scholarly journals Application of an ultrasound semi-quantitative assessment in the degradation of silk fibroin scaffolds in vivo

2021 ◽  
Vol 20 (1) ◽  
Author(s):  
Lihui Cai ◽  
Nan Gao ◽  
TingYu Sun ◽  
Ke Bi ◽  
Xin Chen ◽  
...  

Abstract Background Research on the degradation of silk fibroin (SF) scaffolds in vivo lacks uniform and effective standards and experimental evaluation methods. This study aims to evaluate the application of ultrasound in assessing the degradation of SF scaffolds. Methods Two groups of three-dimensional regenerated SF scaffolds (3D RSFs) were implanted subcutaneously into the backs of Sprague-Dawley rats. B-mode ultrasound and hematoxylin and eosin (HE) staining were performed on days 3, 7, 14, 28, 56, 84, 112, 140, and 196. The cross-sectional areas for two groups of 3D RSFs that were obtained using these methods were semi-quantitatively analyzed and compared to evaluate the biodegradation of the implanted RSFs. Results The 3D RSFs in the SF-A group were wholly degraded at the 28th week after implantation. In contrast, the 3D RSFs in the SF-B group were completely degraded at the 16th week. Ultrasonic examination showed that the echoes of 3D RSFs in both groups gradually decreased with the increase of the implantation time. In the early stages of degradation, the echoes of the samples were higher than the echo of the muscle. In the middle of degeneration, the echoes were equal to the echo of the muscle. In the later stage, the echoes of the samples were lower than that of the muscle. The above changes in the SF-B group were earlier than those in the SF-A group. Semi-quantitative analysis of the cross-sectional areas detected using B-mode ultrasound revealed that the degradations of the two 3D RSF groups were significantly different. The degradation rate of the SF-B group was found to be higher than that of the SF-A group. This was consistent with the semi-quantitative detection results for HE staining. Regression analysis showed that the results of the B-mode ultrasound and HE staining were correlated in both groups, indicating that B-mode ultrasound is a reliable method to evaluate the SF scaffold degradation in vivo. Conclusions This study suggests that B-mode ultrasound can clearly display the implanted SF scaffolds non-invasively and monitor the degradation of the different SF scaffolds after implantation in living organisms in real-time.

2021 ◽  
Author(s):  
Lihui Cai ◽  
Nan Gao ◽  
Tingyu Sun ◽  
Ke Bi ◽  
Xin Chen ◽  
...  

Abstract Background: Research on the degradation of silk fibroin (SF) scaffolds in vivo lacks uniform and effective standards and experimental evaluation methods. This study aims to evaluate the application of ultrasound in assessing the degradation of SF scaffolds.Methods: Two groups of three-dimensional regenerated SF scaffolds (3D RSFs) were implanted subcutaneously into the backs of Sprague-Dawley rats. B-mode ultrasound and hematoxylin and eosin (HE) staining were performed on days 3, 7, 14, 28, 56, 84, 112, 140, and 196. The cross-sectional areas for two groups of 3D RSFs that were obtained using these methods were semi-quantitatively analyzed and compared to evaluate the biodegradation of the implanted RSFs.Results: Semi-quantitative analysis of the cross-sectional areas detected using B-mode ultrasound revealed that the degradations of the two 3D RSF groups were significantly different. The degradation rate of the SF-B group was found to be higher than that of the SF-A group. This was consistent with the semi-quantitative detection results for HE staining. Regression analysis showed that the results of the B-mode ultrasound and HE staining were correlated in both groups, indicating that B-mode ultrasound is a reliable method to evaluate the SF scaffold degradation in vivo. As the SF scaffold degraded, its echo gradually decreased. In the early stages of degradation, the echo of the SF scaffold was higher than that of the muscle. In the middle stage of degradation, the echo was equal to that of the muscle. In the later stage, the echo was lower than that of the muscle.Conclusions: This study suggests that B-mode ultrasound can clearly display the implanted SF scaffolds non-invasively and monitor the degradation of the different SF scaffolds after implantation in living organisms in real-time.


Author(s):  
Sezin Yigit ◽  
Nadia Hallaj ◽  
James Sugarman ◽  
Lester Chong ◽  
Samantha Roman ◽  
...  

Recent studies have demonstrated silk fibroin’s ability to extend the shelf life of foods by mitigating the hallmarks of spoilage, namely oxidation and dehydration. Due to the potential for this protein to become more widespread, its safety was evaluated comprehensively. First, a bacterial reverse mutation test (Ames test) was conducted in five bacterial strains. Second, an in vivo erythrocyte test was conducted with Sprague Dawley rats at doses up to 1,000mg/kg-bw/day. Third, a range-finder study was conducted with Sprague Dawley rats at the highest consumption amount given solubility and oral gavage volume constrains (500mg/kg-bw/day). Fourth, a 28-day study in Sprague Dawley rats was conducted at the 500mg/kg-bw/day amount. Fifth, an in vitro pepsin digestion assay was performed to assess the potential for protein allergenicity. Sixth, allergenic potential was further assessed using liquid chromatography-mass spectroscopy for detection of allergenic insect proteins. Seventh, the protein sequences were subjected to bioinformatic analyses. Together, these studies raise no mutagenic, carcinogenic, toxicological, or allergenic concerns with the oral consumption of silk fibroin.


Author(s):  
Shu-Chieh Hu ◽  
Matthew S Bryant ◽  
Estatira Sepehr ◽  
Hyun-Ki Kang ◽  
Raul Trbojevich ◽  
...  

Abstract The tobacco-specific nitrosamine NNK [4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone] is found in tobacco products and tobacco smoke. NNK is a potent genotoxin and human lung carcinogen; however, there are limited inhalation data for the toxicokinetics (TK) and genotoxicity of NNK in vivo. In the present study, a single dose of 5x10−5, 5x10−3, 0.1, or 50 mg/kg body weight (BW) of NNK, 75% propylene glycol (vehicle control), or air (sham control) was administered to male Sprague-Dawley (SD) rats (9-10 weeks age) via nose-only inhalation (INH) exposure for 1 hour. For comparison, the same doses of NNK were administered to male SD rats via intraperitoneal (IP) injection and oral gavage (PO). Plasma, urine, and tissue specimens were collected at designated timepoints and analyzed for levels of NNK and its major metabolite 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanol (NNAL) and tissue levels of DNA adduct O6-methylguanine by LC/MS/MS. TK data analysis was performed using a non-linear regression program. For the genotoxicity subgroup, tissues were collected at 3 hours post-dosing for comet assay analysis. Overall, the TK data indicated that NNK was rapidly absorbed and metabolized extensively to NNAL after NNK administration via the three routes. The IP route had the greatest systemic exposure to NNK. NNK metabolism to NNAL appeared to be more efficient via INH than IP or PO. NNK induced significant increases in DNA damage in multiple tissues via the three routes. The results of this study provide new information and understanding of the toxicokinetics and genotoxicity of NNK.


Plants ◽  
2021 ◽  
Vol 10 (4) ◽  
pp. 774
Author(s):  
Max Langer ◽  
Thomas Speck ◽  
Olga Speck

Although both the petiole and lamina of foliage leaves have been thoroughly studied, the transition zone between them has often been overlooked. We aimed to identify objectively measurable morphological and anatomical criteria for a generally valid definition of the petiole–lamina transition zone by comparing foliage leaves with various body plans (monocotyledons vs. dicotyledons) and spatial arrangements of petiole and lamina (two-dimensional vs. three-dimensional configurations). Cross-sectional geometry and tissue arrangement of petioles and transition zones were investigated via serial thin-sections and µCT. The changes in the cross-sectional geometries from the petiole to the transition zone and the course of the vascular bundles in the transition zone apparently depend on the spatial arrangement, while the arrangement of the vascular bundles in the petioles depends on the body plan. We found an exponential acropetal increase in the cross-sectional area and axial and polar second moments of area to be the defining characteristic of all transition zones studied, regardless of body plan or spatial arrangement. In conclusion, a variety of terms is used in the literature for describing the region between petiole and lamina. We prefer the term “petiole–lamina transition zone” to underline its three-dimensional nature and the integration of multiple gradients of geometry, shape, and size.


2021 ◽  
Vol 7 (1) ◽  
Author(s):  
Himanshu Kushwah ◽  
Nidhi Sandal ◽  
Meenakshi Chauhan ◽  
Gaurav Mittal

Abstract Background Uncontrolled bleeding is one of the primary reasons for preventable death in both civilian trauma and military battle field. This study evaluates in vitro and in vivo hemostatic potential of four biopolymeric natural gums, namely, gum tragacanth, guar gum, xanthan gum, and gum acacia. In vitro evaluation of whole blood clotting time and erythrocyte agglutination assay were carried out. In vitro cytotoxicity studies with respect to each gum were done in human lymphocytes to ascertain percent cell viability. In vivo hemostatic potential of each gum (as sponge dressing and powder form) was evaluated in Sprague Dawley rats using tail bleeding assay and compared with commercially available hemostatic sponge. Other important parameters like (a) time taken for complete hemostasis, (b) amount of blood absorbed, (c) adherence strength of developed hemostatic dressing(s), (d) incidence of re-bleeding, and (e) survival of animals were also studied. Results Of the four test gums studied, xanthan gum (@3mg/ml of blood) and gum tragacanth (@35mg/ml of blood) were able to clot blood in least time (58.75±6.408 s and 59.00±2.082 s, respectively) and exhibited very good hemostatic potential in vitro. Except for xanthan gum, all other test gums did not exhibit any significant cytotoxicity at different time points till 24 h. In rat tail bleeding experiments, gum tragacanth sponge dressing and powder achieved hemostasis in least time (156.2±12.86 s and 76±12.55 s, respectively) and much earlier than commercially available product (333.3±38.84 s; p˂0.01). Conclusion Results indicate potential of gum tragacanth to be developed into a suitable hemostatic product.


1991 ◽  
Vol 7 (3) ◽  
pp. 125-139 ◽  
Author(s):  
David R. Bevan ◽  
David M. Ruggio

To evaluate health risks associated with exposure to particulates in the environment, it is necessary to quantify the bioavailability of carcinogens associated with the particulates. Direct analysis of bioavailability in vivo is most readily accomplished by adsorbing a radiolabeled form of the carcinogen to the particulate. A sam ple of native diesel particulate collected from an Oldsmobile die sel engine that contained 1.03 μ g benzo[ a] pyrene ( BaP)/ g particulate was supplemented with exogenous [ 3 H]- BaP to pro duce a particulate containing 2.62 μ g BaP/g. To insure that elu tion of BaP from native and [3 H] -BaP-supplemented particulate was similar, in vitro analyses were performed. When using phos pholipid vesicles composed of dimyristoylphosphatidylcholine (DMPC), 1.52% of total BaP was eluted from native particulate into the vesicles in 18 hrs; from [ 3 H] -BaP supplemented particu late, 1.68% was eluted. Using toluene as eluent, 2.55% was eluted from native particulate, and 8.25% from supplemented particulate, in 6 hrs. Supplemented particulate was then instilled intratracheally into male Sprague-Dawley rats and distribution of radioactivity was analyzed at selected times over 3 days. About 50% of radioactivity remained in lungs at 3 days following instil lation, with 30% being excreted into feces and the remainder dis tributed throughout the organs of the rats. To estimate the amount of radioactivity that entered feces through swallowing of a portion of the instilled dose, [3 H] -BaP-supplemented particu late was instilled intratracheally into rats that had a cannula sur gically implanted in the bile duct. Rate of elimination of radio activity into bile was monitored; 10.6% of radioactivity was re covered in 6 hr, an amount slightly lower than the 12.8% ex creted in 6 hrs into feces of animals with intact bile ducts. Our studies provide a quantitative description of the distribution of BaP and its metabolites following intratracheal instillation of diesel particulate. Because rates of elution of BaP in vitro are similar for native diesel particulate and particulate with supple mental [ 3H] -BaP, our results provide a reasonable estimate of the bioavailability in vivo of BaP associated with diesel particu late.


2015 ◽  
Vol 770 ◽  
pp. 156-188 ◽  
Author(s):  
Patricio Winckler ◽  
Philip L.-F. Liu

A cross-sectionally averaged one-dimensional long-wave model is developed. Three-dimensional equations of motion for inviscid and incompressible fluid are first integrated over a channel cross-section. To express the resulting one-dimensional equations in terms of the cross-sectional-averaged longitudinal velocity and spanwise-averaged free-surface elevation, the characteristic depth and width of the channel cross-section are assumed to be smaller than the typical wavelength, resulting in Boussinesq-type equations. Viscous effects are also considered. The new model is, therefore, adequate for describing weakly nonlinear and weakly dispersive wave propagation along a non-uniform channel with arbitrary cross-section. More specifically, the new model has the following new properties: (i) the arbitrary channel cross-section can be asymmetric with respect to the direction of wave propagation, (ii) the channel cross-section can change appreciably within a wavelength, (iii) the effects of viscosity inside the bottom boundary layer can be considered, and (iv) the three-dimensional flow features can be recovered from the perturbation solutions. Analytical and numerical examples for uniform channels, channels where the cross-sectional geometry changes slowly and channels where the depth and width variation is appreciable within the wavelength scale are discussed to illustrate the validity and capability of the present model. With the consideration of viscous boundary layer effects, the present theory agrees reasonably well with experimental results presented by Chang et al. (J. Fluid Mech., vol. 95, 1979, pp. 401–414) for converging/diverging channels and those of Liu et al. (Coast. Engng, vol. 53, 2006, pp. 181–190) for a uniform channel with a sloping beach. The numerical results for a solitary wave propagating in a channel where the width variation is appreciable within a wavelength are discussed.


2016 ◽  
Vol 3 (4) ◽  
pp. 167-175 ◽  
Author(s):  
Jiang Pu ◽  
Yuanyuan Deng ◽  
Xiaoyan Tan ◽  
Gaofeng Chen ◽  
Cong Zhu ◽  
...  

2012 ◽  
Vol 303 (8) ◽  
pp. R850-R860 ◽  
Author(s):  
Miriam Goebel-Stengel ◽  
Andreas Stengel ◽  
Lixin Wang ◽  
Gordon Ohning ◽  
Yvette Taché ◽  
...  

Various molecular forms of CCK reduce food intake in rats. Although CCK-8 is the most studied form, we reported that CCK-58 is the only detectable endocrine peptide form in rats. We investigated the dark-phase rat chow intake pattern following injection of CCK-8 and CCK-58. Ad libitum-fed male Sprague-Dawley rats were intraperitoneally injected with CCK-8, CCK-58 (0.6, 1.8, and 5.2 nmol/kg), or vehicle. Food intake pattern was assessed during the dark phase using an automated weighing system that allowed continuous undisturbed monitoring of physiological eating behavior. Both CCK-8 and CCK-58 dose dependently reduced 1-h, dark-phase food intake, with an equimolar dose of 1.8 nmol being similarly effective (−49% and −44%). CCK-58 increased the latency to the first meal, whereas CCK-8 did not. The intermeal interval was reduced after CCK-8 (1.8 nmol/kg, −41%) but not after CCK-58. At this dose, CCK-8 increased the satiety ratio by 80% and CCK-58 by 160%, respectively, compared with vehicle. When behavior was assessed manually, CCK-8 reduced locomotor activity (−31%), whereas grooming behavior was increased (+59%). CCK-58 affected neither grooming nor locomotor activity. In conclusion, reduction of food intake by CCK-8 and CCK-58 is achieved by differential modulation of food intake microstructure and behavior. These data highlight the importance of studying the molecular forms of peptides that exist in vivo in tissue and circulation of the animal being studied.


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