Effect of different ascorbate supplementations on in vitro cartilage formation in porcine high-density pellet cultures

Ibold, Yvonne; Lübke, Carsten; Pelz, S; Augst, H.; Kaps, Christian; Ringe, Jochen; Sittinger, Michael

doi:10.1016/j.tice.2008.12.001

Cited by 21 publications

(21 citation statements)

References 55 publications

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“…11,29 As presented, our porcine pellet cultures showed a good quality of ECM comparable to humans. A high ECM content of collagen Type II and proteoglycans like aggrecan is typical for differentiated human chondrocytes in vitro and in vivo.…”

Section: Porcine 3d Pellet Culturessupporting

confidence: 62%

Tissue‐engineered cartilage of porcine and human origin as in vitro test system in arthritis research

Göhring

Lübke

Andreas

et al. 2010

Biotechnology Progress

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The increasing prevalence of cartilage destruction during arthritis has entailed an intensified amount for in vitro cartilage models to analyze pathophysiological processes and to screen for antirheumatic drugs. Tissue engineering offers the opportunity to establish highly organized 3D cell cultures facilitating the formation of in vitro models that reflect the human situation. We report the comparison of porcine chondrocyte pellet and alginate bead cultures as model systems for human cartilage and the further development into a human system that was applied in an arthritis model. In porcine pellet and alginate cultures, formation of cartilage matrix similar to human matrix was verified by histology and PCR. As alginate beads could be cultivated batch-wise in one well of a multiwell plate, we further developed this setting into a human system. In contrast, each pellet had to be cultivated individually in one well of a multiwell plate, which is time consuming. Following stimulation of human chondrocyte alginate cultures with conditioned media from human synovial fibroblasts derived from arthritis patients, microarray analysis verified the induction of genes related to cartilage destruction (like MMP10, -12) and inflammation (like IL6, -8 and chemokines). Several genes are coding for proteins that are members of inflammatory and catabolic pathways. Belonging to the most affected pathways, we identified the focal adhesion, cytokine-cytokine receptor interaction, ECM-receptor signalling, Jak-STAT signalling, and toll-like receptor signalling pathways, all relevant in arthritis. Therefore, we demonstrate that engineered cartilage of porcine and human origin represents a powerful in vitro model for cartilage in vivo.

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Section: Porcine 3d Pellet Culturessupporting

confidence: 62%

Tissue‐engineered cartilage of porcine and human origin as in vitro test system in arthritis research

Göhring

Lübke

Andreas

et al. 2010

Biotechnology Progress

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“…100 μM L-ascorbate-2-phosphate (we used 50 μM) resulted in decreased COL-I expression at the beginning of the culture and slightly increased COL-II and aggrecan expression (Ibold et al, 2009). Another study on mesenchymal stem cells revealed that ascorbate supplementation (0-500 μM) had almost no effect on growth and proliferation until passage 3 (Choi et al, 2008).…”

Section: S Pauly Et Al Human Rotator Cuff Tendon Cell Culturesmentioning

confidence: 99%

Characterization of tendon cell cultures of the human rotator cuff

Pauly

Klatte²,

Strobel³

et al. 2010

eCM

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Rotator cuff tears are common soft tissue injuries of the musculoskeletal system that heal by formation of repair tissue and may lead to high retear rates and joint dysfunction. In particular, tissue from chronic, large tendon tears is of such degenerative nature that it may be prone to retear after surgical repair. Besides several biomechanical approaches, biologically based strategies such as application of growth factors may be promising for increasing cell activity and production of extracellular tendon matrix at the tendon-to-bone unit. As a precondition for subsequent experimental growth factor application, the aim of the present study was to establish and characterize a human rotator cuff tendon cell culture.Long head biceps (LHB)-and supraspinatus muscle (SSP)-tendon samples from donor patients undergoing shoulder surgery were cultivated and examined at the RNA level for expression of collagen type-I, -II and -III, biglycan, decorin, tenascin-C, aggrecan, osteocalcin, tenomodulin and scleraxis (by Real-time PCR). Finally, results were compared to chondrocytes and osteoblasts as control cells.An expression pattern was found which may reflect a human rotator cuff tenocyte-like cell culture. Both SSP and LHB tenocyte-like cells differed from chondrocyte cell cultures in terms of reduced expression of collagen type-II (p≤0.05) and decorin while higher levels of collagen type-I were seen (p≤0.05). With respect to osteoblasts, tenocytelike cells expressed lower levels of osteocalcin (p≤0.05) as well as tenascin C, biglycan and collagen type-III. Expression of scleraxis, tenomodulin and aggrecan was similar between all cell types.This study represents a characterization of tenocytelike cells from the human rotator cuff as close as possible. It helps analyzing their biological properties and allows further studies to improve production of tendon matrix and osteofibroblastic integration at the tendon-bone unit following tendon repair.

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“…Ascorbate levels have been modulated over narrow ranges in shortterm pellet culture, 34 and comparisons have been made between chondrocyte-seeded agarose constructs that have been cultured in the presence or absence of ascorbate, 33,35 yet the effects of its concentration over a broad range on tissue-engineered constructs remain to be uncovered.…”

Section: Introductionmentioning

confidence: 99%

Insulin, Ascorbate, and Glucose Have a Much Greater Influence Than Transferrin and Selenous Acid on theIn VitroGrowth of Engineered Cartilage in Chondrogenic Media

Cigáň

Nims

Albro

et al. 2013

Tissue Engineering Part A

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The primary goal of this study was to characterize the response of chondrocyte-seeded agarose constructs to varying concentrations of several key nutrients in a chondrogenic medium, within the overall context of optimizing the key nutrients and the placement of nutrient channels for successful growth of cartilage tissue constructs large enough to be clinically relevant in the treatment of osteoarthritis (OA). To this end, chondrocyte-agarose constructs (ø4 · 2.34 mm, 30 · 10 6 cells/mL) were subjected to varying supplementation levels of insulin (0· to 30· relative to standard supplementation), transferrin (0· to 30·), selenous acid (0· to 10·), ascorbate (0· to 30· ), and glucose (0· to 3·). The quality of resulting engineered tissue constructs was evaluated by their compressive modulus (E -Y ), tensile modulus (E +Y ), hydraulic permeability (k), and content of sulfated glycosaminoglycans (sGAG) and collagen (COL); DNA content was also quantified. Three control groups from two separate castings of constructs (1· concentrations of all medium constituents) were used. After 42 days of culture, values in each of these controls were, respectively, E -Y = 518 -78, 401 -113, 236 -67 kPa; E +Y = 1420 -430, 1140 -490, 1240 -280 kPa; k = 2.3 -0.8 · 10 -3 , 5.4 -7.0 · 10 -3 , 3.3 -1.3 · 10 -3 mm 4 /N$s; sGAG = 7.8 -0.3, 6.3 -0.4, 4.1 -0.5%/ww; COL = 1.3 -0.2, 1.1 -0.3, 1.4 -0.4%/ww; and DNA = 11.5 -2.2, 12.1 -0.6, 5.2 -2.8 mg/disk. The presence of insulin and ascorbate was essential, but their concentrations may drop as low as 0.3· without detrimental effects on any of the measured properties; excessive supplementation of ascorbate (up to 30·) was detrimental to E -Y , and 30· insulin was detrimental to both E +Y and E -Y . The presence of glucose was similarly essential, and matrix elaboration was significantly dependent on its concentration ( p < 10 -6

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Effect of different ascorbate supplementations on in vitro cartilage formation in porcine high-density pellet cultures

Cited by 21 publications

References 55 publications

Tissue‐engineered cartilage of porcine and human origin as in vitro test system in arthritis research

Tissue‐engineered cartilage of porcine and human origin as in vitro test system in arthritis research

Characterization of tendon cell cultures of the human rotator cuff

Insulin, Ascorbate, and Glucose Have a Much Greater Influence Than Transferrin and Selenous Acid on theIn VitroGrowth of Engineered Cartilage in Chondrogenic Media

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