Collagen and Proteoglycan Production by Bovine Fetal and Adult Chondrocytes Under Low Levels of Calcium and Zinc Ions

Koyano, Yasuhiko; Hejna, Michael J.; Flechtenmacher, Johannes; Schmid, Thomas; Thonar, Eugene J.‐M.A.; Mollenhauer, Jürgen

doi:10.3109/03008209609000700

Cited by 22 publications

(27 citation statements)

References 36 publications

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“…Collagen production was reported to be the lowest at [Ca 2+ ] = 1 to 2mM in a chondrocyte-monolayer study, and an increase of [Ca 2+ ] from 2mM stimulated collagen production [12]. Other studies show that a lower calcium level can stimulate the Type X collagen synthesis and suppress the Type I collagen synthesis while leaving the Type II collagen unchanged [46,47].…”

Section: Discussionmentioning

confidence: 98%

“…These ions bind to proteoglycans by electrostatic force to maintain structure and function of the tissue [11]. Thus calcium metabolism and the changes in calcium ion concentration are important during the cartilage development and under pathological conditions [12]. In particular, the calcium level in synovial fluid of human knee joints has been reported to be 4mM or higher [13], while (inexplicably) most previous studies have been performed in culture medium with a [Ca 2+ ] of 1.8mM.…”

Section: Introductionmentioning

confidence: 94%

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Calcium Concentration Effects on the Mechanical and Biochemical Properties of Chondrocyte-Alginate Constructs

et al. 2008

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Alginate gel crosslinked by calcium ions (Ca 2+ ) has been widely used in cartilage tissue engineering. However, most studies have been largely performed in vitro in medium with a calcium concentration ([Ca 2+ ]) of 1.8mM, while the calcium level in the synovial fluid of the human knee joints, for example, has been reported to be 4mM or even higher. To simulate the synovial environment, the two studies in this paper were designed to investigate how the alginate scaffold alone, as well as the chondrocytes seeded alginate gel responds to variations in medium [Ca 2+ ]. In Study A, the mechanical properties of 2% alginate hydrogel were tested in 0.15M NaCl and various [Ca 2+ ] (1.0mM, 1.8mM, and 4mM). In Study B, primary bovine chondrocytes was seeded in alginate gel, and biochemical contents and mechanical properties were determined after incubation for 28 days in three [Ca 2+ ] (1.8mM, 4mM, and 8mM). For both studies, it was found that the magnitude of the complex shear modulus (|G*|) at 1Hz doubled and the corresponding phase angle shift angle (δ) increased > 2° as a result of the approximate 4-fold change in [Ca 2+ ]. At high [Ca 2+ ], the chondrocyte glycosaminogylcan (GAG) production inside the chondrocyte-alginate constructs was suppressed significantly. This is likely due to a decrease in the porosity of the chondrocyte-alginate constructs as a result of compaction in structure caused by an increased crosslinking density with [Ca 2+ ]. These may be important considerations in the eventual successful implementation of cartilage tissueengineered constructs in the clinical setting.

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Section: Discussionmentioning

confidence: 98%

Section: Introductionmentioning

confidence: 94%

Calcium Concentration Effects on the Mechanical and Biochemical Properties of Chondrocyte-Alginate Constructs

et al. 2008

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“…CaCl 2 levels have been shown to be important in maintaining the cartilage phenotype. 48,49 CaCl 2 was therefore removed from the cultures for the last 14 days and gels maintained stability, which might be attributed to the high cell proliferation and matrix synthesis.…”

Section: Discussionmentioning

confidence: 99%

Chondrocyte Culture in Three Dimensional Alginate Sulfate Hydrogels Promotes Proliferation While Maintaining Expression of Chondrogenic Markers

Mhanna

Kashyap

Palazzolo

et al. 2014

Tissue Engineering Part A

103

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The loss of expression of chondrogenic markers during monolayer expansion remains a stumbling block for cell-based treatment of cartilage lesions. Here, we introduce sulfated alginate hydrogels as a cartilage biomimetic biomaterial that induces cell proliferation while maintaining the chondrogenic phenotype of encapsulated chondrocytes. Hydroxyl groups of alginate were converted to sulfates by incubation with sulfur trioxidepyridine complex (SO 3 /pyridine), yielding a sulfated material cross-linkable with calcium chloride. Passage 3 bovine chondrocytes were encapsulated in alginate and alginate sulfate hydrogels for up to 35 days. Cell proliferation was five-fold higher in alginate sulfate compared with alginate ( p = 0.038). Blocking beta1 integrins in chondrocytes within alginate sulfate hydrogels significantly inhibited proliferation ( p = 0.002). Sulfated alginate increased the RhoA activity of chondrocytes compared with unmodified alginate, an increase that was blocked by b1 blocking antibodies ( p = 0.017). Expression and synthesis of type II collagen, type I collagen, and proteoglycan was not significantly affected by the encapsulation material evidenced by quantitative reverse transcription polymerase chain reaction (qRT-PCR) and immunohistochemistry. Alginate sulfate constructs showed an opaque appearance in culture, whereas the unmodified alginate samples remained translucent. In conclusion, alginate sulfate provides a three dimensional microenvironment that promotes both chondrocyte proliferation and maintenance of the chondrogenic phenotype and represents an important advance for chondrocyte-based cartilage repair therapies providing a material in which cell expansion can be done in situ.

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“…On the other hand, it was reported that lowering molecular weight. In contrast, pretreatment of anti-cFos antibody did not reduce the signals of DNA-binding the calcium concentration of the culture medium did not alter the DNA synthesis [18], and that after confluence proteins bound to AP-1 sequences ( fig. 14).…”

Section: Bmentioning

confidence: 98%

A Low Calcium Environment Enhances AP-1 Transcription Factor-Mediated Gene Expression in the Development of Osteoblastic MC3T3-E1 Cells

Deyama

Deyama²,

Matsumoto³

et al. 1999

Mineral Electrolyte Metab

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Animals fed a low calcium diet develop hypocalcemia and osteoporotic bone. Earlier we conjectured that a low calcium environment might be one of the factors causing abnormalities in hard tissues. Osteoblastic MC3T3-E1 cells (E1 cells) undergo a process of proliferation and differentiation and then produce small mineralized nodules. In this study, we examined the effects of a low calcium environment on osteoblast-like cells cultured with 10% fetal bovine serum and ascorbic acid. Under the culture condition, nodules with characteristics of normal bone appeared by day 30 regardless of the calcium conditions. However, the low calcium environment enhanced the mRNA expressions of c-fos, c-jun and osteocalcin, a specific marker of the osteoblast phenotype. And the exposure to the low calcium medium inhibited the formation of bone nodules. We further studied the differential expressions of c-fos and c-jun in relation to their responses to serum as a function of phenotypic development in the low calcium environment. Both c-fos and c-jun expressions were highly activated by treatment with epidermal growth factor (EGF), but the magnitude of activation was significantly larger under the low calcium condition than the normal condition at each stage. In addition, DNA-binding activities of activating protein-1 (AP-1), Fos/Jun family dimers, were also accelerated by EGF treatment in the low calcium environment. Our findings suggested that osteocalcin, a bone formation marker, c-fos and c-jun genes, and family protein products (AP-1) interacted to restore the normal cell function which deteriorated in the low calcium environment.

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Collagen and Proteoglycan Production by Bovine Fetal and Adult Chondrocytes Under Low Levels of Calcium and Zinc Ions

Cited by 22 publications

References 36 publications

Calcium Concentration Effects on the Mechanical and Biochemical Properties of Chondrocyte-Alginate Constructs

Calcium Concentration Effects on the Mechanical and Biochemical Properties of Chondrocyte-Alginate Constructs

Chondrocyte Culture in Three Dimensional Alginate Sulfate Hydrogels Promotes Proliferation While Maintaining Expression of Chondrogenic Markers

A Low Calcium Environment Enhances AP-1 Transcription Factor-Mediated Gene Expression in the Development of Osteoblastic MC3T3-E1 Cells

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