Cyclic Mechanical Stretch Stress Increases the Growth Rate and Collagen Synthesis of Nucleus Pulposus Cells In Vitro

Matsumoto, Taihei; Kawakami, Mamoru; Kuribayashi, Koichi; Takenaka, Toru; Tamaki, Toru

doi:10.1097/00007632-199902150-00002

Cited by 91 publications

(50 citation statements)

References 24 publications

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“…The device consisted of a computer-controlled vacuum unit, a culture well plate with a flexiblepolystyrene culture well bottom coated with type I collagen (Flex I, Flexercell International Corp., McKeesport, PA, USA), and another culture well plate with a non-deformable culture well bottom made of the same materials as Flex I (Flex 11, Flexercell International Corp., McKeesport, PA, USA). Application of a vacuum provides a hemispherically downward deformation to the flexible Flex I bottom, resulting in a non-homogenous strain profile with a maximum at the Table 2 Sequences of RT-PCR primers used in the present study periphery and a geometric decline toward zero at the center or the culture well bottom [2,4,9,19,21]. The cells were placed in Flex I and Flex I1 culture well plates at a density of 1 x 10' cellslwell.…”

Section: Methodsmentioning

confidence: 99%

“…Seventy-two h after the cells were cultured, the culture medium was replaced with DMEM supplemented with 0.5'%1 FBS, 50 yglml L-ascorbic acid (Sigma Chemical, St. Louis, MO, USA), penicillin G (100 Ulml). and streptomycin (100 pglml) to keep the cells quiescent for 24 h. At the beginning of the present study, the cells in the Flex I plates were subjected to 10-s cycles of 20%) elongation and 10 s of relaxation at 37 "C in a 5% COz incubator by referring to a previously described protocol [21]. This preliminary study revealed that the cells could not survive under this condition over 96 h. The stretching protocol was, then, modified.…”

Section: Methodsmentioning

confidence: 99%

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Mechanical stretching force promotes collagen synthesis by cultured cells from human ligamentum flavum via transforming growth factor‐β1

Nakatani

Marui

Hitora

et al. 2002

Journal Orthopaedic Research

133

125

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Although mechanical stress as a result of spinal instability is known to cause hypertrophy of the ligameiitum flavum resulting in degenerative spinal canal stenosis, the mechanism of the ligament hypertrophy is not well understood. In the present study, we investigated the effect of mechanical stretching force on collagen synthesis and transforming growth factor-/3 1 (TGF-PI) production using ligament cells isolated from human ligamentum flavum in vitro. Ligamenturn flavum cells (LFCs) were isolated from human ligamenturn flavum obtained from patients who underwent lumbar spine surgery. The LFCs were subjected to a mechanical stretching force using a cominercially available stretching device that physically deformed the cells. Collagen synthesis and TGF-PI production levels in the LFCs were then examined. Notable increases were observed in the gene expressions of collagen types I, 111, and V in LFCs subjected to mechanical stretching force. The increase in collagen gene expression of LFCs was inhibited in the presence of anti-TGF-pl antibodies. Production of TGF-PI by the LFCs also increased significantly by the mechanical stretching force. Exogenous application of TGF-PI was confirmed to increase collagen synthesis of the LFCs. This data indicated that mechanical stretching force can promote TGF-Dl production by LFCs, resulting in hypertrophy of the ligament.

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Mechanical stretching force promotes collagen synthesis by cultured cells from human ligamentum flavum via transforming growth factor‐β1

Nakatani

Marui

Hitora

et al. 2002

Journal Orthopaedic Research

133

125

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“…Over the last few decades, several studies have shown that changes in the mechanical environment of the disc can result in remodeling, breakdown, and reorganization of the extracellular matrix [ 1, 7,13,[18][19][20][21][22]29,32,33,35,41,44], leading in some cases to signs of accelerated disc degeneration. Such work forms the *Corresponding author.…”

Section: Introductionmentioning

confidence: 99%

Anabolic and catabolic mRNA levels of the intervertebral disc vary with the magnitude and frequency of in vivo dynamic compression

MacLean

Lee

Alini

et al. 2004

Journal Orthopaedic Research

168

181

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The goal of this study was to characterize the anabolic and catabolic mRNA response of the disc to dynamic loading to determine if variations in the magnitude and/or frequency of loading could elicit different cellular responses. Sixty-eight Wistar rats were instrumented with an Ilizarov-type device spanning caudal disc 8-9. Seventy-two hours after surgery, animals were anesthetized and loaded at either 1 or 0.2 MPa at a frequency of 1, 0.2 or 0.01 Hz for 2 h (6 groups). The surgical control (Sham) animals underwent anesthesia with no loading. Loaded (c8-9) and internal-control discs (c6-7 and c10-11) were dissected and annulus and nucleus tissue were separately analyzed by real-time RT-PCR for levels of anabolic (collagen-1 A1 , collagen-2A1, aggrecan) and catabolic (MMP-3, MMP-13, ADAMTs-4) mRNA. In the nucleus, a frequency-dependent response was seen at 1 MPa with anabolic genes stimulated at 0.01 Hz and catabolic genes at 1 Hz. In the annulus all frequencies resulted in significant up-regulation of catabolic mRNA at 1 MPa loading. In general loading at 0.2 MPa or 0.2 Hz had little effect on gene expression. The results suggest that gene expression of the annulus appears to be more dependent on the magnitude of applied stress, while the nucleus is both magnitude-and frequency-dependent.

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“…The fine balance between synthesis of components and the breakdown of tissue determines the composition of the matrix; tissue integrity is thus dependent on the activity of the nucleus pulposus cells [53]. In vivo [20,43] and in vitro [14] environmental factors such as mechanical stress [30,34] and nutrient levels have been found to affect matrix composition, presumably by affecting macromolecule biosyntheAbstract It has long been known that very acidic conditions can be found in degenerate discs. The effect of these acid conditions on matrix turnover are, however, unknown.…”

Section: Introductionmentioning

confidence: 99%

The effect of extracellular pH on matrix turnover by cells of the bovine nucleus pulposus

Razaq

Wilkins

Urban

2003

European Spine Journal

169

164

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IntroductionNucleus pulposus cells are solely responsible for the production and maintenance of the extracellular matrix of the nucleus throughout life [31]. The cells have no direct contact with each other and rely on cell-matrix interactions and signals from the environment to maintain the function of the tissue. The cells synthesize all the matrix components [10,37,38,50] and also produce degradative enzymes including metalloproteinases (MMPs) and their inhibitors (tissue inhibitors of metalloproteinases or TIMPs), involved in matrix turnover [7,45,53]. The fine balance between synthesis of components and the breakdown of tissue determines the composition of the matrix; tissue integrity is thus dependent on the activity of the nucleus pulposus cells [53]. In vivo [20,43] and in vitro [14] environmental factors such as mechanical stress [30,34] and nutrient levels have been found to affect matrix composition, presumably by affecting macromolecule biosyntheAbstract It has long been known that very acidic conditions can be found in degenerate discs. The effect of these acid conditions on matrix turnover are, however, unknown. This study aimed to examine the effect of acidity on production of matrix components and on agents which break down the matrix in order to gain insight into the effect of pathological values of pH on matrix turnover. Cells were isolated from the nucleus of bovine discs and from bovine articular cartilage, embedded in alginate beads and cultured at pH levels maintained within the ranges seen in normal and pathological discs: pH 7.4-pH 6.3 for 48 h. Rates of sulphated glycosaminoglycan (GAG) and protein synthesis were measured, as well as rates of production of some agents involved in matrix breakdown, i.e. total and activated matrix metalloproteinases (MMPs) and their inhibitors (TIMPs). The results showed that acid conditions had a profound effect on cell matrix turnover; at pH 6.4, total production of most species measured was inhibited by more than 50% compared to production at pH 7.2; production of sulphated GAGs and of TIMP-1 fell by >90%. However production of active metalloproteinases by disc cells was relatively insensitive to pH, with activity at pH 6.3 not statistically different from that at pH 7.2. These findings indicate that exposure to acid conditions appears particularly deleterious for the disc matrix, as it inhibits the disc cells from synthesising functionally important molecules such as the sulphated GAGs but does not prevent the production of agents able to degrade matrix components. The low values of pH seen in some degenerate discs are thus likely to be involved in breakdown of the disc matrix.

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Cyclic Mechanical Stretch Stress Increases the Growth Rate and Collagen Synthesis of Nucleus Pulposus Cells In Vitro

Abstract: Mechanical stress on nucleus pulposus cells promotes the proliferation of cells and alters the properties of intervertebral disc cells. This study may reflect the adaptation of the intervertebral disc to increased motion and stress.

Cited by 91 publications

References 24 publications

Mechanical stretching force promotes collagen synthesis by cultured cells from human ligamentum flavum via transforming growth factor‐β1

Mechanical stretching force promotes collagen synthesis by cultured cells from human ligamentum flavum via transforming growth factor‐β1

Anabolic and catabolic mRNA levels of the intervertebral disc vary with the magnitude and frequency of in vivo dynamic compression

The effect of extracellular pH on matrix turnover by cells of the bovine nucleus pulposus

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