF/P(LLA-CL) NGC was used as a bridge implanted across a 10 mm defect in the sciatic nerve of rats and the outcome in terms of of regenerated nerve at 4 and 8 weeks was evaluated by a combination of electrophysiological assessment and histological and immunohistological analysis, as well as electron microscopy. The electrophysiological examination showed that functional recovery of the regenerated nerve in the SF/P(LLA-CL) NGC group was superior to that in the P(LLA-CL) NGC group. The morphological analysis also indicated that the regenerated nerve in the SF/P(LLA-CL) NGC was more mature. All the results demonstrated that the aligned SF/P(LLA-CL) NGC promoted peripheral nerve regeneration significantly better in comparison with the aligned P(LLA-CL) NGC, thus suggesting a potential application in nerve regeneration.

Synergic effects of crypt-like topography and ECM proteins on intestinal cell behavior in collagen based membranes

The basement membrane of small intestinal epithelium possesses complex topography at multiple scales ranging from the mesoscale to nanoscale. Specifically, intestinal crypt-villus units are comprised of hundred-micron-scale well-like invaginations and finger-like projections; intestinal cell phenotype is related to location on this crypt-villus unit. A biomimetic intestinal cell culture system composed of type I collagen based permeable cell culture membranes incorporating both micron-scale intestinal crypt-like topography and nanometer scale topography was fabricated. Membranes were pre-incubated with either laminin (Ln) or fibronectin (Fn), inoculated with intestinal epithelial Caco-2 cells and cultured for 1–21 days to study the relative significance of influence of crypt-like topography and biomimetic substrate chemistry on cell phenotype. Crypt-like topography inhibited Caco-2 differentiation during early culture, as evidenced by slower cell spreading and lower brush border enzyme activity. For example, alanine aminopeptidase activity was lower on Ln-coated patterned collagen (∼3.4 ± 0.24 mU/mg) compared to flat collagen (10.84 ± 0.55 mU/mg) at day 7. Caco-2 cultured on Fn-coated collagen started to spread earlier (1 day vs 3 days) and formed longer protrusions than on Ln-coated collagen. Pre-coating of Ln enhanced cell differentiation, as the maximum activity of a cell differentiation marker (alkaline phosphatase) was 2–3 times higher than on Fn-coated collagen, and maintained differentiated phenotype in long term (up to 21 days) culture. In general, compared to substrate topography, coating with ECM protein had more prominent and longer effect on cell behavior. Crypt-like topography affected Caco-2 spreading and differentiation during early culture, however the effect diminished as culture progressed. This information will benefit intestinal tissue engineering scaffold design, and modification of in vitro intestinal cell models.