Visual Histological Grading System for the Evaluation of<i>in Vitro</i>–Generated Neocartilage

Grogan, Shawn P.; Barbero, Andrea; Winkelmann, Verena; Rieser, Franz; Fitzsimmons, James S.; O’Driscoll, Shawn W.; Martin, Ivan; Mainil‐Varlet, Pierre

doi:10.1089/ten.2006.12.2141

Cited by 193 publications

(164 citation statements)

References 39 publications

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“…Normal articular cartilage was obtained at notchplasty in patients (13,18,19, and 21 years of age) undergoing anterior cruciate ligament reconstructions (n 0 4). Unnecessary parts of loose fragments in knee OCD (15,15,15, and 21 years of age) were obtained at arthroscopy (n 0 4).…”

Section: Tissues Cells and Cell Culturementioning

confidence: 99%

“…Proteoglycans in pellets were observed with safranin O staining. Redifferentiation potential was assessed by the Bern pellet histology score (range, 0-9) [18,19]. To assess the effect of tissuederived mesenchymal stem cells (MSCs), we performed in vitro differentiation analysis for the other mesenchymal lineages.…”

Section: In Vitro Differentiation and Histological Analysesmentioning

confidence: 99%

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Comparison between normal and loose fragment chondrocytes in proliferation and redifferentiation potential

Sakata

Furumatsu

Miyazawa

et al. 2012

International Orthopaedics (SICOT)

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Purpose Loose fragments in osteochondritis dissecans (OCD) of the knee require internal fixation. On the other hand, loose fragments derived from spontaneous osteonecrosis of the knee (SONK) are usually removed. However, the difference in healing potential between OCD-and SONK-related loose fragments has not been elucidated. In this study, we investigated proliferative activity and redifferentiation potential of normal cartilage-derived and loose fragment-derived chondrocytes. Methods Cells were prepared from normal articular cartilages and loose fragment cartilages derived from knee OCD and SONK. Cellular proliferation was compared. Redifferentiation ability of pellet-cultured chondrocytes was assessed by real-time PCR analyses. Mesenchymal differentiation potential was investigated by histological analyses. Positive ratio of a stem cell marker CD166 was evaluated in each cartilaginous tissue. Results Normal and OCD chondrocytes showed a higher proliferative activity than SONK chondrocytes. Chondrogenic pellets derived from normal and OCD chondrocytes produced a larger amount of safranin O-stained proteoglycans compared with SONK-derived pellets. Expression of chondrogenic marker genes was inferior in SONK pellets. The CD166-positive ratio was higher in normal cartilages and OCD loose fragments than in SONK loose fragments. Conclusions The OCD chondrocytes maintained higher proliferative activity and redifferentiation potential compared with SONK chondrocytes. Our results suggest that chondrogenic properties of loose fragment-derived cells and the amount of CD166-positive cells may affect the repair process of osteochondral defects.

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Section: Tissues Cells and Cell Culturementioning

confidence: 99%

Section: In Vitro Differentiation and Histological Analysesmentioning

confidence: 99%

Comparison between normal and loose fragment chondrocytes in proliferation and redifferentiation potential

Sakata

Furumatsu

Miyazawa

et al. 2012

International Orthopaedics (SICOT)

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“…A histological grading system for in vitro engineered neocartilage (Bern Score) was used to evaluate and compare the difference in chondrogenesis from the pellet-cultured MSCs. 19 The scoring was performed by a pathologist and two researchers who were blinded to the contents of the pellet. Their average score was used as the final one.…”

Section: Histological Analysismentioning

confidence: 99%

Chondrogenic differentiation of adipose tissue‐derived mesenchymal stem cells: Greater doses of growth factor are necessary

Kim

2008

Journal Orthopaedic Research

112

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There have been controversies regarding the chondrogenic potential of adipose tissue-derived mesenchymal stem cells (ATMSCs) compared with bone marrow-derived mesenchymal stem cells (BMMSCs). The purpose of this study was to confirm the hypothesis that chondrogenesis can be achieved from ATMSCs comparable to that from BMMSCs by using greater dose of currently known chondrogenic growth factors. Chondrogenesis was induced from ATMSCs by culturing them in pellets under the following conditions: #1 without growth factors (negative control); #2 5 ng/mL of TGF-b 2 ; #3 5 ng/mL of TGF-b 2 and 100 ng/mL of IGF-I; #4 15 ng/mL of TGF-b 2 ; #5 15 ng/mL of TGF-b 2 and 300 ng/mL of IGF-I; #6 25 ng/mL of TGF-b 2 ; #7 25 ng/mL of TGF-b 2 and 500 ng/mL of IGF-I. After 4 weeks of in vitro culture, the pellets were harvested for DNA quantification, analysis of the glycosaminoglycan content, reverse transcription, and real-time PCR for collagen type I (COL1A1), collagen type II (COL2A1), and Sox-9. Safranin-O and immunohistochemical staining for type II collagen also were carried out, and histological grading was performed based on the findings. A combination of 25 ng/mL TGF-b 2 and 500 ng/mL IGF-I produced results comparable to the positive control (BMMSCs treated with 5 ng/mL TGF-b 2 ) as demonstrated by DNA amount, GAG analysis, real-time PCR, and histological findings. Although ATMSCs have lower chondrogenic potentials than BMMSCs, chondrogenesis comparable to BMMSCs can be induced from ATMSCs using a greater dose combination of growth factors. These results lend a further support to the application of ATMSCs for cartilage tissue engineering. ß

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“…We have used "The Bern Score" with a few modifications as a semi-quantitative scoring method to evaluate the chondrogenicity of 3D spheroids, based on the staining of matrix proteoglycans with Alcian blue (Grogan et al, 2006). The Bern score evaluates the cartilage formation by three categories and each of the categories have a scoring range from 0 to 3 with an overall score of 9 for each spheroid (Table 1).…”

Section: Quantitative Analysis Of Histologymentioning

confidence: 99%

Mesenchymal stromal cells from human umbilical cords display poor chondrogenic potential in scaffold-free three dimensional cultures

Islam

Hansen²,

Mennan³

et al. 2016

eCM

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Many researchers world over are currently investigating the suitability of stromal cells harvested from foetal tissues for allogeneic cell transplantation therapies or for tissue engineering purposes. In this study, we have investigated the chondrogenic potential of mesenchymal stromal cells (MSCs) isolated from whole sections of human umbilical cord or mixed cord (UCSCs-MC), and compared them with cells isolated from synovial membrane (SMSCs), Hoffa's fat pad (HFPSCs) and cartilage. All MSCs were positive for surface markers including CD73, CD90, CD105, CD44, CD146 and CD166, but negative for CD11b, CD19, CD34, CD45 and HLA-DR in addition to CD106 and CD271. Chondrogenic potential of all cell sources was studied using 3D pellet cultures incubated in the presence of different combinations of anabolic substances such as dexamethasone, IGF-1, TGF-β1, TGF-β3, BMP-2 and BMP-7. BMP-2 and dexamethasone in combination with TGF-β1 or TGF-β3 excelled at inducing chondrogenesis on SMSCs, HFPSCs and chondrocytes, as measured by glycosaminoglycans and collagen type II staining of pellets, quantitative glycosaminoglycan expression, quantitative PCR of cartilage signature genes and electron microscopy. In contrast, none of the tested growth factor combinations was sufficient to induce chondrogenesis on UCSCs-MC. Moreover, incubation of UCSCs-MC spheroids in the presence of cartilage pieces or synovial cells in co-cultures did not aid chondrogenic induction. In summary, we show that in comparison with MSCs harvested from adult joint tissues, UCSCs-MC display poor chondrogenic abilities. This observation should alert researchers at the time of considering UCSCs-MC as cartilage forming cells in tissue engineering or repair strategies.

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Visual Histological Grading System for the Evaluation ofin Vitro–Generated Neocartilage

Cited by 193 publications

References 39 publications

Comparison between normal and loose fragment chondrocytes in proliferation and redifferentiation potential

Comparison between normal and loose fragment chondrocytes in proliferation and redifferentiation potential

Chondrogenic differentiation of adipose tissue‐derived mesenchymal stem cells: Greater doses of growth factor are necessary

Mesenchymal stromal cells from human umbilical cords display poor chondrogenic potential in scaffold-free three dimensional cultures

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