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2.
Acta Parasitol ; 68(3): 705-710, 2023 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-37531009

RESUMEN

PURPOSE: This study aims to develop and evaluate a cost-effective, user-friendly multiplex quantitative real-time polymerase chain reaction (qPCR) method for detecting multiple tick-borne pathogens associated with human and veterinary diseases. METHODS: In silico PCR was performed to design and evaluate primer sequences reported for amplifying Rickettsia spp., Borrelia spp., and Ehrlichia spp. Single and multiplex qPCR assays were then standardized to detect individual pathogens and multiple pathogens in a single reaction. Positive controls were generated to determine the dynamic range of the methods. In the validation phase, a total of 800 samples were screened for the presence of tick-borne pathogens. RESULTS: Identification in a single qPCR reaction (multiplex) of Ehrlichia spp., and Borrelia spp. with a limit of detection of 10 copies and Rickettsia spp. with 100 copies, a PCR efficiency (E) of 90-100% and a coefficient of correlation (R2) of 0.998-0.996 for all pathogens. CONCLUSION: The ability to detect three significant pathogens (Ehrlichia spp., Rickettsia spp., and Borrelia spp.) in a single qPCR reaction offers a significant advantage in the field of molecular diagnostics for tick-borne diseases. This advancement has a profound impact on public health as it facilitates the selection of appropriate treatment protocols, thereby reducing complications associated with disease progression. The streamlined approach provided by this method simplifies the diagnostic process and enables timely intervention, ultimately improving patient outcomes and mitigating the potential risks associated with untreated or misdiagnosed tick-borne infections.


Asunto(s)
Borrelia , Rickettsia , Enfermedades por Picaduras de Garrapatas , Garrapatas , Animales , Humanos , Garrapatas/microbiología , Reacción en Cadena en Tiempo Real de la Polimerasa , Rickettsia/genética , Ehrlichia/genética , Enfermedades por Picaduras de Garrapatas/diagnóstico , Enfermedades por Picaduras de Garrapatas/veterinaria , Borrelia/genética
3.
Life (Basel) ; 13(8)2023 Aug 11.
Artículo en Inglés | MEDLINE | ID: mdl-37629580

RESUMEN

BACKGROUND: The presence of side effects and low bioavailability of rhein has limited its use in the treatment of osteoarthritis. We aimed to evaluate the in vitro response of human articular chondrocytes to the presence of the combination of platelet-rich plasma (PRP) and rhein. METHODS: Solutions of rhein were prepared to assess solubility and select a working concentration. A stimulus with interleukin-1ß (IL-ß, 10 ng/mL) was induced for 24 h on human chondrocytes. Five treatment groups were established: control, IL-ß control, PRP, rhein, and PRP + rhein. Cell viability, cell migration, nitric oxide (NO) production, tumor necrosis factor-α (TNF-α), and gene expression analyses were carried out. RESULTS: A concentration of 50 mg/L was selected after a dose-response curve assay. Both NO and tumor TNF-α production significantly decreased after PRP and PRP + rhein treatments at 24 and 48 h. The wound healing assay revealed a significant stimulation of migration after 72 h with the PRP and PRP + rhein treatments. Expression of IL-1ß, IL-6, MMP-13, and ADAMTS-5 was significantly downregulated, particularly after treatment with the combination of PRP + rhein. CONCLUSIONS: Much of the determinations denoted a better performance of the combination of PRP and rhein in decreasing the levels of the different targets evaluated; however, this was not great enough to detect a significant difference in comparison with the PRP treatment alone.

4.
Curr Issues Mol Biol ; 44(12): 5827-5838, 2022 Nov 22.
Artículo en Inglés | MEDLINE | ID: mdl-36547057

RESUMEN

Background: Achilles-tendon rupture prevails as a common tendon pathology. Adipose-derived mesenchymal stem cells (ADMSCs) are multipotent stem cells derived from adipose tissue with attractive regeneration properties; thus, their application in tendinopathies could be beneficial. Methods: Male rabbit ADMSCs were obtained from the falciform ligament according to previously established methods. After tenotomy and suture of the Achilles tendon, 1 × 106 flow-cytometry-characterized male ADMSCs were injected in four female New Zealand white rabbits in the experimental group (ADMSC group), whereas four rabbits were left untreated (lesion group). Confirmation of ADMSC presence in the injured site after 12 weeks was performed with quantitative sex-determining region Y (SRY)-gene RT-PCR. At Week 12, histochemical analysis was performed to evaluate tissue regeneration along with quantitative RT-PCR of collagen I and collagen III mRNA. Results: Presence of male ADMSCs was confirmed at Week 12. No statistically significant differences were found in the histochemical analysis; however, statistically significant differences between ADMSC and lesion group expression of collagen I and collagen III were evidenced, with 36.6% and 24.1% GAPDH-normalized mean expression, respectively, for collagen I (p < 0.05) and 26.3% and 11.9% GAPDH-normalized mean expression, respectively, for collagen III (p < 0.05). The expression ratio between the ADMSC and lesion group was 1.5 and 2.2 for collagen I and collagen III, respectively. Conclusion: Our results make an important contribution to the understanding and effect of ADMSCs in Achilles-tendon rupture.

5.
Exp Ther Med ; 22(5): 1282, 2021 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-34630637

RESUMEN

Joint cartilage damage affects 10-12% of the world's population. Medical treatments improve the short-term quality of life of affected individuals but lack a long-term effect due to injury progression into fibrocartilage. The use of mesenchymal stem cells (MSCs) is one of the most promising strategies for tissue regeneration due to their ability to be isolated, expanded and differentiated into metabolically active chondrocytes to achieve long-term restoration. For this purpose, human adipose-derived MSCs (Ad-MSCs) were isolated from lipectomy and grown in xeno-free conditions. To establish the best differentiation potential towards a stable chondrocyte phenotype, isolated Ad-MSCs were sequentially exposed to five differentiation schemes of growth factors in previously designed three-dimensional biphasic scaffolds with incorporation of a decellularized cartilage matrix as a bioactive ingredient, silk fibroin and bone matrix, to generate a system capable of being loaded with pre-differentiated Ad-MSCs, to be used as a clinical implant in cartilage lesions for tissue regeneration. Chondrogenic and osteogenic markers were analyzed by reverse transcription-quantitative PCR and cartilage matrix generation by histology techniques at different time points over 40 days. All groups had an increased expression of chondrogenic markers; however, the use of fibroblast growth factor 2 (10 ng/ml) followed by a combination of insulin-like growth factor 1 (100 ng/ml)/TGFß1 (10 ng/ml) and a final step of exposure to TGFß1 alone (10 ng/ml) resulted in the most optimal chondrogenic signature towards chondrocyte differentiation and the lowest levels of osteogenic expression, while maintaining stable collagen matrix deposition until day 33. This encourages their possible use in osteochondral lesions, with appropriate properties for use in clinical patients.

6.
Int J Mol Sci ; 20(20)2019 Oct 17.
Artículo en Inglés | MEDLINE | ID: mdl-31627374

RESUMEN

In cartilage tissue engineering, biphasic scaffolds (BSs) have been designed not only to influence the recapitulation of the osteochondral architecture but also to take advantage of the healing ability of bone, promoting the implant's integration with the surrounding tissue and then bone restoration and cartilage regeneration. This study reports the development and characterization of a BS based on the assembly of a cartilage phase constituted by fibroin biofunctionalyzed with a bovine cartilage matrix, cellularized with differentiated autologous pre-chondrocytes and well attached to a bone phase (decellularized bovine bone) to promote cartilage regeneration in a model of joint damage in pigs. BSs were assembled by fibroin crystallization with methanol, and the mechanical features and histological architectures were evaluated. The scaffolds were cellularized and matured for 12 days, then implanted into an osteochondral defect in a porcine model (n = 4). Three treatments were applied per knee: Group I, monophasic cellular scaffold (single chondral phase); group II (BS), cellularized only in the chondral phase; and in order to study the influence of the cellularization of the bone phase, Group III was cellularized in chondral phases and a bone phase, with autologous osteoblasts being included. After 8 weeks of surgery, the integration and regeneration tissues were analyzed via a histology and immunohistochemistry evaluation. The mechanical assessment showed that the acellular BSs reached a Young's modulus of 805.01 kPa, similar to native cartilage. In vitro biological studies revealed the chondroinductive ability of the BSs, evidenced by an increase in sulfated glycosaminoglycans and type II collagen, both secreted by the chondrocytes cultured on the scaffold during 28 days. No evidence of adverse or inflammatory reactions was observed in the in vivo trial; however, in Group I, the defects were not reconstructed. In Groups II and III, a good integration of the implant with the surrounding tissue was observed. Defects in group II were fulfilled via hyaline cartilage and normal bone. Group III defects showed fibrous repair tissue. In conclusion, our findings demonstrated the efficacy of a biphasic and bioactive scaffold based on silk fibroin and cellularized only in the chondral phase, which entwined chondroinductive features and a biomechanical capability with an appropriate integration with the surrounding tissue, representing a promising alternative for osteochondral tissue-engineering applications.


Asunto(s)
Regeneración Ósea , Ingeniería de Tejidos/métodos , Animales , Cartílago , Diferenciación Celular , Condrocitos , Fibroínas , Porcinos , Andamios del Tejido
7.
Stem Cells Int ; 2019: 9792369, 2019.
Artículo en Inglés | MEDLINE | ID: mdl-31149016

RESUMEN

Articular cartilage injuries remain as a therapeutic challenge due to the limited regeneration potential of this tissue. Cartilage engineering grafts combining chondrogenic cells, scaffold materials, and microenvironmental factors are emerging as promissory alternatives. The design of an adequate scaffold resembling the physicochemical features of natural cartilage and able to support chondrogenesis in the implants is a crucial topic to solve. This study reports the development of an implant constructed with IGF1-transduced adipose-derived mesenchymal stem cells (immunophenotypes: CD105+, CD90+, CD73+, CD14-, and CD34-) embedded in a scaffold composed of a mix of alginate/milled bovine decellularized knee material which was cultivated in vitro for 28 days (3CI). Histological analyses demonstrated the distribution into isogenous groups of chondrocytes surrounded by a de novo dense extracellular matrix with balanced proportions of collagens II and I and high amounts of sulfated proteoglycans which also evidenced adequate cell proliferation and differentiation. This graft also shoved mechanical properties resembling the natural knee cartilage. A modified Bern/O'Driscoll scale showed that the 3CI implants had a significantly higher score than the 2CI implants lacking cells transduced with IGF1 (16/18 vs. 14/18), representing high-quality engineering cartilage suitable for in vivo tests. This study suggests that this graft resembles several features of typical hyaline cartilage and will be promissory for preclinical studies for cartilage regeneration.

8.
Biomed Mater ; 13(3): 035001, 2018 02 20.
Artículo en Inglés | MEDLINE | ID: mdl-29461975

RESUMEN

Ceramics and bioceramics, such as hydroxyapatite and zirconium, are used in bone tissue engineering. Hydroxyapatite has chemical properties similar to bone while zirconium offers suitable mechanical properties. The aim of this article is to evaluate the ability to support cell growth and osteoblastic mineralization of hydroxyapatite-zirconium obtained by a new system based on different low temperatures, such as 873 K (HZ600), 923 K (HZ650) and 973 K (HZ700). Hydroxyapatite-zirconia obtained by this new system was examined in terms of thermogravimetric features and x-ray diffractograms. Furthermore, the ability for supporting osteoblast growth and mineralization were analyzed. By x-ray diffraction analysis, we clearly demonstrated that no high-temperature processing was required. Moreover, it is possible to form tetragonal-zirconium at 923 K. Proliferation assays showed that osteoblast growth was not influenced by any of the composite evaluated. Regarding the osteogenic marker Col1, a 2-fold increase in expression was observed for HZ650 compared to HZ600 and HZ700. Interestingly, osteoblasts grown on HZ650 showed globular accretions covered with collagen bundles and calcium-rich extracellular matrix whereas HZ600 and HZ700 showed no phosphate or calcium deposits. This study demonstrated that at 923 K it is possible to generate stable tetragonal-zirconium and the resulting HZ650 composite is able to promote a suitable osteoblast mineralization process.


Asunto(s)
Durapatita/química , Osteoblastos/citología , Circonio/química , Huesos/metabolismo , Calcio/química , Diferenciación Celular , Proliferación Celular , Frío , Colágeno/química , Matriz Extracelular/metabolismo , Femenino , Humanos , Osteoblastos/metabolismo , Placenta/metabolismo , Embarazo , Temperatura , Andamios del Tejido , Difracción de Rayos X
9.
Histol Histopathol ; 33(6): 609-618, 2018 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-29313321

RESUMEN

OBJECTIVE: To determine the effects of autologous leukocyte-poor platelet-rich plasma (LP-PRP) on the expression of markers involved in cartilage-extracellular matrix production and inflammation in cartilage explants bearing osteoarthritis. MATERIALS AND METHODS: Cartilage explants and LP-PRP were obtained from 10 patients who underwent total knee arthroplasty. The explants were cultured in spinner flasks for 28 days in the presence of interleukin (IL)-1ß and/or LP-PRP. The gene expression of catabolic (MMP13, ADAMTS5, and IL1ß) and anabolic factors (COL2A1, ACAN, and SOX9) was quantified. A histological assessment was performed according to a modified Mankin score, and quantification of type II and I collagen deposition. RESULTS: The gene expression of catabolic factors and the Mankin score were lower in LP-PRP- and LP-PRP/IL-1ß- than in IL-1ß-treated explants, suggesting less matrix degradation in explants cultured in the presence of LP-PRP. Higher expression of genes involved in cartilage matrix restoration was observed in LP-PRP and LP-PRP/IL-1ß- when compared to IL-1ß-treated explants. The explants treated with LP-PRP and LP-PRP/IL-1ß exhibited a higher deposition of type II collagen as well as a lower deposition of type I collagen and also better surface integrity and a significant increase in the number of chondrocytes. CONCLUSION: LP-PRP treatment favored restoration in early osteoarthritic cartilage and reduced the pro-inflammatory effect of IL-1ß. LP-PRP is a promising therapy for early osteoarthritis, as it promotes extracellular matrix repair, reduces inflammation, and slows cartilage degeneration.


Asunto(s)
Cartílago Articular/metabolismo , Osteoartritis , Plasma Rico en Plaquetas , Condrocitos/metabolismo , Matriz Extracelular , Femenino , Humanos , Masculino , Persona de Mediana Edad , Técnicas de Cultivo de Órganos , Osteoartritis/metabolismo , Plasma Rico en Plaquetas/citología
10.
Gen Physiol Biophys ; 36(4): 373-381, 2017 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-28653651

RESUMEN

Melittin is the main component of bee venom consisting of 26 amino acids that has multiple effects, including antibacterial, antiviral and anti-inflammatory in various cell types. This peptide forms pores in biological membranes and triggers cell death. Therefore it has potential as an anti-cancer therapy. However, the therapeutic application of melittin is limited due to its main side effect, hemolysis, which is especially pronounced following intravenous administration. In the present study, we formulated tetrameric melittin-carrying poly-D,L-lactic-co-glycolic acid nanoparticles (PLGA-NPs) and analyzed the lytic activity of this system on liposomes that resembles breast cancer cells. Tetrameric melittin binds avidly to PLGA-NPs with an encapsulation efficiency of 97% and retains its lytic activity demonstrating the effectiveness of PLGA-NPs as nanocarriers for this cytolytic peptide.


Asunto(s)
Preparaciones de Acción Retardada/química , Liposomas/química , Meliteno/química , Fluidez de la Membrana/efectos de los fármacos , Nanocápsulas/química , Nanocápsulas/ultraestructura , Implantes Absorbibles , Difusión , Diseño de Fármacos , Meliteno/administración & dosificación
11.
Cir Cir ; 85(2): 127-134, 2017.
Artículo en Español | MEDLINE | ID: mdl-27633464

RESUMEN

BACKGROUND: A fracture repair involves complex cellular processes. However, despite optimal treatment, some fractures heal slowly or do not repair. These complications support the need for innovative therapies. Electromagnetic stimulation is a non-invasive technology that could have a direct impact on many cellular pathways. OBJECTIVE: To demonstrate the effectiveness of electro-stimulation by alternating current applied during bone elongation to accelerate the consolidation process for 30 days in an animal model. MATERIALS AND METHODS: A device with closed circuit and graduated voltage was designed and kept in contact with the external fixator. Group A was elongated without electro-stimulation and group B was electro-stimulated since the beginning of the distraction. Radiographs were taken at 15 and 30 days post-surgical. Haematoxylin and eosin staining and Masson's trichrome stain were performed. RESULTS: No significant difference were observed in bone density of group A (4.05±3.24, P=0.163). In group B there was a significant difference (61.06±20.17, P=0.03) in bone density. Group A maintained a fibrous tissue repair, with areas of cartilage and bone matrix. Group B had more organised tissue in the stages of bone repair. CONCLUSION: Because there is a significant difference in the growth and callus formation at 15 and 30 days between groups, electro-stimulation could be considered as an adjuvant during bone elongation.


Asunto(s)
Alargamiento Óseo/métodos , Terapia por Estimulación Eléctrica , Fijación de Fractura , Animales , Huesos , Perros , Terapia por Estimulación Eléctrica/métodos , Fijadores Externos , Modelos Animales
12.
Stem Cells Int ; 2016: 7403890, 2016.
Artículo en Inglés | MEDLINE | ID: mdl-27818692

RESUMEN

Adipose-derived mesenchymal stem cells (ADMSCs) are inducible to an osteogenic phenotype by the bone morphogenetic proteins (BMPs). This facilitates the generation of implants for bone tissue regeneration. This study evaluated the in vitro osteogenic differentiation of ADMSCs transduced individually and in combination with adenoviral vectors expressing BMP2 and BMP7. Moreover, the effectiveness of the implant containing ADMSCs transduced with the adenoviral vectors AdBMP2/AdBMP7 and embedded in demineralized bone matrix (DBM) was tested in a model of tibial fracture in sheep. This graft was compared to ewes implanted with untransduced ADMSCs embedded in the same matrix and with injured but untreated animals. In vivo results showed accelerated osteogenesis in the group treated with the AdBMP2/AdBMP7 transduced ADMSC graft, which also showed improved restoration of the normal bone morphology.

13.
Arch Orthop Trauma Surg ; 136(12): 1723-1732, 2016 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-27506585

RESUMEN

INTRODUCTION: Knee osteoarthritis (OA) is a degenerative and progressive articular cartilage disease. Infiltration of autologous platelet-rich plasma (PRP) has been proposed as a therapeutic alternative due to the content of biologically active cytokines in PRP. We aimed to compare the clinical response of acetaminophen and intra-articular leukocyte-poor PRP (LP-PRP) in early knee OA. MATERIALS AND METHODS: A total of 65 patients with clinically and radiographically documented knee OA (grade 1-2) were analyzed. Patients were randomized into two groups: 32 were treated with acetaminophen (500 mg/8 h) over 6 weeks, and 33 received three intra-articular injections of autologous LP-PRP (once every 2 weeks). All patients were evaluated by the Visual Analogue Scale (VAS), the Western Ontario and McMaster Universities (WOMAC) score, and the SF-12 health survey at baseline and 6, 12, and 24 weeks of follow-up. All LP-PRP preparations were analyzed for the platelet, leukocyte, IL-1ra, and TGF-ß concentrations. RESULTS: The decrease in the VAS pain level in the LP-PRP group was greater than that in the acetaminophen group (p < 0.05). Patients treated with LP-PRP showed a sustained improvement in knee function at week 24 (p < 0.01). The SF-12 results only indicated an improvement in quality-of-life in the LP-PRP group at 6, 12, and 24 weeks of follow-up (p < 0.01). Both IL-1ra and TGF-ß were detected in the LP-PRP samples (313.8 ± 231.6 and 21,183.8 ± 8556.3 pg/mL, respectively). CONCLUSIONS: Treatment with LP-PRP injections resulted in a significantly better clinical outcome than did treatment with acetaminophen, with sustained lower EVA and WOMAC scores and improvement in quality-of-life (higher SF-12 score). Therapy with LP-PRP may positively modify the inflammatory joint environment by counteracting IL-1ß action.


Asunto(s)
Acetaminofén/uso terapéutico , Artralgia/terapia , Osteoartritis de la Rodilla/terapia , Plasma Rico en Plaquetas , Analgésicos no Narcóticos/uso terapéutico , Artralgia/diagnóstico , Artralgia/etiología , Femenino , Humanos , Inyecciones Intraarticulares , Articulación de la Rodilla , Masculino , Persona de Mediana Edad , Osteoartritis de la Rodilla/complicaciones , Osteoartritis de la Rodilla/fisiopatología , Dimensión del Dolor , Resultado del Tratamiento
14.
Gac Med Mex ; 152(3): 381-5, 2016.
Artículo en Español | MEDLINE | ID: mdl-27335195

RESUMEN

Since the introduction of bone morphogenetic proteins, their use has become an invaluable ally for the treatment of bone defects. These proteins are potent growth factors, related to angiogenic and osteogenic activity. The osteoinductive capacity of recombinant bone morphogenetic protein (rhBMP) in the formation of bone and cartilage has been confirmed in in vitro studies and evaluated in clinical trials. To obtain a therapeutic effect, administration is systemic, by injection over the physiological dose. Among the disadvantages, ectopic bone formation or high morbidity in cases of spinal fusion is observed. In this review, the roles of bone morphogenetic proteins in bone repair and clinical applications are analyzed. These findings represent advances in the study of bone regeneration and application of growth factors for more predictable results.


Asunto(s)
Enfermedades Óseas/terapia , Proteínas Morfogenéticas Óseas/administración & dosificación , Regeneración Ósea/efectos de los fármacos , Enfermedades Óseas/patología , Proteínas Morfogenéticas Óseas/efectos adversos , Humanos , Inyecciones , Osificación Heterotópica/inducido químicamente , Osteogénesis/efectos de los fármacos , Proteínas Recombinantes/administración & dosificación , Proteínas Recombinantes/efectos adversos , Fusión Vertebral/métodos
15.
Mycopathologia ; 181(1-2): 151-7, 2016 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-26419685

RESUMEN

Coccidioidomycosis is a respiratory fungal infection with occasional systemic dissemination. The disseminated coccidioidomycosis is considered a multifaceted disease. In medicine, disseminated coccidioidomycosis is included within a group of infectious diseases that have been referred as the great imitators. In many cases, malignancies are included in the presumptive diagnosis. In veterinary medicine, disseminated coccidioidomycosis is common in dogs. Nonetheless, despite of being a diagnostic dilemma, disseminated coccidioidomycosis is underestimated and frequently not included into differentials, even in endemic zones. Herein, we describe three cases of granulomatous inflammation caused by Coccidioides spp. which were masquerading malignancies in dogs (0.39 %). The presumptive diagnoses in these cases were osteosarcoma, lymphoma and neurofibroma, respectively. A PCR assay employing tissues in paraffin blocks resulted positive for C. posadasii in one of these cases. A comparative discussion on the ambiguous clinic-pathological presentation of disseminated coccidioidomycosis in dogs and humans is included.


Asunto(s)
Biopsia , Coccidioides/aislamiento & purificación , Coccidioidomicosis/veterinaria , Neoplasias/veterinaria , Animales , Coccidioides/genética , Coccidioidomicosis/diagnóstico , Coccidioidomicosis/patología , Diagnóstico Diferencial , Perros , Neoplasias/diagnóstico , Neoplasias/patología , Patología Molecular , Reacción en Cadena de la Polimerasa
17.
Stem Cells Dev ; 24(21): 2577-90, 2015 Nov 01.
Artículo en Inglés | MEDLINE | ID: mdl-26230358

RESUMEN

Cellular adhesion enables communication between cells and their environment. Adhesion can be achieved throughout focal adhesions and its components influence osteoblast differentiation of human mesenchymal stem cells (hMSCs). Because cell adhesion and osteoblast differentiation are closely related, this article aimed to analyze the expression profiles of adhesion-related proteins during osteoblastic differentiation of two hMSCs subpopulations (CD105(+) and CD105(-)) and propose a strategy for assembling bone grafts based on its adhesion ability. In vitro experiments of osteogenic differentiation in CD105(-) cells showed superior adhesion efficiency and 2-fold increase of α-actinin expression compared with CD105(+) cells at the maturation stage. Interestingly, levels of activated ß1-integrin increased in CD105(-) cells during the process. Additionally, the CD105(-) subpopulation showed 3-fold increase of phosphorylated FAK(Y397) compared to CD105(+) cells. Results also indicate that ERK1/2 was activated during CD105(-) bone differentiation and participation of mitogen-activated protein kinase (MAPK)-p38 in CD105(+) differentiation through a focal adhesion kinase (FAK)-independent pathway. In vivo trial demonstrated that grafts containing CD105(-) showed osteocytes embedded in a mineralized matrix, promoted adequate graft integration, increased host vascular infiltration, and efficient intramembranous repairing. In contrast, grafts containing CD105(+) showed deficient endochondral ossification and fibrocartilaginous tissue. Based on the expression of α-actinin, FAKy,(397) and ERK1/2 activation, we define maturation stage as critical for bone graft assembling. By in vitro assays, CD105(-) subpopulation showed superior adhesion efficiency compared to CD105(+) cells. Considering in vitro and in vivo assays, this study suggests that integration of a scaffold with CD105(-) subpopulation at the maturation stage represents an attractive strategy for clinical use in orthopedic bioengineering.


Asunto(s)
Diferenciación Celular/fisiología , Quinasas MAP Reguladas por Señal Extracelular/metabolismo , Células Madre Mesenquimatosas/citología , Osteoblastos/citología , Osteogénesis/fisiología , Adhesión Celular/fisiología , Células Cultivadas , Femenino , Proteína-Tirosina Quinasas de Adhesión Focal/metabolismo , Humanos , Fosforilación , Transducción de Señal/fisiología , Proteínas Quinasas p38 Activadas por Mitógenos/metabolismo
18.
Int. j. morphol ; 32(4): 1199-1206, Dec. 2014. ilus
Artículo en Español | LILACS | ID: lil-734659

RESUMEN

La osteoartrosis es un padecimiento del aparato locomotor con una prevalencia elevada y en crecimiento, paralela al envejecimiento de la población. La infiltración intraarticular de sustancias para aliviar la sintomatología de la osteoartrosis es una práctica común en el consultorio médico de los especialistas que atienden esta enfermedad. Aunque la sintomatología mejora con la infiltración de anestésicos locales, corticoesteroides y suplementos viscosantes, es aún incierto el efecto que estas sustancias tienen sobre la integridad del cartílago articular. Este estudio explora a nivel macroscópico e histológico el efecto de la infiltración de ropivacaína, metilprednisolona y ácido hialurónico sobre el cartílago articular en un modelo de osteoartrosis química en conejos (n=24). Nuestros resultados indican que en los grupos infiltrados con metilprednisolona (n=8) y ropivacaína (n=8) la estructura del cartílago articular presento alteraciones más severas con respecto a su grupo control, además de una disminución importante en la síntesis de matriz extracelular. En el grupo infiltrado con ácido hialurónico (n=8), las alteraciones macroscópicas e histológicas del cartílago articular mejoraron con respecto a su grupo control, presentando una estructura integra y síntesis de matriz extracelular normal.


Osteoarthritis is a musculoskeletal condition with a high prevalence, increasing with the aging of population. The intraarticular infiltration of substances to relieve the symptoms of osteoarthritis is a common practice in medical practice. Although symptoms improved with the infiltration of local anesthetics, corticosteroids and supplements, it is still uncertain what effect these substances have on the integrity of articular cartilage. This study explores the macroscopic and histological effects of infiltration of Ropivacaine, Methylprednisolone and Hyaluronic Acid on articular cartilage in a model of chemical osteoarthritis in rabbits (n=24). Our results indicate that in the infiltrated groups with Methylprednisolone (n=8) and Ropivacaine (n=8) the structure of articular cartilage present more severe alterations with respect to its control group and an important decrease in the synthesis of extracellular matrix. In-group infiltrated with hyaluronic acid (n=8), macroscopic and histological changes of articular cartilage improved with respect to its control group, presenting a normal structure and normal extracellular matrix synthesis.


Asunto(s)
Animales , Masculino , Conejos , Metilprednisolona/administración & dosificación , Cartílago Articular/efectos de los fármacos , Cartílago Articular/patología , Osteoartritis de la Rodilla , Amidas/administración & dosificación , Ácido Hialurónico/administración & dosificación , Metilprednisolona/farmacología , Modelos Animales de Enfermedad , Amidas/farmacología , Ácido Hialurónico/farmacología
19.
Cir Cir ; 82(2): 163-9, 2014.
Artículo en Español | MEDLINE | ID: mdl-25312315

RESUMEN

BACKGROUND: in the treatment of early osteoartrosis, analgesics and non-steroidal anti-inflammatory drugs are frequently used to relieve pain. Currently, platelet rich plasma is used as an alternative in the treatment of osteoartrosis. The aim of this study was to evaluate the effect of platelet rich plasma compared to paracetamol as a treatment for patients with knee osteoartrosis grade I. METHODS: we evaluated 42 patients who were randomized into two groups. Group one was treated with 5 mL of platelet rich plasma in two applications, while group two was treated with 1 gr of oral paracetamol every 8 hours for 30 days. Both patient groups received supervised physical rehabilitation during the 6 month observation period. Peripheral blood samples were taken to measure plasma IL-1ß, TNF-a and TGF-ß1 levels at day 0 and at 6 months post-treatment. Clinical evaluation was conducted using the KOOS at the start of the study and for every subsequent month during the study period. RESULTS: the Knee injury and Osteoarthritis Outcome Score (KOOS) for group one at the start of the treatment was measured at 30.1 points, whereas at the end, it was measured at 48.2 points, showing a clinical improvement of 60%. There were no statistically significant differences in IL-1ß and TNF-a levels between groups treated either with platelet rich plasma or paracetamol. CONCLUSIONS: Our patients treated with platelet rich plasma showed a statistically significant increase in the serum levels of TGF-ß1, which was associated with an improvement in the clinical evaluation used (KOOS).


Antecedentes: la osteoartrosis se caracteriza por dolor, rigidez articular y crepitación, con datos radiológicos específicos. El dolor se controla con analgésicos y antiinflamatorios no esteroides. En la actualidad, el plasma rico en plaquetas es una alternativa de tratamiento de esta enfermedad. Objetivo: comparar el efecto del plasma rico en plaquetas versus paracetamol en el tratamiento de pacientes con osteoartrosis de rodilla grado I. Material y métodos: ensayo prospectivo y experimental en el que se estudiaron 42 pacientes que se asignaron al azar a dos grupos. Al grupo 1 (experimental) se le indicaron 5 mL de plasma rico en plaquetas, en dos aplicaciones; el grupo 2 (control) se trató con 1 g de paracetamol oral cada 8 horas durante 30 días. Ambos grupos recibieron terapia física supervisada, con un tiempo de seguimiento total de seis meses. Para determinar las concentraciones de sangre periférica se tomaron muestras de IL-1ß, TNF-a y TGF-ß en plasma el día 0 y a los 6 meses. La valoración clínica se realizó con el Knee injury and Osteoarthritis Outcome Score (KOOS) al inicio y mensualmente durante el estudio. Resultados: los resultados de IL-1ß y TNF-a no mostraron diferencia significativa entre los grupos tratados con plasma rico en plaquetas y paracetamol. La escala KOOS para el grupo 1 al inicio del tratamiento fue de 30.1 puntos y al finalizar el tratamiento 48.2 puntos, con mejoría clínica de 60%. Conclusiones: los pacientes tratados con plasma rico en plaquetas tuvieron un incremento significativo en las concentraciones séricas de TGF-ß1 que se asoció con la mejoría clínica relacionada con el KOOS.


Asunto(s)
Acetaminofén/uso terapéutico , Osteoartritis de la Rodilla/terapia , Plasma Rico en Plaquetas , Acetaminofén/administración & dosificación , Administración Oral , Adulto , Animales , Biomarcadores , Femenino , Humanos , Interleucina-1beta/sangre , Recuento de Leucocitos , Masculino , Persona de Mediana Edad , Osteoartritis de la Rodilla/sangre , Osteoartritis de la Rodilla/tratamiento farmacológico , Osteoartritis de la Rodilla/rehabilitación , Estudios Prospectivos , Factor de Crecimiento Transformador beta1/sangre , Resultado del Tratamiento , Factor de Necrosis Tumoral alfa/análisis
20.
Nat Prod Commun ; 9(8): 1217-21, 2014 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-25233608

RESUMEN

In this review, the composition, actions, and clinical applications of acemannan in medicine and its effectiveness as an adjunct in the treatment of diseases are presented. An electronic literature search was performed up to January 2014 for studies and research presenting data to validate the efficacy of acemannan. A total of 50 titles, abstracts and full-text studies were selected and reviewed. Acemannan has various medicinal properties like osteogenic, anti-inflammatory, and antibacterial, which accelerate healing of lesions. Also, acemannan is known to have antiviral and antitumor activities in vivo through activation of immune responses. It was concluded that Aloe vera has immense potential as a therapeutic agent. Even though the plant is a promising herb with various clinical applications in medicine and dentistry, more clinical research needs to be undertaken to validate and explain the action of acemannan in healing, so that it can be established in the field of medicine and a more precise understanding of the biological activities of these is required to develop Aloe vera as a pharmaceutical source.


Asunto(s)
Aloe/química , Mananos/farmacología , Extractos Vegetales/farmacología , Animales , Antiinfecciosos/química , Antiinfecciosos/farmacología , Antiinflamatorios/química , Antiinflamatorios/farmacología , Humanos , Mananos/química , Extractos Vegetales/química
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