Effect of Extracellular Matrix Protein on the Rate of Proteoglycan Synthesis in Rabbit Intervertebral Disc Cells

Periostin, an extracellular matrix protein, is widely expressed in a variety of tissues and cells. It has many biological functions and is related to many diseases: for example, it promotes cell proliferation and differentiation in osteoblasts, which are closely related to osteoporosis, and mediates cell senescence and apoptosis in chondrocytes, which are involved in osteoarthritis. Furthermore, it also plays an important role in mediating inflammation and reconstruction during bronchial asthma, as well as in promoting bone development, reconstruction, repair, and strength. Therefore, periostin has been explored as a potential biomarker for various diseases. Recently, periostin has also been found to be expressed in intervertebral disc cells as a component of the intervertebral extracellular matrix, and to play a crucial role in the maintenance and degeneration of intervertebral discs. This article reviews the biological role of periostin in bone marrow-derived mesenchymal stem cells, osteoblasts, osteoclasts, chondrocytes, and annulus fibrosus and nucleus pulposus cells, which are closely related to spinal degenerative diseases. The study of its pathophysiological effects is of great significance for the diagnosis and treatment of spinal degeneration, although additional studies are needed.

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Effect of Age and Frequency on Intervertebral Disc Cell Response to Dynamic Compression

ASME 2007 Summer Bioengineering Conference ◽

10.1115/sbc2007-176595 ◽

2007 ◽

Author(s):

Casey L. Korecki ◽

Catherine K. Kuo ◽

Rocky S. Tuan ◽

James C. Iatridis

Keyword(s):

Extracellular Matrix ◽

Intervertebral Disc ◽

Mechanical Stimulation ◽

Matrix Protein ◽

Dynamic Compression ◽

Model Systems ◽

Extracellular Matrix Protein ◽

Tissue Culture Cell ◽

Protein Accumulation

The intervertebral disc (IVD) is a unique orthopaedic tissue consisting of at least two cell types: fibroblast-like annulus fibrosus (AF) cells and chondrocyte-like nucleus pulposus (NP) cells. Culture of cells in 3D gel matrices (such as alginate or agarose), maintains the normal morphology and ECM molecule production of chondrocytes for extended periods of time and also allows the application of various forms of mechanical stimulation, such as hydrostatic or compressive loading. In vivo studies have shown IVD cells to be responsive to frequency, duration, and amplitude of mechanical load [1]. IVD literature on mechanobiology uses varying methodologies to apply dynamic loads (compression, hydrostatic forces), with different times of mechanical stimulation, differences in model systems (in vivo, tissue culture, cell culture), species, and ages, and an optimal loading protocol to stimulate extracellular matrix protein accumulation is unknown. The overall goal of this work is to evaluate the potential, and perhaps even feasibility, of mechanical stimulation for extracellular matrix (ECM) regeneration using intervertebral disc cells. Also of interest is whether cells from mature tissue are capable of serving as a potential cell source for future IVD regeneration [2,3].

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1280: Increased Susceptibility to Genitovaginal Prolapse Associated with a Polymorphism in the Promoter of the Extracellular Matrix Protein LAMC1

The Journal of Urology ◽

10.1016/s0022-5347(18)31494-0 ◽

2007 ◽

Vol 177 (4S) ◽

pp. 421-422

Author(s):

Ganka Nikolova ◽

Christian O. Twiss ◽

Hane Lee ◽

Nelson Stanley ◽

Janet Sinsheimer ◽

...

Keyword(s):

Extracellular Matrix ◽

Matrix Protein ◽

Extracellular Matrix Protein ◽

Increased Susceptibility

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Homogenous overexpression of the extracellular matrix protein Netrin-1 in a hollow fiber bioreactor

Applied Microbiology and Biotechnology ◽

10.1007/s00253-021-11438-0 ◽

2021 ◽

Author(s):

Aniel Moya-Torres ◽

Monika Gupta ◽

Fabian Heide ◽

Natalie Krahn ◽

Scott Legare ◽

...

Keyword(s):

Extracellular Matrix ◽

Hollow Fiber ◽

Mammalian Cells ◽

Matrix Protein ◽

Continuous Production ◽

Extracellular Matrix Protein ◽

Close Monitoring ◽

High Quality ◽

Biolayer Interferometry ◽

Hollow Fiber Bioreactor

Abstract The production of recombinant proteins for functional and biophysical studies, especially in the field of structural determination, still represents a challenge as high quality and quantities are needed to adequately perform experiments. This is in part solved by optimizing protein constructs and expression conditions to maximize the yields in regular flask expression systems. Still, work flow and effort can be substantial with no guarantee to obtain improvements. This study presents a combination of workflows that can be used to dramatically increase protein production and improve processing results, specifically for the extracellular matrix protein Netrin-1. This proteoglycan is an axon guidance cue which interacts with various receptors to initiate downstream signaling cascades affecting cell differentiation, proliferation, metabolism, and survival. We were able to produce large glycoprotein quantities in mammalian cells, which were engineered for protein overexpression and secretion into the media using the controlled environment provided by a hollow fiber bioreactor. Close monitoring of the internal bioreactor conditions allowed for stable production over an extended period of time. In addition to this, Netrin-1 concentrations were monitored in expression media through biolayer interferometry which allowed us to increase Netrin-1 media concentrations tenfold over our current flask systems while preserving excellent protein quality and in solution behavior. Our particular combination of genetic engineering, cell culture system, protein purification, and biophysical characterization permitted us to establish an efficient and continuous production of high-quality protein suitable for structural biology studies that can be translated to various biological systems. Key points • Hollow fiber bioreactor produces substantial yields of homogenous Netrin-1 • Biolayer interferometry allows target protein quantitation in expression media • High production yields in the bioreactor do not impair Netrin-1 proteoglycan quality Graphical abstract

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