Investigation of cell-accelerated corrosion (CAC) on the CoCrMo alloy with segregation banding: Hip implant applications.

Metal alloy microstructure plays a crucial role in corrosion associated with total hip replacement (THR). THR is a prominent strategy that uses metal implants such as cobalt-chromium-molybdenum (CoCrMo) alloys due to their advantageous biological and mechanical properties. Despite all benefits, these implants undergo corrosion and wear processes in-vivo in a synergistic manner called tribocorrosion. Also, the implant retrieval findings reported that fretting corrosion occurred in-vivo, evidenced by the damage patterns that appeared on the THR junction interfaces. There is no scientific data on the studies reporting the fretting corrosion patterns of CoCrMo microstructures in the presence of specific biological treatments to date. In the current study, Flat-on-flat fretting corrosion set-up was customized and used to study the tribocorrosion patterns of fretting corrosion to understand the role of alloy microstructure. Alloy microstructural differences were created with the implant stock metal's longitudinal and transverse cutting orientations. As a result, the transverse created the non-banded, homogenous microstructure, whereas the longitudinal cut resulted in the banded, non-homogenous microstructure on the surface of the alloy (in this manuscript, the terms homogenous and banded were used). The induced currents were monitored using a three-electrode system. Three different types of electrolytes were utilized to study the fretting corrosion patterns with both homogeneous and banded microstructures: 1. Control media 2. Spent media (the macrophage cell cultured media) 3. Challenged media (media collected after the macrophage was treated with CoCrMo particles). From the electrochemical results, in the potentiostat conditions, the banded group exhibited a higher induced current in both challenged and spent electrolyte environments than in control due to the synergistic activity of CoCrMo particles and macrophage demonstrating more corrosion loss. Additionally, both Bode and Nyquist plots reported a clear difference between the banded and homogeneous microstructure, especially with challenged electrolytes becoming more corrosion-resistant post-fretting than pre-fretting results. The banded microstructure showed a unique shape of the fretting loop, which may be due to tribochemical reactions. Therefore, from the electrochemical, mechanical, and surface analysis data results, the transverse/homogenous/non-banded alloy microstructure groups show a higher resistance to fretting-corrosion damage.

Current advancements in bio-ink technology for cartilage and bone tissue engineering.

In tissue engineering, the fate of a particular organ/tissue regeneration and repair mainly depends on three pillars - 3D architecture, cells used, and stimulus provided. 3D cell supportive structure development is one of the crucial pillars necessary for defining organ/tissue geometry and shape. In recent years, the advancements in 3D bio-printing (additive manufacturing) made it possible to develop very precise 3D architectures with the help of industrial software like Computer-Aided Design (CAD). The main requirement for the 3D printing process is the bio-ink, which can act as a source for cell support, proliferation, drug (growth factors, stimulators) delivery, and organ/tissue shape. The selection of the bio-ink depends upon the type of 3D tissue of interest. Printing tissues like bone and cartilage is always challenging because it is difficult to find printable biomaterial that can act as bio-ink and mimic the strength of the natural bone and cartilage tissues. This review describes different biomaterials used to develop bio-inks with different processing variables and cell-seeding densities for bone and cartilage 3D printing applications. The review also discusses the advantages, limitations, and cell bio-ink compatibility in each biomaterial section. The emphasis is given to bio-inks reported for 3D printing cartilage and bone and their applications in orthopedics and orthodontists. The critical/important performance and the architectural morphology requirements of desired bone and cartilage bio-inks were compiled in summary.

Dynamic microfluidic bioreactor-Hip simulator (DMBH) system for implant toxicity monitoring.

The generation of degradation products (DPs) like ions and organo-metallic particles from corroding metallic implants is an important healthcare concern. These DPs generate local and systemic toxicity. The impact on local toxicity is well documented, however, little is known about systemic toxicity. This is mainly due to the limited scope of the current microtiter plate-based (static) toxicity assay techniques. These methods do not mimic the systemic (dynamic) conditions. In this study, it is hypothesized that DPs incubated with cells in static conditions might provide improper systemic toxicity results, as there is no movement mimicking the blood circulation around cells. This study reports the development of a three-chambered prototype microfluidic system connected to the operational hip implant simulator to test the cellular response induced by the DPs. This setup is called a dynamic microfluidic bioreactor-hip simulator system. We hypothesize that a dynamic microfluidic system will provide a realistic toxicology response induced by DPs than a static cell culture plate. To prove the hypothesis, Neuro2a (N2a) cells were used as representative cells to study systemic neurotoxicity by the implant DPs. The microfluidic bioreactor system was validated by comparing the cell toxicity against the traditional static system and using COMSOL modeling for media flow with DPs. The hip implant simulator used in this study was a state-of-the-art sliding hip simulator developed in our lab. The results suggested that static toxicity was significantly more compared to dynamic microfluidic-based toxicity. The newly developed DMBH system tested for in situ systemic toxicity on N2a cells and demonstrated very minimum toxicity level (5.23%) compared to static systems (31.16%). Thus, the new DMBH system is an efficient tool for in situ implant metal systemic toxicity testing.

Development and Characterization of Conducting-Polymer-Based Hydrogel Dressing for Wound Healing

Objectives: Normal and chronic wound healing is a global challenge. Electrotherapy has emerged as a novel and efficient technique for treating such wounds in recent decades. Hydrogel applied to the wound to uniformly distribute the electric current is an important component in wound healing electrotherapy. This study reports the development and wound healing efficacy testing of vitamin D entrapped polyaniline (PANI)-chitosan composite hydrogel for electrotherapy. Materials and Methods: To determine the morphological and physicochemical properties, techniques like scanning electron microscopy (SEM); differential scanning calorimetry; X-ray diffraction; fourier-transform infrared spectroscopy were used. Moreover, pH, conductance, viscosity, and porosity were measured to optimize and characterize the vitamin D entrapped PANI-chitosan composite hydrogel. The biodegradation was studied using lysozyme, whereas the water uptake ability was studied using phosphate buffer. Ethanolic phosphate buffer was used to perform the vitamin D entrapment and release study. Cell adhesion, proliferation, and electrical stimulation experiments were conducted by seeding dental pulp stem cells (DPSC) into the scaffolds and performing (3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide) assay; SEM images were taken to corroborated the proliferation results. The wound healing efficacy of electrotherapy and the developed hydrogel were studied on excision wound healing model in rats, and the scarfree wound healing was further validated by histopathology analysis. Results: The composition of the developed hydrogel was optimized to include 1% w/v PANI and 2% w/v of chitosan composite. This hydrogel showed 1455 µA conduction, 98.97% entrapment efficiency and 99.12% release of vitamin D in 48 hrs. The optimized hydrogel formulation showed neutral pH of 6.96 and had 2198 CP viscosity at 26°C. The hydrogel showed 652.4% swelling index and 100% degradation in 4 weeks. The in vitro cell culture studies performed on hydrogel scaffolds using DPSC and electric stimulation strongly suggested that electrical stimulation enhances the cell proliferation in a three-dimensional (3D) scaffold environment. The in vivo excision wound healing studies also supported the in vitro results suggesting that electrical stimulation of the wound in the presence of the conducting hydrogel and growth factors like vitamin D heals the wound much faster (within 12 days) compared to non-treated control wounds (requires 21 days for complete healing). Conclusion: The results strongly suggested that the developed PANI-chitosan composite conducting hydrogel acts effectively as an electric current carrier to distribute the current uniformly across the wound surface. It also acted as a drug delivery vehicle for delivering vitamin D to the wound. The hydrogel provided a moist environment, a 3D matrix for free migration of the cells, and antimicrobial activity due to chitosan, all of which contributed to the electrotherapy's faster wound healing mechanism, confirmed through the in vitro and in vivo experiments.

The role of Vitamin E in hip implant-related corrosion and toxicity: Initial outcome.

In orthopedic healthcare, Total Hip Replacement (THR) is a common and effective solution to hip-related bone and joint diseases/fracture; however, corrosion of the hip implant and the release of degradation metal ions/particles can lead to early implant failure and pose potential toxicity risk for the surrounding tissues. The main objective of this work was to investigate the potential role of Vitamin E to minimize corrosion-related concerns from CoCrMo hip implants. The study focused on two questions (i) Can Vitamin E inhibit CoCrMo corrosion? and (ii) Does Vitamin E moderate the toxicity associated with the CoCrMo implant particles? In the study (i) the electrochemical experiments (ASTM G61) with different concentrations of Vitamin E (1, 2, 3 mg/ml against the control) were performed using normal saline and simulated synovial fluid (Bovine calf serum-BCS, 30 g/L protein, pH 7.4) as electrolytes. The polished CoCrMo disc (Ra 50 nm) was the working electrode. The findings suggested that both Vitamin E-Saline (45 ± 0.9%) and Vitamin E-BCS (91 ± 3%) solutions protected against implant corrosion at a Vitamin E concentration of 3 mg/ml, but Vitamin E-BCS showed protection at all Vitamin E (1-3 mg/ml) concentration levels. These results suggested that the Vitamin E and the protein present in the BCS imparted additive effects towards the electrochemical inhibition. In the study (ii) the role of Vitamin E in cytotoxicity inhibition was studied using a mouse neuroblastoma cell line (N2a) for CoCrMo particles and Cr ions separately. The CoCrMo particles were generated from a custom-built hip simulator. The alamarBlue assay results suggested that Vitamin E provides significant protection (85% and 75% proliferation) to N2a cells against CoCrMo particles and Cr ions, respectively at 1 µg/ml concentration, as compared to the control group. However, the results obtained from ROS expression and DNA fiber staining suggest that Vitamin E is only effective against CoCrMo degradation particles and not against Cr ions. In summary, the findings show that Vitamin E can minimize the corrosion processes and play a role in minimizing the potential toxicity associated with implants.

The use of negative oxygen ion clusters [O2-(H2O)n] and bicarbonate ions [HCO3-] as the supportive treatment of COVID-19 infections: A possibility.

The COVID-19 or novel coronavirus SARS-CoV-2 pandemic is challenging worldwide healthcare system and severely affecting global economy. Furious efforts to end the pandemic including prevention of spread of SARS-CoV-2, use of antiviral drugs, symptomatic treatments and vaccination are underway. But there are no effective treatments available to save the dying patient in stage 2 (pulmonary) and stage 3 (hyperinflammation) of the infection. The detailed genetic and phenotypical analysis of SARS-CoV-2 revealed that the spike protein (S1) has increased positive charges (compared to SARS-CoV) on them and are responsible for attachment to human angiotensin-converting enzyme 2 (ACE2) receptor and infection by the virus. In addition, it was also reported that the inflammation in the tissue rendered the lung environment more acidic supporting the fusion of SARS-CoV-2 with the cells. We hypothesize that the intermittent use of the oxygen ionizer generating negative oxygen ion clusters [O2-(H2O)n] and sodium bicarbonate nebulizer (generating HCO3-); when connected to ventilator inlet or oxygen concentrator will neutralize the spike protein of the virus in respiratory tract and lungs and change the lung environment to neutral/alkaline condition respectively facilitating improved oxygen pressure in blood. These physical changes can effectively reduce the viral burden and help the patient recover from the infection faster.

Systemic toxicity eliciting metal ion levels from metallic implants and orthopedic devices - A mini review.

The metal/metal alloy-based implants and prostheses are in use for over a century, and the rejections, revisions, and metal particle-based toxicities were reported concurrently. Complications developed due to metal ions, metal debris, and organo-metallic particles in orthopedic patients have been a growing concern in recent years. It was reported that local and systemic toxicity caused by such released products from the implants is one of the major reasons for implant rejection and revision. Even though the description of environmental metal toxicants and safety limits for their exposure to humans were well established in the literature, an effort was not adequately performed in the case of implant-based metal toxicology. Since the metal ion concentration in serum acts as a possible indicator of the systemic toxicity, this review summarizes the reported human serum safe limits, toxic limits, and concentration range (µg/L, ppb, etc.) for mild to severe symptoms of six (cardiac, hepatic, neuro, nephron, dermal and endocrine) systemic toxicities for twelve most commonly used metallic implants. It also covers the widely used metal ion quantification techniques and systemic toxicity treatments reported.

The role of fretting-frequency on the damage modes of THR modular junction: In-vitro study.

According to the National Center for Health Statistics, currently, more than 250,000 total hip replacements annually in the US alone, with an estimated increase to 500,000 by the year 2030. The usage of tapered junctions between the femoral neck and head gives the surgeon flexibility in implant assembly. However, these modular junctions are subjected to micro-motion that may cause chemical and fretting-corrosion at the modular junction. Therefore, it is imperative to study these forces to mitigate their effects. The current study aims to understand the effects of fretting-corrosion as a function of fretting frequencies caused by common physical activities in an in-vitro model of hip modular junctions. The fretting system has a tribological contact condition of flat-on-flat, mounted to a load frame. CoCrMo pins were polished and immersed in a synovial fluid-like electrolyte solution (Bovine calf serum 30 g/l). Electrochemical measurements were made using a potentiostat. Samples then undergo 3600 cycles at 50 µm (to simulate gross slips), with a horizontal load at 200 N, and a frequency of 0.5 Hz, 0.7 Hz, 1 Hz, and 1.5 Hz to simulate Sit Down-Stand Up, Stair Climb, Walking, and Jogging, respectively. Worn surfaces were then examined under optical and scanning electron microscopy. The evolution of free potential as a function of time for tested frequencies shows the initial potential drop and stabilized trend in the potential evolution. The sample group at a higher frequency displays a higher tendency of corrosion than a lower frequency; however, the dissipation energy decreases as a function of fretting frequency. Both electrochemical and mechanical responses correlate to the variation in the fretting frequencies. Organometallic complexes were found on the surfaces of the samples that were subjected to a slower frequency of fretting, whereas mechanical grooving was noticed on samples with a faster frequency. Hence, these preliminary studies suggest that implant failure rates may be altered based on fretting-frequencies induced by physical activity. Further studies will be required to verify the findings and explore the potential role of fretting frequency in the damage modes of the modular junction.

Development of Polylactic Acid and Bovine Serum Albumin-layered-coated Chitosan Microneedles Using Novel Bees Wax Mould

Objectives: This work illustrates a novel method of fabrication of polymeric microneedle (MN) construct using bees wax as mould and development of coated polymeric MNs for drug delivery. Materials and Methods: A novel method of MN fabrication using bees wax as mould was established. The porous chitosan MN arrays were fabricated and coated with polylactic acid (PLA). The optimized MN arrays were coated with bovine serum albumin (BSA). The MNs were subjected to physiochemical and tensile strength characterization, followed by drug release study. The skin penetration and irritation study were performed in vivo in Wistar Albino rats. Results: The constructed MN arrays contain MNs with 0.9 mm length, 600 µm width at the base, 30-60 µm diameter at the tip, and 1.5 mm distance between 2 needles. These MNs patch was having good mechanical strength (0.72 N/needle) and tensile strength 15.23 Mpa. The MN array patch had 6.26% swelling index and 98.5% drug release was observed on the 50th hr. Good penetration and no skin irritation was observed for optimized MN batch. Conclusion: Polymeric MN arrays were successfully developed using bees wax mould and were successfully coated with PLA to deliver the BSA through skin epidermis layer.

Assessing the potential use of chitosan scaffolds for the sustained localized delivery of vitamin D.

Vitamin D is a commonly used bone modulator in regenerative medicine. Several modalities have been explored for the delivery of vitamin D including nanoparticles and scaffold. The present study aimed to assess the potential use of a bio-degradable chitosan scaffold for the delivery of vitamin D. The objectives included fabrication of a bio-degradable chitosan scaffold, integration of vitamin D into the scaffold, characterization of the vitamin D integrated scaffold. Characterization was carried out using, X-ray diffraction, Fourier transform infrared spectroscopy, and differential scanning calorimetry. The structure of the scaffold was assessed by scanning electron microscopy. The scaffold was placed in phosphate buffer saline and the release duration of vitamin D was observed using UV spectrophotometry. Dental pulp mesenchymal stem cells were added to the scaffold to study the scaffold associated toxicity and the functionality of the scaffold released vitamin D. The vitamin D release period from the scaffold was estimated to be for 80 hrs. MTT assay of the stem cells was comparable to that of the control group (stem cells cultured in media) inferring that the scaffold is not toxic towards the stem cells. The positive alizarin red S staining, a higher expression of alkaline phosphatase, osteocalcin, and RunX2 confirmed the functional capability (osteogenic differentiation of the stem cells) of the released vitamin D. Based on the data from the present study, it can be inferred that chitosan scaffold can be used for the sustained delivery of functional vitamin D for 3-5 days.

Efficacy of scaffold-mediated localized chemotherapy in cancer: A systematic review of current research.

OBJECTIVE: To assess the efficacy of scaffold-mediated localized chemotherapy in cancer. METHODS: Databases including PubMed, Cochrane Library, and SCOPUS were searched for articles reporting the use of scaffold-mediated localized drug delivery in cancer. Essential data including scaffold fabrication material and methods, drug dosage and release duration and its effect on the cancer cells were extracted. RESULTS: 15 articles out of 60 screened, fulfilled the eligibility criteria. Among the 15 studies, 5 studies included only cell lines and 2 studies were on mouse models, while 8 studies involved a combination of cell lines and mouse models. Scaffold materials included both synthetic polymers such as poly-lactide, polycaprolactone and natural materials including d-periosteum and human micro-fragmented adipose tissueA wide number of other variables included the fabrication procedure, drugs used, and the methods used to assess the effects on cancer. As a result, it was not possible to make any direct comparison of the efficacy of the therapeutic strategy used in each of these studies. CONCLUSION: Irrespective of the many variables, a common consensus in all the included studies was that scaffold mediated localized drug delivery effectively reduced cancer cell viability by increasing drug bioavailability to the target tissue, while its localized effect reduced the risk of systemic toxicity.

Low-intensity current (LIC) stimulation of subcutaneous adipose derived stem cells (ADSCs) - A missing link in the course of LIC based wound healing.

Millions of people die as a result of fatal injuries accounting for 9% of the total global annual deaths. Non fatal injuries generally result in variety of wounds. The normal wound healing process is slow and takes weeks to months, depending on the type of wound. In last two decades, electrotherapy called low-intensity currents (LIC) for the treatment became popular for faster wound healing, as well as in management of nonresponding and ulcerative wounds. It was reported that LIC mimics 'the current of injury' which is generated by body on wounding and helps in faster wound healing. Researchers have also studied the migration of localized cell and other bio-molecules under the influence of LIC helping the wound to heal faster. Literature review has also suggested that, electrical stimulation of isolated adipose tissue derived stem cells (ADSCs) releases growth factors and differentiates in to specialized cells like fibroblasts and keratinocytes in laboratory conditions. These research areas are well explored and emerged as independent state-of-the-arts therapies and technologies. Considering the fact, that adipose tissue (along with ADSCs) is present subcutaneously, a new hypothesis is proposed which states that 'low intensity current (LIC) stimulation of wound stimulates subcutaneous adipose tissue containing ADSCs which releases different growth factors and also differentiates into certain cells like fibroblasts, neurons and keratinocytes. These cells easily migrate to wound site due to lipolysis and loosening of fat tissue, resulting in faster wound healing'. Thus this hypothesis provides a missing link between two state of the art technologies; first one is 'LIC based electrotherapy' and second one is 'in-vitro LIC stimulation of ADCSs' where role and significance of in-situ ADCSs were never studied.

Customized Peptide Biomaterial Synthesis via an Environment-Reliant Auto-Programmer Stigmergic Approach.

Stigmergy, a form of self-organization, was employed here to engineer a self-organizing peptide capable of forming a nano- or micro-structure and that can potentially be used in various drug delivery and biomedical applications. These self-assembling peptides exhibit several desirable qualities for drug delivery, tissue engineering, cosmetics, antibiotics, food science, and biomedical surface engineering. In this study, peptide biomaterial synthesis was carried out using an environment-reliant auto-programmer stigmergic approach. A model protein, α-gliadin (31, 36, and 38 kD), was forced to attain a primary structure with free –SH groups and broken down enzymatically into smaller fragments using chymotrypsin. This breakdown was carried out at different environment conditions (37 and 50 °C), and the fragments were allowed to self-organize at these temperatures. The new peptides so formed diverged according to the environmental conditions. Interestingly, two peptides (with molecular weights of 13.8 and 11.8 kD) were isolated when the reaction temperature was maintained at 50 °C, while four peptides with molecular weights of 54, 51, 13.8, and 12.8 kD were obtained when the reaction was conducted at 37 °C. Thus, at a higher temperature (50 °C), the peptides formed, compared to the original protein, had lower molecular weights, whereas, at a lower temperature (37 °C), two peptides had higher molecular weights and two had lower molecular weights.

Synthesis and Evaluation of a Sodium Alginate-4-Aminosalicylic Acid Based Microporous Hydrogel for Potential Viscosupplementation for Joint Injuries and Arthritis-Induced Conditions.

A microporous hydrogel was developed using sodium alginate (alg) and 4-aminosalicylic acid (4-ASA). The synthesized hydrogel was characterized using various analytical techniques such as Fourier transform infrared spectroscopy (FTIR), Carbon-13 nuclear magnetic resonance (13C-NMR), X-ray powder diffraction (XRD), scanning electron microscopy (SEM), and differential scanning calorimetry (DSC). Additonal carboxyl and hydroxyl functional groups of 4-ASA provided significant lubrication and stress-triggered sol-gel transition to the conjugated hydrogel. In addition, cytotoxicity analysis was undertaken on the conjugated hydrogel using human dermal fibroblast-adult (HDFa) cells, displaying non-toxic characteristics. Drug release profiles displaying 49.6% in the first 8 h and 97.5% within 72 h, similar to the native polymer (42.8% in first 8 h and 90.1% within 72 h). Under applied external stimuli, the modified hydrogel displayed significant gelling properties and structure deformation/recovery behaviour, confirmed using rheological evaluation (viscosity and thixotropic area of 8095.3 mPas and 26.23%, respectively). The modified hydrogel, thus, offers great possibility for designing smart synovial fluids as a biomimetic aqueous lubricant for joint-related injuries and arthritis-induced conditions. In addtion, the combination of thixotropy, non-toxicity, and drug release capabilities enables potential viscosupplementation for clinical application.

A composite chitosan-gelatin bi-layered, biomimetic macroporous scaffold for blood vessel tissue engineering.

A composite chitosan-gelatin macroporous hydrogel-based scaffold with bi-layered tubular architecture was engineered by solvent casting-co-particulate leaching. The scaffold constituted an inner macroporous layer concealed by a non-porous outer layer mimicking the 3D matrix of blood vessels with cellular adhesion and proliferation. The scaffold was evaluated for its morphological, physicochemical, physicomechanical and biodurability properties employing SEM, FTIR, DSC, XRD, porositometry, rheology and texture analysis. The fluid uptake and biodegradation in the presence of lysozymes was also investigated. Cellular attachment and proliferation was analysed using human dermal fibroblasts (HDF-a) seeded onto the scaffold and evaluated by MTT assay, SEM, and confocal microscopy. Results demonstrated that the scaffold had a desirable tensile strength=95.81±11kPa, elongation at break 112.5±13%, porosity 82% and pores between 100 and 230µm, 50% in vitro biodegradation at day 16 and proliferated fibroblasts over 20 days. These results demonstrate that scaffold may be an excellent tubular archetype for blood vessel tissue engineering.

Intestinal Targeting of Ganciclovir Release Employing a Novel HEC-PAA Blended Lyomatrix.

A hydroxyethylcellulose-poly(acrylic acid) (HEC-PAA) lyomatrix was developed for ganciclovir (GCV) intestine targeting to overcome its undesirable degradation in the stomach. GCV was encapsulated within the HEC-PAA lyomatrix prepared by lyophilization. Conventional tablets were also prepared with identical GCV concentrations in order to compare the GCV release behavior from the lyomatrix and tablets. GCV incorporation (75.12%) was confirmed using FTIR, DSC, and TGA. The effect of GCV loading on the microstructure properties of the lyomatrix was evaluated by SEM, AFM, and BET surface area measurements. The in vitro drug release study showed steady and rapid release profiles from the GCV-loaded lyomatrix compared with the tablet formulation at identical pH values. Minimum GCV release was observed at acidic pH (≤40%) and maximum release occurred at intestinal pH values (≥90%) proving the intestinal targeting ability of the lyomatrix. Kinetic modeling revealed that the GCV-loaded lyomatrix exhibited zero-order release kinetics (n = 1), while the tablets were best described via the Peppas model. Textural analysis highlighted enhanced matrix resilience and rigidity gradient (12.5%, 20 Pa) for the GCV-loaded lyomatrix compared to the pure (7%, 9.5 Pa) HEC-PAA lyomatrix. Bench-top MRI imaging was used to confirm the mechanism of GCV release behavior by monitoring the swelling and erosion rates. The swelling and erosion rate of the tablets was not sufficient to achieve rapid zero-order GCV release as with the lyomatrix. These combined results suggest that the HEC-PAA lyomatrix may be suitable for GCV intestinal targeting after oral administration.

A bio-injectable algin-aminocaproic acid thixogel with tri-stimuli responsiveness.

In this article a novel bio-injectable algin-aminocaproic acid (Alg-ACA) tri-stimuli responsive thixogel system is reported. The designed soft thixotrophic hydrogel (thixogel) was characterized using various analytical techniques such as FT-IR, NMR, SEM, AFM and DSC. The soft thixogel system was further investigated for stress responsiveness using different rheological studies which confirmed the thixotropic nature of the gel [Thixotropic area (Ar) of Alg-ACA (1:0.5), Alg-ACA (1:1) and Alg-ACA (1:2), were 23.5%, 43.1%, and 27.59%, respectively, which were higher than that of Na-Alg (2.08%)]. The thixogel also demonstrated temperature and ultrasonication responsiveness. This tri-stimuli responsive soft thixogel system was rendered flowable (fluid) on applying the described physical stimuli and recovered its "rigid" gel structure upon removal of the applied stimuli. This approach of synthesizing a thixogels may be applicable to a broad variety of other natural polymers and has the potential for use in biomedical applications.

Microwave-assisted facile synthesis of a new tri-block chitosan conjugate with improved mucoadhesion.

A new chitosan-based tri-block conjugate, O-PEG-chitosan-N-cysteine was synthesized using microwave irradiation. For synthesis of this derivative, chitosan was modified to a PEG-chitosan conjugate followed by PEG-chitosan-cysteine using 6-O PEGylation and 2-N-thiolation, respectively. The synthesized derivative was characterized using various analytical techniques such as FT-IR and (1)H NMR spectroscopy. The conjugate was also analyzed for its biochemical, biodegradation and mucoadhesive properties. The modified chitosan conjugate exhibited improved mucoadhesion behavior (14.0 h) with greater biodegradation compared to the parent polymer (6.3h). The in silico modeling corroborated with the in vitro study demonstrating a stable complex between mucin and O-PEG-chitosan-N-cysteine conjugate (ΔE=-60.100 kcal/mol) compared to mucin and chitosan conjugate. The synthesis proposed herein, involves the use of microwave irradiation which causes a substantial reduction in the reaction time (approximately 2.30 h) compared to conventional method (35 h).

Spectrophotometric bioassay method for urokinase.

INTRODUCTION: Urokinase is a potent plasminogen activator. Present Bio assay methods of Urokinase are very tedious. So a new simple spectrophotometric Bio assay method was developed to estimate thrombolytic activity of Urokinase. METHODS AND RESULTS: This Bio assay is designed for the quantitative in vitro spectrophotometric determination of Urokinase activity by utilizing its thrombolytic activity to carry out the lysis of plasma clots. The initial concentration of the plasma clots was adjusted to 0.2+/-0.01 absorbance and the linear decrease in the absorbance by addition of Urokinase concentration from 200 to 1200 IU/ml at lambda(max) 530 nm was studied. The activity of sample Urokinase can be quantified by comparing the absorbance of sample Urokinase with Urokinase standard Bio assay calibration curve and predicted in IU. The r(2) value of standard calibration curve was found out to be 0.993. The repeatability of the Bio assay was studied by performing the experiment six times with fresh plasma samples and the results showed significant similarity. DISCUSSION: We can conclude that the novel Bio assay method was found to be simple, economical, reproducible and accurate than the present Bio assay methods.

Purification and partial characterization of thermostable serine alkaline protease from a newly isolated Bacillus species HSRB08 from hotspring.

The purpose of the research was to study the purification and partial characterization of thermostable serine alkaline protease from a newly isolated Bacillus species HSRB08, which was isolated from hotspring. The enzyme was purified in a 2-step procedure involving ammonium sulfate precipitation and Sephadex G-200 chromatography. The enzyme was shown to have molecular weight of 66 kD by sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) and Gelatin Zymogram and was purified 15.3-fold with a yield of 7.5%. It was most active at 45 degrees C, pH 9.0, with casein as substrate. It was strongly activated by metal ions such as Ca2+, Mg2+, and Mn2+. Enzyme activity was inhibited strongly by phenylmethyl sulphonyl fluoride (PMSF) but was not inhibited by ethylene diamine tetra acetic acid (EDTA), while a slight inhibition was observed with beta-mercaptoethanol (beta-ME). The compatibility of the enzyme was studied with commercial and local detergents in the presence of 10 mM CaCl2. The addition of 10 mM CaCl2 individually and in combination, was found to be very effective in improving the enzyme stability. This enzyme improved the cleansing power of various detergents. It removed blood stains completely when used with detergents in the presence of 10 mM CaCl2.