Comparative evaluation of surface-modified zirconia for the growth of bone cells and early osseointegration.

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.

Evaluation of Osteogenic Potential of a Polysaccharide-Based Hydrogel Coating on Titanium.

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.

TRPV4 acts as a mitochondrial Ca2+-importer and regulates mitochondrial temperature and metabolism.

TRPV4 is associated with the development of neuropathic pain, sensory defects, muscular dystrophies, neurodegenerative disorders, Charcot Marie Tooth and skeletal dysplasia. In all these cases, mitochondrial abnormalities are prominent. Here, we demonstrate that TRPV4, localizes to a subpopulation of mitochondria in various cell lines. Improper expression and/or function of TRPV4 induces several mitochondrial abnormalities. TRPV4 is also involved in the regulation of mitochondrial numbers, Ca2+-levels and mitochondrial temperature. Accordingly, several naturally occurring TRPV4 mutations affect mitochondrial morphology and distribution. These findings may help in understanding the significance of mitochondria in TRPV4-mediated channelopathies possibly classifying them as mitochondrial diseases.

Modified tamarind kernel polysaccharide-based matrix alters neuro-keratinocyte cross-talk and serves as a suitable scaffold for skin tissue engineering.

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.

Bio-extract amalgamated sodium alginate-cellulose nanofibres based 3D-sponges with interpenetrating BioPU coating as potential wound care scaffolds.

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.

Carbohydrate-Coated Gold-Silver Nanoparticles for Efficient Elimination of Multidrug Resistant Bacteria and in Vivo Wound Healing.

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.

A Modified Polysaccharide-Based Hydrogel for Enhanced Osteogenic Maturation and Mineralization Independent of Differentiation Factors.

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.

A carboxy methyl tamarind polysaccharide matrix for adhesion and growth of osteoclast-precursor cells.

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.

TRPV4-mediated channelopathies.

Transient receptor potential vanilloid sub type 4 (TRPV4) is a member of non-selective cation channel that is important for sensation of several physical and chemical stimuli and also involved in multiple physiological functions. Recently it gained immense medical and clinical interest as several independent studies have demonstrated that mutations in the TRPV4 gene can results in genetic disorders like Brachyolmia, Charcot-Marie-Tooth disease type 2C, Spinal Muscular Atrophy and Hereditary Motor and Sensory Neuropathy type 2. Close analysis of the data obtained from these naturally occurring as well as other TRPV4 mutants suggest that it is not the altered channel activity of these mutants per se, but the involvement and interaction of other factors that seem to modulate oligomerization, trafficking and degradation of TRPV4 channels. Also, these factors can either enhance or reduce the activity of TRPV4. In addition, there are some potential signaling events that can also be involved in these genetic disorders. In this review, we analyzed how and what extent certain cellular and molecular functions like oligomerization, surface expression, ubiquitination and functional interactions might be affected by these mutations.