Nanohydroxyapatite incorporated photocrosslinked gelatin methacryloyl/poly(ethylene glycol)diacrylate hydrogel for bone tissue engineering.

The development of novel strategies that aim to augment the regenerative potential of bone is critical for devising better treatment options for bone defects or injuries. Facilitation of bone repair and regeneration utilizing composite hydrogels that simulates bone matrix is emerging as a viable approach in bone tissue engineering. The present study aimed to develop nanohydroxyapatite-incorporated gelatin methacryloyl (GelMA)/poly(ethylene glycol) diacrylate (PEGDA) hydrogel (GMPH hydrogel). A facile blending and photocrosslinking approach was employed to incorporate nanohydroxyapatite into the inter-crosslinked polymeric hydrogel network to obtain an ECM mimicking matrix for assisting bone tissue regeneration. Chemical characterization of GelMA and the GMPH hydrogel was carried out using FTIR and 1H NMR. Physical properties of GMPH, such as gelation, swelling and degradation ratios, and internal morphology, signified the suitability of GMPH hydrogel for tissue engineering. Cell viability assay demonstrated a healthy proliferation of MG63 osteoblast cells in GMPH hydrogel extracted growth medium, indicating the hydrogel's cytocompatibility and suitability for bone tissue engineering. Our study documented the fabrication of a novel GelMA/PEGDA-nanohydroxyapatite hydrogel that possesses ideal physicochemical and biological properties for bone tissue engineering.

Sulfated polysaccharide ascophyllan from Padina tetrastromatica enhances healing of burn wounds by ameliorating inflammatory responses and oxidative damage.

Sulfated polysaccharide ascophyllan from marine brown algae has been identified to have burn wound healing properties. Thus, we examined the effects of ascophyllan fraction (AF3) on the inflammatory response and oxidative damage in burn wounds. Full-thickness burn wounds in rats were then treated twice per day with topical AF3 ointment (5%), while control groups were treated with 10% povidone-iodine (positive control) and petroleum jelly-based ointment (negative control). The activity of cyclooxygenase-2 and myeloperoxidase and levels of C-reactive protein, nitric oxide, and proinflammatory cytokines (tumor necrosis factor-α, interleukin-6, and interleukin-1ß) were observed to have significantly decreased in peripheral blood mononuclear cells, serum, and wound tissue of the group treated with AF3 ointment on day 8 after wounding. The expression of inducible nitric oxide synthase, endothelial nitric oxide synthase, and vascular endothelial growth factor at the mRNA level was determined to be upregulated in the wound tissue of the AF3 ointment-treated group. After treatment with AF3 ointment, the antioxidant enzyme activity and level of reduced glutathione were upregulated, whereas the content of thiobarbituric acid reactive substances decreased. Treatment of burn wounds using 5% AF3 ointment decreases oxidative damage associated with inflammation deceptively via inhibition of inflammatory enzymes, regulation of proinflammatory cytokines, upregulation of angiogenesis, and activity of antioxidant enzymes.

Sulfated polysaccharides from the edible marine algae Padina tetrastromatica attenuates isoproterenol-induced oxidative damage via activation of PI3K/Akt/Nrf2 signaling pathway - An in vitro and in vivo approach.

The reactive oxygen species (ROS) induced oxidative stress is an inevitable factor for the pathogenesis of cardiovascular diseases. The edible marine algae-derived sulfated polysaccharides gained special attention as novel bioactive compounds having potential pharmacological activities. The present study evaluated in vitro and in vivo cardioprotective properties of sulfated polysaccharides from the edible brown marine algae Padina tetrastromatica (PSPS) against isoproterenol (ISO) induced cardiac damage. The cardioprotective properties of PSPS were first evaluated in H9c2 cardiac myoblasts and the results were confirmed by in vivo studies conducted in male Sprague-Dawley rats. The biochemical parameters, histopathological analysis, mRNA expressions, and ELISA studies indicated that PSPS significantly decreased (p < 0.05) the cardiac damage induced by ISO by reducing lipid peroxidation and improving antioxidant status, both in vitro and in vivo, via modulating PI3k/Akt/Nrf2 signaling pathway. The histopathological evidence further reinforced our findings and highlighted the promising cardioprotective activities offered by PSPS.

The efficacy of sulfated polysaccharides from Padina tetrastromatica in modulating the immune functions of RAW 264.7 cells.

Immunomodulation is a collective term of immunostimulation and immunosuppression. Immunotherapy by means of immunomodulation is gaining additional significance as the frequency of mutant microbes as well as cancer cases are increasing in the present time. A wide population of marine vegetation has contributed to the traditional and modern therapeutic regimens owing to the abundance of bioactive molecules. Among the seaweed born macromolecules, the sulfated polysaccharides (SPS) from marine macro algae were reported to exhibit excellent biological activities in addition to their structural and nutritional roles. Interestingly, SPS from the marine brown algae Padina tetrastromatica is not yet explored for their immunomodulatory potential. In the present study, SPS were extracted by ethanol precipitation, purified using DEAE cellulose column chromatography, and named as Ethanolic Sulfated Polysaccharide-Column Purified (ESPs-CP). The study includes, evaluation of macrophage proliferation, prostaglandin and nitric oxide production, COX-2, 5-LOX, and iNOS estimation and gene expression studies in RAW 264.7 cells. ESPs-CP strongly stimulated macrophage proliferation and production of prostaglandin and nitric oxide. They also increased COX-2, 5-LOX, and iNOS concentration in macrophages, which was comparable to that of LPS stimulated macrophages. Increase of prostaglandin and NO production may be due to increased expression of COX-2 and iNOS as observed in gene expression studies. The mRNA expression of pro- inflammatory cytokines such as IL-6, IL-1ß, TNF-α and anti-inflammatory cytokines IL-10 and TGF-ß were also enhanced by ESPs-CP. The evaluations signify the possibilities of SPS as potent immunostimulators during immune deficiencies.

An efficient protocol for the Agrobacterium-mediated genetic transformation of microalga Chlamydomonas reinhardtii.

Algal-based recombinant protein production has gained immense interest in recent years. The development of algal expression system was earlier hindered due to the lack of efficient and cost-effective transformation techniques capable of heterologous gene integration and expression. The recent development of Agrobacterium-mediated genetic transformation method is expected to be the ideal solution for these problems. We have developed an efficient protocol for the Agrobacterium-mediated genetic transformation of microalga Chlamydomonas reinhardtii. Pre-treatment of Agrobacterium in TAP induction medium (pH 5.2) containing 100 µM acetosyringone and 1 mM glycine betaine and infection of Chlamydomonas with the induced Agrobacterium greatly improved transformation frequency. This protocol was found to double the number of transgenic events on selection media compared to that of previous reports. PCR was used successfully to amplify fragments of the hpt and GUS genes from transformed cells, while Southern blot confirmed the integration of GUS gene into the genome of C. reinhardtii. RT-PCR, Northern blot and GUS histochemical analyses confirm GUS gene expression in the transgenic cell lines of Chlamydomonas. This protocol provides a quick, efficient, economical and high-frequency transformation method for microalgae.

Effect of lycopene from Chlorella marina on high cholesterol-induced oxidative damage and inflammation in rats.

Even though the role of all-trans lycopene from tomato in controlling atherosclerosis was reported, but no report is available on the cis-isomer of lycopene obtained from an easily available source green algae Chlorella marina. So in this study, Sprague Dawley rats fed with high-cholesterol diet were given standard drug lovastatin; algal lycopene (AL) (cis/all-trans 40:60) and tomato all-trans lycopene (TL) and the following parameters were studied. Total cholesterol, low-density lipoprotein, triglycerides were decreased significantly and the high-density lipoprotein levels were increased on treatment with AL. The activities of antioxidant enzymes catalase, superoxide dismutase, glutathione peroxidase, and glutathione reductase were found to be increased, whereas thiobarbituric acid reactive substances levels were decreased in AL when compared to the drug and TL-treated rats. The activities of inflammatory marker enzymes like cyclooxygenase, 15-lipoxygenase in monocytes and myeloperoxidase, C-reactive protein and ceruloplasmin levels in serum were found to be decreased on treatment with AL. Histopathological studies revealed that lycopene from this alga could reduce fatty liver and aortic plaque when compared to the drug and TL. Algal lycopene showed very significant antioxidant and anti-inflammatory effect in high-cholesterol fed rats. Therefore, AL from C. marina would be recommended for the treatment of hyperlipidemia.

Effect of ascophyllan from brown algae Padina tetrastromatica on inflammation and oxidative stress in carrageenan-induced rats.

Sulfated polysaccharide ascophyllan was isolated from the brown algae Padina tetrastromatica and purified by ion-exchange chromatography. Anti-inflammatory effect of ascophyllan fraction against carrageenan-induced paw edema in rats was studied. Paw edema in rats was induced by injecting 0.1 ml, 1 % carrageenan suspension in 0.9 % NaCl solution into the sub-plantar tissue of the right hind paw. Carrageenan caused a significant increase in the activity of inflammatory marker enzymes like lipoxygenases and cyclooxygenase in peripheral blood mononuclear cells and paw tissue and also increased the concentration of prostaglandin E2 (PGE2) and myeloperoxidase (MPO) in paw tissue. When compared to the reference drug diclofenac, ascophyllan fraction-3 (AF3) treatment significantly reduced the activities of anti-inflammatory enzymes, concentration of PGE2 and MPO. AF3 treatment decreased the mRNA level expression of TNF-α and IL-6. Concentration of thiobarbituric acid reactive substances was decreased. Activities of antioxidant enzymes and reduced glutathione level were increased on treatment with AF3. Histopathology of paw tissue showed decreased edema formation and cellular infiltration on supplementation with AF3. Thus the results demonstrated the potential beneficiary effect of ascophyllan fraction on carrageenan-treated rats.

Anti-inflammatory and antioxidant potential of alginic acid isolated from the marine algae, Sargassum wightii on adjuvant-induced arthritic rats.

The present study evaluated the anti-inflammatory and antioxidant potential of alginic acid isolated from brown algae Sargassum wightii in arthritic rats. Arthritis was induced in male Sprague-Dawley rats by intradermal injection of complete Freund's adjuvant into the right hind paw, produce inflammation of the joint tissue. Paw edema volume, enzymes linked to inflammation such as cyclooxygenase, lipoxygenase and myeloperoxidase, and the level of ceruloplasmin, C-reactive protein and rheumatoid factor were evaluated in all the experimental groups. Oxidative stress during inflammation was analyzed by estimating lipid peroxidation and the activities of antioxidant enzymes such as superoxide dismutase, catalase, glutathione peroxidase, glutathione reductase and non-enzymatic antioxidant, reduced glutathione. Alginic acid treatment (100 mg/kg) in arthritic rats exhibited reduced paw edema volume along with reduced activities of enzymes such as cyclooxygenase, lipoxygenase and myeloperoxidase. Reduction in the level of C-reactive protein, ceruloplasmin and rheumatoid factor were also observed in arthritic rats treated with alginic acid along with reduced lipid peroxidation and enhanced activities of antioxidant enzymes, which suggest the antioxidant potential of the compound. Histopathological analysis of paw tissue showed that alginic acid treatment reduced paw edema and inflammatory infiltration in arthritic rats. Overall results suggest that alginic acid isolated from Sargassum wightii exhibits potent anti-inflammatory and antioxidant activity, and can develop this marine alga as an alternative source for therapy and can be used as a drug candidate for the development of anti-inflammatory agent.

Alginic acid isolated from Sargassum wightii exhibits anti-inflammatory potential on type II collagen induced arthritis in experimental animals.

The present study evaluated the anti-inflammatory potential of alginic acid isolated from the brown algae Sargassum wightii in type II collagen induced arthritic rats, a well established arthritic model that resembles more closely to human rheumatoid arthritis in its clinical, pathological, immunological and histological aspects. Type II collagen induced arthritic rats showed increased activities of inflammatory marker enzymes like cycloxygenase-2 (COX-2), lipoxygenase (5-LOX), xanthine oxidase (XO) and myeloperoxidase (MPO) along with increased concentration of rheumatoid factor (RF), ceruloplasmin and C-reactive protein (CRP). Treatment with alginic acid significantly reduced the activities of COX-2 and 5-LOX along with reduction in MPO, XO, RF and CRP. Alginic acid treatment reverted to the altered levels of hematological parameters like RBC count, WBC count and ESR in arthritic rats. Concentrations of proinflammatory cytokines like IL-1 ß, TNF α and IL-6 were significantly higher in arthritic rats which were reduced on treatment with alginic acid. Increased activities of lysosomal enzymes that manifest the systemic damage during arthritis were significantly reduced by the treatment with alginic acid which indicates the reduction in the rupture and degradation of connective tissue. Histopathology of knee joint tissues showed that extensive bone degradation and synovial hyperplasia along with infiltrating cells and treatment with alginic acid reversed the histopathological changes which indicate the protective potential of alginic acid in rheumatoid arthritis.

Antioxidant activity of Aulosira fertilisima on CCl4 induced hepatotoxicity in rats.

Free radicals cause cell injury, when they are generated in excess or when the antioxidant defense is impaired. Carbon tetrachloride (CCl4) is used as a model for liver injury. In this study antioxidant activity of ethanol extract of A. fertilisima (EEA) was investigated using CCl4 intoxicated rat liver as the experimental model. Oral administration of EEA at a dose of 100 mg/kg body weight, for 14 consecutive days, the rate of the production of antioxidant enzymes like super oxide dismutase, catalase, glutathione peroxidase and glutathione transferase in rats compared to the CCl4 treated group without any supporting treatment. Liver damage is detected by the measurement of the activities of serum enzymes like aspartate aminotransferase, alanine aminotransferase, gamma glutamyl transpeptidase and alkaline phosphatase which were released in to the blood from damaged cells. The normalization of these enzymes levels was observed in rats treated with EEA (100 mg/kg body weight) by reducing the leakage of the above enzymes in to the blood. The findings provide a rationale for further studies on isolation of active principles and its pharmacological evaluation. Protection offered by silymarin (standard reference drug) seemed relatively greater.