Revista complutense de educación

Resumen basado en el de la publicaciónSe lleva a cabo una revisión general del procedimiento cloze, procedimiento que es ampliamente conocido y utilizado como instrumento de evaluación de la lectura en los países de habla inglesa pero que apenas es conocido y empleado en España. Dicha revisión hace referencia tanto a los aspectos metodológicos relacionados con dicho procedimiento como a los distintos usos para los que puede emplearse en el campo de la evaluación de la lectura.ES

TNFα promotes proliferation of human synovial MSCs while maintaining chondrogenic potential

<div><p>Synovial mesenchymal stem cells (MSCs) are a candidate cell source for cartilage and meniscus regeneration. If we can proliferate synovial MSCs more effectively, we can expand clinical applications to patients with large cartilage and meniscus lesions. TNFα is a pleiotropic cytokine that can affect the growth and differentiation of cells in the body. The purpose of this study was to examine the effect of TNFα on proliferation, chondrogenesis, and other properties of human synovial MSCs. Passage 1 human synovial MSCs from 2 donors were cultured with 2.5 x 10⁻¹²~10⁻⁷ g/ml, 10 fold dilution series of TNFα for 14 days, then the cell number and colony number was counted. The effect of the optimum dose of TNFα on proliferation was also examined in synovial MSCs from 6 donors. Chondrogenic potential of synovial MSCs pretreated with TNFα was evaluated in 6 donors. The expressions of 12 surface antigens were also examined in 3 donors.2.5 ng/ml and higher concentration of TNFα significantly increased cell number/dish and cell number/colony in both donors. The effect of 25 ng/ml TNFα was confirmed in all 6 donors. There was no significant difference in the weight, or amount of glycosaminoglycan and DNA of the cartilage pellets between the MSCs untreated and MSCs pretreated with 25 ng/ml TNFα. TNFα decreased expression rate of CD 105 and 140b in all 3 donors. TNFα promoted proliferation of synovial MSCs with increase of cell number/ colony. Pretreatment with TNFα did not affect chondrogenesis of synovial MSCs. However, TNFα affected some properties of synovial MSCs.</p></div

Comparison of chondrogenic potential of synovial MSCs pretreated with or without TNFα.

<p>(A) Experimental design. Synovial MSCs pretreated with 25 ng/ml TNFα or without TNFα (Control) were harvested, pelleted, and cultured in the chondrogenic medium without TNFα for 21 days. (B) Macroscopic and histological features of cartilage pellets. For histology, the sections were stained with safranin-o (C) Diameter and weight of the cartilage pellets. Average values are shown (n = 6, *p<0.05 by Wilcoxon signed-ranks test).</p

The effect of 25 ng/ml TNFα on proliferation of synovial MSCs.

<p>(A) Experimental design. Synovial MSCs were cultured with 2.5 x 10^-8g/ml TNFα or without TNFα (Control) for 14 days. (B) Representative dishes stained with crystal violet (6 donors). (C) Representative cell colonies stained with crystal violet. Cell number/dish, colony number/dish, and cell number/colony. Average values are shown (n = 6, *p<0.05 by Wilcoxon signed-ranks test).</p

Surface antigen expression of synovial MSCs treated with TNFα.

<p>(A) Experimental design. Synovial MSCs were cultured with or without TNFα (Control) for 14 days for flow cytometry analysis. (B) Surface epitope expression. Average values are shown (n = 3).</p

Comparison of adipogenic and calcification potential of synovial MSCs pretreated with or without TNFα.

<p>(A) Experimental design. Synovial MSCs were pretreated with 25 ng/ml TNFα or without TNFα (Control) for 14 days, then the medium was changed and the cells were cultured in adipogenic medium or calcification medium for further 21 days. (B) Representative cells stained with oil red-o for adipogenesis. (C) Representative cells stained with alizarin red for calcification.</p

Purified Human Synovium Mesenchymal Stem Cells as a Good Resource for Cartilage Regeneration

<div><p>Mesenchymal stem cells (MSCs) have the ability to differentiate into a variety of lineages and to renew themselves without malignant changes, and thus hold potential for many clinical applications. However, it has not been well characterized how different the properties of MSCs are depending on the tissue source in which they resided. We previously reported a novel technique for the prospective MSC isolation from bone marrow, and revealed that a combination of cell surface markers (LNGFR and THY-1) allows the isolation of highly enriched MSC populations. In this study, we isolated LNGFR⁺ THY-1 ⁺ MSCs from synovium using flow cytometry. The results show that the synovium tissue contained a significantly larger percentage of LNGFR ⁺ THY-1 ⁺ MSCs. We examined the colony formation and differentiation abilities of bone marrow-derived MSCs (BM-MSCs) and synovium-derived MSCs (SYN-MSCs) isolated from the same patients. Both types of MSCs exhibited a marked propensity to differentiate into specific lineages. BM-MSCs were preferentially differentiated into bone, while in the SYN-MSC culture, enhanced adipogenic and chondrogenic differentiation was observed. These data suggest that the tissue from which MSCs are isolated should be tailored according to their intended clinical therapeutic application.</p></div

OPN Full levels in synovial fluid are positively correlated with joint damage in lateral tibial plateau.

<p>Severity of articular cartilage damage was scored according to the protocol described by Asano et al <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0049014#pone.0049014-Asano1" target="_blank">[32]</a> with minor modification (<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0049014#pone.0049014.s001" target="_blank">Table S1</a>). Open bar; intact articular cartilage (score = 1) Closed bar; damaged articular cartilage (score = 2∼6). Number of samples is indicated below each column. Data are indicated mean+/− SEM. *; p<0.05.</p

Kinetics of OPN Full and N-half in synovial fluid after joint injury.

<p>(Left panels) Time course changes of OPN Full (upper panel) and OPN N-half (lower panel) protein levels in synovial fluid after ACL rupture. Open bar: within 1 month after rupture n = 14, Closed bar: greater than 1 month after rupture n = 68. (Right panels) OPN Full (upper panel) and OPN N-half (lower panel) protein levels in synovial fluid collected from pre (n = 23) and post (n = 93) ACL reconstruction surgery. Data are indicated mean+/− SEM. **; p<0.01.</p

High chondrogenic differentiation ability of SYN-MSCs.

<p>(A) Representative images of chondrocyte pellets on day 21 derived from BM-MSCs and SYN-MSCs (scale bar = 1 mm). (B) Comparison of chondrocyte pellet size between BM-MSCs and SYN-MSCs (n = 4, *p < 0.05). (C) Micrographs of chondrocyte pellets derived from BM-MSCs and SYN-MSCs on day 21 stained with Toluidine blue and Safranin O (scale bar = 200 μm). (D) Expression ratio of <i>SOX9</i>, <i>AGGRECAN</i>, <i>and COL11A1</i> mRNA to <i>β-ACTIN</i> mRNA in chondrocyte pellets (n = 3). Control: non-induced MSCs, Induced: induced MSCs for chondrogenic lineage.</p