Comparative chondrogenesis of human cells in a 3D integrated experimental–computational mechanobiology model

Raimondi, Manuela Teresa; Bonacina, E; Candiani, Gabriele; Laganà, Matteo; Rolando, Elena; Talò, Giuseppe; Pezzoli, Daniele; D’Anchise, Roberto; Pietrabissa, Riccardo; Moretti, Matteo

doi:10.1007/s10237-010-0232-8

Cited by 18 publications

(13 citation statements)

References 28 publications

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“…This result is particularly striking considering the pathological origin of the articular chondrocytes used in this study, since the ability of HAC from OA joints to redifferentiate has been found to be impaired in other culture conditions [29]. Our findings are consistent with other studies showing that perfusion inhibits type I collagen expression in human chondrocytes cultured in pellets [30] and in cartilage explants [31]. The molecular mechanisms underlying the inhibitory effect of perfusion on type I collagen in chondrocytes are still unknown.…”

Section: Discussionsupporting

confidence: 90%

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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Section: Discussionsupporting

confidence: 90%

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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“…The stereo microscope setup (Fig. 3(b)), as implemented in this work, provides an overall field of view of 6.14 × 4.07 mm 2 , and has a spatial resolution of 1.2 μm/pixel. The procedure for measurement of surface topography is given as follows.…”

Section: Fluorescent Stereo Microscopy System and Validationmentioning

confidence: 99%

“…For example, the investigation in mechanobiology will benefit from a technique to measure the 3D topography and deformation of biofilms and tissues [1][2][3]. Traditional 3D profilometry techniques can hardly be employed for the surface profile measurement for biological cells or soft materials at the microscale.…”

Section: Introductionmentioning

confidence: 99%

Fluorescent stereo microscopy for 3D surface profilometry and deformation mapping

Hu¹,

Luo²,

Du³

et al. 2013

Opt. Express

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Abstract:Recently, mechanobiology has received increased attention. For investigation of biofilm and cellular tissue, measurements of the surface topography and deformation in real-time are a pre-requisite for understanding the growth mechanisms. In this paper, a novel threedimensional (3D) fluorescent microscopic method for surface profilometry and deformation measurements is developed. In this technique a pair of cameras are connected to a binocular fluorescent microscope to acquire micrographs from two different viewing angles of a sample surface doped or sprayed with fluorescent microparticles. Digital image correlation technique is used to search for matching points in the pairing fluorescence micrographs. After calibration of the system, the 3D surface topography is reconstructed from the pair of planar images. When the deformed surface topography is compared with undeformed topography using fluorescent microparticles for movement tracking of individual material points, the full field deformation of the surface is determined. The technique is demonstrated on topography measurement of a biofilm, and also on surface deformation measurement of the biofilm during growth. The use of 3D imaging of the fluorescent microparticles eliminates the formation of bright parts in an image caused by specular reflections. The technique is appropriate for non-contact, full-field and real-time 3D surface profilometry and deformation measurements of materials and structures at the microscale. Kormos, B. P. Griffith, H. Koyanagi, and J. F. Antaki, "High-resolution fluorescent particle-tracking flow visualization within an intraventricular axial flow left ventricular assist device," Artif. Organs 20(5), 534-540 (1996). 7. J. Sakakibara, K. Hishida, and M. Maeda, "Vortex structure and heat transfer in the stagnation region of an impinging plane jet (simultaneous measurements of velocity and temperature fields by digital particle image velocimetry and laser-induced fluorescence)," Int. J. Heat Mass Tran. 40(13), 3163-3176 (1997). 278-288 (2004). 38. M. A. Sutton, X. Ke, S. M. Lessner, M. Goldbach, M. Yost, F. Zhao, and H. W. Schreier, "Strain field measurements on mouse carotid arteries using microscopic three-dimensional digital image correlation," J.

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“…Nonetheless, the poor ability in delivering the genetic material to the target cells as compared to viruses has hardly limited the employment of non-viral gene delivery vectors in clinical trials thus far. In addition to therapeutic applications, they have become a highly widespread technique in genomics (16) and, in general, in all bio-laboratories in fundamental and applied research (17)(18)(19).…”

Section: Non-viral Vectorsmentioning

confidence: 99%

We still have a Long Way to go to Effectively Deliver Genes!

Pezzoli

Chiesa

Nardo

et al. 2012

Journal of Applied Biomaterials & Functional Materials

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Gene therapy is emerging as a revolutionary alternative to conventional therapeutic approaches. However, its clinical application is still hampered by the lack of safe and effective gene delivery techniques. Among the plethora of diverse approaches used to ferry nucleic acids into target cells, non-viral vectors represent promising and safer alternatives to viruses and physical techniques. Both cationic lipids and polymers spontaneously wrap and shrink the genetic material in complexes named lipoplexes and polyplexes, respectively, thereby protecting it and shielding its negative charges. The development of non-viral vectors commenced more than two decades ago. Since then, some major classes of interesting molecules have been identified and modified to optimize their properties. However, the way towards the final goal of gene delivery, i.e. protein expression or gene silencing, is filled with obstacles and current non-viral carriers still have concerns about their overall efficiency. We strongly believe that the future of non-viral gene delivery relies on the development of multifunctional vectors specifically tailored with diverse functionalities that act more like viruses. Although these vectors are still a long way from clinical practice they are the ideal platform to effectively shuttle the genetic material to target cells in a safe and controlled way. In this review, after briefly introducing the basis of gene delivery and therapeutic applications we discuss the main polymeric and lipidic vectors utilized for gene delivery, focusing on the strategies adopted to overcome the major weaknesses inherent to their still limited activity, on the way towards ideal multifunctional vectors.

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Comparative chondrogenesis of human cells in a 3D integrated experimental–computational mechanobiology model

Cited by 18 publications

References 28 publications

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

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

Fluorescent stereo microscopy for 3D surface profilometry and deformation mapping

We still have a Long Way to go to Effectively Deliver Genes!

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