Exosome-Functionalized, Drug-Laden Bone Substitute along with an Antioxidant Herbal Membrane for Bone and Periosteum Regeneration in Bone Sarcoma.

Developing advanced methods for effective bone reconstructive strategies in case of critical bone defects caused by tumor resection, trauma, and other implant-related complications remains a challenging problem in orthopedics. In the clinical management of bone diseases, there is a paradigm shift in using local drugs at the injury site; however, the dead space created during the surgical debridement of necrotic bone and soft tissues (periosteum and underlying muscle) leads to ineffective bone formation, thereby leading to secondary complications, and thus calls for better regenerative approaches. In this study, we have utilized an exosome-functionalized doxorubicin-loaded biodegradable nanocement (NC)-based carrier along with a Cissus quadrangularis (CQ) extract-laden antioxidant herbal membrane for simultaneously managing the periosteum as well as bone formation in the tumor resection model of osteosarcoma. We initially evaluated the efficacy of scaffolds for in vitro mineralization and bone formation. To examine the in vivo effectiveness, we developed a human osteosarcoma cell line (Saos-2)-induced tumor xenograft model with a critical-sized bone defect. The findings revealed that doxorubicin released from NC was successful in killing the tumor cells and was present even after 30 days of implantation. Additionally, the incorporation of exosomes aided the bone formation, resulting in around a 2.6-fold increase in the bone volume compared to the empty group as evaluated by micro-CT. The herbal membrane assisted in the development of periosteum and mineralizing bone callous as validated through histological and immunofluorescence analysis. Thus, our findings describe a one-step biomaterial-based cell-free approach to regenerate bone in osteosarcoma and prevent further fracture due to the complete development of periosteum and lost bone.

Advent of phytobiologics and nano-interventions for bone remodeling: a comprehensive review.

Bone metabolism constitutes the intricate processes of matrix deposition, mineralization, and resorption. Any imbalance in these processes leads to traumatic bone injuries and serious disease conditions. Therefore, bone remodeling plays a crucial role during the regeneration process maintaining the balance between osteoblastogenesis and osteoclastogenesis. Currently, numerous phytobiologics are emerging as the new therapeutics for the treatment of bone-related complications overcoming the synthetic drug-based side effects. They can either target osteoblasts, osteoclasts, or both through different mechanistic pathways for maintaining the bone remodeling process. Although phytobiologics have been widely used since tradition for the treatment of bone fractures recently, the research is accentuated toward the development of osteogenic phytobioactives, constituent-based drug designing models, and efficacious delivery of the phytobioactives. To achieve this, different plant extracts and successful isolation of their phytoconstituents are critical for osteogenic research. Hence, this review emphasizes the phytobioactives based research specifically enlisting the plants and their constituents used so far as bone therapeutics, their respective isolation procedures, and nanotechnological interventions in bone research. Also, the review enlists the vast array of folklore plants and the newly emerging nano-delivery systems in treating bone injuries as the future scope of research in the phytomedicinal orthopedic applications.

Effect of Withania somnifera hydroalcoholic extract and other dietary interventions in improving muscle strength in aging rats.

BACKGROUND: Aging leads to loss of skeletal muscle, diminished muscle strength, and decline in physical functions. OBJECTIVE: This study evaluates Withania somnifera and some dietary interventions to combat muscle weakness in aging rats. MATERIALS AND METHODS: Rats (12-13 months old) corresponding to a human age of 60-65 years were assigned to various groups and given orally a standardized W. somnifera extract (WSE, 500 mg/kg) or a protein cocktail comprising soybean (1.5 g/kg) and quinoa (1 g/kg) or a combination of WSE and the protein cocktail or whey protein (1 g/kg) as a reference standard or only resistance exercise for 60 days. Grip strength and blood glucose levels were monitored weekly. At the end of the treatment, total protein, inflammatory markers (CRP, IL-6 and TNF-α), AMPK, malondialdehyde, glutathione, antioxidant enzymes and apoptotic regulator genes (Bax and Bcl-2) were assayed. The biceps brachii muscle of all animals was subjected to histomorphological study. RESULTS: All treatments successfully attenuated aging-elevated glucose, CRP, IL-6, TNF-α, AMPK, malondialdehyde, and Bax levels. A significant restoration of the aging-depleted total protein levels, glutathione, superoxide dismutase, catalase, and Bcl-2 was noted in the treatment groups. An increase in grip strength and greater biceps mass with all treatments indicated regaining of the frail aging muscle's strength and functionality. The WSE + protein treatment elicited the best results among all treatment groups to optimize muscle strength. CONCLUSION: All the interventions curbed muscle loss and strengthened the skeletal muscle by reducing inflammation, oxidative stress and apoptosis, and increasing ATP availability to the muscle.

An Ayurvedic formulation of Emblica officinalis and Curcuma longa alleviates insulin resistance in diabetic rats: Involvement of curcuminoids and polyphenolics.

BACKGROUND: Nishamalaki is an Ayurvedic herbal formulation used to treat type 2 diabetes mellitus (T2DM), comprises Emblica officinalis and Curcuma longa. OBJECTIVE(S): One of the main cause of T2DM is Insulin Resistance (IR) hence, this study was planned to evaluate IR lowering effect of a standardized Nishamalaki extract "EmbliQur" in high-fat diet (HFD) and streptozotocin (STZ) induced T2DM rats. MATERIALS AND METHODS: Curcuminoids (23.89% w/w), gallic acid (5.27% w/w) and tannins (25.44% w/w) were quantified from EmbliQur. Rats were fed HFD throughout the study of 45 days and received STZ (40 mg/kg, i.p) on the 15th day of the study. Rats with more than 250 mg/dl of fasting blood glucose level (FBGL) were considered diabetic and selected for administration of EmbliQur (500 mg and 1000 mg/kg) or the standard drug metformin (120 mg/kg, p.o) from the 18th day of the study for the next 27 days. FBGL and insulin levels of all rats were measured weekly and an oral glucose tolerance test (OGTT) was done at the end of the study. The values of FBGL and insulin were used to calculate IR by the HOMA-IR, QUICKI and Matsuda methods. RESULTS: Rats treated with STZ/HFD had significantly higher than normal FBGL and insulin levels throughout the study and exhibited skewed IR indices in the above three methods of IR assessment. EmbliQur treatment successfully lowered the HFD/STZ-elevated BGL and insulin levels, and ameliorated IR in all models of IR evaluation. CONCLUSION: EmbliQur 1000 mg/kg was noted to be more effective than EmbliQur 500 mg/kg in alleviating IR.

c-Phycocyanin primed silver nano conjugates: Studies on red blood cell stress resilience mechanism.

Green synthesis of metal-encased nutraceutical nano-hybrids has been a target for research over the last few years. In the present investigation, we have reported temperature dependent facile synthesis of silver nanoparticles using FDA approved c phycocyanin (cPC). The cPC conjugated silver nanoparticles (AgcPCNPs) were characterized by TEM, Zeta Potential, UV-vis, XPS, FTIR, and CD Spectroscopy. The temperature optimization studies suggested the synthesis of stable AgcPCNPs at 40 °C while at higher temperature system shows aggregated appearance. Molecular docking studies predicted the exclusive interaction of C, D, I, and J chains of cPC with the surface of AgNPs. Moreover, AgcPCNPs significantly (p < 0.1 %) counteract the toxic nature of AgNPs on red blood cell by measuring parameters like total RBC count, % hemolysis, % hematocrit, coagulation time, pH, electrolyte concentrations and degree of blood cell lipid peroxidation by the anti-oxidation mechanism. Skin fibroblast in vitro cell migration result suggeststhat AgcPCNPs enhanced the degree of cell movement towards the wound area. Data obtained collectively demonstrate that AgcPCNPs can be a better agent in the dermal wound healing with reduced toxicity with the bi-phasic advantage of cPC as a wound healer and Ag nano-metal as an anti-bacterial agent.

Enhanced bone mineralization using hydroxyapatite-based ceramic bone substitute incorporating Withania somnifera extracts.

Withania somnifera (ashwagandha) is used in Indian traditional medicine for its various health benefits. Withaferin-A, a steroidal lactone present in this herb, has shown proteosomal inhibition-based enhancement of bone mineralization. In the present work, chitosan microparticles blended with total methanolic root extract of W. somnifera were incorporated as a porogen in calcium phosphate-based hydroxyapatite bone filler. The controlled release of bioactive molecules enabled enhanced proliferation and differentiation of pre-osteoblasts. Microparticle percentages were optimized to have a minimum effect on the setting time, mechanical strength and degradability of hydroxyapatite bone filler. In vitro cell adhesion, proliferation and differentiation were evaluated to determine the biocompatibility of the composites. On the basis of the desirable results obtained, we provide a preliminary rationale for the use of methanolic extract-blended chitosan microparticle-impregnated calcium phosphate filler for enhanced bone regeneration.

AMP-Activated Protein Kinase Restricts Zika Virus Replication in Endothelial Cells by Potentiating Innate Antiviral Responses and Inhibiting Glycolysis.

Viruses are known to perturb host cellular metabolism to enable their replication and spread. However, little is known about the interactions between Zika virus (ZIKV) infection and host metabolism. Using primary human retinal vascular endothelial cells and an established human endothelial cell line, we investigated the role of AMP-activated protein kinase (AMPK), a master regulator of energy metabolism, in response to ZIKV challenge. ZIKV infection caused a time-dependent reduction in the active phosphorylated state of AMPK and of its downstream target acetyl-CoA carboxylase. Pharmacological activation of AMPK using 5-aminoimidazole-4-carboxamide ribonucleotide (AICAR), metformin, and a specific AMPKα activator (GSK621) attenuated ZIKV replication. This activity was reversed by an AMPK inhibitor (compound C). Lentivirus-mediated knockdown of AMPK and the use of AMPKα-/- mouse embryonic fibroblasts provided further evidence that AMPK has an antiviral effect on ZIKV replication. Consistent with its antiviral effect, AMPK activation potentiated the expression of genes with antiviral properties (e.g., IFNs, OAS2, ISG15, and MX1) and inhibited inflammatory mediators (e.g., TNF-α and CCL5). Bioenergetic analysis showed that ZIKV infection evokes a glycolytic response, as evidenced by elevated extracellular acidification rate and increased expression of key glycolytic genes (GLUT1, HK2, TPI, and MCT4); activation of AMPK by AICAR treatment reduced this response. Consistent with this, 2-deoxyglucose, an inhibitor of glycolysis, augmented AMPK activity and attenuated ZIKV replication. Thus, our study demonstrates that the anti-ZIKV effect of AMPK signaling in endothelial cells is mediated by reduction of viral-induced glycolysis and enhanced innate antiviral responses.

Orthobiologics with phytobioactive cues: A paradigm in bone regeneration.

Bone injuries occur due to various traumatic and disease conditions. Healing of bone injury occurs via a multi-stage intricate process. Body has the potential to rectify most of the bone injuries but some severe traumatic cases with critical size defects may require interventions. Autografts are still considered the "gold standard" for fracture healing but due to limitations associated with it, new alternatives are warranted. The field of orthobiologics has provided novel approaches using scaffolds, bioactive molecules, stem cells for the treatment of bone defects. Phyto-bioactives have been widely used in alternative medicine and folklore practices for curing bone ailments. It is believed that different bioactive constituents in plants work synergistically to give the therapeutic efficacy. Bioactives in plants extracts act upon different signal transduction pathways aiding in bone healing. The present review focuses on the use, chemical composition, mode of delivery, mechanism of action, and possible future strategies of three medicinal plants popularly used in traditional medicine for bone healing: Cissus quadrangularis, Withania somnifera and Tinospora cordifolia. Plants extracts seem to be a natural and non-toxic therapeutic alternative in treating bone injuries. Most of the studies on bone healing for these plants have reported oral administration of the extracts and presented them as a safe alternative without any side effects despite giving higher doses. Forthcoming studies could be directed towards the local delivery of extracts at the defect site. Unification of herbal extracts and orthobiologics could be an interesting direction in the field of bone healing in future. The present review intends to provide a bird's eye view of different strategies used in bone healing, mechanisms involved and future direction of advancements using phytobioactives and orthobiologics.

Plant latex: a promising antifungal agent for post harvest disease control.

Bioactive compounds from plant latex are potential source of antifungic against post harvest pathogens. Latex from a total of seven plant species was investigated for its phytochemical and antifungal properties. Six fungi namely Aspergillus fumigatus, A. niger, A. terreus, F. solani, P. digitatum and R. arrhizus were isolated from infected fruits and vegetables and tested against various solvent extracts of latex. Analysis of latex extracts with phytochemical tests showed the presence of alkaloids, flavonoids, glycosides, phenols, saponins, steroids, tannins and terpenoids. Antifungal assay revealed the potential inhibitory activity of petroleum ether extracts against the postharvest fungal isolates. Various degree of sensitivity was observed irrespective of plant species studied with A. terreus and P. digitatum as the most susceptible ones. F. solani and A. fumigatus were moderately sensitive to the latex extracts tested. Among the plants, latex of Thevetia peruviana (75.2%) and Artocarpus heterophyllus (64.8%) were having potential antifungal activity against the isolates followed by Manilkara zapota (51.1%). In conclusion, use of plant latex makes interest to control postharvest fungal diseases and is fitting well with the concept of safety for human health and environment.