Ethanolamines -- Properties, Rapid prototyping -- Methods, Tissue engineering -- Research, and Hyaluronic acid -- Properties

Introduction AS WORLD POPULATION AND the average human lifespan increases, the global medical need for donor organs will also increase. (1,2) For example, as of March 2010 there were over [...]
Bioprinting by the codeposition of cells and biomaterials is constrained by the availability of printable materials. Herein we describe a novel macromonomer, a new two-step photocrosslinking strategy, and the use of a simple rapid prototyping system to print a proof-of-concept tubular construct. First, we synthesized the methacrylated ethanolamide derivative of gelatin (GE-MA). Second, partial photochemical cocrosslinking of GE-MA with methacrylated hyaluronic acid (HA-MA) gave an extrudable gel-like fluid. Third, the new HA-MA:GE-MA hydrogels were biocompatible, supporting cell attachment and proliferation of HepG2 C3A, Int-407, and NIH 3T3 cells in vitro. Moreover, hydrogels injected subcutaneously in nude mice produced no inflammatory response. Fourth, using the Fab@Home printing system, we printed a tubular tissue construct. The partially crosslinked hydrogels were extruded from a syringe into a designed base layer, and irradiated again to create a firmer structure. The computer-driven protocol was iterated to complete a cellularized tubular construct with a cell-free core and a cell-free structural halo. Cells encapsulated within this printed construct were viable in culture, and gradually remodeled the synthetic extracellular matrix environment to a naturally secreted extracellular matrix. This two-step photocrosslinkable biomaterial addresses an unmet need for printable hydrogels useful in tissue engineering.

Spinal cord injuries -- Physiological aspects, Spinal cord injuries -- Care and treatment, and Morphometrics (Biology) -- Methods

Introduction Spinal Cord Injury (SCI) results in a devastating condition with enormous financial, (1-4) social, and personal costs.5 It is estimated that the annual incidence of SCI in the United [...]
Spinal cord injury (SCI) represents a significant health and social problem, and therefore it is vital to develop novel strategies that can specifically target it. In this context, the objective of the present work was to develop a new range of three-dimensional (3D) tubular structures aimed at inducing the regeneration within SCI sites. Up to six different 3D tubular structures were initially developed by rapid prototyping: 3D bioplotting-based on a biodegradable blend of starch. These structures were then further complemented by injecting Gellan Gum, a polysaccharide-based hydrogel, in the central area of structures. The mechanical properties of these structures were assessed using dynamic mechanical analysis, under both dry and wet conditions, and their morphologies/ porosities were analyzed using micro-computed tomography and scanning electron microscopy. Biological evaluation was carried out to determine their cytotoxicity, using both minimum essential medium (MEM) extraction and MTS tests, as well as by encapsulation of an oligodendrocyte-like cell (M03-13 cell line) within the hydrogel phase. The histomorphometric analysis showed a fully interconnected network of pores with porosity ranging from 70% to 85%. Scaffolds presented compressive modulus ranging from 17.4 to 62.0 MPa and 4.42 to 27.4 MPa under dry and wet conditions, respectively. Cytotoxicity assays revealed that the hybrid starch/poly-[epsilon]-caprolactone/Gellan Gum scaffolds were noncytotoxic, as they did not cause major alterations on cell morphology, proliferation, and metabolic viability. Moreover, preliminary cell encapsulation assays showed that the hybrid scaffolds could support the in vitro culture of oligodendrocyte-like cells. Finally, preliminary in vivo studies conducted in a hemisection rat SCI model revealed that the above-referred structures were well integrated within the injury and did not trigger chronic inflammatory processes. The results herein presented indicate that these 3D systems might be of use in future SCI regeneration approaches.

3D technology, Three-dimensional display systems -- Usage, Tissue engineering -- Physiological aspects, and Tissue engineering -- Usage

Introduction SYNTHETIC HYDROLYTICALLY DEGRADABLE POLYMERS have been tested, some used in the field of regenerative medicine for the last four decades, because they are free from most of the shortcomings [...]
Synthetic polymers have attracted much attention in tissue engineering due to their ability to modulate biomechanical properties. This study investigated the feasibility of processing poly(e-caprolactone) (PCL) homopolymer, PCL-poly(ethylene glycol) (PEG) diblock, and PCL-PEG-PCL triblock copolymers into three-dimensional porous scaffolds. Properties of the various polymers were investigated by dynamic thermal analysis. The scaffolds were manufactured using the desktop robot-based rapid prototyping technique. Gross morphology and internal three-dimensional structure of scaffolds were identified by scanning electron microscopy and micro-computed tomography, which showed excellent fusion at the filament junctions, high uniformity, and complete interconnectivity of pore networks. The influences of process parameters on scaffolds' morphological and mechanical characteristics were studied. Data confirmed that the process parameters directly influenced the pore size, porosity, and, consequently, the mechanical properties of the scaffolds. The in vitro cell culture study was performed to investigate the influence of polymer nature and scaffold architecture on the adhesion of the cells onto the scaffolds using rabbit smooth muscle cells. Light, scanning electron, and confocal laser microscopy showed cell adhesion, proliferation, and extracellular matrix formation on the surface as well as inside the structure of both scaffold groups. The completely interconnected and highly regular honeycomb-like pore morphology supported bridging of the pores via cell-to-cell contact as well as production of extracellular matrix at later time points. The results indicated that the incorporation of hydrophilic PEG into hydrophobic PCL enhanced the overall hydrophilicity and cell culture performance of PCL-PEG copolymer. However, the scaffold architecture did not significantly influence the cell culture performance in this study.

Cardiovascular system -- Research, Nanoparticles -- Physiological aspects, Nanoparticles -- Research, and Tissue engineering -- Usage

Introduction SCAFFOLD DESIGN IN tissue engineering (TE) plays an important role in modulating tissue growth and development. Various scaffold fabrication techniques, including rapid prototyping, solvent casting, salt leaching, and electrospinning, [...]
Despite the attractive features of nanofibrous scaffolds for cell attachment in tissue-engineering (TE) applications, impeded cell ingrowth has been reported in electrospun scaffolds. Previous findings have shown that the scaffold can function as a sieve, keeping cells on the scaffold surface, and that cell migration into the scaffold does not occur in time. Because fiber diameter is directly related to the pore size of an electrospun scaffold, the objective of this study was to systematically evaluate how cell delivery can be optimized by tailoring the fiber diameter of electrospun poly([epsilon]-caprolactone) (PCL) scaffolds. Five groups of electrospun PCL scaffolds with increasing average fiber diameters (3.4-12.1 [micro]m) were seeded with human venous myofibroblasts. Cell distribution was analyzed after 3 days of culture. Cell penetration increased proportionally with increasing fiber diameter. Unobstructed delivery of cells was observed exclusively in the scaffold with the largest fiber diameter (12.1 [micro]m). This scaffold was subsequently evaluated in a 4-week TE experiment and compared with a poly(glycolic acid)-poly(4-hydroxybutyrate) scaffold, a standard scaffold used successfully in cardiovascular tissue engineering applications. The PCL constructs showed homogeneous tissue formation and sufficient matrix deposition. In conclusion, fiber diameter is a crucial parameter to allow for homogeneous cell delivery in electrospun scaffolds. The optimal electrospun scaffold geometry, however, is not generic and should be adjusted to cell size.