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1.
Acta Ortop Mex ; 37(1): 30-35, 2023.
Artículo en Español | MEDLINE | ID: mdl-37857395

RESUMEN

INTRODUCTION: osteoarthritis is one of the most prevalent chronic diseases in the world and is defined as the gradual loss of cartilage in the joints, mainly that of the knee. It is considered a cause of disability in older adults and is characterized by pain, stiffness and loss of mobility. MATERIAL AND METHODS: observational study to evaluate the effect of the combination of non-hydrolyzed type II native collagen (CII-NH), omega-3 (Om-3) and astaxanthin (AX), in a population of 182 patients with knee osteoarthritis grade I/II. Measurements of thigh circumference, arcs of movement and pain were obtained through international scales such as the visual analogue pain scale (VAS), the Lequesne index and the Western Ontario McMaster Universities Osteoarthritis Index (WOMAC) scale. Medical check-ups were performed every 30 days for three months. The Statgraphics software (Statgraphics Technologies, Virginia) was used, the evaluation of the variables and the statistical significance were determined by t Student test and the results are shown as a mean. RESULTS: it was shown that daily consumption increases mobility, decreases knee pain and inflammation in patients within three months. Additionally, there was a reduction in the consumption of nonsteroidal anti-inflammatory drugs (NSAIDs) by the study subjects. CONCLUSION: the fixed combination of non-hydrolyzed type II collagen, omega-3 and astaxanthin, generates, in the short term, a decrease in inflammation and stiffness in patients with osteoarthritis.


INTRODUCCIÓN: la osteoartritis es una de las enfermedades crónicas con mayor prevalencia en el mundo y se define como la pérdida gradual de cartílago en las articulaciones, principalmente la de rodilla. Es considerada como una causa de discapacidad en adultos mayores y se caracteriza por dolor, rigidez y pérdida de la movilidad. MATERIAL Y MÉTODOS: estudio observacional para evaluar el efecto de la combinación de Sólo debe decir, colágeno nativo tipo II, omega-3 (Om-3) y astaxantina (AX), en una población de 182 pacientes con artrosis de rodilla grado I/II. Se obtuvieron las mediciones de circunferencia del muslo, arcos de movimiento y dolor a través de las escalas internacionales como la escala visual analógica de dolor (EVA), el índice de Lequesne y escala Western Ontario McMaster Universities Osteoarthritis Index (WOMAC). Las revisiones médicas se realizaron cada 30 días durante tres meses. Se utilizó el software Statgraphics (Statgraphics Technologies, Virginia), la evaluación de las variables y la significancia estadística fueron determinadas por la prueba t de Student y los resultados se muestran como media. RESULTADOS: se demostró que el consumo diario incrementa la movilidad, disminuye el dolor y la inflamación de rodilla en los pacientes en un lapso de tres meses. Adicionalmente se registró una reducción en el consumo de antiinflamatorios no esteroideos (AINE) por parte de los sujetos de estudio. CONCLUSIÓN: la combinación fija colágeno nativo tipo II, omega-3 y astaxantina, genera, en el corto plazo, disminución de la inflamación y la rigidez en pacientes con osteoartritis.


Asunto(s)
Osteoartritis de la Rodilla , Humanos , Anciano , Osteoartritis de la Rodilla/tratamiento farmacológico , Colágeno Tipo II , Resultado del Tratamiento , Dolor , Inflamación
2.
Poult Sci ; 102(10): 102916, 2023 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-37499613

RESUMEN

This study aimed to assess the influence of glycosaminoglycan (chondroitin and glucosamine sulfates) supplementation in the diet of broilers on the expression of matrix metallopeptidase 9 (MMP-9) and metallopeptidase inhibitor 2 (TIMP-2) genes, the synthesis of proteoglycans, collagen type II and chondrocytes, bone and cartilage macroscopy, bone mineral densitometry, bone breaking strength and mineral profile. A completely randomized design was carried out in a 3 × 3 factorial scheme (3 levels of chondroitin sulfate: 0.00, 0.05, and 0.10%; and 3 levels of glucosamine sulfate: 0.00, 0.15, and 0.30%), totaling 9 treatments. At 21 and 42 d of age, broilers were slaughtered, and tibias and femurs were collected for evaluation. There was an interaction (P < 0.05) of sulfates for the expression of MMP-9 and its inhibitor TIMP-2 in femur articular cartilage, as well as for the number of chondrocytes, collagen type II and proteoglycans in tibia articular cartilage, bone and cartilage macroscopy and mineral profile (P < 0.05), with better results obtained with the inclusion of chondroitin and/or glucosamine sulfates in the feed. In conclusion, chondroitin and glucosamine sulfates can be used in broiler diets in order to favor the development of the structure of the locomotor system (bones and joints), thus preventing locomotion problems.


Asunto(s)
Cartílago Articular , Glicosaminoglicanos , Animales , Glicosaminoglicanos/metabolismo , Inhibidor Tisular de Metaloproteinasa-2/metabolismo , Inhibidor Tisular de Metaloproteinasa-2/farmacología , Pollos , Colágeno Tipo II/metabolismo , Colágeno Tipo II/farmacología , Metaloproteinasa 9 de la Matriz/metabolismo , Metaloproteinasa 9 de la Matriz/farmacología , Proteoglicanos/genética , Proteoglicanos/metabolismo , Sulfatos de Condroitina/metabolismo , Sulfatos de Condroitina/farmacología , Glucosamina/metabolismo , Glucosamina/farmacología , Minerales/metabolismo , Sulfatos/metabolismo
3.
Int. j. morphol ; 41(2): 583-590, abr. 2023. ilus
Artículo en Inglés | LILACS | ID: biblio-1440339

RESUMEN

SUMMARY: Rheumatoid arthritis (RA) that affects the synovial knee joint causes swelling of the synovial membrane and tissue damage. Interleukin-17A (IL-17A) and the enzyme glycogen synthase kinase-3β (GSK3β) are involved in the pathogenesis of RA. The link between IL-17A, GSK3β, the oxidative stress, and the profibrogenic marker alpha-smooth muscle actin (α-SMA) with and without TDZD-8, GSK3β inhibitor has not been studied before. Consequently, active immunization of rats was performed to induce RA after three weeks using collagen type II (COII) injections. The treated group received daily injection of 1 mg/kg TDZD-8 for 21 days following the immunization protocol (COII+TDZD-8). Blood and synovium tissue samples were harvested at the end of the experiment. RA development was confirmed as corroborated by a substantial increase in blood levels of the highly specific autoantibody for RA, anti-citrullinated protein antibody as well as augmentation of reactive oxidative species (ROS) levels measured as lipid peroxidation. RA induction also increased synovium tissue levels of IL-17A and the profibrogenic marker, α-SMA. All these parameters seemed to be significantly (p<0.0001) ameliorated by TDZD-8. Additionally, a significant correlation between IL-17A, ROS, and α-SMA and biomarkers of RA was observed. Thus, knee joint synovium RA induction augmented IL-17A/GSK3β/ROS/α-SMA axis mediated arthritis in a rat model of RA, which was inhibited by TDZD-8.


La artritis reumatoide (AR) que afecta la articulación sinovial de la rodilla provoca inflamación de la membrana sinovial y daño tisular. La interleucina-17A (IL-17A) y la enzima glucógeno sintasa quinasa-3β (GSK3β) están involucradas en la patogenia de la AR. No se ha estudiadol vínculo entre IL-17A, GSK3β, el estrés oxidativo y el marcador profibrogénico actina de músculo liso alfa (α-SMA) con y sin inhibidor de TDZD-8, GSK3β. En consecuencia, se realizó una inmunización activa de ratas para inducir la AR después de tres semanas usando inyecciones de colágeno tipo II (COII). El grupo tratado recibió una inyección diaria de 1 µg/ kg de TDZD-8 durante 21 días siguiendo el protocolo de inmunización (COII+TDZD-8). Se recogieron muestras de sangre y tejido sinovial al final del experimento. El desarrollo de AR se confirmó como lo corroboró el aumento sustancial en los niveles sanguíneos del autoanticuerpo altamente específico para AR, el anticuerpo antiproteína citrulinada, así como el aumento de los niveles de especies oxidativas reactivas (ROS) medidos como peroxidación lipídica. La inducción de AR también aumentó los niveles de tejido sinovial de IL-17A y el marcador profibrogénico, α-SMA. Todos estos parámetros parecían mejorar significativamente (p<0,0001) con TDZD-8. Además, se observó una correlación significativa entre IL- 17A, ROS y α-SMA y biomarcadores de AR. Por lo tanto, la inducción de AR en la sinovial de la articulación de la rodilla aumentó la artritis mediada por el eje IL-17A/GSK3β/ROS/α-SMA en un modelo de rata de AR, que fue inhibida por TDZD-8.


Asunto(s)
Animales , Ratas , Artritis Reumatoide , Tiadiazoles/administración & dosificación , Fibrosis , Inmunohistoquímica , Western Blotting , Actinas , Inmunización , Especies Reactivas de Oxígeno , Ratas Wistar , Interleucina-17 , Colágeno Tipo II/administración & dosificación , Modelos Animales de Enfermedad , Glucógeno Sintasa Quinasa 3 beta
4.
Int. j. morphol ; 40(1): .84-90, feb. 2022.
Artículo en Inglés | LILACS | ID: biblio-1385595

RESUMEN

SUMMARY: Rheumatoid arthritis (RA), an inflammatory autoimmune disease that causes cartilage degradation and tissue destruction, can affect synovial joints such as the knee joint. The link between the nitrosative stress enzyme inducible nitric oxide synthase (iNOS) and the cytokine interleukin-1 (IL-1β) in RA-induced knee joint synovial membrane damage with and without the incorporation of the GSK3β inhibitor TDZD-8 has never been studied. As a result, we used active immunization method with collagen type II (COII) for twenty one days to induce RA in rats. TDZD-8 (1 mg/kg; i.p.) was given daily into matched immunized rats for three weeks after day 21 (COII+TDZD-8). Blood and tissue samples were taken 42 days after immunization. A dramatic increase in rheumatoid factor (RF) blood levels, as well as considerable synovial tissue damage and inflammatory cell infiltration of the synovial membrane, were used to validate the onset of RA following COII immunization. COII immunization increased tissue levels of iNOS protein and IL- 1β mRNA and protein expression, which TDZD-8 suppressed considerably (p<0.0001). Furthermore, there was a significantly (p<0.001) positive correlation between iNOS, inflammatory biomarkers, and RF. We concluded that TDZD-8 reduced RA-induced IL-1β -iNOS axis-mediated arthritis in the rat knee joint synovium.


RESUMEN: La artritis reumatoide (AR), es una enfermedad autoinmune inflamatoria que causa la degradación del cartílago y la destrucción del tejido, pudiendo afectar las articulaciones sinoviales, como la articulación de la rodilla. No se ha estudiado el vínculo entre la óxido nítrico sintasa inducible por la enzima del estrés nitrosativo (iNOS) y la citocina interleucina-1 (IL-1β) en el daño de la membrana sinovial de la articulación de la rodilla provocado por AR con y sin la incorporación del inhibidor de GSK3β TDZD-8. Utilizamos el método de inmunización activa con colágeno tipo II (COII) durante veintiún días para inducir AR en ratas. Se administró TDZD-8 (1 mg/kg; i.p.) diariamente a ratas inmunizadas emparejadas durante tres semanas después del día 21 (COII+TDZD- 8). Se tomaron muestras de sangre y tejido 42 días después de la inmunización. Se observó un gran aumento de los niveles sanguíneos del factor reumatoideo (FR), así como un daño considerable del tejido sinovial e infiltración de células inflamatorias en la membrana sinovial, para validar la aparición de la AR después de la inmunización con COII. La inmunización con COII aumentó los niveles tisulares de la proteína iNOS y la expresión de proteína y ARNm de IL-1β, que TDZD-8 suprimió considerablemente (p<0,0001). Además, hubo una correlación positiva significativa (p<0,001) entre iNOS, biomarcadores inflamatorios y FR. Concluimos que TDZD- 8 redujo la artritis mediada por el eje IL-1β-iNOS inducida por la AR en la sinovial de la articulación de la rodilla de rata.


Asunto(s)
Animales , Ratas , Artritis Reumatoide/inmunología , Tiadiazoles/administración & dosificación , Glucógeno Sintasa Quinasa 3 beta/antagonistas & inhibidores , Artritis Reumatoide/inducido químicamente , Inmunohistoquímica , Ratas Wistar , Colágeno Tipo II/administración & dosificación , Modelos Animales de Enfermedad , Interleucina-1beta , Glucógeno Sintasa Quinasa 3 beta/administración & dosificación , Estrés Nitrosativo/efectos de los fármacos , Inflamación
5.
J Ethnopharmacol ; 286: 114928, 2022 Mar 25.
Artículo en Inglés | MEDLINE | ID: mdl-34929309

RESUMEN

ETHNOPHARMACOLOGICAL RELEVANCE: Herbs have been commonly used for the treatment of rheumatoid arthritis (RA). It has been verified that Erteng Tongbi Decoction has good therapeutic effects on RA, while, relatively few studies on the relationship between its components and anti-rheumatoid efficacy were carried out. AIM OF THE STUDY: To discuss the anti-RA effects of Erteng Tongbi Decoction on collagen-induced arthritis (CIA) in mice and the influence of T cell differentiation and cytokines balance. MATERIALS AND METHODS: Separate researches on the two traditional Chinese medicines of the Erteng Tongbi Decoction were conducted. First, a murine peritoneal macrophage model was established, and then the cytokines levels and macrophage maturity were measured to select the best extraction solvent. Furthermore, ethanol extracts were partitioned successively with four kinds of solvents, and the anti-inflammatory parts were selected by the same vitro model. Subsequently, mice were arbitrarily divided into control, CIA model, positive control, effective parts alone or in combination. After 20 days of oral administration, the weight, hind paw volume, rheumatism index value, and the pathological changes were checked to assess the obvious level of arthritis. Furthermore, the levels of IL-6, TNF-α, IL-10, and IL-17A in serum and the balance of Th17/Treg and Th1/Th2 cells in spleen and mesenteric lymph nodes (MLN) was detected. Finally, the major active constituents were identified. RESULTS: In vitro, the anti-inflammatory effects of ethanol extracts was much better than water extract. In addition, the effective parts of Celastrus orbiculatus Thunb. ethanol extract were petroleum ether parts and dichloromethane parts. The effective parts of Spatholobus suberectus Dunn. ethanol extracts was petroleum ether parts and ethyl acetate parts screened. In vivo, effective parts compatibility could inhibit the progression of inflammation by modulating T cell differentiation and cytokines balance. Constituent analysis revealed that effective parts contained sesquiterpenes alkaloids, phenolic acids, and flavanols. CONCLUSIONS: Erteng Tongbi Decoction could notably ameliorate CIA mice by modulating T cell differentiation and cytokines balance and support its application in folk medicine.


Asunto(s)
Antirreumáticos/farmacología , Artritis Experimental/tratamiento farmacológico , Artritis Reumatoide/tratamiento farmacológico , Medicamentos Herbarios Chinos/farmacología , Animales , Antiinflamatorios/administración & dosificación , Antiinflamatorios/aislamiento & purificación , Antiinflamatorios/farmacología , Antirreumáticos/administración & dosificación , Antirreumáticos/aislamiento & purificación , Artritis Experimental/patología , Artritis Reumatoide/patología , Diferenciación Celular/efectos de los fármacos , Colágeno Tipo II , Citocinas/metabolismo , Medicamentos Herbarios Chinos/administración & dosificación , Inflamación/tratamiento farmacológico , Inflamación/patología , Masculino , Ratones , Ratones Endogámicos C57BL , Ratones Endogámicos DBA , Linfocitos T/citología
6.
Exp Mol Pathol ; 123: 104689, 2021 12.
Artículo en Inglés | MEDLINE | ID: mdl-34592200

RESUMEN

The aim of this study was to analyze the expression of mBD4, mBD3 and CRAMP in joint of mice with type II collagen-induced arthritis/CIA and to explore its possible association with IL-10, IL-4, IFN-γ, IL-17, MMP3, RANK/RANKL/OPG and histological parameters. METHODS: CIA was induced in 44 DBA/1 J mice. The joints from mice were classified into the onset, peak and remission phase of CIA. Histological sections were stained with hematoxylin-eosin and safranin O. The expression of CRAMP, mBD-3, mBD-4, and MMP-3 was evaluated using reverse transcription polymerase chain reaction (RT-PCR) and immunohistochemistry. The expression of IL-10, IL-4, IFN-γ, IL-17, RANK/RANKL/OPG was analyzed by RT-PCR. RESULTS: We observed that inflammation and immunostained cells for CRAMP increased in the peak and remission phases compared to the control group. In addition, increments in relative expressions of CRAMP were detected for the remission phase and in IL-4 and IL-17 in the peak phase compared to the control and onset phase. In addition, an increase in IL-10 in a peak phase compared to the control, as well as the relative expression of IFN-γ in remission phase was higher than in the onset phase. This was accompanied by an increase in cartilage damage in the peak phase compared to the control. Cells immunostained to MMP3 increased in the peak phase compared to the onset and control group, and relative expression of MMP3 was detected in the peak phase compared to the onset, remission, and control group. We observed that the relative expression of RANK and RANKL in the peak phase was higher than in control and onset phase. Finally, the relative expression of OPG in the peak phase compared to the onset, remission, and control group was detected. Regarding CRAMP behavior in the different phases studied, it was positively correlated with IL-4 and RANK, and showed a negative correlation with IFN-γ, IL-17, IL-10, RANKL, OPG and RANKL/OPG ratio in the control group. Also was positively correlated with IFN-γ, IL-17, IL-4, IL-10, as well as with RANK, RANKL, and OPG in the onset and peak phases of the CIA. In the peak phase, CRAMP showed a positive association with MMP3, and we observed a direct correlation between CRAMP and IFN-γ and RANKL/OPG ratio in remission phase. mBD3 correlates positively with IFN-γ, IL-17, IL-10, RANKL, OPG and RANKL/OPG ratio, and showed a negative correlation with CRAMP, MMP3, and RANK in the control group. Also, it was directly associated with IFN-γ, IL-17, IL-4, IL-10 and RANKL in the onset phase while it was inversely associated with CRAMP, MMP-3, RANK, RANKL, and OPG in the peak phase. Finally, mBD3 was inversely correlated with MMP3 in the remission phase and was directly associated with CRAMP, IFN-γ and RANKL/OPG ratio in this phase. mBD4 was directly associated with CRAMP, IFN-γ, IL-17, IL-4, IL-10, RANKL / OPG in the onset phase, and with CRAMP, IFN-γ, IL-17, IL-4, IL-10, MMP3, RANK, RANKL and OPG in the peak phase. Finally, mBD4 was positively associated with mBD3, IFN-γ, IL-17, IL-10, RANK, RANKL OPG and RANKL/OPG in the CIA remission phase. CONCLUSIONS: Our results demonstrate that CRAMP plays an important role in CIA progress and suggest that its abundance is associated with local pro- and anti-inflammatory status. This makes us propose CRAMP as a possible contributor of bone reconstruction in the last stage of CIA.


Asunto(s)
Artritis/genética , Remodelación Ósea/genética , Catelicidinas/genética , Proteínas de Unión al ADN/genética , Factores de Transcripción/genética , beta-Defensinas/genética , Animales , Artritis/inducido químicamente , Artritis/patología , Colágeno Tipo II/toxicidad , Regulación de la Expresión Génica/genética , Humanos , Inflamación/genética , Inflamación/patología , Ratones
7.
Int. j. morphol ; 39(1): 311-317, feb. 2021. ilus, graf
Artículo en Inglés | LILACS | ID: biblio-1385290

RESUMEN

SUMMARY: Rheumatoid arthritis (RA) is considered an autoimmune disease distinguished by chronic synovial membrane inflammation, degraded cartilage, as well as bone destruction, which lead to joints pain and stiffness. The pathogenesis of RA involved at least two mechanisms: Cellular proliferation and activation of glycogen synthase kinase-3β (GSK3β) enzyme. Thus, we tested the hypothesis that the GSK3binhibitor, TDZD-8, can treat the synovial tissue toward collagen type II (COII)-mediated RA linked to apoptosis induction and biomarker suppression of inflammation. Wistar rats were immunized with COII (the model group) for 21 days. Matched immunized rats were daily injected with TDZD-8 (1 mg/kg; i.p) for three additional weeks (COII+TDZD- 8).After 42 days of post-immunization, blood and tissues were collected. Histology (H&E) and immunohistochemistry (CD45; leukocyte common antigen) images showed that COII induced RA was demonstrated by profound damage to the synovial tissue and infiltration of the inflammatory cells, which were substantially ameliorated with TDZD-8. In addition, COII immunization caused the induction of rheumatoid factor (RF), tumor necrosis factor-alpha (TNF-α), interleukin-6 (IL-6), and interleukin 1 beta (IL-1β) that were substantially (p<0.05) suppressed by TDZD-8. Whereas, TDZD-8 augmented the apoptotic biomarker, Bcl-2-associated X protein (Bax), which was significantly (p<0.05) ameliorated by RA. We also showed a substantial relationship (p<0.001) between the blood levels of RF and the synovial tissue levels of TNF-α (r = 0.759), IL-1b (r = 0.969), IL-6 (r = 0.749), and Bax (r = - 0.914). These results indicate effective treatment of the injured synovial tissue by TDZD-8 against COII-induced RA in rats, which also decreases inflammatory biomarkers and augmentation of apoptosis.


RESUMEN: La artritis reumatoide (AR) es una enfermedad autoinmune que se distingue por la inflamación crónica de la membrana sinovial, el cartílago degradado y la destrucción de los huesos, lo que provoca dolor y rigidez en las articulaciones. La patogenia de la AR involucra al menos dos mecanismos: la proliferación celular y la activación de la enzima glucógeno sintasa quinasa-3b (GSK3β) Por lo tanto, probamos la hipótesis de que el inhibidor de GSK3β, TDZD-8, puede tratar el tejido sinovial hacia el colágeno tipo II (COII) - AR mediada por inducción de apoptosis y supresión de biomarcadores de inflamación. Se inmunizaron ratas Wistar con COII (el grupo modelo) durante 21 días. Se inyectaron diariamente ratas emparejadas inmunizadas con TDZD-8 (1 mg / kg; i.p) durante tres semanas adicionales (COII + TDZD-8). Después de 42 días de post-inmunización, se recolectó sangre y tejidos. Las imágenes de histología (H&E) e inmunohistoquímica (CD45; antígeno común de leucocitos) mostraron que la AR inducida por COII presentaba un daño profundo en el tejido sinovial e infiltración de las células inflamatorias, las que mejoraron con TDZD-8. Además, la inmunización con COII provocó la inducción de factor reumatoide (FR), factor de necrosis tumoral alfa (TNF-α), interleucina-6 (IL-6) e interleucina 1 beta (IL-1β) que fueron suprimidos por TDZD-8 de manera significativa (p < 0.05). Considerando que TDZD-8 aumentó el biomarcador apoptótico, la proteína X asociada a Bcl-2 (Bax), que fue mejorado (p <0,05) por RA. También se observó una relación sustancial (p <0,001) entre los niveles sanguíneos de RF y los niveles de tejido sinovial de TNF-α (r = 0,759), IL-1β (r = 0,969), IL-6 (r = 0,749), y Bax (r = -0,914). Estos resultados indicaron un tratamiento eficaz del tejido sinovial lesionado por TDZD-8 contra la AR inducida por COII en ratas, que también disminuye los biomarcadores inflamatorios y el aumento de la apoptosis.


Asunto(s)
Animales , Masculino , Ratas , Artritis Reumatoide/tratamiento farmacológico , Tiadiazoles/administración & dosificación , Colágeno Tipo II/efectos adversos , Artritis Experimental/tratamiento farmacológico , Tiadiazoles/farmacología , Inmunohistoquímica , Western Blotting , Ratas Wistar , Apoptosis , Modelos Animales de Enfermedad , Interleucina-1beta , Inflamación
8.
Braz J Phys Ther ; 25(1): 62-69, 2021.
Artículo en Inglés | MEDLINE | ID: mdl-32151525

RESUMEN

OBJECTIVE: Considering the osteoarthritis (OA) model that integrates the biological, mechanical, and structural components of the disease, the present study aimed to investigate the association between urinary C-Telopeptide fragments of type II collagen (uCTX-II), knee joint moments, pain, and physical function in individuals with medial knee OA. METHODS: Twenty-five subjects radiographically diagnosed with knee OA were recruited. Participants were evaluated through three-dimensional gait analysis, uCTX-II level, the WOMAC pain and physical function scores, and the 40m walk test. The association between these variables was investigated using Pearson's product-moment correlation, followed by a hierarchical linear regression, controlled by OA severity and body mass index (BMI). RESULTS: No relationship was found between uCTX-II level and knee moments. A significant correlation between uCTX-II level and pain, physical function, and the 40m walk test was found. The hierarchical linear regression controlling for OA severity and BMI showed that uCTX-II level explained 9% of the WOMAC pain score, 27% of the WOMAC physical function score, and 7% of the 40m walk test. CONCLUSION: Greater uCTX-II level is associated with higher pain and reduced physical function and 40m walk test performance in individuals with medial knee OA.


Asunto(s)
Colágeno Tipo II/química , Colágeno Tipo I/química , Articulación de la Rodilla/fisiopatología , Osteoartritis de la Rodilla/fisiopatología , Dolor/fisiopatología , Péptidos/química , Biomarcadores , Colágeno Tipo I/orina , Humanos , Péptidos/orina
9.
Cartilage ; 13(2_suppl): 839S-846S, 2021 12.
Artículo en Inglés | MEDLINE | ID: mdl-31441318

RESUMEN

OBJECTIVE: We sought to evaluate the effect of different concentrations of ethanol on phenotype and activity of articular chondrocyte synthesis of neonatal rats in 2-dimensional (2D) and 3-dimensional (3D) culture. METHODS: Chondrocytes were cultured in chondrogenic medium with different concentrations of ethanol: 0.0% v/v (control); 0.05% v/v (8.6 mM); 0.25% v/v (42.9 mM), and 0.5% v/v (85.7 mM). Chondrocytes under 2D culture were subjected to MTT assay, while chondrocytes under 3D culture were processed for paraffin inclusion and stained by periodic acid Schiff (PAS) to evaluate mean chondrocyte diameter and percentages of cells, nucleus, cytoplasm, well-differentiated matrix, and PAS+ areas. The expression of gene transcripts for aggrecan, Sox9, and type II collagen was evaluated by real-time quantitative polymerase chain reaction. RESULTS: There was no difference between groups by the MTT assay. PAS staining revealed that chondrocytes treated with 0.5% v/v ethanol had higher percentages of cytoplasm and nuclear areas, but with a reduction in PAS+ matrix area. The mean diameter of chondrocytes was similar between groups. The expression of aggrecan in the group treated with 0.5% v/v ethanol was lower in comparison to that in the control. In the groups treated with 0.25% v/v and 0.5% v/v ethanol, the percentage of differentiated cartilage was lower in comparison with that in the control. The group treated with 0.05% v/v ethanol was similar to the control in all parameters. CONCLUSIONS: Ethanol acted directly on in vitro cultured articular chondrocytes of newborn rats, altering the chondrocyte phenotype and its synthesis activity, and these effects were dose dependent.


Asunto(s)
Cartílago Articular , Condrocitos , Animales , Cartílago Articular/metabolismo , Condrocitos/metabolismo , Colágeno Tipo II/metabolismo , Etanol/metabolismo , Etanol/farmacología , Fenotipo , Ratas
10.
Knee Surg Sports Traumatol Arthrosc ; 29(6): 1990-1999, 2021 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-32852566

RESUMEN

PURPOSE: To examine healing adaptations over 17 weeks post Achilles tendon (AT) rupture in the injured region (IR) compared to an uninjured region (UIR) of the AT. METHODS: Twenty-four rats were subjected to a complete right-sided AT rupture, while the left side served as a control. ATs were harvested at 1, 2, 8 and 17 weeks post-rupture and stained with antibodies specific to Collagen type I (Col I) and II (Col II) as well as Alcian Blue and Picrosirius Red staining techniques. Histopathological changes, proteoglycan content, collagen alignment and immunoexpression were assessed. RESULTS: Both regions examined, IR and UIR, exhibited over weeks 1-17 similar healing adaptations of increasing collagen alignment, decreasing Col I immunoexpression, as well as increasing proteoglycan content and Col II occurrence. Increased proteoglycan content was found already at week 2 in the UIR, while it first increased at week 8 in the IR. The area positive to Col II was increased compared to controls at week 8 in the UIR, whereas it first raised at week 17 in the IR. Collagen disorganization successively declined to reach control levels at week 17 in the UIR, but was still higher in the IR. CONCLUSION: This study demonstrated that uninjured areas of the AT remote from the rupture site also undergo pronounced remodeling, although with time-span differences relative to injured AT portions. These changes including the pathologic heterotopic mineralization and chondrogenic differentiation observed in both regions may have implications in the choice of rehabilitation regimes in order to prevent secondary rupture.


Asunto(s)
Tendón Calcáneo/lesiones , Tendón Calcáneo/fisiopatología , Cicatrización de Heridas/fisiología , Tendón Calcáneo/patología , Animales , Condrogénesis , Colágeno Tipo I/metabolismo , Colágeno Tipo II/metabolismo , Femenino , Modelos Animales , Proteoglicanos/metabolismo , Ratas Sprague-Dawley , Rotura/patología , Rotura/fisiopatología
11.
Nutrients ; 12(10)2020 Oct 09.
Artículo en Inglés | MEDLINE | ID: mdl-33050122

RESUMEN

Food consumption has significant positive effects on an individual's health status, including the reduction of symptoms associated with musculoskeletal pain. However, specific food groups indicated for the treatment of pain are not yet determined. Hence, this review aimed to analyze the effects of nutritional interventions with specific diets, oils and/or fatty acids, and foodstuffs in natura in the reduction of musculoskeletal pain. An integrative review was conducted in the following databases: Embase, PubMed, LILACS, and Google Scholar. Clinical trials written in English, Spanish, and Portuguese and published between 2000 and March 2020 were included in this review. Seventeen studies were included. Among these, a reduction of musculoskeletal pain with different types of nutritional interventions, such as vegan and Mediterranean diets and the consumption of blueberry, strawberry, passion fruit peel extract, argan oil, fish oil (omega-3), olive oil, and undenatured type II collagen and vitamin D gel capsules, was observed in 14 studies. Eight studies evaluated the profiles of several inflammatory markers, and of these, decreased interleukin (IL)-6, IL-1ß, and tumor necrosis factor-α levels were observed in two studies. This review suggests that different nutritional interventions with specific diets, oils and/or fatty acids, and foodstuffs in natura reduce musculoskeletal pain, specifically in adults with osteoarthritis. Besides pain improvement, nutritional interventions, including the consumption of strawberry and vitamin D gel capsules, decrease the levels of several inflammatory markers.


Asunto(s)
Dieta Saludable , Dolor Musculoesquelético/dietoterapia , Fenómenos Fisiológicos de la Nutrición/fisiología , Colágeno Tipo II , Dieta Mediterránea , Dieta Vegana , Femenino , Aceites de Pescado , Fragaria , Humanos , Mediadores de Inflamación/metabolismo , Interleucina-1beta/metabolismo , Interleucina-6/metabolismo , Masculino , Dolor Musculoesquelético/metabolismo , Aceite de Oliva , Factor de Necrosis Tumoral alfa/metabolismo , Vitamina D
12.
Int J Exp Pathol ; 101(1-2): 55-64, 2020 02.
Artículo en Inglés | MEDLINE | ID: mdl-32459025

RESUMEN

Rheumatoid arthritis is a disabling autoimmune disease with a high global prevalence. Treatment with disease-modifying anti-arthritic drugs (DIMARDs) has been routinely used with beneficial effects but with adverse long-term consequences; novel targeted biologics and small-molecule inhibitors are promising options. In this study, we investigated whether purified omega unsaturated fatty acids (ω-UFAs) and dialysable leukocyte extracts (DLEs) prevented the development of arthritis in a model of collagen-induced arthritis (CIA) in mice. We also investigated whether the transcription factor NF-κB and the NLRP3 inflammasome were involved in the process and whether their activity was modulated by treatment. The development of arthritis was evaluated for 84 days following treatment with nothing, dexamethasone, DLEs, docosahexaenoic acid, arachidonic acid, and oleic acid. Progression of CIA was monitored by evaluating clinical manifestations, inflammatory changes, and histological alterations in the pads' articular tissues. Both DLEs and ω-UFAs led to an almost complete inhibition of the inflammatory histopathology of CIA and this was concomitant with the inhibition of NF-kB and the inhibition of the activation of NLRP3. These data suggest that ω-UFAs and DLEs might have NF-κB as a common target and that they might be used as ancillary medicines in the treatment of arthritis.


Asunto(s)
Antiinflamatorios/farmacología , Antirreumáticos/farmacología , Artritis Experimental/prevención & control , Cartílago Articular/efectos de los fármacos , Extractos Celulares/farmacología , Ácidos Grasos Insaturados/farmacología , Leucocitos , Animales , Ácido Araquidónico/farmacología , Artritis Experimental/inducido químicamente , Artritis Experimental/metabolismo , Artritis Experimental/patología , Cartílago Articular/metabolismo , Cartílago Articular/patología , Colágeno Tipo II , Diálisis , Ácidos Docosahexaenoicos/farmacología , Femenino , Inflamasomas/efectos de los fármacos , Inflamasomas/metabolismo , Ratones Endogámicos BALB C , Ratones Endogámicos DBA , FN-kappa B/metabolismo , Proteína con Dominio Pirina 3 de la Familia NLR/metabolismo , Ácido Oléico/farmacología
13.
PLoS One ; 15(3): e0230228, 2020.
Artículo en Inglés | MEDLINE | ID: mdl-32163510

RESUMEN

This study was designed to evaluate the anti-inflammatory effects of a curcumin treatment on the knee of rats with induced osteoarthritis. Fifteen adult rats were used and divided in three groups: the osteoarthritis group (OAG), control group (CG-without induction of osteoarthritis), and curcumin-treated osteoarthritis group (COAG). Osteoarthritis was induced in the right knee of rats in the OAG and COAG by administering an intra-articular injection of 1 mg of zymosan. Fourteen days after induction, 50 mg/kg curcumin was administered by gavage daily for 60 days to the COAG. After the treatment period, rats from all groups were euthanized. Medial femoral condyles were collected for light microscopy and immunohistochemical staining. The expression of SOX-5, IHH, MMP-8, MMP-13, and collagen 2 (Col2) was analyzed. The COAG exhibited an increase in the number of chondrocytes in the surface and middle layers compared with that of the OAG and CG, respectively. The COAG also showed a decrease in the thicknesses of the middle and deep layers compared with those of the OAG, and an increase in Col2 expression was observed in all articular layers (surface, middle, and deep) in the COAG compared with that in the OAG. SOX-5 expression was increased in the surface and deep layers of the COAG compared with those in the OAG and CG. Based on the results of this study, the curcumin treatment appeared to exert a protective effect on cartilage, as it did not result in an increase in cartilage thickness or in MMP-8 and MMP-13 expression but led to increased IHH, Col2, and SOX-5 expression and the number of chondrocytes.


Asunto(s)
Cartílago Articular/efectos de los fármacos , Curcumina/farmacología , Articulación de la Rodilla/efectos de los fármacos , Osteoartritis/tratamiento farmacológico , Animales , Cartílago Articular/metabolismo , Condrocitos/efectos de los fármacos , Condrocitos/metabolismo , Colágeno Tipo II/metabolismo , Inyecciones Intraarticulares/métodos , Articulación de la Rodilla/metabolismo , Masculino , Osteoartritis/metabolismo , Ratas , Ratas Wistar
14.
Genet Test Mol Biomarkers ; 24(2): 105-111, 2020 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-31999490

RESUMEN

Aims: To investigate the possible roles of the single nucleotide polymorphisms (SNPs) MATN3 (rs77245812) and DOT1L (rs12982744) with susceptibility to knee osteoarthritis (KOA) among mestizos from the northeast region of Mexico. In addition, we analyzed the relationship of their urinary levels of carboxy terminal telopeptide of collagen type II (CTX-II) and the radiological grade of disease. Materials and Methods: A total of 223 individuals from a Northeast Mexico Mestizo population were included in this study: 110 patients with primary KOA and 113 healthy controls. Genotyping of the MATN3 (rs77245812) and DOT1L (rs12982744) SNPs was performed by real-time polymerase chain reaction. Results: No association was found between the polymorphisms MATN3 (rs77245812), DOT1L (rs12982744), and the risk of developing KOA (odds ratio [OR] = 1.33, 95% confidence interval [CI] = 0.42-6.48, p = 0.621) (OR = 2.03, 95% CI = 0.35-11.5, p = 0.422). However, urinary CTX-II levels were considerably higher by radiographic grade. Conclusions: An increase in CTX-II per radiographic grade was observed in the case group, but no association was found between MATN3 and DOT1L genes and the risk of KOA in Mexican mestizos.


Asunto(s)
Colágeno Tipo II/orina , N-Metiltransferasa de Histona-Lisina/genética , Osteoartritis de la Rodilla , Fragmentos de Péptidos/orina , Polimorfismo de Nucleótido Simple , Adulto , Femenino , Humanos , Masculino , Proteínas Matrilinas/genética , México , Persona de Mediana Edad , Osteoartritis de la Rodilla/genética , Osteoartritis de la Rodilla/orina
15.
Clin Rheumatol ; 38(10): 2897-2907, 2019 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-31236747

RESUMEN

INTRODUCTION/OBJECTIVES: Articular cartilage is the target tissue of osteoarthritis (OA), and because it lacks capillary networks, the microenvironment is hypoxic. Hypoxia inducible factor-1 alpha (HIF-1α) regulates the homeostasis of this tissue. The aim of this study was to investigate whether genetic polymorphisms of the HIF-1α signaling pathway are involved in the development of knee OA. METHOD: We performed a case-control association study and genotyped 134 knee OA patients and 267 healthy controls. All participants were genotyped in order to evaluate 42 SNPs from 22 genes involved in the HIF-1α signaling pathway using the OpenArray technology. Gene-gene interactions (epistasis) were analyzed using the multifactor dimensionality reduction (MDR) method. RESULTS: The MDR analysis showed epistasis between AKT2 (rs8100018) and IGF1 (rs2288377), AKT2 (rs8100018) and IGF1 (rs35767), IGF1 (rs35767) and COL2A1 (rs1793953), and between GSK3B (rs6438552) and IGF1 (rs35767) polymorphisms, with information gain values of 21.24%, 8.37%, 9.93%, and 5.73%, respectively. Additionally, our model allowed us to identify high- and low-risk genotypes among COL2A1 rs1793953, GSK3B rs6438552, AKT2 rs8100018, and IGF1 rs35767 polymorphisms. CONCLUSIONS: Knowing the interactions of these polymorphisms involved in HIF-1α signaling pathway could provide a new diagnostic support tool to identify individuals at high risk of developing knee OA.


Asunto(s)
Subunidad alfa del Factor 1 Inducible por Hipoxia/genética , Osteoartritis de la Rodilla/genética , Osteoartritis de la Rodilla/fisiopatología , Polimorfismo de Nucleótido Simple , Transducción de Señal , Adulto , Capilares/patología , Estudios de Casos y Controles , Colágeno Tipo II/genética , Epistasis Genética , Femenino , Genotipo , Glucógeno Sintasa Quinasa 3 beta/genética , Haplotipos , Homeostasis , Humanos , Subunidad alfa del Factor 1 Inducible por Hipoxia/metabolismo , Factor I del Crecimiento Similar a la Insulina/genética , Masculino , México , Persona de Mediana Edad , Modelos Genéticos , Proteínas Proto-Oncogénicas c-akt/genética , Riesgo
16.
Vet Clin Pathol ; 48(2): 320-327, 2019 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-31210381

RESUMEN

BACKGROUND: The accumulation of cartilage breakdown products in body fluids has been extensively investigated to assess the accuracy of molecular biomarkers from a diagnostic, prognostic, and therapeutic perspective. Nevertheless, to the authors' knowledge, there is a lack of information about spontaneous models of hip osteoarthritis and the differentiating ability of collagen, noncollagen, and inflammatory biomarkers. OBJECTIVES: We aimed to assess the accuracy of four plasma biomarkers that could differentiate between healthy dogs and dogs with hip dysplasia. METHODS: Twenty-four dogs were used in this institutionally approved study (12 in the mild to severe hip dysplasia group; 12 in the control group). Plasma concentrations of biomarkers were compared. The ability of each marker to differentiate control from diseased dogs was assessed using an independent t-test, logistic regression, and receiving operating characteristics (ROC) analysis. RESULTS: Three biomarkers were significantly different between the two groups. The collagen marker procollagen type II propeptide (PIICP) was useful in differentiating between control and diseased dogs with the best combination of sensitivity and specificity. The four biomarkers showed high area under the curve (AUC) values. CONCLUSIONS: The results indicate that plasma biomarkers can be used as a screening tool for canine hip dysplasia. Although the cutoff values and diagnostic ability of the biomarkers used in this study show promising results, the sources of individual variability should be addressed. Future studies with larger groups of dogs are needed to correlate plasma levels in serum and synovial fluid during clinical disease.


Asunto(s)
Proteínas de Unión al Calcio/sangre , Colágeno Tipo II/sangre , Colágeno/sangre , Displasia Pélvica Canina/sangre , Animales , Biomarcadores/sangre , Perros , Femenino , Displasia Pélvica Canina/diagnóstico , Masculino , Pronóstico , Líquido Sinovial/química
17.
Cells ; 8(4)2019 04 11.
Artículo en Inglés | MEDLINE | ID: mdl-30979017

RESUMEN

Changes in extracellular matrix (ECM) components in the lungs are associated with the progression of respiratory diseases, such as asthma, chronic obstructive pulmonary disease (COPD), and acute respiratory distress syndrome (ARDS). Experimental and clinical studies have revealed that structural changes in ECM components occur under chronic inflammatory conditions, and these changes are associated with impaired lung function. In bronchial asthma, elastic and collagen fiber remodeling, mostly in the airway walls, is associated with an increase in mucus secretion, leading to airway hyperreactivity. In COPD, changes in collagen subtypes I and III and elastin, interfere with the mechanical properties of the lungs, and are believed to play a pivotal role in decreased lung elasticity, during emphysema progression. In ARDS, interstitial edema is often accompanied by excessive deposition of fibronectin and collagen subtypes I and III, which can lead to respiratory failure in the intensive care unit. This review uses experimental models and human studies to describe how inflammatory conditions and ECM remodeling contribute to the loss of lung function in these respiratory diseases.


Asunto(s)
Remodelación de las Vías Aéreas (Respiratorias) , Asma/fisiopatología , Matriz Extracelular/patología , Pulmón/patología , Enfermedad Pulmonar Obstructiva Crónica/fisiopatología , Síndrome de Dificultad Respiratoria/fisiopatología , Animales , Hiperreactividad Bronquial/metabolismo , Colágeno Tipo I/metabolismo , Colágeno Tipo II/metabolismo , Modelos Animales de Enfermedad , Fibronectinas/metabolismo , Humanos
18.
Cartilage ; 10(2): 157-172, 2019 04.
Artículo en Inglés | MEDLINE | ID: mdl-28933195

RESUMEN

OBJECTIVE: Hyaline cartilage degenerative pathologies induce morphologic and biomechanical changes resulting in cartilage tissue damage. In pursuit of therapeutic options, electrical and mechanical stimulation have been proposed for improving tissue engineering approaches for cartilage repair. The purpose of this review was to highlight the effect of electrical stimulation and mechanical stimuli in chondrocyte behavior. DESIGN: Different information sources and the MEDLINE database were systematically revised to summarize the different contributions for the past 40 years. RESULTS: It has been shown that electric stimulation may increase cell proliferation and stimulate the synthesis of molecules associated with the extracellular matrix of the articular cartilage, such as collagen type II, aggrecan and glycosaminoglycans, while mechanical loads trigger anabolic and catabolic responses in chondrocytes. CONCLUSION: The biophysical stimuli can increase cell proliferation and stimulate molecules associated with hyaline cartilage extracellular matrix maintenance.


Asunto(s)
Cartílago Articular/citología , Condrocitos/fisiología , Cartílago Hialino/citología , Osteoartritis/fisiopatología , Estimulación Física/métodos , Agrecanos/fisiología , Animales , Cartílago Articular/fisiopatología , Proliferación Celular/fisiología , Colágeno Tipo II/fisiología , Estimulación Eléctrica/métodos , Terapia por Estimulación Eléctrica/métodos , Matriz Extracelular/fisiología , Glicosaminoglicanos/fisiología , Humanos , Cartílago Hialino/fisiopatología , Ingeniería de Tejidos/métodos
19.
Arch Endocrinol Metab ; 62(4): 438-445, 2018 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-30304108

RESUMEN

OBJECTIVE: This study evaluated the effects of combination therapy of curcumin and alendronate on BMD and bone turnover markers in postmenopausal women with osteoporosis. SUBJECTS AND METHODS: In a randomized, double-blind trial study, 60 postmenopausal women were divided into three groups: control, alendronate, and alendronate + curcumin. Each group included 20 patients. Total body, total hip, lumbar spine and femoral neck BMDs were measured by dual-energy X-ray absorptiometry (DXA) at baseline and after 12 months of therapy. Bone turnover markers such as bone-specific alkaline phosphatase (BALP), osteocalcin and C-terminal cross-linking telopeptide of type I collagen (CTx) were measured at the outset and 6 months later. RESULTS: Patients in the control group suffered a significant decrease in BMD and increased bone turnover markers at the end of study. The group treated with only alendronate showed significantly decreased levels of BALP and CTx and increased levels of osteocalcin compared to the control group. The alendronate group also showed significant increases in the total body, total hip, lumbar spine and femoral neck BMDs at the end of study compared to the control group. In the curcumin + alendronate group, BALP and CTx levels decreased and osteocalcin levels increased significantly at the end of study compared to the control and alendronate groups. BMD indexes also increased in four areas significantly at the end of study compared to the control and alendronate groups. CONCLUSION: The combination of curcumin and alendronate has beneficial effects on BMD and bone turnover markers among postmenopausal women with osteoporosis. Arch Endocrinol Metab. 2018;62(4):438-45.


Asunto(s)
Alendronato/farmacología , Conservadores de la Densidad Ósea/farmacología , Densidad Ósea/efectos de los fármacos , Curcumina/farmacología , Osteoporosis Posmenopáusica/metabolismo , Anciano , Fosfatasa Alcalina/análisis , Fosfatasa Alcalina/efectos de los fármacos , Remodelación Ósea/efectos de los fármacos , Colágeno Tipo II/efectos de los fármacos , Colágeno Tipo II/orina , Método Doble Ciego , Quimioterapia Combinada/métodos , Femenino , Humanos , Persona de Mediana Edad , Osteocalcina/análisis , Osteocalcina/efectos de los fármacos , Fragmentos de Péptidos/efectos de los fármacos , Fragmentos de Péptidos/orina
20.
Arch. endocrinol. metab. (Online) ; 62(4): 438-445, July-Aug. 2018. tab, graf
Artículo en Inglés | LILACS | ID: biblio-950087

RESUMEN

ABSTRACT Objective: This study evaluated the effects of combination therapy of curcumin and alendronate on BMD and bone turnover markers in postmenopausal women with osteoporosis. Subjects and methods: In a randomized, double-blind trial study, 60 postmenopausal women were divided into three groups: control, alendronate, and alendronate + curcumin. Each group included 20 patients. Total body, total hip, lumbar spine and femoral neck BMDs were measured by dual-energy X-ray absorptiometry (DXA) at baseline and after 12 months of therapy. Bone turnover markers such as bone-specific alkaline phosphatase (BALP), osteocalcin and C-terminal cross-linking telopeptide of type I collagen (CTx) were measured at the outset and 6 months later. Results: Patients in the control group suffered a significant decrease in BMD and increased bone turnover markers at the end of study. The group treated with only alendronate showed significantly decreased levels of BALP and CTx and increased levels of osteocalcin compared to the control group. The alendronate group also showed significant increases in the total body, total hip, lumbar spine and femoral neck BMDs at the end of study compared to the control group. In the curcumin + alendronate group, BALP and CTx levels decreased and osteocalcin levels increased significantly at the end of study compared to the control and alendronate groups. BMD indexes also increased in four areas significantly at the end of study compared to the control and alendronate groups. Conclusion: The combination of curcumin and alendronate has beneficial effects on BMD and bone turnover markers among postmenopausal women with osteoporosis. Arch Endocrinol Metab. 2018;62(4):438-45


Asunto(s)
Humanos , Femenino , Persona de Mediana Edad , Anciano , Densidad Ósea/efectos de los fármacos , Osteoporosis Posmenopáusica/metabolismo , Alendronato/farmacología , Curcumina/farmacología , Conservadores de la Densidad Ósea/farmacología , Fragmentos de Péptidos/efectos de los fármacos , Fragmentos de Péptidos/orina , Osteocalcina/análisis , Osteocalcina/efectos de los fármacos , Método Doble Ciego , Remodelación Ósea/efectos de los fármacos , Colágeno Tipo II/efectos de los fármacos , Colágeno Tipo II/orina , Quimioterapia Combinada/métodos , Fosfatasa Alcalina/análisis , Fosfatasa Alcalina/efectos de los fármacos
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