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Treating different types of bone defects is difficult, complicated, time-consuming, and expensive. Here, we demonstrate that transient receptor potential cation channel subfamily V member 4 (TRPV4), a mechanosensitive, thermogated, and nonselective cation channel, is endogenously present in the mesenchymal stem cells (MSCs). TRPV4 regulates both cytosolic Ca 2+ levels and mitochondrial health. Accordingly, the hydrogel made from a natural modified biopolymer carboxymethyl tamarind CMT-Hy and encapsulated with TRPV4-modulatory agents affects different parameters of MSCs, such as cell morphology, focal adhesion points, intracellular Ca 2+ , and reactive oxygen species- and NO-levels. TRPV4 also regulates cell differentiation and biomineralization in vitro . We demonstrate that 4α-10-CMT-Hy and 4α-50-CMT-Hy (the hydrogel encapsulated with 4αPDD, 10 and 50 nM, TRPV4 activator) surfaces upregulate mitochondrial health, i.e., an increase in ATP- and cardiolipin-levels, and improve the mitochondrial membrane potential. The same scaffold turned out to be nontoxic in vivo . 4α-50-CMT-Hy enhances the repair of the bone-drill hole in rat femur, both qualitatively and quantitatively in vivo . We conclude that 4α-50-CMT-Hy as a scaffold is suitable for treating large-scale bone defects at low cost and can be tested for clinical trials.

Introduction: Reducing the healing period after surgical placement of dental implants can facilitate the loading of dental prostheses., Aim: The aim is to compare the osteogenic potential of unmodified titanium disks with titanium disks that were surface-modified or hydrogel-coated., Materials and Methodology: One hundred eight titanium disks (Ø6 × 2-mm) were divided into three groups: (1) unmodified titanium as control (Ti-C); (2) sandblasted and acid-etched (Ti-SLA), and (3) coated with tamarind kernel polysaccharide hydrogel grafted with acrylic acid (Ti-TKP-AA). The osteogenic potential and cytotoxic effect of various groups of titanium were compared using human osteoblasts Saos-2. The surface topography of the titanium disks and morphology of osteoblasts grown on disks were investigated by scanning electron microscopy (n = 3). Cell attachment to the disks and actin expression intensity were investigated by confocal imaging (n = 3). Cytotoxicity was quantified by cell viability assay (n = 9). Osteoblast maturation was determined by alkaline phosphatase assay (n = 9). Cell mineralization was quantified by Alizarin red staining (n = 9). One-way analysis of variance followed by Tukey's multiple comparisons test was used for intergroup comparisons ( α = 0.05)., Results: The surface modifications on Ti-SLA and Ti-TKP-AA support better morphology and proliferation of osteoblasts than Ti-C (P< 0.001) and significantly higher levels of actin cytoskeleton accumulation (P< 0.0001). Ti-TKP-AA showed a significantly higher maturation rate than Ti-C (P< 0.001). Ti-TKP-AA showed > twofold increased mineralization than Ti-C and Ti-SLA (P< 0.001)., Conclusions: TKP-AA hydrogel-coated titanium promotes faster osteoblast proliferation, maturation, and mineralization than SLA-treated or untreated titanium. These advantages can be explored for achieving early osseointegration and prosthetic loading of titanium dental implants., Competing Interests: The authors have declared that no competing interests exist., (Copyright © 2024, Majhi et al.)

Gold nanoparticles (AuNPs) have been reported to modulate bone tissue regeneration and are being extensively utilized in biomedical implementations attributable to their low cytotoxicity, biocompatibility and simplicity of functionalization. Lately, biologically synthesized nanoparticles have acquired popularity because of their environmentally acceptable alternatives for diverse applications. Here we report the green synthesis of AuNPs by taking the biopolymer Carboxymethyl Tamarind (CMT) as a unique reducing as well as a stabilizing agent. The synthesized CMT-AuNPs were analyzed by UV-vis spectrophotometer, DLS, FTIR, XRD, TGA, SEM and TEM. These results suggest that CMT-AuNPs possess an average size of 19.93 ± 8.52 nm and have long-term stability. Further, these CMT-AuNPs promote the proliferation together with the differentiation and mineralization of osteoblast cells in a "dose-dependent" manner. Additionally, CMT-AuNPs are non-toxic to SD rats when applied externally. We suggest that the CMT-AuNPs have the potential to be a suitable and non-toxic agent for differentiation and mineralization of osteoblast cells in vitro and this can be tested in vivo as well., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

In the current work, biomaterial composed of Xanthan gum and Diethylene glycol dimethacrylate with impregnation of graphite nanopowder filler in their matrices was fabricated successfully for their potential usage in the engineering of bone defects. Various physicochemical properties associated with the biomaterial were characterized using FTIR, XRD, TGA, SEM etc. The biomaterial rheological studies imparted the better notable properties associated with the inclusion of graphite nanopowder. The biomaterial synthesized exhibited a controlled drug release. Adhesion and proliferation of different secondary cell lines do not generate ROS on the current biomaterial and thus show its biocompatibility and non-toxic nature. The synthesized biomaterial's osteogenic potential on SaOS-2 cells was supported by increased ALP activity, enhanced differentiation and biomineralization under osteoinductive circumstances. The current biomaterial demonstrates that in addition to the drug-delivery applications, it can also be a cost-effective substrate for cellular activities and has all the necessary properties to be considered as a promising alternative material suitable for repairing and restoring bone tissues. We propose that this biomaterial may have commercial importance in the biomedical field., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier B.V. All rights reserved.)

Bone defects, including bone loss due to increased osteoclast activity, have become a global health-related issue. Osteoclasts attach to the bone matrix and resorb the same, playing a vital role in bone remodeling. Ca 2+ homeostasis plays a pivotal role in the differentiation and maturation of osteoclasts. In this work, we examined the role of TRPV1, a nonselective cation channel, in osteoclast function and differentiation. We demonstrate that endogenous TRPV1 is functional and causes Ca 2+ influx upon activation with pharmacological activators [resiniferatoxin (RTX) and capsaicin] at nanomolar concentration, which enhances the generation of osteoclasts, whereas the TRPV1 inhibitor (5'-IRTX) reduces osteoclast differentiation. Activation of TRPV1 upregulates tartrate-resistant acid phosphatase activity and the expression of cathepsin K and calcitonin receptor genes, whereas TRPV1 inhibition reverses this effect. The slow release of capsaicin or RTX at a nanomolar concentration from a polysaccharide-based hydrogel enhances bone marrow macrophage (BMM) differentiation into osteoclasts whereas release of 5'-IRTX, an inhibitor of TRPV1, prevents macrophage fusion and osteoclast formation. We also characterize several subcellular parameters, including reactive oxygen (ROS) and nitrogen (RNS) species in the cytosol, mitochondrial, and lysosomal profiles in BMMs. ROS were found to be unaltered upon TRPV1 modulation. NO, however, had elevated levels upon RTX-mediated TRPV1 activation. Capsaicin altered mitochondrial membrane potential (ΔΨm) of BMMs but not 5'-IRTX. Channel modulation had no significant impact on cytosolic pH but significantly altered the pH of lysosomes, making these organelles less acidic. Since BMMs are precursors for osteoclasts, our findings of the cellular physiology of these cells may have broad implications in understanding the role of thermosensitive ion channels in bone formation and functions, and the TRPV1 modulator-releasing hydrogel may have application in bone tissue engineering and other biomedical sectors., Competing Interests: The authors declare no competing financial interest., (© 2022 The Authors. Published by American Chemical Society.)

Statement of Problem: Rapid osseointegration between implant and bone tissue for early loading of a prosthesis with sufficient primary stability depends on the surface characteristics of the implant. The development and characterization of suitable surface coatings on dental implants is a major challenge., Purpose: The purpose of this in vitro study was to evaluate and compare the osteogenic potential and cytotoxicity of unmodified zirconia, acid-etched zirconia, bioactive glass-coated zirconia, and tamarind kernel polysaccharide with hydrophilic acrylic acid (TKP-AA) hydrogel-coated zirconia., Material and Methods: Thirty-six disks each of unmodified zirconia, acid-etched, 45S5 bioactive glass-coated, and TKP-AA hydrogel-coated zirconia were evaluated for osteogenic potential and cytotoxic effect by using human osteoblast Saos-2 cells. The surface topography of the disks and the morphology of the cells grown on these surfaces were examined by scanning electron microscopy (n=3). The cell attachment was evaluated by confocal imaging (n=3). The cytotoxic effect was evaluated by cell viability assay (n=9). Osteoblast maturation was assessed by alkaline phosphatase assay (n=9) and cell mineralization by alizarin red staining (n=9). ANOVA and Bonferroni multiple comparison post hoc tests were used to evaluate the statistical significance of the intergroup differences in these characteristics (α=.05)., Results: The surface modifications resulted in distinct changes in the surface morphology of zirconia disks and the growth of Saos-2 cells. Zirconia disks coated with TKP-AA promoted higher proliferation of osteoblasts compared with unmodified disks (P<.001). Similarly, the surface modifications significantly increased the differentiation of mesenchymal stem cells to osteoblasts as compared with uncoated zirconia (P<.001). However, the rate of differentiation to osteoblasts was similar among the surface modifications. Acid-etched and TKP-AA-coated disks promoted mineralization of osteoblasts to the same extent, except bioactive glass coating, which significantly increased the rate of mineralization (P<.001)., Conclusions: Surface modification of zirconia by acid etching and coating with Bioglass or TKP-AA hydrogel resulted in the improved growth and differentiation of osteoblasts. TKP-AA hydrogel coating promoted the proliferation of osteoblasts, whereas Bioglass coating showed better mineralization. TKP-AA hydrogel coating is a promising candidate for improving the osseointegration of dental implants that warrants further investigation., (Copyright © 2021 Editorial Council for the Journal of Prosthetic Dentistry. Published by Elsevier Inc. All rights reserved.)

A major limitation in the bio-medical sector is the availability of materials suitable for bone tissue engineering using stem cells and methodology converting the stochastic biological events towards definitive as well as efficient bio-mineralization. We show that osteoblasts and Bone Marrow-derived Mesenchymal Stem Cell Pools (BM-MSCP) express TRPM8, a Ca 2+ -ion channel critical for bone-mineralization. TRPM8 inhibition triggers up-regulation of key osteogenesis factors; and increases mineralization by osteoblasts. We utilized CMT:HEMA, a carbohydrate polymer-based hydrogel that has nanofiber-like structure suitable for optimum delivery of TRPM8-specific activators or inhibitors. This hydrogel is ideal for proper adhesion, growth, and differentiation of osteoblast cell lines, primary osteoblasts, and BM-MSCP. CMT:HEMA coated with AMTB (TRPM8 inhibitor) induces differentiation of BM-MSCP into osteoblasts and subsequent mineralization in a dose-dependent manner. Prolonged and optimum inhibition of TRPM8 by AMTB released from the gels results in upregulation of osteogenic markers. We propose that AMTB-coated CMT:HEMA can be used as a tunable surface for bone tissue engineering. These findings may have broad implications in different bio-medical sectors.

Advanced technologies like skin tissue engineering are requisite of various disorders where artificially synthesized materials need to be used as a scaffold in vivo, which in turn can allow the formation of functional skin and epidermal layer with all biological sensory functions. In this work, we present a set of hydrogels which have been synthesized by the method utilizing radical polymerization of a natural polymer extracted from kernel of Tamarindus indica, commonly known as Tamarind Kernel Powder (TKP) modified by utilizing the monomer acrylic acid (AA) in different mole ratios. These materials are termed as TKP: AA hydrogels and characterized by Atomic Force Microscopy (AFM), surface charge, and particle size distribution using Dynamic Light Scattering measurements. These materials are biocompatible with mouse dermal fibroblasts (NIH- 3T3) and human skin keratinocytes (HaCaT), as confirmed by MTT and biocompatibility assays. These TKP: AA hydrogels do not induce unwanted ROS signaling as confirmed by mitochondrial functionality determined by DCFDA staining, Mitosox imaging, and measuring the ATP levels. We demonstrate that in the co-culture system, TKP: AA allows the establishment of proper neuro-keratinocyte contact formation, suggesting that this hydrogel can be suitable for developing skin with sensory functions. Skin corrosion analysis on SD rats confirms that TKP: AA is appropriate for in vivo applications as well. This is further confirmed by in vivo compatibility and toxicity studies, including hemocompatibility and histopathology of liver and kidney upon direct introduction of hydrogel into the body. We propose that TKP: AA (1: 5) offers a suitable surface for skin tissue engineering with sensory functions applicable in vitro, in vivo, and ex vivo. These findings may have broad biomedical and clinical importance., (Copyright © 2020 Elsevier B.V. All rights reserved.)

In this work, sodium alginate (SA) based "all-natural" composite bio-sponges were designed for potential application as wound care scaffold. The composite bio-sponges were developed from the aqueous amalgamation of SA and cellulose nanofibres (CNFs) in bio-extracts like Rice water (Rw) and Giloy extract (Ge). These sponges were modified by employing a simple coating strategy using vegetable oil-based bio-polyurethane (BioPU) to tailor their physicochemical and biological properties so as to match the specific requirements of a wound care scaffold. Bio-sponges with shared interpenetrating polymeric network structures were attained at optimized BioPU coating formulation. The interpenetration of BioPU chains within the sponge construct resulted in the formation of numerous micro-networks in the interconnected microporous structure of sponges (porosity ≥75%). The coated sponge showed a superior mechanical strength (compressive strength ~3.8 MPa, compressive modulus ~35 MPa) with appreciable flexibility and recoverability under repeated compressive loading-unloading cycles. A tunable degradation behaviour was achieved by varying BioPU coating concentrations owing to the different degree of polymer chain entanglement within the sponge construct. The physical entanglement of BioPU chains with core structural components of sponge improved their structural stability by suppressing their full fragmentation in water-based medium without affecting its swelling behaviour (swelling ratio > 1000%). The coated sponge surface has provided a suitable moist-adherent physical environment to support the adhesion and growth of skin cells (HaCaT cells). The MTT (3-(4,5-dimethyl thiazolyl-2)-2,5-diphenyltetrazolium bromide) assay and hemolytic assay revealed the non-toxic and biocompatible nature of coated sponges in vitro. Moreover, no signs of skin erythema or edema were observed during in vivo dermal irritation and corrosion test performed on the skin of Sprague Dawley (SD) rats. Our initial observations revealed the credibility of these sponges as functional wound care scaffolds as well as its diverse potential as a suitable substrate for various tissue engineering applications., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Multidrug resistant (MDR) bacteria have emerged as a major clinical challenge. The unavailability of effective antibiotics has necessitated the use of emerging nanoparticles as alternatives. In this work, we have developed carbohydrate-coated bimetallic nanoparticles (Au-AgNP, 30-40 nm diameter) that are nontoxic toward mammalian cells yet highly effective against MDR strains as compared to their monometallic counterparts (Ag-NP, Au-NP). The Au-AgNP is much more effective against Gram-negative MDR Escherichia coli and Enterobacter cloacae when compared to most of the potent antibiotics. We demonstrate that in vivo , Au-AgNP is at least 11000 times more effective than Gentamicin in eliminating MDR Methicillin Resistant Staphylococcus aureus (MRSA) infecting mice skin wounds. Au-AgNP is able to heal and regenerate infected wounds faster and in scar-free manner. In vivo results show that this Au-AgNP is very effective antibacterial agent against MDR strains and does not produce adverse toxicity. We conclude that this bimetallic nanoparticle can be safe in complete skin regeneration in bacteria infected wounds.

This study reports the synthesis of an unprecedented bio-based aquasorbent guargum-g-hyperbranched poly (acrylic acid); bGG-g-HBPAA by employing graft-copolymerization and "Strathclyde methodology" simultaneously in emulsion and its possible use as a sustainable nutrient bed for the effective growth of Anabaena cylindrica and Vigna radiata seedlings. The formation of bGG-g-HBPAA and the presence of hyperbranched architectures was confirmed from XRD, FTIR, 13 C NMR, solubility, intrinsic viscosity, BET surface area/ pore size, SEM and rheology analyses. The synthesized grade with a branching percent of 65.4% and a swelling percentage of 13,300% facilitated maximum growth of the cultured species as compared to guargum and its linear graft. Semi synthetic bGG-g-HBPAA culture medium was optically transparent, dried at a controlled rate, held a huge amount of water for growth, provided sufficient space for unhindered growth and featured dimensional stability., (Copyright © 2019. Published by Elsevier Ltd.)

A potential nanograde antibiotic with hyperbranched architecture was synthesized from melt esterification of poly(ethylene glycol) or PEG and Citric acid or CA with 1:1 mol composition. PEG of different molecular weights, c.a. 4000, 6000 and 20,000 were used during the polyesterification. The polyester molecules of nanometric size were highly water soluble and showed a melting point between 55 and 60 °C. The branching status was established from spectroscopy, flow behaviour (viscosity) and rheological evidences. The extent of branching and flowability, both were reduced as the molecular weight of PEG was increased. During in-vitro pathological study, all the grades showed reasonably strong antibacterial affect (both with gram positive and negative bacteria), high selectivity, biocompatibility and controlled generation of reactive oxygen species or ROS, however, the grade with maximum level of branching and functional chain ends displayed highest therapeutic efficiency, may that be considered further as a potential agent for next level investigation., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Purpose: With increased life expectancy, disorders in lifestyle and other clinical conditions, and the changes in the connective tissues such as in bone, impose diverse biomedical problems. Cells belong to osteogenic lineages are extremely specific for their surface requirements. Therefore, suitable surfaces are the critical bottle neck for successful bone tissue engineering. This study involves assessment of polysaccharide-based hydrogel which effectively allows growth, differentiation and mineralisation of osteogenic cells even in the absence of osteogenic inducing factors., Materials and Methods: Tamarind Kernel Polysaccharide was grafted with acrylic acid at different mole ratio. The critical parameter, surface morphology for bio application was assessed by SEM. MTT assay has been performed with hydrogels on Saos-2 cells. The biocompatibility and adhesion of different cell lines (F-11, Saos-2, Raw 264.7 and MSCs) on hydrogel surface was performed by Phalloidin and DAPI staining. Further the differentiation, mineralization and expression of different osteogenic markers, ALP assay, Alizarin Red staining and q-PCR was performed., Results: The hydrogels show highly porous and interconnected pores. MTT assay demonstrates the hydrogel have no cytotoxicity towards Saos-2 cells and are suitable for proliferation of different lineage of cell lines. ALP, Alizarin red staining and q-PCR assay shows that the hydrogel surface enhances the differentiation, mineralization and expression of different osteogenic genes in Saos-2 cells in the absence of any osteogenic inducing factors. Conclusion Synthesized hydrogel surface triggers signalling events towards osteogenesis even in the absence of added growth factors. We proposed that this material can be used for effective bone tissue engineering in vitro at low cost.

Patho-physiologies related to skin are diverse in nature such as burns, skin ulcers, atopic dermatitis, psoriasis etc. which impose severe bio-medical problems and thus enforce requirement of new and healthy skin prepared through tissues engineering methodologies. However, fully functional and biodegradable matrix for attachment, growth, proliferation and differentiation of the relevant cells is not available. In the present study, we introduce a set of hydrogels synthesized by incorporation of a synthetic monomer (Hydroxyethlmethacryate) with a semi-synthetic polymer backbone (carboxy methyl tamarind, CMT) in different mole ratios. We termed these materials as CMT:HEMA based hydrogels and these were characterized by different physico-chemical techniques, namely by X-Ray Diffraction, SEM and Dynamic Light Scattering. Biocompatibility studies with HaCaT, NIH-3T3 and mouse dermal fibroblasts confirm that this material is biocompatible. MTT assay further confirmed that this material does not have any cytotoxic effects. Assays for mitochondrial functionality such as ATP assay and mitochondrial reactive oxygen (ROS) generation also suggest that this material is safe and does not have any cytotoxicity. Hemolytic assay with red blood cells and acute skin irritation test on SD Rats confirmed that this material is suitable for ex-vivo application in future. We suggest that this hydrogel is suitable for in-vivo applications and may have clinical and commercial importance against skin disorders., (Copyright © 2018. Published by Elsevier Ltd.)

Antibacterial therapy is of paramount importance in treatment of several acute and chronic infectious diseases caused by pathogens. Over the years extensive use and misuse of antimicrobial agents has led to emergence of multidrug resistant (MDR) and extensive drug resistant (XDR) pathogens. This drastic escalation in resistant phenotype has limited the efficacy of available therapeutic options. Thus, the need of the hour is to look for alternative therapeutic approaches to mitigate healthcare concerns caused due to MDR bacterial infections. Nanoparticles have gathered much attention as potential candidates for antibacterial therapy. Equipped with advantages of, wide spectrum bactericidal activity at very low dosage, inhibitor of biofilm formation and ease of permeability, nanoparticles have been considered as leading therapeutic candidates to curtail infections resulting from MDR bacteria. However, substrate non-specificity of efflux pumps, particularly those belonging to resistance nodulation division super family, have been reported to reduce efficacy of many potent antibacterial therapeutic drugs. Previously, we had reported antibacterial activity of polysaccharide-capped silver nanoparticles (AgNPs) toward MDR bacteria. We showed that AgNPs inhibits biofilm formation and alters expression of cytoskeletal proteins FtsZ and FtsA, with minimal cytotoxicity toward mammalian cells. In the present study, we report no reduction in antibacterial efficacy of silver nanoparticles in presence of AcrAB-TolC efflux pump proteins. Antibacterial tests were performed according to CLSI macrobroth dilution method, which revealed that both silver nanoparticles exhibited bactericidal activity at very low concentrations. Further, immunoblotting results indicated that both the nanoparticles modulate the transporter AcrB protein expression. However, expression of the membrane fusion protein AcrA did show a significant increase after exposure to AgNPs. Our results indicate that both silver nanoparticles are effective in eliminating MDR Enterobacter cloacae isolates and their action was not inhibited by AcrAB-TolC efflux protein expression. As such, the above nanoparticles have strong potential to be used as effective and alternate therapeutic candidates to combat MDR gram-negative Enterobacterial pathogens.

Bone related problems are increasing as a consequence of increased life expectancy, disorders in life style, and other medical conditions enforcing the need for functional bones prepared in vitro at affordable cost. Lack of suitable surface which promotes growth of both osteogenic and nonosteogenic cells is a major limitation. Here a novel biomaterial is reported that is synthesized from natural polysaccharide, namely, tamarind kernel polysaccharide (TKP), which is grafted with hydrophilic acrylic acid (AA) by radical polymerization. Modification in surface functionality removes unwanted proteins and alters hydrophilic/hydrophobic balance. TKP-AA is suitable for the growth of different nonosteogenic and osteogenic cells. This material is suitable for osteoblasts and promotes in vitro mineralization and differentiation without the addition of exogenous growth factors. TKP-AA can be used for the growth of mesenchymal stem cell-derived osteoblasts. It is suggested that TKP-AA can potentially be used as a scaffold for diverse cell types and particularly for bone tissue engineering at low cost., (© 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

Development of effective anti-microbial therapeutics has been hindered by the emergence of bacterial strains with multi-drug resistance and biofilm formation capabilities. In this article, we report an efficient green synthesis of silver nanoparticle (AgNP) by in situ reduction and capping with a semi-synthetic polysaccharide-based biopolymer (carboxymethyl tamarind polysaccharide). The CMT-capped AgNPs were characterized by UV, DLS, FE-SEM, EDX and HR-TEM. These AgNPs have average particle size of ~20-40 nm, and show long time stability, indicated by their unchanged SPR and Zeta-potential values. These AgNPs inhibit growth and biofilm formation of both Gram positive (B. subtilis) and Gram negative (E. coli and Salmonella typhimurium) bacterial strains even at concentrations much lower than the minimum inhibitory concentration (MIC) breakpoints of antibiotics, but show reduced or no cytotoxicity against mammalian cells. These AgNPs alter expression and positioning of bacterial cytoskeletal proteins FtsZ and FtsA. CMT-capped AgNPs can effectively block growth of several clinical isolates and MDR strains representing different genera and resistant towards multiple antibiotics belonging to different classes. We propose that the CMT-capped AgNPs can have potential bio-medical application against multi-drug-resistant microbes with minimal cytotoxicity towards mammalian cells.

Transient Receptor Potential cation channel, subfamily Melastatin, member 8 (TRPM8) is involved in detection of cold temperature, different noxious compounds and in execution of thermo- as well as chemo-sensitive responses at cellular levels. Here we explored the molecular evolution of TRPM8 by analyzing sequences from various species. We elucidate that several regions of TRPM8 had different levels of selection pressure but the 4th-5th transmembrane regions remain highly conserved. Analysis of synteny suggests that since vertebrate origin, TRPM8 gene is linked with SPP2, a bone morphogen. TRPM8, especially the N-terminal region of it, seems to be highly variable in human population. We found 16,656 TRPM8 variants in 1092 human genomes with top variations being SNPs, insertions and deletions. A total of 692 missense mutations are also mapped to human TRPM8 protein of which 509 seem to be delateroiours in nature as supported by Polyphen V2, SIFT and Grantham deviation score. Using a highly specific antibody, we demonstrate that TRPM8 is expressed endogenously in the testis of rat and sperm cells of different vertebrates ranging from fish to higher mammals. We hypothesize that TRPM8 had emerged during vertebrate evolution (ca 450 MYA). We propose that expression of TRPM8 in sperm cell and its role in regulating sperm function are important factors that have guided its molecular evolution, and that these understandings may have medical importance.

Remodeling of bone by tissue engineering is a realistic option for treating several bone-related pathophysiological ailments such as osteoporosis, bone tumor, bone cancer or abnormal bone development. But, these possibilities are hindered due to lack of proper natural and biodegradable surface on which bone precursor cells can adhere efficiently and grow further. Here we describe the synthesis and characterization of a new hydrogel as an effective surface which can acts as a material for bone tissue engineering. This hydrogel has been prepared by chemically grafting a semi-synthetic polymer with a synthetic monomer, namely hydroxyethyl methacrylate (HEMA). Carboxy methyl tamarind (CMT) was selected as the semi-synthetic polymer. The hydrogel was prepared at different mole ratios and at the ratio of 1:10 (CMT:HEMA) yielded the best hydrogel as characterized by several physico-chemical analysis such as UV spectroscopy, FT-IR spectroscopy and swelling properties. We further demonstrate that this material is suitable for effective adhesion, growth and further clustering of bone precursor cells (RAW 264.7). This material is also compatible for growing other sensitive cells such as neuronal cells (Neuro2a) and human umbilical vein endothelial cells (HUVEC) demonstrating that this surface does not possess any cytotoxicity and is compatible for primary human cells too. We conclude that the hydrogel made of CMT:HEMA at a ratio of 1:10 can be suitable for bone tissue engineering and thus may have clinical as well as commercial application in future., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

Sperm cells exhibit extremely high sensitivity in response to slight changes in temperature, osmotic pressure and/or presence of various chemical stimuli. In most cases throughout the evolution, these physico-chemical stimuli trigger Ca (2+)-signaling and subsequently alter structure, cellular function, motility and survival of the sperm cells. Few reports have recently demonstrated the presence of Transient Receptor Potential (TRP) channels in the sperm cells from higher eukaryotes, mainly from higher mammals. In this work, we have explored if the sperm cells from lower vertebrates can also have thermo-sensitive TRP channels. In this paper, we demonstrate the endogenous presence of one specific thermo-sensitive ion channel, namely Transient Receptor Potential Vanilloid family member sub type 1 (TRPV1) in the sperm cells collected from fresh water teleost fish, Labeo rohita. By using western blot analysis, fluorescence assisted cell sorting (FACS) and confocal microscopy; we confirm the presence of this non-selective cation channel. Activation of TRPV1 by an endogenous activator NADA significantly increases the quality as well as the duration of fish sperm movement. The sperm cell specific expression of TRPV1 matches well with our in silico sequence analysis. The results demonstrate that TRPV1 gene is conserved in various fishes, ranging from 1-3 in copy number, and it originated by fish-specific duplication events within the last 320 million years (MY). To the best of our knowledge, this is the first report demonstrating the presence of any thermo-sensitive TRP channels in the sperm cells of early vertebrates as well as of aquatic animals, which undergo external fertilization in fresh water. This observation may have implications in the aquaculture, breeding of several fresh water and marine fish species and cryopreservation of fish sperms.

Green, hydrophobic device for controlled transdermal release of diclofenac sodium was designed from in situ nanosilica/acrylic acid grafted guargum membranes. Best grafting condition was assigned and nanocomposites were formed in situ using varying proportions of aqueous nanosilica sol. Nanocomposite/drug conjugates were formed by bringing down the medium pH from 9.0 to 7.0. The conjugates were characterized through infrared and solid state NMR spectroscopy, electron microscopy, hydro-swelling, surface contact angle, viscometry and biocompatibility. Most balanced property was exhibited by the membrane containing 1wt% nanosilica. It also had shown the highest encapsulation efficacy vis-à-vis slowest release as compared to others during experimentation in a Franz diffusion cell., (Copyright © 2012 Elsevier Ltd. All rights reserved.)

Though stress is an integrated part of the modern life, defining stress in biological systems is difficult. Anxiety, medication, metabolic disorder, neuro-endocrinological abnormalities, immunological responses, neuro-immune interaction and several other internal and external factors are important which induce stress and pain in higher organisms. Stress and pain are often synonymous and overlapping to a large extent, but these two responses are different at the behavioral, cellular and molecular levels. Importance of Transient Receptor Potential (TRP) group of non-selective cation channels in the development and regulation of different forms of pain is well established. However, recent studies confirmed that TRPs can regulate neuroplastic changes through neuro-endocrine signaling, neuro-immune interactions and psychological state variables suggesting that abnormalities in TRP-signaling can indeed affect the hypothalamic-pituitary-adrenal (HPA) axis and several other metabolic pathways and thus may generate stress at various levels. Therefore, TRPs are important factors that can link stress with pain. This review summarizes the role of TRPs, their effects and clinical implications in the context of different types of pain which can be relevant for stress too.

TRP channels are localized at specialized sub-cellular compartments like filopodial tips, ciliary structures, growth cones and spines that have importance in the context of several sensory functions. Several motor proteins largely regulate these localizations. Recent studies indicate that both physical and genetic interactions exist between TRP channels with actin and microtubule-based motor proteins. These two groups of proteins share specialized and fine regulation underlying physiological functions. Indeed, mutations causing loss of these interactions and regulations result in development of pathophysiological disorders and syndromes. In this review we analyze the recent progress made in cell-biological, biochemical, electrophysiological and genetic studies and summarize the multi-dimensional crosstalk between TRP channels with different motor proteins.

Structural and regulatory components in the spinous processes neurons are still not fully understood. While the presence, regulation and function of actin cytoskeleton are now well established, involvement of microtubule cytoskeleton is only now being studied. In spite of biochemical, genetic and anatomical evidence, neurobiologists have experienced difficulties to even establish the physical presence and to demonstrate the functional relevance of "filamentous microtubules" in the spines. However, recent studies have demonstrated that filamentous microtubules regularly invade most spines. Moreover, functional aspects like stabilization and maturation of spines can be directly correlated with the presence of these microtubules; low doses of microtubule stabilizers can induce as many as 50% more spines. Clinically, knowledge of microtubule active drugs is extremely important, especially in the area of oncology, where they are commonly used as anti-neoplastic agents. Unfortunately, a major dose-limiting side effect of this class of drugs is a painful peripheral neuropathy. We review the recent concept of microtubules in the spine, and also critically analyze their role in chemotherapy-induced neuropathic pain., (Copyright 2010 Elsevier Ltd. All rights reserved.)

The mechanism of active stress generation in tension wood is still not fully understood. To characterize the functional interdependency between the G-layer and the secondary cell wall, nanostructural characterization and mechanical tests were performed on native tension wood tissues of poplar (Populus nigra x Populus deltoids) and on tissues in which the G-layer was removed by an enzymatic treatment. In addition to the well-known axial orientation of the cellulose fibrils in the G-layer, it was shown that the microfibril angle of the S2-layer was very large (about 36 degrees). The removal of the G-layer resulted in an axial extension and a tangential contraction of the tissues. The tensile stress-strain curves of native tension wood slices showed a jagged appearance after yield that could not be seen in the enzyme-treated samples. The behaviour of the native tissue was modelled by assuming that cells deform elastically up to a critical strain at which the G-layer slips, causing a drop in stress. The results suggest that tensile stresses in poplar are generated in the living plant by a lateral swelling of the G-layer which forces the surrounding secondary cell wall to contract in the axial direction.

The feasibility of Fourier transform infrared (FT-IR) microscopy to monitor in situ the enzymatic degradation of wood was investigated. Cross-sections of poplar wood were treated with cellulase Onozuka RS within a custom-built fluidic cell. Light-optical micrographs and FT-IR spectra were acquired in situ from normal and tension wood fibers. Light-optical micrographs showed almost complete removal of the gelatinous (G) layer in tension wood. No structural and spectral changes were observed in the lignified cell walls. The accessibility of cellulose within the lignified cell wall was found to be the main limiting factor, whereas the depletion of the enzyme due to lignin adsorption could be ruled out. The fast, selective hydrolysis of the crystalline cellulose in the G-layer, even at room temperature, might be explained by the gel-like structure and the highly porous surface. Young plantation grown hardwood trees with a high proportion of G-fibers thus represent an interesting resource for bioconversion to fermentable sugars in the process to bioethanol.