RESUMEN

Biomaterials serve as an integral component of tissue engineering. They are designed to provide architectural framework of native extracellular matrix so as to encourage cell growth and eventual tissue regeneration. Naturally occurring biopolymers as scaffolds offer options for cartilage tissue engineering due to anti-inflammatory, biocompatibility, biodegradability, low toxicity of degradation by-products and plasticity in processing into a variety of material formats. Here we studied in vitro anti-inflammatory potential of marine macromolecules cross-linked bio-composite scaffold composed of hydroxyapatite, alginate, chitosan and fucoidan named as HACF on LPS stimulated RAW 264.7 macrophage cells. The effects of HACF on the viability of RAW264.7 cells, nitrite level, intracellular ROS as well as the mRNA levels of NF-κB, iNOS, COX-2, TNF-α, IL-1ß and IL-6 were examined in LPS induced RAW264.7 macrophage cells. The results revealed that HACF hydrogel scaffold exerts anti-inflammatory effect by inhibiting the production of ROS, suppress NF-kB translocation to the nucleus and thereby inhibiting the production of inflammatory mediators. Hence, our results confirm that HACF has a strong anti-oxidant capacity to inhibit inflammation associated gene expression by suppressing NF-kB signaling pathway. It clearly reveals the anti-oxidant and anti-inflammatory effect of HACF hydrogel scaffold on LPS induced RAW 264.7 cells.

Asunto(s)

Lipopolisacáridos , Ingeniería de Tejidos , Animales , Antiinflamatorios/farmacología , Cartílago , Macrófagos , Ratones , FN-kappa B , Óxido Nítrico , Células RAW 264.7

RESUMEN

There is an intense interest in developing innovative biomaterials which support the invasion and proliferation of living cells for potential applications in tissue engineering and regenerative medicine. Present study demonstrated the in vivo biocompatibility and toxicity of a macromolecules cross-linked biocomposite scaffold composed of hydroxyapatite, alginate, chitosan and fucoidan abbreviated as HACF. The in vivo biocompatibility and toxicity of HACF scaffold were tested by comparing them with those of a biocompatible surgical metal implant (SMI) in a subcutaneous rat model. Following the implantation, animals were sacrificed and the scaffolds were resected at 1st, 4th, and 8th weeks; the surrounding tissue along with the implant was removed to evaluate its biocompatibility. The effects of implanted biomaterial scaffolds on vital organ systems such as liver, kidney, etc., have been studied by hematology and serum biochemistry. The activities of pro-inflammatory marker enzymes such as COX, 5-LOX, 15-LOX, and NOS were normal in rats implanted with HACF scaffold. Hematological parameters, antioxidant and lipid peroxidation status were also found to be normal in implanted rats same as that of control and SMI. The modulatory effect of implanted scaffold over inflammatory and stress signaling cascades were confirmed by the normalized mRNA expressions of NF-κB, TNF-α and IL-6. The histopathological analysis of liver, kidney and tissue support our results. Taken together, these results demonstrated that HACF biocomposite scaffold signifies its suitability for further research as a scaffold material for cartilage tissue engineering applications.

Asunto(s)

Materiales Biocompatibles/química , Materiales Biocompatibles/farmacología , Cartílago/citología , Ensayo de Materiales , Ingeniería de Tejidos , Alginatos/química , Animales , Antioxidantes/metabolismo , Cartílago/efectos de los fármacos , Quitosano/química , Durapatita/química , Glutatión/metabolismo , Mediadores de Inflamación/metabolismo , Riñón/efectos de los fármacos , Riñón/metabolismo , Peroxidación de Lípido/efectos de los fármacos , Hígado/efectos de los fármacos , Hígado/enzimología , Hígado/metabolismo , Masculino , Monocitos/efectos de los fármacos , Monocitos/metabolismo , Polisacáridos/química , Ratas

RESUMEN

Marine biopolymer composite materials provide a technological platform for launching biomedical applications. Biomaterials demand good biocompatibility without the possibility of inflammation or foreign body reactions. In this study, we prepared two biocomposite hydrogels namely; HAC (hydroxyapatite, alginate & chitosan) and HACF (hydroxyapatite, alginate, chitosan & fucoidan) followed by calcium chloride cross linking. The prepared scaffolds were characterized by Fourier transform infrared spectroscopy and scanning electron microscopy. Porosity measurement, swelling, biodegradation, hemolysis, RBC aggregation, plasma protein adsorption and cytotoxicity studies were also done. The hydrogel scaffold HACF possessed a well-defined porous architecture, sufficient water holding capacity, better hemocompatibility and biodegradability. The biocompatibility was confirmed through in vitro cytotoxicity studies such as MTT assay, Neutral red uptake, DAPI staining, Trypan blue dye exclusion test and direct contact assay in L929 mouse fibroblast cells. In addition, immunomodulatory and anti-inflammatory properties of both of these scaffolds were revealed by the mRNA expressions of major inflammatory marker genes in cytotoxic condition such as TNF-α, IL-6 and NF-κB. The physiochemical characterization and biological responses of HACF hydrogel signifies its suitability for various tissue engineering applications.

Asunto(s)

Materiales Biocompatibles , Hidrogeles , Ingeniería de Tejidos/métodos , Andamios del Tejido , Alginatos/química , Animales , Materiales Biocompatibles/síntesis química , Materiales Biocompatibles/química , Materiales Biocompatibles/toxicidad , Cloruro de Calcio/química , Línea Celular , Supervivencia Celular , Quitosano/química , Reactivos de Enlaces Cruzados/química , Durapatita/química , Ácido Glucurónico/química , Ácidos Hexurónicos/química , Hidrogeles/química , Hidrogeles/toxicidad , Interleucina-6/metabolismo , Ratones , Polisacáridos/química , Porosidad , Factor de Necrosis Tumoral alfa/metabolismo

RESUMEN

The health benefits of omega-3 polyunsaturated fatty acids (ω-3 PUFA), mainly eicosapentaenoic acid (EPA 20:5) and docosahexaenoic acid (DHA, 22:6), have been long known. Although various studies have demonstrated the health benefits of ω-3 PUFA, the mechanisms of action of ω-3 PUFAs are still not completely understood. While the major commercial source is marine fish oil, in this study we suggest the marine micro algae, Dunaliella salina as an alternate source of omega-3 fatty acids. Treatment with this algal omega-3 fatty acid concentrate (Ds-ω-3 FA) resulted in significant down-regulation of LPS-induced production of TNF-α and IL-6 by peripheral blood mononuclear cells (PBMCs). The concentrate was also found to be a potent blocker of cyclooxygenase (COX-2) and matrix metalloproteinase (MMP-2 and MMP-9) expression. The present study reveals the anti-inflammatory properties of Ds-ω-3 FA concentrate including the inhibition of NF-κB translocation.

Asunto(s)

Antiinflamatorios/farmacología , Chlorophyta/química , Ácidos Grasos Omega-3/farmacología , Leucocitos Mononucleares/efectos de los fármacos , FN-kappa B/metabolismo , Transporte Activo de Núcleo Celular/efectos de los fármacos , Antiinflamatorios/aislamiento & purificación , Western Blotting , Técnicas de Cultivo de Célula , Células Cultivadas , Ciclooxigenasa 2/biosíntesis , Ácidos Grasos Omega-3/aislamiento & purificación , Humanos , Inmunohistoquímica , Interleucina-6/biosíntesis , Leucocitos Mononucleares/inmunología , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Factor de Necrosis Tumoral alfa/biosíntesis

RESUMEN

Even though the role of lycopene from tomato (trans form) in controlling prostate cancer was reported, lycopene (cis and trans 60:40) isolated from green algae Chlorella marina was not reported so far. The present study aimed to assess the anti-proliferative and apoptotic effect of lycopene from a new source and to compare the activity with available trans lycopene by using androgen-independent human prostate cancer cell lines. Exposure of PC-3 and DU-145 cell lines to algal lycopene (AL) at a dose of 20 and 50 µM significantly inhibited the growth and colony formation, and the percentage of inhibition was higher than tomatal lycopene (TL)-treated groups. The stability of AL in cell culture medium was high, when compared to TL under standard cell culture conditions. The level of lycopene was not detected in PC-3 cell lines cultured in medium lacking lycopene. Staining cells with acridine orange and ethidium bromide, the PC-3 control cells showed largely non-fragmented intact nucleoid. Stronger apoptosis signal was induced with higher concentrations (50 µM) of algal lycopene. Increased DNA damage was observed in AL- and TL-treated cells which appear as comet during single-cell gel electrophoresis. Flow cytometry results revealed that AL caused PC-3 cells to accumulate in the G0/G1 phase and to undergo apoptosis. The effect was higher in AL groups than TL-treated groups. Algal lycopene showed very significant anti-proliferative and apoptotic effect in human prostate cancer cell lines. Therefore, algal lycopene from C.marina would be recommended for the treatment of prostate cancer.

Asunto(s)

Anticarcinógenos/farmacología , Apoptosis/efectos de los fármacos , Carotenoides/farmacología , Proliferación Celular/efectos de los fármacos , Chlorella/química , Neoplasias de la Próstata/tratamiento farmacológico , Neoplasias de la Próstata/patología , Western Blotting , Ciclo Celular/efectos de los fármacos , Cromatografía Líquida de Alta Presión , Ensayo Cometa , Daño del ADN/efectos de los fármacos , Citometría de Flujo , Humanos , Licopeno , Masculino , Células Tumorales Cultivadas , Ensayo de Tumor de Célula Madre

RESUMEN

The role of commercially available lycopene (all-trans) from tomato in controlling arthritis has been reported. Even though many reports are available that the cis form of lycopene is more biologically active, no report seems to be available on lycopene (cis and trans) isolated from an easily available and culturable sources. In the present study, the anti-arthritic effect of lycopene (cis and trans) from the algae Chlorella marina (AL) has been compared with lycopene (all-trans) from tomato (TL) and indomethacin (Indo). Arthritis (CIA) was developed in male Sprague dawley rats by collagen and the following parameters were studied. The activities of inflammatory marker enzymes like cyclooxygenase (COX), lipoxygenase (LOX) and myeloperoxidase (MPO) were found to be decreased on treatment with AL when compared to TL and Indo. Changes in Erythrocyte sedimentation rate (ESR), white blood cell (WBC) count, red blood cells (RBC) count, hemoglobin (Hb), C-reactive protein (CRP), rheumatoid factor (RF), and ceruloplasmin levels observed in the blood of arthritic animals were brought back to normal by AL when compared to TL and Indo. Histopathology of paw and joint tissues showed marked reduction in edema on supplementation of AL. Thus these results indicate the potential beneficiary effect of algal lycopene on collagen induced arthritis in rats when compared to TL and even to the commonly used anti-inflammatory drug indomethacin. Therefore lycopene from C. marina would be recommended as a better natural source with increased activity and without side effects in the treatment of anti-inflammatory diseases.

Asunto(s)

Antiinflamatorios/farmacología , Artritis Experimental/tratamiento farmacológico , Carotenoides/farmacología , Chlorella/química , Extractos Vegetales/farmacología , Animales , Artritis Experimental/sangre , Artritis Experimental/metabolismo , Proteína C-Reactiva/metabolismo , Carotenoides/aislamiento & purificación , Ceruloplasmina/farmacología , Colágeno Tipo II , Edema/tratamiento farmacológico , Edema/metabolismo , Recuento de Eritrocitos/métodos , Eritrocitos/efectos de los fármacos , Eritrocitos/metabolismo , Hemoglobinas/efectos de los fármacos , Hemoglobinas/metabolismo , Indometacina/farmacología , Articulaciones/efectos de los fármacos , Articulaciones/metabolismo , Recuento de Leucocitos/métodos , Licopeno , Solanum lycopersicum/química , Masculino , Prostaglandina-Endoperóxido Sintasas/metabolismo , Ratas , Ratas Sprague-Dawley , Factor Reumatoide/metabolismo

RESUMEN

L-asparaginase isolated in our laboratory fromAeromonas has been found to be antileukaemic. In the present study changes in the levels of serum enzymes in leukaemic mice and under treatment withAeromonas L-asparaginase has been compared. A significant increase in the levels of serum lactate dehydrogenase with tumour growth and a decrease during therapy was observed. A significant decrease in alanine transaminase activity during tumour growth and an increase during treatment was noticed. Increased levels of aspartate transaminase and alkaline phosphatase was observed during enzyme therapy. Total acid phosphatase was found to be increased during tumour growth and decreased considerably during treatment.