Artificial cavernosa-like tissue based on multibubble Matrigel and a human corpus cavernous fibroblast scaffold

Ex vivo tissue culture of the human corpus cavernosum(cc)can be used to explore the tissue structural changes and complex signaling networks.At present,artificial Cc-like tissues based on acellular or three-dimensional(3D)-printed scaffolds are used to solve the scarcity of primary penis tissue samples.However,inconvenience and high costs limit the wide application of such methods.Here,we describe a simple,fast,and economical method of constructing artificial CC-like tissue.Human Cc fibroblasts(FBs),endothelial cells(ECs),and smooth muscle cells(SMCs)were expanded in vitro and mixed with Matrigel in specific proportions.A large number of bubbles were formed in the mixture by vortexing combined with pipette blowing,creating a porous,spongy,and spatial structure.The Cc FBs produced a variety of signaling factors,showed multidirectional differentiation potential,and grew in a 3D grid in Matrigel,which is necessary for CC-like tissue to maintain a porous structure as a cell scaffold.Within the C-like tissue,ECs covered the surface of the lumen,and SMCs were located inside the trabeculae,similar to the structure of the primary Cc.Various cell components remained stable for 3 days in vitro,but the EC content decreased on the 7th day.Wingless/integrated(WNT)signaling activation led to lumen atrophy and increased tissue fibrosis in CC-like tissue,inducing the same changes in characteristics as in the primary Cc.This study describes a preparation method for human artificial cC-like tissue that may provide an improved experimental platform for exploring the function and structure of the cC and conducting drug screening for erectile dysfunction therapy.

Application of digital image correlation to full-field measurement of shrinkage strain of dental composites

Objectives: Polymerization shrinkage of dental composites remains a major concern in restorative dentistry because it can lead to micro-cracking of the tooth and debonding at the tooth-restoration interface. The aim of this study was to measure the full-field polymerization shrinkage of dental composites using the optical digital image correlation (DIC) method and to evaluate how the measurement is influenced by the factors in experiment setup and image analysis. Methods: Four commercial dental composites, Premise Dentine, Z100, Z250 and Tetric EvoCeram, were tested. Composite was first placed into a slot mould to form a bar specimen with rectangular-section of 4 mm mm, followed by the surface painting to create irregular speckles. Curing was then applied at one end of the specimen while the other part were covered against curing light for simulating the clinical curing condition of composite in dental cavity. The painted surface was recorded by a charge-coupled device (CCD) camera before and after curing. Subsequently, the volumetric shrinkage of the specimen was calculated with specialist DIC software based on image cross correlation. In addition, a few factors that may influence the measuring accuracy, including the subset window size, speckle size, illumination light and specimen length, were also evaluated. Results: The volumetric shrinkage of the specimen generally decreases with increasing distance from the irradiated surface with a conspicuous exception being the composite Premise Dentine as its maximum shrinkage occurred at a subsurface distance of about 1 mm instead of the irradiated surface. Z100 had the greatest maximum shrinkage strain, followed by Z250, Tetric EvoCeram and then Premise Dentine. Larger subset window size made the shrinkage strain contour smoother. But the cost was that some details in the heterogeneity of the material were lost. Very small subset window size resulted in a lot of noise in the data, making it difficult to discern the general pattern in the strain distribution. Speckle size did not seem to have obvious effect on the volumetric shrinkage strain along specimen length; however, larger speckles resulted in higher level of noise or heterogeneity in the shrinkage distribution. Compared with bright illumination, dimmer lighting produced larger standard deviations in the measured shrinkage, indicating a higher level of noise. The longer the specimen, the greater was the rate of reduction with distance from the irradiated surface, especially for the longitudinal strain. Significance: The image correlation method is capable of producing full-field polymerization shrinkage of dental composites. The accuracy of the measurements relies on selection of optimal parameters in experimental setup and DIC analysis.

Proteomic profiling of the hemolymph at the fifth instar of the silkworm Bombyx mori

Two-dimensional gel electrophoresis （2-DE） followed by matrix-assisted laser desorption ionization- time-of-flight/time-of-flight mass spectrometry （MS） analysis were used to charaterize the hemolymph proteomic profiles of the silkworm, Bombyx mori. At days 4 （V4） and 5 （VS） of the fifth （final） instar, when the larvae were at the fast-growing stage, we found dramatic changes in spots representing proteins having an approximate molecular weight （MW） of 30 kDa. Of these spots, four 30K proteins were highly up- regulated, implying a close association with the growth and development ofB. mori larvae. To understand the molecular basis and underlying mechanisms involved in development and metamorphosis, the proteome of whole hemolymph at V5 was analyzed using shotgun liquid chromatography tandem mass spectrometry with an LTQ-Orbitrap. A total of 108 proteins were identified without any false discovery hits. These proteins were involved in a variety of cellular functions, including metabolism, development, nutrient transport and reserve, and defense response. Gene ontology analysis showed that 3.4% of these proteins had nutrient reservoir activities and 5.7% were involved in the response to stimulus. Pathway analysis revealed that 22 proteins with common targets were involved in various cellular processes such as immunity, differentiation, proliferation and metamorphosis. These results suggested that some key factors such as the 30K proteins in hemolymph play important roles in B. mori growth and development. Moreover, the multiple functions of hemolymph may be operated by a complex biological network.

Mesenchymal stem cells-extracellular vesicles alleviate pulmonary fibrosis by regulating immunomodulators

BACKGROUND Pulmonary fibrosis(PF)is a chronic interstitial lung disease characterized by fibroblast proliferation and extracellular matrix formation,causing structural damage and lung failure.Stem cell therapy and mesenchymal stem cells-extracellular vesicles(MSC-EVs)offer new hope for PF treatment.AIM To investigate the therapeutic potential of MSC-EVs in alleviating fibrosis,oxidative stress,and immune inflammation in A549 cells and bleomycin(BLM)-induced mouse model.METHODS The effect of MSC-EVs on A549 cells was assessed by fibrosis markers[collagen I andα-smooth muscle actin(α-SMA),oxidative stress regulators[nuclear factor E2-related factor 2(Nrf2)and heme oxygenase-1(HO-1),and inflammatory regu-lators[nuclear factor-kappaB(NF-κB)p65,interleukin(IL)-1β,and IL-2].Similarly,they were assessed in the lungs of mice where PF was induced by BLM after MSC-EV transfection.MSC-EVs ion PF mice were detected by pathological staining and western blot.Single-cell RNA sequencing was performed to investigate the effects of the MSC-EVs on gene expression profiles of macrophages after modeling in mice.RESULTS Transforming growth factor(TGF)-β1 enhanced fibrosis in A549 cells,significantly increasing collagen I andα-SMA levels.Notably,treatment with MSC-EVs demonstrated a remarkable alleviation of these effects.Similarly,the expression of oxidative stress regulators,such as Nrf2 and HO-1,along with inflammatory regulators,including NF-κB p65 and IL-1β,were mitigated by MSC-EV treatment.Furthermore,in a parallel manner,MSC-EVs exhibited a downregulatory impact on collagen deposition,oxidative stress injuries,and inflammatory-related cytokines in the lungs of mice with PF.Additionally,the mRNA sequencing results suggested that BLM may induce PF in mice by upregulating pulmonary collagen fiber deposition and triggering an immune inflammatory response.The findings collectively highlight the potential therapeutic efficacy of MSC-EVs in ameliorating fibrotic processes,oxidative stress,and inflammatory responses associated with PF.CONCLUSION MSC-EVs could ameliorate fibrosis in vitro and in vivo by downregulating collagen deposition,oxidative stress,and immune-inflammatory responses.

Distinctive impacts of atmospheric intraseasonal oscillations on the net ecosystem exchange of the southeastern China forest between spring and summer

The southeastern China(SEC)forest is an important terrestrial biospheric carbon sink in the global carbon cycle,with its total net ecosystem exchange(NEE)accounting for about 3.2%of the global forest NEE.The prevailing atmospheric intraseasonal oscillations(ISOs)over East Asia strongly modulate climatic conditions over the SEC during spring and summer,thus leading to significant ISOs in the NEE of the SEC forest.As atmospheric ISOs show strong seasonality,this study examined the distinctive impacts of atmospheric ISOs on the NEE of the SEC forest between spring and summer.During spring,the vertical coupling of 10–30-d atmospheric ISOs in the lower and upper troposphere leads to strong 10‒30-d ISOs of solar radiation and temperature over the SEC.The 10‒30-d ISOs of solar radiation and temperature further result in the 10‒30-d ISOs of gross primary productivity(GPP)and terrestrial ecosystem respiration(TER).With the covariation in GPP and TER,the NEE of the SEC forest exhibits significant 10‒30-d ISOs.In contrast,the intraseasonal variations in climatic conditions over the SEC are associated with the 15‒60-d tropical atmospheric ISO during summer.While the induced 15‒60-d ISO of solar radiation leads to that of GPP,the induced 15‒60-d ISO of temperature is small and less effective;thus,the 15‒60-d ISO of NEE mainly originates from that of GPP.