ECM-derived Scaffolds Research Articles

Crosslinking strategies for preparation of extracellular matrix-derived cardiovascular scaffolds.

Heart valve and blood vessel replacement using artificial prostheses is an effective strategy for the treatment of cardiovascular disease at terminal stage. Natural extracellular matrix (ECM)-derived materials (decellularized allogeneic or xenogenic tissues) have received extensive attention as the cardiovascular scaffold. However, the bioprosthetic grafts usually far less durable and undergo calcification and progressive structural deterioration. Glutaraldehyde (GA) is a commonly used crosslinking agent for improving biocompatibility and durability of the natural scaffold materials. However, the nature ECM and GA-crosslinked materials may result in calcification and eventually lead to the transplant failure. Therefore, studies have been conducted to explore new crosslinking agents. In this review, we mainly focused on research progress of ECM-derived cardiovascular scaffolds and their crosslinking strategies.

Controlled release of transforming growth factor-β3 from cartilage-extra-cellular-matrix-derived scaffolds to promote chondrogenesis of human-joint-tissue-derived stem cells

The objective of this study was to develop a scaffold derived from cartilaginous extracellular matrix (ECM) that could be used as a growth factor delivery system to promote chondrogenesis of stem cells. Dehydrothermal crosslinked scaffolds were fabricated using a slurry of homogenized porcine articular cartilage, which was then seeded with human infrapatellar-fat-pad-derived stem cells (FPSCs). It was found that these ECM-derived scaffolds promoted superior chondrogenesis of FPSCs when the constructs were additionally stimulated with transforming growth factor (TGF)-β3. Cell-mediated contraction of the scaffold was observed, which could be limited by the additional use of 1-ethyl-3-3dimethyl aminopropyl carbodiimide (EDAC) crosslinking without suppressing cartilage-specific matrix accumulation within the construct. To further validate the utility of the ECM-derived scaffold, we next compared its chondro-permissive properties to a biomimetic collagen–hyaluronic acid (HA) scaffold optimized for cartilage tissue engineering (TE) applications. The cartilage-ECM-derived scaffold supported at least comparable chondrogenesis to the collagen–HA scaffold, underwent less contraction and retained a greater proportion of synthesized sulfated glycosaminoglycans. Having developed a promising scaffold for TE, with superior chondrogenesis observed in the presence of exogenously supplied TGF-β3, the final phase of the study explored whether this scaffold could be used as a TGF-β3 delivery system to promote chondrogenesis of FPSCs. It was found that the majority of TGF-β3 that was loaded onto the scaffold was released in a controlled manner over the first 10days of culture, with comparable long-term chondrogenesis observed in these TGF-β3-loaded constructs compared to scaffolds where the TGF-β3 was continuously added to the media. The results of this study support the use of cartilage-ECM-derived scaffolds as a growth factor delivery system for use in articular cartilage regeneration.

Oriented cartilage extracellular matrix-derived scaffold for cartilage tissue engineering

The structure of a cartilage scaffold is required to mimic native articular cartilage, which has an oriented structure associated with its mechanical function. In this study, an oriented cartilage extracellular matrix (ECM)-derived scaffold was fabricated composed of microtubules arranged in parallel in vertical section. The mechanical property was higher than that of a typical non-oriented scaffold (p<0.05). Oriented and non-oriented scaffolds were seeded with chondrogenic-induced bone mesenchymal stem cells and cell-scaffold constructs were implanted subcutaneously in the dorsa of nude mice. At 4 weeks, all samples stained positive for safranin O, toluidine blue, and collagen type II, but negative for collagen type I. Oriented-structure constructs contained numerous parallel giant bundles of densely packed collagen fibers with chondrocyte-like cells aligned along the fibers. Total DNA, glycosaminoglycans and collagen contents increased with time and these values were similar in the two groups. Compared with the native articular cartilage, the Young's modulus of the tissue-engineered (TE) cartilage reached 42.9%, 23.0% in oriented and non-oriented scaffolds respectively, at 4 weeks. These results indicate that oriented ECM-derived scaffolds enhance the biomechanical property of TE cartilage and thus represent a promising approach to cartilage tissue engineering.

Fabrication a novel cartilage ECM-derived 3-D porous acellular matrix scaffold and in vivo cartilage tissue engineering with PKH26-labeled chondrogenic bone marrow-derived mesenchymal stem cells.

Objective The objective of this study is non-destructive evaluation of tissue engineering constructs in vivo using PKH26 and molecular light imaging system, and exploring the feasibility for tissue engineering cartilage using canine chondrocytes and porous cartilage acellular matrix scaffold. Methods After induced by chondrogenic medium, BMSCs were seeded into the CEDPS scaffolds, cell attachment was confirmed by SEM and the viability of attached cells on the scaffold was confirmed by a live/dead assess-ment. Chondrogenically induced BMSCs labeled with fluorescent dye PKH26 were then grown on scaffolds in vitro and implanted subcutaneously into nude mice. Then in vivo fluorescent imaging system was used for e-valuating the cell-scaffold constructs. After 4 weeks, the constructs was analyzed by histology, immunohisto-chemistry and immunofluorescence examnation. Results SEM showed a large mount of extraceilular matrix around the cells as time grown. Dead/Live staining in the confocal microscopy of cell-scaffold constructs re-vealed cells with green fluorescence (live cell). Four weeks later, cartilage-like tissue formed in nude mice, with positive staining for Safranin O, tuoluidine blue and collagen Ⅱ. Cells in the samples seemed to confirm that they originated from the labeled BMSCs, as confirmed by in vivo fluorescent imaging and immunofluo-rescence examination. Conclusion Cartilage ECM-derived scaffolds can be used for effective cartilage tis-sue engineering both in vitro and in vivo. As well, PKH26 fluorescent labeling and in vivo fluorescent imag-ing can be useful for cell tracking and analyzing cell-scaffold constructs in vivo. Key words: Cartilage; Tissue engineering; Mesenchymal stem cells; Bone marrow; Fluorescence

Hybrid nanofibrous scaffolds from electrospinning of a synthetic biodegradable elastomer and urinary bladder matrix

Synthetic materials can be electrospun into submicron or nanofibrous scaffolds to mimic extracellular matrix (ECM) scale and architecture with reproducible composition and adaptable mechanical properties. However, these materials lack the bioactivity present in natural ECM. ECM-derived scaffolds contain bioactive molecules that exert in vivo mimicking effects as applied for soft tissue engineering, yet do not possess the same flexibility in mechanical property control as some synthetics. The objective of the present study was to combine the controllable properties of a synthetic, biodegradable elastomer with the inherent bioactivity of an ECM derived scaffold. A hybrid electrospun scaffold composed of a biodegradable poly(ester-urethane)urea (PEUU) and a porcine ECM scaffold (urinary bladder matrix, UBM) was fabricated and characterized for its bioactive and physical properties both in vitro and in vivo. Increasing amounts of PEUU led to linear increases in both tensile strength and breaking strain while UBM incorporation led to increased in vitro smooth muscle cell adhesion and proliferation and in vitro mass loss. Subcutaneous implantation of the hybrid scaffolds resulted in increased scaffold degradation and a large cellular infiltrate when compared with electrospun PEUU alone. Electrospun UBM/PEUU combined the attractive bioactivity and mechanical features of its individual components to result in scaffolds with considerable potential for soft tissue engineering applications.