Catechin, epicatechin, curcumin, garlic, pomegranate peel and neem extracts of Indian origin showed enhanced anti-inflammatory potential in human primary acute and chronic wound derived fibroblasts by decreasing TGF-ß and TNF-α expression.

BACKGROUND: Although chronic wounds are devastating and can cause burden at multiple levels, chronic wound research is still far behind. Chronic wound treatment is often less efficient due to delay in diagnosis and treatment, non-specific treatment mainly due to lack of knowledge of wound healing or healing resistance genes. It's known that chronic wounds do not progress towards healing, because it gets stalled in inflammatory phase of wound healing. OBJECTIVE: We aimed to use phytoextracts possessing excellent anti-inflammatory properties to regulate the unbalanced levels of cytokines responsible for increased inflammation. METHODS: Evaluation of anti-inflammatory activity of selected phytoextracts namely, Camellia sinensis (L.) Kuntze, Acacia catechu (L.f) Willd., Curcuma longa (L.), Allium sativum (L.), Punica granatum (L.) and Azadirachta indica A. hereafter, called as catechin, epicatechin, curcumin, garlic, pomegranate and neem extracts, respectively in Acute wound fibroblasts (AWFs) and Chronic wound fibroblasts (CWFs) using flow cytometry. RESULTS: The phytoextracts exhibited no cytotoxicity below 100 µg/ml on normal Human Dermal fibroblasts (HDFs), while garlic extract showed highest cell viability followed by catechin, epicatechin, curcumin, pomegranate peel and neem based on IC50 value. Garlic, catechin and epicatechin extracts showed highest anti-inflammatory activities for both TGF-ß and TNF-α in both AWFs and CWFs treated cells. After treatment of AWFs with catechin, epicatechin and garlic extracts, TGF-ß and TNF-α expression was significantly reduced compared to untreated AWFs and reached to almost normal HDFs level. Also, after treatment of CWFs with catechin, epicatechin and garlic extracts, TGF-ß and TNF-α expression was significantly reduced compared to untreated CWFs and was lesser than untreated AWFs. CONCLUSION: The present findings reveal the potential of catechin, epicatechin and garlic extracts for the treatment of acute and chronic wounds with excellent anti-inflammatory properties.

Standardized ready-to-use laboratory protocol to isolate primary human wound associated fibroblasts from infected wound samples for clinical applications.

Owing to the importance of fibroblasts in healing of wounds, it is necessary to isolate and culture them under in vitro conditions for the purpose of understanding the wound biology, drug discovery and development of personalized treatment. Although, several fibroblast cell lines are commercially available, they fail to represent the patient associated parameters. However, establishing a primary fibroblast culture, especially from infected wound samples, is challenging as the sample is more prone to contamination and number of live cells will be minimum in heterogeneous population. Also, it takes lot of efforts and resources for optimization of the protocol to get good quality cell lines from wound samples necessitating multiple trials, resulting in large number of clinical samples to be processed. To the best of our knowledge, for the first time we are reporting the standardized protocol to isolate primary human fibroblasts from acute and chronic wound samples. In this study, various parameters such as explant size (1-2 mm), explant drying time (2 min), transportation and growth culture media (antibiotics (working concentrations 1-3) and serum concentration (10%)) have been optimised. This can be altered for specific needs of cell in terms of both quality and quantity. Outcome of the work provides a ready-to-use protocol, which is very useful to those who want to initiate primary fibroblasts cell culture from infected wound samples either for clinical or research purpose. Further, these cultured primary wound associated fibroblasts have various clinical and biomedical applications in tissue grafting, treatment of burns and scars and wound regeneration especially in non-healing chronic wounds.

Biomolecule conjugated inorganic nanoparticles for biomedical applications: A review.

Last decade has witnessed impressive progress in the fields of medicine and bioengineering with the aid of nanomaterials. Nanomaterials are favoured for their improved bio-chemical as well as mechanical properties with tremendous applications in biomedical domains such as disease diagnosis, targeted drug delivery, medical imaging, in vitro diagnostics, designing innovatory cross-functional implants and regenerative tissue engineering. The current situation insists upon crafting nanotools that are capable of catering to biological needs and construct more efficient biomedical strategies. In the recent years, surface functionalization and capping with biomolecules has initiated substantial interest towards research. In this regard, search of suitable biofunctionalized nanoparticles seem to be like finding pearls from ocean. Conjugating biological molecules with inorganic materials has paved the way for unravelling innovative functional materials with dramatically improved properties and a wide range of uses. Inorganic nanoparticles such as metals, metal oxides, as well as quantum dots have been hybridised or conjugated with biomolecules such as proteins, peptides, carbohydrates, and nucleic acids. The present review reports on various biomolecule functionalized inorganic nanomaterials highlighting the biomolecule-inorganic nanoparticle interaction studies, the mechanism of functionalization, antimicrobial efficacy of the functionalised nanoconjugates and its use in various biomedical applications.

Recent advances in pomegranate peel extract mediated nanoparticles for clinical and biomedical applications.

Manufacturing new materials at the nanoscale level is a field that is rapidly expanding with widespread application in advanced science and MMT is effectively used for the technology. Nanoparticles (NP), the building blocks of nanotechnology, exhibit improved properties than the larger counterparts and can be prepared from a variety of metals, including silver, copper, gold, zinc, and others. Phytonanotechnology is gaining major attention as various clinical researches have focused on the excellent properties (physicochemical and biological) of nanoscale phytochemicals and its applications in biological systems. In recent developments, pomegranate (Punica granatum L.) has gained major attention due to the phenolic compounds like apigenin, caffeic acid, chlorogenic acid, cyanidin, ellagic acid, gallic acid, granatin A, granatin B, pelargonidin, punicalagin, punicalin and quercetin found in its peel. Pomegranate Peel Extract (PPE) that aid the synthesis of PPE mediated nanoparticles (PPE-MNPs) like PPE-MAuNPs, PPE-MAgNPs, PPE-MZnONPs, PPE-MCuNPs, PPE-MPtNPs and PPE-MFeNPs has yielded plethora of beneficial properties in both plants and humans. In the current review, we discuss in detail the recent advances in synthesis and characterization of various nanoparticles from PPE. Moreover, the multitude biological properties of PPE-MNPs make up the long list of clinical uses. In addition, we discuss the pharmacokinetics, current advantages, and limitations of PPE-MNPs which can further help in development of more efficient therapeutics. Despite some of the challenges, PPE-MNPs hold a lot of potential for drug delivery and are always a better choice. The convergence of science and engineering has created new hopes, in which phytomedicines will have more efficacy, bioavailability, and less toxicity.

Human primary chronic wound derived fibroblasts demonstrate differential pattern in expression of fibroblast specific markers, cell cycle arrest and reduced proliferation.

INTRODUCTION: Although wound refers to simple cut in the skin, most wounds don't heal because of the various local and systemic factors that lead to its complexity and chronicity. Thus, prior understanding of the status of the wound is necessary and methods that can differentiate between the healing and non-healing wounds at a much earlier stage is crucial for a successful treatment. METHODS: The current study aims at differentiating Acute Wound Fibroblasts (AWFs) and Chronic Wound Fibroblasts (CWFs) based on differential expression of fibroblast specific markers such as Vimentin and Alpha Smooth Muscle Actin (α-SMA) and compare its cell cycle and proliferation. RESULTS: Immunostaining and western blotting analysis showed that, AWFs and CWFs differentially expressed vimentin and α-SMA, with AWFs and CWFs showing higher expression of vimentin and α-SMA respectively. AWFs showed higher distributions in G0/G1 (67.43% vs. 62.16%), S phase (22.61% vs. 8.51%) compared to CWFs. However, AWFs showed decreased distributions compared to CWFs in G2 + M phase (8.14% vs. 10.6%). Thus, it was observed that CWFs showed cell cycle arrest in the G1/G0 phase and inhibited DNA synthesis, which was further confirmed by reduced proliferation of CWFs. We suggest that, differential expression of the cell specific markers can be attributed to its pathophysiological status and chronicity of the wound and reduced proliferation rate of CWFs is due to lesser expression of vimentin, which is a key protein for in vitro cell proliferation. CONCLUSIONS: Outcome of the study serve as an immunological tool to guide the chronicity of the wound, which helps to understand the wound towards design of personalized care. The findings also represent a promising opportunity to gain insight into how cell cycle arrest can impact on wound healing and clinical outcomes.

Risks of mucormycosis in the current Covid-19 pandemic: a clinical challenge in both immunocompromised and immunocompetent patients.

Mucormycosis, also called "Black Fungus", is a new cause for worry in the current Coronavirus disease 2019 (covid-19) pandemic. Mucormycosis is devasting due to its high rate of morbidity and mortality which is a great cause of concern. Mucormycosis, in general, affects immunocompromised patients including diabetic, people with malignancies, organ and stem cell transplants and people affected with pandemic diseases like covid-19. Diagnosis of Mucormycosis is often delayed either due to clinical complications or misdiagnosed as symptoms of other diseases, especially covid-19. This could delay the treatment protocol which results in the failure of treatment. Mortality rate due to secondary infections in covid-19 patients with uncontrolled diabetics and who are on steroid therapy can soon reach 100% if diagnosis and treatment doesn't happen on timely basis. Risk of Mucormycosis is not just in immunosuppressed patients, but immunocompetent people with late diagnosis are also prone to infection. In view of this, we present a comprehensive review on risks of Mucormycosis in immunocompromised and immunocompetent patients highlighting the epidemiology, forms of Mucormycosis, immune response against Mucorales, difficulties in diagnosis and challenges in treatment of Mucormycosis, with emphasis on covid-19 associated Mucormycosis. Importantly, we have discussed the precautions and care to effectively manage Mucormycosis in immunocompromised and immunocompetent patients. Thus, current review helps clinicians in understanding various risk factors in both immunocompromised (especially covid-19 patients) and immunocompetent patients which is critical in managing Mucormycosis in current covid-19 pandemic.

Modeling and analysis of nanoscale interaction forces between Acidithiobacillus ferrooxidans and AFM tip.

The specificity exhibited by Acidithiobacillus ferrooxidans, a chemolithoautotrophic acidophile, in their attachment to different sulphide minerals has resulted in effective physical separation of these minerals. This can be explained in terms of surface forces of interaction between the cells and substrates using AFM. In the light of this, the present study reports interaction studies with A. ferrooxidans cells and an AFM silicon nitride tip. The aim of the present investigation is to probe the nanoscale interactions between A. ferrooxidans cells and silicon nitride tip of an AFM, model the approach and retraction forces, and elucidate the effects of pH, ionic strength and surface biopolymers on interfacial forces.

Soft-particle model analysis of effect of LPS on electrophoretic softness of Acidithiobacillus ferrooxidans grown in presence of different metal ions.

The effect of surface lipopolysaccharides (LPS) on the electrophoretic softness and fixed charge density in the ion-penetrable layer of Acidithiobacillus ferrooxidans cells grown in presence of copper or arsenic ions have been discussed. The electrophoretic mobility data were analyzed using the soft-particle electrophoresis theory. Cell surface potentials of all the strains based on soft-particle theory were lower than those estimated using the conventional Smoluchowski theory. Exposure to metal ions increased the surface electrophoretic softness with decrease in the fixed charge density. Effect of cell surface lipopolysaccharides on the model parameters are investigated and discussed.

Selective separation of pyrite and chalcopyrite by biomodulation.

Selective separation of pyrite from other associated ferrous sulphides at acidic and neutral pH has been a challenging problem. This paper discusses the utility of Acidithiobacillus ferrooxidans for the selective flotation of chalcopyrite from pyrite. Consequent to interaction with bacterial cells, pyrite remained depressed even in the presence of potassium isopropyl xanthate collector while chalcopyrite exhibited significant flotability. However, when the minerals were conditioned together, the selectivity achieved was poor due to the activation of pyrite surface by the copper ions in solution. The selectivity was improved when the sequence of conditioning with bacterial cells and collector was reversed, since the bacterial cells were able to depress collector interacted pyrite effectively, while having negligible effect on chalcopyrite. The observed behaviour is analysed and discussed in detail. The separation obtained was significant both at acidic and alkaline pH. This selectivity achieved was retained when the minerals were interacted with both bacterial cells and collector simultaneously.

Selective separation of arsenopyrite from pyrite by biomodulation in the presence of Acidithiobacillus ferrooxidans.

Effective methods for selective separation using flotation or flocculation of arsenopyrite from pyrite by biomodulation using Acidithiobacillus ferrooxidans are presented here. Adhesion of the bacterium to the surface of arsenopyrite was very slow compared to that to pyrite, resulting in a difference in surface modification of the minerals subsequent to interaction with cells. The cells were able to effectively depress pyrite flotation in presence of collectors like potassium isopropyl xanthate and potassium amyl xanthate. On the other hand the flotability of arsenopyrite after conditioning with the cells was not significantly affected. The activation of pyrite by copper sulfate was reduced when the minerals were conditioned together, resulting in better selectivity. Selective separation could also be achieved by flocculation of biomodulated samples.

Modeling and analysis of biooxidation of gold bearing pyrite-arsenopyrite concentrates by Thiobacillus ferrooxidans.

The results of modeling the biooxidation of a mixed sulfidic concentrate by Thiobacillus ferrooxidans is reported here. A kinetic model, which accounts for the dissolution of sulfide matrix due to both bacterial attachment onto the mineral surface and indirect leaching, has been proposed. A comprehensive system approach is employed for modeling the complex biooxidation process by (a) the decomposition of the complete system into several subsystems, (b) modeling individual systems, and (c) integrating the subsystems model in a final system model. The model for subsystems was developed by writing mass balance equations for the different species involved. The bacterial balance accounts for its growth, both on solid substrate and in solution, and for the attachment to and detachment from the surface. The kinetic parameters of the model were determined by designing the experiments in such a manner that only one subsystem was operational. This model was tested in both laboratory scale batch and continuous biooxidation processes. The model predictions agreed with the experimental data reasonably well. A further analysis of the model was carried out to predict the conditions for efficient biooxidation. Studies on the effect of residence time and pulp density on steady-state behavior showed that there is a critical residence time and pulp density below which washout conditions occur. Operation at pulp densities lower than 5% and residence times lower than 72 h was found unfavorable for efficient leaching.