Comparative Analysis of Collagen and Chitosan-based Dressing for Haemostatic and Wound Healing Application.

Collagen and chitosan have haemostatic, tissue fix and wound healing properties but the poor mechanical property limits their application. Therefore, various concentrations of collagen (1-6%) and chitosan (1-2%) were used to develop biopolymer-coated gauzes, with and without glycerol as plasticiser. Glycerol-treated gauzes showed desired mechanical and adhesive property in comparison to polymer-coated gauzes alone. Developed gauzes were characterized using differential scanning calorimetry, thermal gravimetric analysis and Fourier transform infrared spectrophotometry to confirm the biopolymer coating and stability. Scanning electron microscopy showed multilayer coating of the biopolymer and faster clotting in chitosan gauzes in comparison to collagen. Surface plasmon resonance assay confirmed that chitosan exhibited more binding affinity of 65 RU in comparison to collagen, which showed 55 RU with erythrocytes. Decrease in the value of plateletcrit and mean platelet volume confirmed platelet adhesion and aggregation over the surface of polymer-coated dressings. Gamma scintigraphy studies showed 85 ± 2% formulation retention up to 12 h at the wound site in comparison to 40 ± 3% retention of the radiopharmaceutical alone. Collagen and chitosan-coated gauze showed 226 ± 15 s and 179 ± 12 s haemostasis time, respectively, which was significantly less from 506 ± 15 s in standard gauze. Chitosan gauze showed faster wound healing in comparison to the collagen-coated gauze. Chitosan and collagen-coated gauzes showed 55 ± 4% wound contraction on day seven in comparison to 25 ± 2% in the control group, while chitosan gauzes showed complete wound contraction on day fourteenth, while the collagen-coated gauze showed 90 ± 3% on the same day.

Alpha-ketoglutarate mediated hepatoprotection against alcohol induced toxicity: in vivo functional observation studies in Sprague Dawley rats using gamma scintigraphy.

Alcohol is the most abused psychoactive substance and known hepatotoxicant. Present study elucidates possible therapeutic effect of oral alpha-ketoglutarate (AKG) supplementation against alcohol induced hepatic dysfunction, using biochemical, histopathological and most importantly, in vivo functional imaging approaches. Animals were divided into three groups of 6 animals each. Group-I (control): Normal saline; Group-II: 20% (v/v) solution of ethanol (5 ml/day) intragastrically using oral gavage for 2 months. Group-III: ethanol treatment as in group-II along with AKG supplementation (2g/kg/bw; intragastrically using oral gavage for 2 months). In vivo hepatobiliary scintigraphy was performed in all animals using 99mTc-mebrofenin (99mTc-MEB) as radiotracer to determine changes in (a) Hepatic extraction fraction (HEF), for quantification of radiotracer uptake, (b) Time to reach maximum hepatic uptake (Tpeak), and (c) Time for hepatic uptake to reduce by 50% (T1/2peak). Biochemical (alanine aminotransferase, aspartate aminotransferase, reduced glutathione, superoxide dismutase, catalase, and lipid peroxidation) and histological parameters were also studied. Hepatic uptake and excretion kinetics using 99mTc-MEB scintigraphy showed prompt 99mTc-MEB clearance from liver in control group (HEF: 91.26 ± 2.32; Tpeak: 143 ± 23 sec; T1/2peak: 434 ± 41 sec), while it was significantly abnormal in ethanol group and showed less efficient radiotracer accumulation (HEF: 62.72 ± 5.6; Tpeak: 201 ± 33 sec; T1/2peak: 542 ± 52 sec). Supplementation of AKG along with ethanol significantly improved liver function (HEF: 76.42 ± 5.3; Tpeak: 155 ± 34 sec; T1/2peak: 455 ± 22 sec). Biochemical and histopathology parameters were correlative to findings of functional imaging study. Results strongly indicate hepatoprotective potential of AKG against alcohol-induced hepatic injury. Study further proposes the use of in vivo hepatobiliary scintigraphy for high throughput screening of other hepatoprotectants.

Decontamination of rat and human skin experimentally contaminated with 99mTc, 201Tl and 131I radionuclides using "Dermadecon" - a skin decontamination kit: an efficacy study.

Radioactive skin contamination is one of the most likely risks which occurs after accidental or occupational radiological accidents apart from internal contamination. In such cases where the radioactive contamination has occurred, the person who is contaminated should be decontaminated as early as possible to reduce the damaging health effects of radiation. In the present study, the decontamination efficiency of a developed skin decontamination kit "dermadecon" has been evaluated in animal models and human subjects using gamma scintigraphy. Decontamination efficiency (percentage of the radioactive contaminant removed) was calculated for each radioactive isotope of the study and compared with control where general washing procedure was followed using liquid and soap. The effectiveness of the kit was calculated in animal model with respect to 99mTc-sodium-pertechnetate (99mTcO4-), 201TlCl and 131I and was found 92.84 ± 4.9%, 91.18 ± 3.23% and 94.67 ± 2.92%, respectively. Whereas, in case of human skin, the decontamination efficiency for 99mTcO4- was observed to be 95.00 ± 3.21%. On the basis of findings from the study, it can be concluded that the decontamination agents of the used skin decontamination kit are effective for removal of localized radioactive contaminants from skin, as compared with normal decontamination using soap and water.

Pulmonary delivery of nanosized alendronate for decorporation of inhaled heavy metals: formulation development, characterization and gamma scintigraphic evaluation.

CONTEXT: Medical management of heavy metal toxicity including radioactive ones is the cause of concern because of their increased use in energy production, healthcare and mining. As inhalation is one of the primary routes for internalization, a formulation is needed to trap metal(s) at the portal of entry itself. OBJECTIVE: Objective was to formulate and characterize a nanonized dry powder inhaler (DPI) formulation of alendronate sodium as potential inhalable antidote for chelating metal toxicants. METHODS: In vitro binding studies of alendronate with respect to seven non-radioactive heavy metals were carried out using UV-spectroscopy and HPLC. Nanonizing of alendronate particles was achieved by antisolvent precipitation using Pluronic-F68 as stabilizer. Characterization was done with the help of SEM, TEM FT-IR, XRD, DSC, NMR spectroscopy and PSD studies. In vitro and in vivo pulmonary deposition studies were carried out using gamma scintigraphy, followed by a limited pharmacokinetic study in humans. RESULTS: In vitro binding studies confirmed the chelating action of alendronate. Anderson cascade impaction showed that nano-alendronate exhibited significantly higher respirable fraction (58.25 ± 1.32%) compared to the micronized form (28.7 ± 0.59%). Scintigraphy results showed significant increase in the alveolar deposition of drug post-nanonizing. CONCLUSION: Results strongly indicate the role of nano-alendronate DPI as potential inhalable antidote for neutralizing heavy metal toxicity, including radio-metal contamination.

Fabrication, characterization and evaluation of the efficacy of gelatin/hyaluronic acid microporous scaffolds suffused with aloe-vera in a rat burn model.

Burn induced injuries are commonly encountered in civilian and military settings, leading to severe morbidity and mortality. Objective of this study was to construct microporous bioactive scaffolds of gelatin-hyaluronic acid suffused with aloe-vera gel (Gela/HA/AvG), and to evaluate their efficacy in healing partial-thickness burn wounds. Scaffolds were characterized using Fourier transform-infrared spectroscopy, Scanning electron microscopy, and Thermo-gravimetric analysis to understand intermolecular interactions and morphological characteristics. In-vitro fluid uptake ability and hemolytic index of test scaffolds were also determined. In-vitro collagenase digestion was done to assess biodegradability of scaffolds. Wound retraction studies were carried out in Sprague Dawley rats inflicted with partial-thickness burn wounds to assess and compare efficacy of optimized scaffolds with respect to negative and positive control groups. In-vivo gamma scintigraphy using Technetium-99m labeled Immunoglobulin-G (99mTc-IgG) as imaging agent was also performed to validate efficacy results. Histological and immunohistochemical comparison between groups was also made. Scaffolds exhibited mircoporous structure, with pore size getting reduced from 41.3 ± 4.3 µm to 30.49 ± 5.7 µm when gelatin conc. was varied from 1% to 5%. Optimized test scaffolds showed sustained in-vitro swelling behavior, were biodegradable and showed hemolytic index in range of 2.4-4.3%. Wound retraction study along with in-vivo gamma scintigraphy indicated that Gela/HA/AvG scaffolds were not only able to reduce local inflammation faster but also accelerated dermis regeneration. Immunohistochemical analysis, in terms of expression levels of epidermal growth factor and fibroblast growth factor-2 also corroborated in-vivo efficacy findings. Gela/HA/AvG scaffolds, therefore, can potentially be developed into an effective dermal regeneration template for partial-thickness burn wounds.

In-vitro and in-vivo functional observation studies to establish therapeutic potential of alpha-ketoglutarate against methotrexate induced liver injury.

BACKGROUND: Methotrexate (MTX) is widely used in chemotherapy but its associated hepatotoxicity is a major complication, limiting its use. This study evaluates possible therapeutic effect of oral alpha-ketoglutarate (AKG) supplementation against MTX-induced hepatotoxicity. METHODS: HepG2 cells were used to evaluate in-vitro cyto-protection conferred by AKG against MTX induced cytotoxicity. For in-vivo animal study, rats were divided into three groups. Group-I served as control. Group-II animals were administered single intraperitoneal injection of MTX (20 mg/kg/body weight), while Group-III received MTX as in group-II followed by oral AKG (2 gm/kg body weight) for 5 days. 99mTc-Mebrofenin hepatobiliary study was performed under a gamma camera to determine real time functional status of rats' livers. Multiple parameters concerning hepatic mebrofenin uptake and excretion, including Tpeak and T1/2 peak in control and treated animals were determined. Biochemical analysis of the liver homogenate in terms of hepatic enzyme activities in serum, antioxidant status, tissue factor activity, tissue collagen content and histological analysis of the liver tissue were also done. RESULTS: AKG supplementation significantly reversed MTX induced derangement in activities of serum liver enzymes [ALT and ALP (p = 0.003); AST (p = 0.005)], antioxidant status [LPO and GSH (p = 0.005); CAT (p = 0.004)], tissue factor activity (p = 0.005) and tissue collagen content (p = 0.005). Functional imaging confirmed that hepatic retention and fractional biliary excretion were significantly abnormal in MTX treated group (Tpeak: 234 s ± 40 s; T1/2 peak: 846sec ± 32sec) as compared to AKG supplemented group (Tpeak: 144 s ± 35sec; T1/2peak: 468sec ± 27sec). Hepatic extraction fraction (HEF) was 92.2 ± 1.8%, 48.7 ± 2.6% and 69.8 ± 4.3% in control, MTX and AKG supplemented rats respectively. CONCLUSION: 99mTc-mebrofenin imaging strongly suggests therapeutic action of AKG in protecting liver damage by MTX in rats. Functional imaging parameters correlated well with biochemical and histopathological findings.

Facile functionalization of Teriflunomide-loaded nanoliposomes with Chondroitin sulphate for the treatment of Rheumatoid arthritis.

This research aims to coat Teriflunomide (TEF) loaded conventional nanoliposomes (CON-TEF-LIPO) with Chondroitin sulphate (CS) to produce CS-TEF-LIPO for the effective treatment of Rheumatoid arthritis (RA). Both CON-TEF-LIPO and CS-TEF-LIPO were produced, characterized and evaluated for their active targeting potential towards CD44 receptors. Cell cytotoxicity, cell viability and intracellular uptake study on differentiated U937 and MG-63 cells demonstrated the active targeting of CS-TEF-LIPO towards CD44 receptors. Furthermore, in vivo pharmacodynamic, biochemical, radiological and histopathological studies performed in adjuvant induced arthritic (AIA) rat model showed a significant (P < 0.05) reduction in inflammation in arthritic rat paw in CS-TEF-LIPO group compared to TEF and CON-TEF-LIPO groups. Moreover, liver toxicity study revealed that CS-TEF-LIPO showed no signs of toxicity and biodistribution study revealed the accumulation of CS-TEF-LIPO in synovial region of arthritic rat. Taken together, results suggest that CS-TEF-LIPO could provide a new insight for an effective treatment of RA.

Role of 99mTc-Tetrofosmin delayed scintigraphy and color Doppler sonography in characterization of solitary thyroid nodules.

OBJECTIVES: A study was undertaken to evaluate the diagnostic efficiency of Tc-Tetrofosmin scan and color Doppler in the characterization of benign and malignant solitary thyroid nodules. METHODS: Fifty-two patients found to have a cold solitary thyroid nodule on Tc-pertechnetate scintigraphy were included in this study. All patients underwent a single-injection dual-phase (30 min and 120 min) Tc-Tetrofosmin scan. The intranodular vascularity was measured using color Doppler sonography. Fine-needle aspiration cytology was performed on all the patients. In the following days and weeks all patients underwent surgery. RESULTS: Thirteen out of 15 patients with thyroid cancer showed delayed retention of radiotracer (on 120 min images as compared to the initial 30 min image). Thirty-six out of 37 patients harboring benign solitary nodules showed significant washout of tracer on delayed images. Sensitivity, specificity, positive predictive value and negative predictive value of delayed Tc-Tetrofosmin scintigraphy were found to be 86.6, 97.2, 92.8 and 94.7%, respectively. The Doppler study was able to demonstrate increased vascularity in the center of 8 of the 15 malignant nodules. Thirty-two patients harboring a benign solitary nodule showed normal or increased peripheral vascularity on Doppler study. Sensitivity, specificity, positive predictive value and negative predictive value of color Doppler were found to be 53.5, 86.4, 61.5 and 82%, respectively. CONCLUSION: Delayed Tc-Tetrofosmin scintigraphy is a highly sensitive and specific method for characterizing solitary thyroid nodules, while color Doppler has a low sensitivity but relatively high specificity in differentiating benign from malignant thyroid lesions.

Tc-99m ciprofloxacin scans for detection of tubercular bone infection.

Tc-99m MDP and Tc-99m ciprofloxacin scans were performed in 14 patients with suspected tubercular bone disease and in 2 cases of nontubercular bone infection. In 5 patients the findings were true negative. There were no false positives and 1 was false negative. The sensitivity, specificity, and positive predictive value were found to be 93, 71, and 87.5%, respectively, for detection of bone tubercular lesions. However, the test does not distinguish TB osteomyelitis from other types of osteomyelitis. Delayed 24 hour scans were found useful to differentiate between inflammatory and infective lesions. The aim of this study was to evaluate the usefulness of the Tc-99m ciprofloxacin scan as a means to detect tubercular bone disease.

Development and clinical study of submicronic-atropine sulphate respiratory fluid as a novel organophosphorous poisoning antidote.

CONTEXT: Increased use of organophosphate insecticides (OPI) and possibility of terror groups using stocks of nerve agents underscore the need to develop effective and safe antidotes. While intramuscular administration of antidotes like atropine sulphate (AS) has certain lacunae, intravenous route may not be always feasible in emergency field conditions. OBJECTIVE: Objective was (a) to develop a novel inhalable submicronic-AS respiratory fluid as potential antidote for OPI poisoning, (b) in-vitro and in-vivo evaluation in terms of respiratory fraction, and (c) clinical study to assess drug bioavailability in blood and atropinization pattern post-inhalation. METHODS: Formulation was optimized on the basis of particle size of aerosolized droplets and in-vitro nebulization rate. Anderson cascade impaction (ACI) studies were carried out to validate the advantage of test formulation in terms of respirable fraction. Six healthy volunteers were inhaled the test formulation and blood bioavailability and atropinization were noted serially. Gamma scintigraphy was used to quantify total and regional lung deposition of nebulized AS in-vivo. RESULTS: The formulation was optimized using 30% ethanol-saline with particle size in the range of 350-500 nm. In-vitro ACI data showed high respirable fraction (82.6 ± 3.1%) for the test formulation. In-vivo scintigraphy suggested whole lung deposition of 80.2 ± 6.8% of the total inhaled dose. Early blood bioavailability and atropinization pattern confirmed that therapeutic concentration of the drug in blood was reached within 5 min. CONCLUSIONS: 3% submicronic-AS respiratory fluid might be used as potential prophylactic/therapeutic option against OPI poisoning with several advantages over intramuscular injection, including early blood bioavailability and atropinization.

Preliminary evaluation of technetium-99m-labeled ceftriaxone: infection imaging agent for the clinical diagnosis of orthopedic infection.

OBJECTIVE: In this study we sought to assess the efficacy of a technetium-99m (Tc-99m)-labeled third-generation cephalosporin as an infection imaging agent in the accurate detection of the sites of bacterial infection in vivo. DESIGN: Ceftriaxone (CRO) was formulated into a ready-to-use single-vial cold kit with a shelf-life of over 6 months and was successfully labeled with technetium. The radiolabeled drug, Tc-99m-CRO, was subjected to the following preclinical evaluations: radiochemical purity, in vitro and in vivo stability, bacterial binding assay, and pharmacokinetic studies in animals and in human patients. RESULTS: The kit formulation exhibited excellent radiolabeling efficiency (∼99%) and high in vitro and in vivo stability. The radiolabeled drug exhibited slow blood clearance (12% at 4 h), and the high protein binding and excretion pattern of the labeled formulation mimics the reported pharmacokinetic profile of the drug alone. In the animal model, scintigraphy scans showed higher uptake of the radiopharmaceutical in infectious lesions, even at 1 h post-administration, in comparison to inflammatory lesions. The clinical evaluation of Tc-99m-labeled CRO showed a diagnostic accuracy of 83.3%, and a sensitivity and specificity of 85.2% and 77.8%, respectively. CONCLUSIONS: This kit formulation has the potential for imaging bacterial infections with much higher sensitivity and specificity as compared to other Tc-99m-labeled antibiotics available as convenient ready-to-use kits in routine clinical practice.

A new method for radiolabeling of human immunoglobulin-G and its biological evaluation.

BACKGROUND: Radiolabeled human Immunoglobulin-G (hIgG) has demonstrated its utility in inflammation and infection imaging. However, the present method of radiolabeling hIgG is time-consuming and complex. OBJECTIVE: To develop a simplified method of radiolabeling hIgG with technetium-99m ((99m)Tc) via a nicotinyl hydrazine derivative ((99m)Tc-HYNIC-hIgG) and its biological evaluation. RESULTS: In vitro and in vivo studies showed that (99m)Tc-hIgG prepared by this method was fairly stable in physiological saline and human serum till 24 h. Only 4.3% degradation of the radiolabeled drug was seen till 24 h. Blood clearance pattern of the radiopharmaceutical exhibited biphasic exponential pattern. Biodistribution of (99m)Tc-HYNIC-hIgG in mice was observed up to 24 h. Significant accumulation of the radiotracer was found in liver (4.93 %), kidney (3.67%) and intestine (2.12 %) at 4 h interval by 24 h interval, it was reduced to 1.99%, 2.18% and 1.93 % respectively. Significant amount of radioactivity in liver, kidney and intestine suggest hepatobilliary as well as renal route of clearance for (99m)Tc-HYNIC-hIgG. The anterior whole body and spot scintigraphy images showed increased uptake of 99mTc-HYNIC-hIgG, with the area seen as a focal hot spot, indicating good localization of the radiolabeled hIgG at the site of infection. CONCLUSION: The present findings indicate that (99m)Tc-HYNIC-hIgG holds great potential for the scintigraphy localization of inflammation. The shelf life of the developed kit, when stored at (-) 20°C was found to be at least 3 months.

Synthesis, characterization, and in vivo skeletal localization of a new (99m)Tc-based multidentate phosphonate chelate: 5-Amino-1,3-bis(ethylamine-(N,N dimethyl diphosphonic acid) acetamido) benzene.

The tetraphosphonate ligand, 5-amino-1,3-bis(ethylamine-(N,N-dimethyl diphosphonic acid) acetamido) benzene (IPTMP) used in the present study was prepared from 5-nitroisophthalate dimethylester to label with radionuclide for targeted diagnosis and therapy. The synthesized multidentate phosphonate ligand was characterized on the basis of spectroscopic techniques, which exhibited good metal ion control properties when complexed to (99m)Tc with high in vitro and in vivo stability. Excellent quality bone images of rabbit were imaged showing rapid clearance of background activity and visualization of skeleton at 1h.