<i>In Vitro</i>Characterization and<i>In Vivo</i>Behavior of Human Nucleus Pulposus and Annulus Fibrosus Cells in Clinical-Grade Fibrin and Collagen-Enriched Fibrin Gels

Colombini, Alessandra; Lopa, Silvia; Ceriani, Cristina; Lovati, Arianna B.; Croiset, Samantha Jennifer; Giancamillo, A. Di; Lombardi, Giovanni; Banfi, Giuseppe; Moretti, Matteo

doi:10.1089/ten.tea.2014.0279

Cited by 21 publications

(31 citation statements)

References 32 publications

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“…Therefore, in the first part of this study a suitable AF differentiation medium for hASCs in two‐dimensional (2D) culture was defined. The importance of TGF‐β1 or TGF‐β3 to maintain the AF cell phenotype in vitro has been reported in several studies (Colombini et al , ; Guillaume et al , ). However, the present study appears to be the first to compare the effects of TGF‐β1 and TGF‐β3 or their combination on hASC proliferation and differentiation towards AF tissue.…”

Section: Discussionmentioning

confidence: 86%

“…In addition to the composition of the differentiation medium, different cell seeding strategies have been shown to have a major effect on stem cell fate (Ameer et al , ; Colombini et al , ). However, an efficient cell seeding strategy that supports mesenchymal stem cell differentiation towards AF tissue has not been reported previously.…”

Section: Discussionmentioning

confidence: 99%

“…Human AF cells (ScienCell Research Laboratories, Carlsbad, CA, USA) seeded with fibrin gel (human AF cells; ScienCell) were used as a reference cell type in order to verify the phenotype of the differentiated hASCs towards AF tissue. Fibrin gel‐seeded AF cells have previously been shown to maintain the typical features of these cell populations (Colombini et al , ). Human AF cells were expanded according to the manufacturer's protocol in a commercially available medium (NPCM; ScienCell) at 37°C, 5% CO 2 .…”

Section: Methodsmentioning

confidence: 99%

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Differentiation of adipose stem cells seeded towards annulus fibrosus cells on a designed poly(trimethylene carbonate) scaffold prepared by stereolithography

Blanquer

Gebraad

Miettinen

et al. 2016

J Tissue Eng Regen Med

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Cell‐based therapies could potentially restore the biomechanical function and enhance the self‐repair capacity of annulus fibrosus (AF) tissue. However, choosing a suitable cell source and scaffold design are still key challenges. In this study, we assessed the in vitro ability of human adipose stem cells (hASCs), an easily available cell source to produce AF‐like matrix in novel AF‐mimetic designed scaffolds based on poly(trimethylene carbonate) and built by stereolithography. To facilitate efficient differentiation of hASCs towards AF tissue, we tested different culture medium compositions and cell seeding techniques. This is the first study to report that medium supplementation with transforming growth factor (TGF)‐β3 is essential to support AF differentiation of hASCs while TGF‐β1 has negligible effect after 21 days of culture. Fibrin gel seeding resulted in superior cell distribution, proliferation and AF‐like matrix production of hASCs compared to direct and micromass seeding under TGF‐β3 stimulation. Not only the production of sulphated glycosaminoglycans (sGAG) and collagen was significantly upregulated, but the formed collagen was also oriented and aligned into bundles within the designed pore channels. The differentiated hASCs seeded with fibrin gel were also found to have a comparable sGAG:collagen ratio and gene expression profile as native AF cells demonstrating the high potential of this strategy in AF repair. Copyright © 2016 John Wiley & Sons, Ltd.

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Section: Discussionmentioning

confidence: 86%

Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

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Differentiation of adipose stem cells seeded towards annulus fibrosus cells on a designed poly(trimethylene carbonate) scaffold prepared by stereolithography

Blanquer

Gebraad

Miettinen

et al. 2016

J Tissue Eng Regen Med

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“…Surgical procedures were performed as described in previous studies [21, 22]. Briefly, animals were anesthetized by intraperitoneal injection of ketamine chloride (80 mg/kg, Imalgene, Merial) and medetomidine hydrochloride (1 mg/kg, Domitor, Pfizer) and surgeries were performed under a laminar flow hood in sterile conditions.…”

Section: Methodsmentioning

confidence: 99%

Interstitial Perfusion Culture with Specific Soluble Factors Inhibits Type I Collagen Production from Human Osteoarthritic Chondrocytes in Clinical-Grade Collagen Sponges

Mayer¹,

Lopa²,

Talò³

et al. 2016

PLoS ONE

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Articular cartilage has poor healing ability and cartilage injuries often evolve to osteoarthritis. Cell-based strategies aiming to engineer cartilaginous tissue through the combination of biocompatible scaffolds and articular chondrocytes represent an alternative to standard surgical techniques. In this context, perfusion bioreactors have been introduced to enhance cellular access to oxygen and nutrients, hence overcoming the limitations of static culture and improving matrix deposition. Here, we combined an optimized cocktail of soluble factors, the BIT (BMP-2, Insulin, Thyroxin), and clinical-grade collagen sponges with a bidirectional perfusion bioreactor, namely the oscillating perfusion bioreactor (OPB), to engineer in vitro articular cartilage by human articular chondrocytes (HACs) obtained from osteoarthritic patients. After amplification, HACs were seeded and cultivated in collagen sponges either in static or dynamic conditions. Chondrocyte phenotype and the nature of the matrix synthesized by HACs were assessed using western blotting and immunohistochemistry analyses. Finally, the stability of the cartilaginous tissue produced by HACs was evaluated in vivo by subcutaneous implantation in nude mice. Our results showed that perfusion improved the distribution and quality of cartilaginous matrix deposited within the sponges, compared to static conditions. Specifically, dynamic culture in the OPB, in combination with the BIT cocktail, resulted in the homogeneous production of extracellular matrix rich in type II collagen. Remarkably, the production of type I collagen, a marker of fibrous tissues, was also inhibited, indicating that the association of the OPB with the BIT cocktail limits fibrocartilage formation, favoring the reconstruction of hyaline cartilage.

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“…Colombini et al in 2015 performed an in vitro characterization of annulus fibrosus cells in collagen-enriched fibrin gels (Colombini et al, 2015). Specifically, they seeded 600,000 human annulus fibrosus cells/scaffold onto either fibrin gels or collagen-enriched fibrin gels.…”

Section: Other Fibrocartilage: Knee Meniscus and Annulus Fibrosusmentioning

confidence: 99%

A review of in-vitro fibrocartilage tissue engineered therapies with a focus on the temporomandibular joint

Lowe

Almarza

2017

Archives of Oral Biology

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The inability of fibrocartilage, specifically the temporomandibular joint (TMJ) disc, to regenerate and remodel following injury presents a unique problem for clinicians. Tissue engineering then offers a potential regenerative therapy. In vitro testing provides a valuable screening tool for potential tissue engineered solutions. The conclusions drawn for TMJ in vitro research were compared against state of the art fibrocartilage studies in the knee meniscus, and annulus fibrosus of the intervertebral disc (IVD). For TMJ disc regeneration, in vitro tissue engineered approaches, focused on cellular therapies with fibrochondrocytes, have displayed an inability to produce enough collagen, as well as an inability to recapitulate native mechanical properties. Biomaterial approaches have recapitulated the native properties of the TMJ disc, but their in vivo efficacy has yet to be determined. By comparison, the knee meniscus field is the most progressive in the use of stem cells as a cell source. The knee meniscus field has moved away from measuring mechanical properties, and are instead more focused on biochemistry and gene expression. IVD studies mainly use electrospun scaffolds, and have produced the best success in mechanical properties. The TMJ field, in comparison to knee meniscus and IVD, needs to employ stem cell therapies, new biomaterials and manufacturing techniques, and cutting edge molecular assays, in future in vitro approaches to screen for viable technologies to move to in vivo studies.

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In VitroCharacterization andIn VivoBehavior of Human Nucleus Pulposus and Annulus Fibrosus Cells in Clinical-Grade Fibrin and Collagen-Enriched Fibrin Gels

Cited by 21 publications

References 32 publications

Differentiation of adipose stem cells seeded towards annulus fibrosus cells on a designed poly(trimethylene carbonate) scaffold prepared by stereolithography

Differentiation of adipose stem cells seeded towards annulus fibrosus cells on a designed poly(trimethylene carbonate) scaffold prepared by stereolithography

Interstitial Perfusion Culture with Specific Soluble Factors Inhibits Type I Collagen Production from Human Osteoarthritic Chondrocytes in Clinical-Grade Collagen Sponges

A review of in-vitro fibrocartilage tissue engineered therapies with a focus on the temporomandibular joint

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