Development and Evaluation of Biomimetic 3D Coated Composite Scaffold for Application as Skin Substitutes

Skin injuries are an urgent health issue, which raises a great concern in the clinic. Although numerous strategies have been proposed to fabricate skin substitutes for treatment of wounds over the past several decades, fabricating an ideal skin substitute to replace the damaged one can still be a problem. In this study, a novel biomimetic 3D composite skin scaffold is fabricated by combining electrohydrodynamic (EHD) jetting, electrospinning, and coating techniques. Here, the first polycaprolactone (PCL) porous structure is produced by the EHD jetting. Next, the second polylactic acid (PLA) membrane consisted of nanoscale fibers is prepared on the PCL porous structure via the electrospinning. The PCL porous structure and PLA fibers membrane can mimic the dermis and epidermis layer, respectively. Furthermore, gelatin is used as coating solution to enhance the biocompatibility of the scaffold. The structure and morphology of the fabricated scaffolds are analyzed, and the mechanical properties are investigated as well. Moreover, the in vitro and in vivo experiments demonstrate the biocompatibility of the materials and the fabrication process. In conclusion, these results demonstrate that the composite scaffold is effective and holds great potential for skin regeneration in the clinic.     

Self‐Detecting and Self‐Healing Reinforce Elastomer Doped with Aggregation‐Induced Emission Molecules

Most of elastomers for fabrication of comfortable epidermal devices and smart actuators produce responsive signals by the stimuli‐induced deformation. Herein, a dynamic visualization of external stimuli rather than the deformation through synthesis of a self‐healing poly(dimethylsiloxane) (PDMS)‐based elastomer doped with aggregation‐induced emission (AIE) molecules is reported. The self‐healing PDMS‐based elastomer is designed and synthesized through molecule integration of reversible multi‐strength H‐bonds and permanent covalent crosslink sites. The adjustment of the weight ratio of elastic cross‐linker offers tunable mechanical properties of the resultant elastomer. After doping such an elastomer with AIE molecules of 1,1,2,2‐tetrakis(4‐nitrophenyl)ethane, the elastomer composite displays strong on–off fluorescence depending upon mechanical damage and temperatures, which can be used to detect the breaking and self‐healing performances, as well as the temperature change. The strategy described here provides another way to develop smart polymeric elastomers for practical applications.     

水稳碎石材料细观修复行为及最佳修复时机

为了探究水稳碎石(CSM)材料的细观修复行为及最佳修复时机,基于损伤-愈合模型对其修复效果影响因素进行了分析。首先,基于离散元法(DEM)建立CSM的二维半圆弯曲(SCB)数值试件,引入损伤因子模拟材料的局部损伤与愈合效应,进而构建细观尺度的损伤-愈合数值模型。其次,通过虚拟SCB疲劳试验实现不同程度的荷载损伤,基于愈合变量来表征损伤修复过程,模拟局部损伤愈合行为。然后,基于损伤-愈合模型开展不同损伤状态下的修复效果评估,分析初始损伤状态、愈合程度对修复效果的影响规律,并进一步研究了CSM材料的最佳修复时机。结果表明：所提出的损伤-愈合模型能够再现CSM材料在细观尺度上的损伤与愈合行为,初始损伤状态和愈合程度对修复效果的影响具有非线性,在70%～85%的疲劳寿命阶段,局部损伤修复对结构强度的提升作用较为显著。基于对损伤修复效果的数值分析,推荐将70%～85%的疲劳寿命作为CSM材料的最佳修复时机,此外,选择强度优良、数量充足的愈合剂可以最大程度地提升局部损伤修复效果。     

Electrospun PCL Scaffolds as Drug Carrier for Corneal Wound Dressing Using Layer-by-Layer Coating of Hyaluronic Acid and Heparin

Due to its ability to reduce scarring and inflammation, human amniotic membrane is a widely used graft for wound dressings after corneal surgery. To overcome donor dependency and biological variances in the donor tissue, artificial nanofibrous grafts acting as drug carrier systems are promising substitutes. Electrospun nanofibrous scaffolds seem to be an appropriate approach as they offer the properties of permeable scaffolds with a high specific surface, the latter one depending on the fiber diameter. Electrospun scaffolds with fiber diameter of 35 nm, 113 nm, 167 nm and 549 nm were manufactured and coated by the layer-by-layer (LbL) technology with either hyaluronic acid or heparin for enhanced regeneration of corneal tissue after surgery. Studies on drug loading capacity and release kinetics defined a lower limit for nanofibrous scaffolds for effective drug loading. Additionally, scaffold characteristics and resulting mechanical properties from the application-oriented characterization of suture pullout from suture retention tests were examined. Finally, scaffolds consisting of nanofibers with a mean fiber diameter of 113 nm were identified as the best-performing scaffolds, concerning drug loading efficiency and resistance against suture pullout.     

Quantitative Proteomics Reveals Molecular Network Driving Stromal Cell Differentiation: Implications for Corneal Wound Healing

The differentiation of keratocytes to fibroblasts and myofibroblasts is an essential requisite during corneal wound closure. The aim of this study is to uncover factors involved in differentiation-dependent alteration in the protein profile of human corneal stromal cells using quantitative proteomics. Human corneal fibroblasts were cultured and differentiated into keratocytes in serum-free media and myofibroblasts through treatment with TGF-β. The protein cell lysates from the donors were tryptic and were digested and labeled using a 3-plex iTRAQ kit. The labeled peptides were subjected to LCMS analysis. Biological functional analysis revealed a set of crucial proteins involved in the differentiation of human corneal stromal cells which were found to be significantly enriched. The selected proteins were further validated by immunohistochemistry. Quantitative proteomics identified key differentially expressed proteins which are involved in cellular signaling pathways. Proteins involved in integrin signaling (Ras-RAP1b, TLN and FN) and SLIT-ROBO pathways (PFN1, CAPR1, PSMA5) as well as extracellular matrix proteins (SERPINH1, SPARC, ITGβ1, CRTAP) showed enhanced expression in corneal fibroblasts and myofibroblasts compared to keratocytes, indicating their possible role in wound healing. Corneal stromal cell differentiation is associated with the activation of diverse molecular pathways critical for the repair of fibroblasts and myofibroblasts. Identified proteins such as profilin 1 and talin could play a tentative role in corneal healing and serve as a potential target to treat corneal fibrosis.     

Extracellular Vesicles Secreted by Corneal Myofibroblasts Promote Corneal Epithelial Cell Migration

Corneal epithelial wound healing is a multifaceted process that encompasses cell proliferation, migration, and communication from the corneal stroma. Upon corneal injury, bidirectional crosstalk between the epithelium and stroma via extracellular vesicles (EVs) has been reported. However, the mechanisms by which the EVs from human corneal keratocytes (HCKs), fibroblasts (HCFs), and/or myofibroblasts (HCMs) exert their effects on the corneal epithelium remain unclear. In this study, HCK-, HCF-, and HCM-EVs were isolated and characterized, and human corneal epithelial (HCE) cell migration was assessed in a scratch assay following PKH26-labeled HCK-, HCF-, or HCM-EV treatment. HCE cells proliferative and apoptotic activity following EV treatment was assessed. HCF-/HCM-EVs were enriched for CD63, CD81, ITGAV, and THBS1 compared to HCK-EV. All EVs were negative for GM130 and showed minimal differences in biophysical properties. At the proteomic level, we showed HCM-EV with a log >two-fold change in CXCL6, CXCL12, MMP1, and MMP2 expression compared to HCK-/HCF-EVs; these proteins are associated with cellular movement pathways. Upon HCM-EV treatment, HCE cell migration, velocity, and proliferation were significantly increased compared to HCK-/HCF-EVs. This study concludes that the HCM-EV protein cargo influences HCE cell migration and proliferation, and understanding these elements may provide a novel therapeutic avenue for corneal wound healing.     

采后苯丙噻重氮处理促进梨果实的愈伤

研究采后苯丙噻重氮（benzothiadiazole,BTH）处理对梨果实愈伤的促进作用,探讨苯丙烷代谢及抗氧化 酶在愈伤中的作用。以‘玉露香’梨果实为试材,人工模拟损伤后,用100 g/L BTH处理,在常温（20～25 ℃）、 相对湿度80%～85%的黑暗条件下进行愈伤,通过测定质量损失率和损伤接种梨果实的发病率来评价愈伤效果,比 较BTH处理组和对照组梨果实愈伤期间苯丙氨酸解氨酶、超氧化物歧化酶、过氧化物酶和多酚氧化酶的活力,以及 总酚、类黄酮和木质素含量的差异。结果表明：BTH处理对梨果实愈伤期间的质量损失率没有显著影响;愈伤期 间,BTH处理组和对照组梨果实的发病率均降低,处理组和对照组之间存在显著性差异（P＜0.05）,由此表明, BTH处理降低了梨果实的采后发病率;BTH处理明显提高了苯丙烷代谢途径关键酶苯丙氨酸解氨酶、抗氧化酶多酚 氧化酶、过氧化物酶和超氧化物歧化酶的活力;此外,BTH处理还有效促进了梨果实总酚、类黄酮和木质素的积 累。相关性分析结果表明,处理组梨果实的发病率和苯丙氨酸解氨酶活力、总酚含量和类黄酮含量之间呈显著负相 关,和过氧化物酶及多酚氧化酶活力之间呈极显著负相关。采后BTH处理可促进梨果实的愈伤,苯丙烷代谢以及过 氧化物酶和多酚氧化酶在愈伤组织形成中发挥了重要作用。     

Preparation of acetylated chitosan sponges (chitin sponges)

Acetylated chitosan sponges (chitin sponges) were prepared according to acetylation time (25, 50, 75, and 100 h). As the acetylation time increased, the degree of acetylation increased, and a 75‐h acetylation time produced the highest degree of acetylation (DA). The surface morphologies of samples were examined by scanning electron microscopy. Sponge samples were shown by a water uptake ability test to have higher water absorption abilities. An in vitro biodegradation test showed that sponges with a higher DA were more susceptible to lysozyme hydrolysis. Acetylated chitosan sponges were further shown by an in vitro fibroblast proliferation test to have a higher degree of cell viability on increasing the DA, with 75 h exhibiting the maximum effect. The results showed that the wound healing effect of chitosan sponges can be controlled by the DA. © 2010 Wiley Periodicals, Inc. J Appl Polym Sci, 2010     

超高面板坝淤填自愈型止水结构可行性的初步研究

通过有限元计算和复变函数推导，系统研究了面板接缝的渗流规律，给出了接缝渗流量和渗透比降的解析计算公式，可用于面板堆石坝接缝渗流控制的设计。同时结合233m高的水布垭面板坝，对淤填自愈型止水的渗流控制效果及在超高面板坝中应用的可行性进行了计算分析。