Bilayered constructs aimed at osteochondral strategies: The influence of medium supplements in the osteogenic and chondrogenic differentiation of amniotic fluid-derived stem cells

Wake Forest Institute for Regenerative Medicine, Wake Forest University School of Medicine, Winston-Salem, NC 27157, USA.
Acta biomaterialia (Impact Factor: 6.03). 04/2012; 8(7):2795-806. DOI: 10.1016/j.actbio.2012.04.013
Source: PubMed


The development of osteochondral tissue engineered interfaces would be a novel treatment for traumatic injuries and aging associated diseases that affect joints. This study reports the development of a bilayered scaffold, which consists of both bone and cartilage regions. On the other hand, amniotic fluid-derived stem cells (AFSCs) could be differentiated into either osteogenic or chondrogenic cells, respectively. In this study we have developed a bilayered scaffolding system, which includes a starch/polycaprolactone (SPCL) scaffold for osteogenesis and an agarose hydrogel for chondrogenesis. AFSC-seeded scaffolds were cultured for 1 or 2 weeks in an osteochondral-defined culture medium containing both osteogenic and chondrogenic differentiation factors. Additionally, the effect of the presence or absence of insulin-like growth factor-1 (IGF-1) in the culture medium was assessed. Cell viability and phenotypic expression were assessed within the constructs in order to determine the influence of the osteochondral differentiation medium. The results indicated that, after osteogenic differentiation, AFSCs that had been seeded onto SPCL scaffolds did not require osteochondral medium to maintain their phenotype, and they produced a protein-rich, mineralized extracellular matrix (ECM) for up to 2 weeks. However, AFSCs differentiated into chondrocyte-like cells appeared to require osteochondral medium, but not IGF-1, to synthesize ECM proteins and maintain the chondrogenic phenotype. Thus, although IGF-1 was not essential for creating osteochondral constructs with AFSCs in this study, the osteochondral supplements used appear to be important to generate cartilage in long-term tissue engineering approaches for osteochondral interfaces. In addition, constructs generated from agarose-SPCL bilayered scaffolds containing pre-differentiated AFSCs may be useful for potential applications in regeneration strategies for damaged or diseased joints.

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Available from: Rui L. Reis, May 06, 2015
    • "A number of different strategies have been developed to engineer osteochondral constructs. These include the development of bi-phasic or multi-layered scaffolds (Mano and Reis, 2007; Martin et al., 2007; Rodrigues et al., 2012; Sheehy et al., 2013), physical conditioning of tissues through the use of novel bioreactors (Grayson et al., 2010; Wang et al., 2004; Wendt et al., 2005) and spatial growth factor or gene delivery systems (Chen et al., 2011; Mason et al., 1998; Santo et al., 2013). It has also been possible to engineer scaffolds and grafts mimicking the geometrical form of articular surfaces (Alhadlaq et al., 2004; Ding et al., 2013; Hung et al., 2003; Lee et al., 2010; Lee et al., 2009). "
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    ABSTRACT: Arthroplasty is currently the only surgical procedure available to restore joint function following articular cartilage and bone degeneration associated with diseases such as osteoarthritis (OA). A potential alternative to this procedure would be to tissue-engineer a biological implant and use it to replace the entire diseased joint. The objective of this study was therefore to tissue-engineer a scaled-up, anatomically shaped, osteochondral construct suitable for partial or total resurfacing of a diseased joint. To this end it was first demonstrated that a bone marrow derived mesenchymal stem cell seeded alginate hydrogel could support endochondral bone formation in vivo within the osseous component of an osteochondral construct, and furthermore, that a phenotypically stable layer of articular cartilage could be engineered over this bony tissue using a co-culture of chondrocytes and mesenchymal stem cells. Co-culture was found to enhance the in vitro development of the chondral phase of the engineered graft and to dramatically reduce its mineralisation in vivo. In the final part of the study, tissue-engineered grafts (~ 2 cm diameter) mimicking the geometry of medial femorotibial joint prostheses were generated using laser scanning and rapid prototyped moulds. After 8 weeks in vivo, a layer of cartilage remained on the surface of these scaled-up engineered implants, with evidence of mineralisation and bone development in the underlying osseous region of the graft. These findings open up the possibility of a tissueengineered treatment option for diseases such as OA.
    European cells & materials 09/2015; 30:163-186. · 4.89 Impact Factor
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    • "sulfate esters, methyl ethers, pyruvate acid ketals) (Matsuhashi, 1990). Unlike alginate or ulvan, agar does not require additives (such as salts) to form gels which makes it more challenging to process in other forms besides gels (Khanarian, Haney, Burga, & Lu, 2012; Rodrigues et al., 2012; Santoro et al., 2011; Yamada et al., 2012). The use of agar nanofibers could potentiate new and exciting properties of the polymer, not seen at greater scales (Greiner & Wendorff, 2007). "
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    ABSTRACT: Very recently our group has produced novel agar-based fibers by an electrospinning technique using water as solvent and polyvinyl alcohol (PVA) as co-blending polymer. Here, we tested the deep eutectic solvent (DES), (2-hydroxyethyl)trimethylammonium chloride/urea prepared at 1:2 molar ratio, as an alternative solvent medium for agar electrospinning. The electrospun materials were collected with an ethanol bath adapted to a previous electrospinning set-up. 1 wt% agar-in-DES showed improved viscoelasticity and hence, spinnability, when compared to 1% wt agar-in-water and pure agar nanofibers were successfully electrospun if working above the temperature of sol-gel transition (∼80°C). By changing the solvent medium we decreased the PVA concentration (5 wt% starting solution) and successfully produced composite fibers with high agar contents (50/50 agar/PVA). Best composite fibers were formed with the 50/50 and 30/70 agar/PVA solutions. These fibers were mechanically resistant, showed tailorable surface roughness and diverse size distributions, with most of the diameters falling in the sub-micron range. Both nano- and micro- forms of agar fibers (used separately or combined) may have potential for the design of new and highly functional agar-based materials. Copyright © 2015. Published by Elsevier B.V.
    International journal of biological macromolecules 06/2015; DOI:10.1016/j.ijbiomac.2015.06.034 · 2.86 Impact Factor
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    • "We and others have demonstrated that under appropriate inductive conditions human AFSCs [2], [6], [7] and hASCs can be directed into several skeletal tissue-related lineages, such as bone [2], [6]–[8] and cartilage [2], [6], [8]. "
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    ABSTRACT: Current treatments for tendon injuries often fail to fully restore joint biomechanics leading to the recurrence of symptoms, and thus resulting in a significant health problem with a relevant social impact worldwide. Cell-based approaches involving the use of stem cells might enable tailoring a successful tendon regeneration outcome. As growth factors (GFs) powerfully regulate the cell biological response, their exogenous addition can further stimulate stem cells into the tenogenic lineage, which might eventually depend on stem cells source. In the present study we investigate the tenogenic differentiation potential of human- amniotic fluid stem cells (hAFSCs) and adipose-derived stem cells (hASCs) with several GFs associated to tendon development and healing; namely, EGF, bFGF, PDGF-BB and TGF-β1. Stem cells response to biochemical stimuli was studied by screening of tendon-related genes (collagen type I, III, decorin, tenascin C and scleraxis) and proteins found in tendon extracellular matrix (ECM) (Collagen I, III, and Tenascin C). Despite the fact that GFs did not seem to influence the synthesis of tendon ECM proteins, EGF and bFGF influenced the expression of tendon-related genes in hAFSCs, while EGF and PDGF-BB stimulated the genetic expression in hASCs. Overall results on cellular alignment morphology, immunolocalization and PCR analysis indicated that both stem cell source can be biochemically induced towards tenogenic commitment, validating the potential of hASCs and hAFSCs for tendon regeneration strategies.
    PLoS ONE 12/2013; 8(12):e83734. DOI:10.1371/journal.pone.0083734 · 3.23 Impact Factor
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