Phenotypic Drift in Human Tenocyte Culture

Yao, Lishuang; Bestwick, Charles; Bestwick, L. A.; Maffulli, Nicola; Aspden, Richard M.

doi:10.1089/ten.2006.12.1843

Cited by 228 publications

(163 citation statements)

References 40 publications

(60 reference statements)

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“…Another important fact was the short distance between isolation and use of the tenocytes (2-4 days). The dediVerentiation of tenocytes due to the long cultivation periods or 2-D cultivation is a severe problem (Stoll et al 2010;Yao et al 2006).…”

Section: Discussionmentioning

confidence: 99%

Platelet-released growth factors can accelerate tenocyte proliferation and activate the anti-oxidant response element

Tohidnezhad

Varoga

Wruck

et al. 2011

Histochem Cell Biol

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Little is know about the pathophysiology of acute and degenerative tendon injuries. Although most lesions are uncomplicated, treatment is long and unsatisfactory in a considerable number of cases. Besides the common growth factors that were shown to be relevant for tendon integrity more recently protection against oxidative stress was shown to promote tendon healing. To improve tendon regeneration, many have advocated the use of platelet-rich plasma (PRP), a thrombocyte concentrate that can serve as an autologous source of growth factors. In this study, we investigated the effect of platelet-released growth factors (PRGF) on tenocytes. Tenocytes were isolated from the Achilles tendon of postnatal rats. Tenocyte cell cultures were stimulated with PRGF. We used a CyQuant assay and WST assay to analyse tendon cell growth and viability in different concentrations of PRGF. Migration and proliferation of cells grown in PRGF were assessed by a scratch test. A dual-luciferase assay was used to demonstrate the activation of the anti-oxidant response element (ARE) in tenocytes. A positive effect of PRGF could be shown on tendon cell growth and migratory capacity. PRGF activated the Nrf2-ARE pathway in a dose-dependent manner. Here, we provide evidence of a biological effect of PRGF on tenocytes by the promotion of tenocyte growth and activation of the Nrf2-ARE pathway. This is a novel aspect of the action of platelet concentrates on tendon growth.

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Section: Discussionmentioning

confidence: 99%

Platelet-released growth factors can accelerate tenocyte proliferation and activate the anti-oxidant response element

Tohidnezhad

Varoga

Wruck

et al. 2011

Histochem Cell Biol

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“…Each cell source has its limitations including inferior proliferation, differentiation and collagen production abilities of tenocytes [12,13,14,15], risk of teratoma formation of ESCs as well as risk of ectopic bone [3] and tumor induction [16] of BMSCs when using a certain scaffold. For this reason, lineage direction of stem cells is recommended prior to their use in cell-based therapies [9,10].…”

Section: Introductionmentioning

confidence: 99%

Tendon Derived Stem Cells Promote Platelet-Rich Plasma Healing in Collagenase-Induced Rat Achilles Tendinopathy

Chen

Liu

Tang

et al. 2014

Cell Physiol Biochem

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Background/Aims: Tendon injuries are common, difficult to cure and usually healed with fibrosis and scar tissue. The aim of this study was to evaluate tendon derived stem cells (TDSCs) and platelet rich plasma (PRP) in the treatment of collagenase induced Achilles tendinopathy in rat. Methods: Four and 8 weeks (n=18) after TDSCs, PRP, PRP with TDSC or PBS (control) injection into collagenase or saline (sham) injected rat Achilles tendon, tendon tissue was harvested and tendon quality was evaluated by histology and biomechanical testing. TDSCs were cultured and treated by 10% PRP, and the FAK/ERK1/2 signaling pathway and tenocyte-related genes were detected by western blot analysis. Results: Compared to the control, PRP treatment resulted in better healing of injured tendons with improved histological outcomes and biomechanical functions. The addition of TDSCs to PRP treatment significantly enhanced the effects of PRP treatment alone. TDSC injection alone had little effect on tendon healing. PRP and PRP with TDSC treatments of collagenase induced tendon injuries also increased the mRNA and protein expression of tenocyte-related genes (type I collagen, SCX, Tenascin C) and activated the focal adhesion kinase (FAK) and extracellular-regulated kinase (ERK) 1/2 signaling pathways. Treatment of TDSCs in vitro with 10% PRP significantly increased the phosphorylation levels of FAK and ERK1/2 and the protein levels of tenocyte-related genes (Col I, SCX and Tenascin C). Inhibition of the FAK and ERK1/2 signaling pathways abolished the effect of PRP. Conclusion: This study concludes that PRP combined with TDSCs is potentially effective for the treatment of tendinopathy. The PRP induced, FAK and ERK1/2 dependent activation of tenocyte related genes in TDSCs in vitro suggests that the beneficial healing effect of the PRP with TDSC combination might occur by means of an improved TDSC differentiation toward the tenocyte lineage. Thus, a PRP with TDSC combination therapy may be clinically useful.

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“…Nevertheless, cells did appear to take longer to reach confluence in later passages and were less packed in by Passage 5, which was consistent with the findings of decreased cell confluence number through passages observed for Achille's Tendon fibroblasts. 18 There are still some limitations of this study as the protein production was not quantified. Future studies involving longer culture periods and higher number of cells would be required to produce enough matrix to detect meaningful differences.…”

Section: Discussionmentioning

confidence: 97%

“…While these in vitro studies have shed light on the mechanisms of the difference in their healing capabilities, the results have not been based on a single cell passage of choice, when recent evidence suggests that passage has a great impact on cell behavior. 2,18 Recently, dramatic effects of cell passage have been documented in Achilles' tendon fibroblasts 18 where it was observed that cell density at confluency and Collagen Type I (main matrix constituent of tendon) content both decrease with passage while Collagen Type III increases. 18 Cartilage chondrocytes have similar behavior to Achilles' tendon fibroblasts as it was observed that the gene expression of their main matrix constituent (Collagen Type II) also disappeared just after a couple of passages.…”

Section: Introductionmentioning

confidence: 99%

“…2,18 Recently, dramatic effects of cell passage have been documented in Achilles' tendon fibroblasts 18 where it was observed that cell density at confluency and Collagen Type I (main matrix constituent of tendon) content both decrease with passage while Collagen Type III increases. 18 Cartilage chondrocytes have similar behavior to Achilles' tendon fibroblasts as it was observed that the gene expression of their main matrix constituent (Collagen Type II) also disappeared just after a couple of passages. 2 It should be noted that the effect of cell passage on the gene expression of matrix proteins has not been well characterized for fibroblasts from knee ligaments and tendons.…”

Section: Introductionmentioning

confidence: 99%

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Changes in Gene Expression of Matrix Constituents with Respect to Passage of Ligament and Tendon Fibroblasts

2008

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Trauma to the knee joint often results in injury to one or more supporting soft tissue structures, such as the medial collateral (MCL) and anterior cruciate (ACL) ligaments. Also, a portion of the patellar tendon (PT) is frequently used as a replacement graft for the ACL, resulting in a PT defect. The healing responses of these tissues are dramatically different and range from spontaneous healing to little or no healing. Studies have suggested that native cell behavior could be responsible for differences in healing potential. However, it is difficult to make comparisons as the reported results are based on different cellular passages which could have a dramatic effect on their potential to form healing tissues. Therefore, the objective of this study was to quantify the gene expression of collagen and other matrix constituents of fibroblasts from the MCL, ACL, and PT to document how they change with cell passage. We hypothesized that MCL fibroblasts would possess higher potential for matrix production through passages than ACL and PT cells because the MCL mounts a robust healing response unlike the ACL and PT. These differences in matrix expression would be dependent on passage because at earlier passages all cells would mostly be proliferating while at later passages they would tend to become senescent. Cells were isolated from the MCL, ACL, and PT of three rats and passaged a total of five times (Passage 1 to Passage 5). Using real time RT-PCR, expression of all genes of interest (Collagen Type I (ligament/tendon's main matrix constituent), Collagen Type III, Fibronectin, Metalloprotease-13 [MMP-13], and Tissue Inhibitor of Metallopreotease-1 [TIMP-1]) were quantitatively assessed. It was found that cell number for all three fibroblast types remained high from Passage 1 to Passage 5. There was a statistically significant increase in Collagen Type I of rat MCL fibroblasts throughout passage (p < 0.05). This was evident in the higher relative abundance (to GAPDH) at Passages 3 and 4 (14.5 +/- 2.2 fold and 15.3 +/- 6.9 fold, respectively) than at Passage 1 (3.3 +/- 2.6 fold) (p < 0.05). On the other hand, Collagen Type I expression for ACL and PT fibroblasts were lower than that of MCL fibroblasts and remained at 2.5 +/- 2.0 fold and 1.7 +/- 0.8 fold, respectively. Interestingly, the gene expressions of Collagen Type III, Fibronectin, MMP-13, and TIMP-1 for MCL, ACL, and PT fibroblasts were all relatively constant throughout passage and were not significantly different from one another. The findings of this study indicate that passage does affect the Collagen Type I gene expression of rat MCL fibroblasts and further show that for in vitro ligament tissue engineering efforts, MCL fibroblasts have a more robust potential for ligament remodeling and repair due to the increase in collagen gene expression.

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Phenotypic Drift in Human Tenocyte Culture

Cited by 228 publications

References 40 publications

Platelet-released growth factors can accelerate tenocyte proliferation and activate the anti-oxidant response element

Platelet-released growth factors can accelerate tenocyte proliferation and activate the anti-oxidant response element

Tendon Derived Stem Cells Promote Platelet-Rich Plasma Healing in Collagenase-Induced Rat Achilles Tendinopathy

Changes in Gene Expression of Matrix Constituents with Respect to Passage of Ligament and Tendon Fibroblasts

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