Dataset Information

Chitin Nanofiber Micropatterned Flexible Substrates for Tissue Engineering†

ABSTRACT: Engineered tissues require enhanced organization of cells and extracellular matrix (ECM) for proper function. To promote cell organization, substrates with controlled micro- and nanopatterns have been developed as supports for cell growth, and to induce cellular elongation and orientation via contact guidance. Micropatterned ultra-thin biodegradable substrates are desirable for implantation in the host tissue. These substrates, however, need to be mechanically robust to provide substantial support for the generation of new tissues, to be easily retrievable, and to maintain proper handling characteristics. Here, we introduce ultra-thin (<10 ?m), self-assembled chitin nanofiber substrates micropatterned with replica molding for engineering cell sheets. These substrates are biodegradable, mechanically strong, yet flexible, and easily manipulated into the desired shape. As a proof-of-concept, fibroblast cell proliferation, elongation, and alignment were studied on the developed substrates with different pattern dimensions. On the optimized substrates, the majority of the cells aligned (<10°) along the major axis of micropatterned features. With the ease of fabrication and mechanical robustness, the substrates presented herein can be utilized as versatile system for the engineering and delivery of ordered tissue in applications such as myocardial repair.

SUBMITTER: Hassanzadeh P

PROVIDER: S-EPMC3809896 | biostudies-literature | 2013 Sep

REPOSITORIES: biostudies-literature

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Chitin Nanofiber Micropatterned Flexible Substrates for Tissue Engineering<sup>†</sup>

Hassanzadeh Pegah P Kharaziha Mahshid M Nikkhah Mehdi M Shin Su-Ryon SR Jin Jungho J He Simeiqi S Sun Wei W Zhong Chao C Dokmeci Mehmet R MR Khademhosseini Ali A Khademhosseini Ali A Rolandi Marco M

Journal of materials chemistry. B 20130901 34

Engineered tissues require enhanced organization of cells and extracellular matrix (ECM) for proper function. To promote cell organization, substrates with controlled micro- and nanopatterns have been developed as supports for cell growth, and to induce cellular elongation and orientation via contact guidance. Micropatterned ultra-thin biodegradable substrates are desirable for implantation in the host tissue. These substrates, however, need to be mechanically robust to provide substantial suppo ...[more]

PMID: 24179675

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Project description:Tissue engineering aims to capture the structural and functional aspects of diverse tissue types in vitro. However, most approaches are limited in their ability to produce complex 3D geometries that are essential for tissue function. Tissues, such as the vasculature or chambers of the heart, often possess curved surfaces and hollow lumens that are difficult to recapitulate given their anisotropic architecture. Cell-sheet engineering techniques using thermoresponsive substrates provide a means to stack individual layers of cells with spatial control to create dense, scaffold-free tissues. In this study, we developed a novel method to fabricate complex 3D structures by layering multiple sheets of aligned cells onto flexible scaffolds and casting them into hollow tubular geometries using custom molds and gelatin hydrogels. To enable the fabrication of 3D tissues, we adapted our previously developed thermoresponsive nanopatterned cell-sheet technology by applying it to flexible substrates that could be folded as a form of tissue origami. We demonstrated the versatile nature of this platform by casting aligned sheets of smooth and cardiac muscle cells circumferentially around the surfaces of gelatin hydrogel tubes with hollow lumens. Additionally, we patterned skeletal muscle in the same fashion to recapitulate the 3D curvature that is observed in the muscles of the trunk. The circumferential cell patterning in each case was maintained after one week in culture and even encouraged organized skeletal myotube formation. Additionally, with the application of electrical field stimulation, skeletal myotubes began to assemble functional sarcomeres that could contract. Cardiac tubes could spontaneously contract and be paced for up to one month. Our flexible cell-sheet engineering approach provides an adaptable method to recapitulate more complex 3D geometries with tissue specific customization through the addition of different cell types, mold shapes, and hydrogels. By enabling the fabrication of scaled biomimetic models of human tissues, this approach could potentially be used to investigate tissue structure-function relationships, development, and maturation in the dish.

Project description:There is a need for an artificial salivary gland as a long-term remedy for patients suffering from salivary hypofunction, a leading cause of chronic xerostomia (dry mouth). Current salivary gland tissue engineering approaches are limited in that they either lack sufficient physical cues and surface area needed to facilitate epithelial cell differentiation, or they fail to provide a mechanism for assembling an interconnected branched network of cells. We have developed highly-ordered arrays of curved hemispherical "craters" in polydimethylsiloxane (PDMS) using wafer-level integrated circuit (IC) fabrication processes, and lined them with electrospun poly-lactic-co-glycolic acid (PLGA) nanofibers, designed to mimic the three-dimensional (3-D) in vivo architecture of the basement membrane surrounding spherical acini of salivary gland epithelial cells. These micropatterned scaffolds provide a method for engineering increased surface area and were additionally investigated for their ability to promote cell polarization. Two immortalized salivary gland cell lines (SIMS, ductal and Par-C10, acinar) were cultured on fibrous crater arrays of various radii and compared with those grown on flat PLGA nanofiber substrates, and in 3-D Matrigel. It was found that by increasing crater curvature, the average height of the cell monolayer of SIMS cells and to a lesser extent, Par-C10 cells, increased to a maximum similar to that seen in cells grown in 3-D Matrigel. Increasing curvature resulted in higher expression levels of tight junction protein occludin in both cell lines, but did not induce a change in expression of adherens junction protein E-cadherin. Additionally, increasing curvature promoted polarity of both cell lines, as a greater apical localization of occludin was seen in cells on substrates of higher curvature. Lastly, substrate curvature increased expression of the water channel protein aquaporin-5 (Aqp-5) in Par-C10 cells, suggesting that curved nanofiber substrates are more suitable for promoting differentiation of salivary gland cells.

Project description:Objective: The avascular inner regions of the knee menisci cannot self-heal. As a prospective treatment, functional replacements can be generated by cell-based 3D bioprinting with an appropriate cell source and biomaterial. To that end, human meniscus fibrochondrocytes (hMFC) from surgical castoffs of partial meniscectomies as well as cellulose nanofiber-alginate based hydrogels have emerged as a promising cell source and biomaterial combination. The objectives of the study were to first find the optimal formulations of TEMPO (2,2,6,6-tetramethylpiperidine-1-oxyl)-oxidized cellulose nanofiber/alginate (TCNF/ALG) precursors for bioprinting, and then to use them to investigate redifferentiation and synthesis of functional inner meniscus-like extracellular matrix (ECM) components by expanded hMFCs. Methods: The rheological properties including shear viscosity, thixotropic behavior recovery, and loss tangent of selected TCNF/ALG precursors were measured to find the optimum formulations for 3D bioprinting. hMFCs were mixed with TCNF/ALG precursors with suitable formulations and 3D bioprinted into cylindrical disc constructs and crosslinked with CaCl2 after printing. The bioprinted constructs then underwent 6 weeks of in vitro chondrogenesis in hypoxia prior to analysis with biomechanical, biochemical, molecular, and histological assays. hMFCs mixed with a collagen I gel were used as a control. Results: The TCNF/ALG and collagen-based constructs had similar compression moduli. The expression of COL2A1 was significantly higher in TCNF/ALG. The TCNF/ALG constructs showed more of an inner meniscus-like phenotype while the collagen I-based construct was consistent with a more outer meniscus-like phenotype. The expression of COL10A1 and MMP13 were lower in the TCNF/ALG constructs. In addition, the immunofluorescence of human type I and II collagens were evident in the TCNF/ALG, while the bovine type I collagen constructs lacked type II collagen deposition but did contain newly synthesized human type I collagen.

Dataset Information

Chitin Nanofiber Micropatterned Flexible Substrates for Tissue Engineering†

Publications

Chitin Nanofiber Micropatterned Flexible Substrates for Tissue Engineering<sup>†</sup>

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