Control the fate of human umbilical cord mesenchymal stem cells with dual‐enzymatically cross‐linked gelatin hydrogels for potential applications in nerve regeneration

Stem‐cell‐based therapy is a promising strategy to treat challenging neurological diseases, while its application is hindered primarily by the low viability and uncontrolled differentiation of stem cell. Hydrogel can be properly engineered to share similar characteristics with the target tissue, thus promoting cell viability and directing cell differentiation. In this study, we proposed a new dual‐enzymatically cross‐linked and injectable gelatin hydrogel for regulating survival, proliferation, and differentiation of human umbilical cord mesenchymal stem cells (hUC‐MSCs) in a three‐dimensional matrix. This injectable gelatin hydrogel was formed by oxidative coupling of gelatin–hydroxyphenyl acid conjugates catalyzed by hydrogen horseradish peroxidase (HRP) and choline oxidase (ChOx). Modulus and H₂O₂ release can be well controlled by ChOx activity. Results from calcein‐AM/PI staining and Ki67 immunofluorescence tests demonstrated that the survival and proliferation behavior of hUC‐MSCs were highly enhanced in HRP_1UChOx_0.25U hydrogel with lower modulus and less H₂O₂ release compared with other groups. Attractively, the expression of neuron‐specific markers β‐III tubulin, neurofilament light chain (NFL), and synapsin‐1 was significantly increased in HRP_1UChOx_0.25U hydrogel as well. Additionally, in vitro hemolysis test and in vivo HE staining data highlighted the good biocompatibility. Undoubtedly, this injectable gelatin hydrogel's ability to control hUC‐MSCs' fate holds enormous potentials in nervous disorders' therapy and nerve regeneration.     

丹银海绵对糖尿病大鼠皮肤溃疡创面愈合的影响

目的:探讨丹银海绵对糖尿病大鼠皮肤溃疡创面愈合的影响。方法:将雄性Wistar大鼠随机分为8组。除空白对照组外,其余7组大鼠,通过手术在其背部形成一10mm×10mm相同大小,深达肌层的溃疡,分别给予抗生素、生长因子凝胶、纳米银、丹黄散,以及不同浓度的丹银海绵处理溃疡面。给药28d后处死大鼠,取溃疡部位新生肉芽组织,HE染色观察组织形态的变化;WesternBlot检测大鼠创面组织中AGEs、hs-CRP、IL-1、IL-6、bFGF、PDGF、VEGF表达水平。结果:病理切片观察显示适宜浓度的丹银海绵可以促进血管新生并伴扩张,同时生成大量新生成纤维细胞。WesternBlot检测显示高浓度的丹银海绵可以显著上调创面组织中生长因子表达(P<0.05),降低AGEs水平及炎症因子表达(P<0.05)。结论:适宜浓度的丹银海绵对于糖尿病大鼠皮肤溃疡的愈合有明显的促进作用,提高治疗效果。     

Using online content uniformity measurements for rapid automated process development exemplified via an X-ray system

This paper addresses the relevance of automated content testing for the rapid automated process development (RAPD). Our previous work demonstrated that RAPD allowed a fast and efficient development of a continuous capsule-filling process. Target was the mean weight and the relative standard deviation of the weight. Likewise important are the content and the content uniformity. However, an implementation demands a certain level of automation. In general, technology is available that can detect active pharmaceutical ingredient (API) inside the capsules but the final application is linked to additional development and investment in machinery. To eliminate doubts regarding the benefits of an automated content check within the RAPD we present an application example. First, an X-ray system was used to detect barium sulfate accurately inside capsules. Second, a process was developed where barium sulfate was filled. The concentration of excipients was modified in the experiments, as well as the setting of the process parameter. The obtained model provided an explicit understanding of the process. Subsequently, the content uniformity model was compared to a model of the capsule weight relative standard deviation, confirming the benefits of an automated content check in the RAPD. Moreover, we presented another example illustrating the advantages of a connected continuous filling process, which permits evaluation of all process steps and their interactions (i.e. evaluation of the entire process).     

Biodegradable Polydioxanone Microspheres for Transcatheter Arterial Embolization: Proof of Principle

PurposeTo evaluate feasibility, embolization success, biodegradability, reperfusion, and biocompatibility of biodegradable microspheres (MS) made from polydioxanone (PDO) for transcatheter arterial embolization.Materials and MethodsUnilateral selective renal embolization of a segmental artery was performed in 16 New Zealand White rabbits with PDO-MS (100–150 μm and 90–315 μm). Animals were randomly assigned to different observation periods and underwent control digital subtraction angiography (DSA) and MR imaging immediately (n = 3), 1 week (n = 2), 4 weeks (n = 2), 8 weeks (n = 2), 12 weeks (n = 5), and 16 weeks (n = 2) after embolization. Kidneys were harvested for macroscopic and histologic analysis of embolization success, biodegradability, and biocompatibility.ResultsEmbolization was technically successful in 15 of 16 animals. One animal died of anesthesia-related circulatory failure. The 100–150 μm MS were injected easily through 3-F catheters; the 90–315 μm MS tended to clog with intermittent catheter obstruction. DSA and MR imaging showed successful target embolization in 13 of 15 animals. In 2 animals, the entire kidney was affected owing to catheter clogging, including a reflux of MS while flushing. Control DSA and MR imaging showed increasing vascular reperfusion with time. Macroscopic and histologic analysis revealed necrosis/infarction in areas in which embolization was achieved. MS were extensively degraded after 16 weeks, and overall inflammatory reaction was mild.ConclusionsBiodegradable PDO-MS induced effective embolization of target vessels while demonstrating good biocompatibility. MS increasingly dissolved at 16 weeks, partial reperfusion started at week 1, and complete reperfusion started at week 8, thus offering possible advantages as a temporary embolic agent.     

超高效液相色谱-三重四级杆质谱法检测阿归养血糖浆中阿胶及牛皮源成分

目的：建立阿归养血糖浆中阿胶及掺杂牛皮源成分的专属性检测方法。方法：用胰蛋白酶对阿归养血糖浆处方中胶类成分进行酶解，采用超高效液相色谱-三重四极杆质谱（UPLC-QQQ/MS），选择阿胶特征分子离子峰m/z 540.0（双电荷）→612.2、m/z 540.0（双电荷）→924.4及牛皮源特征离子对m/z 641.3（双电荷）→726.2、m/z 641.3（双电荷）→783.3作为检测离子对，离子化模式为ESI⁺，进行多反应监测。结果：26批阿归养血糖浆中未检出阿胶的有13批，其中3批检出牛皮源成分，1批检出牛皮源成分但未超出限度，9批均未检出阿胶及牛皮源成分；26批样品检出阿胶的有13批，其中同时检出牛皮源成分的有1批，未检出牛皮源成分的为12批。结论：该检测方法专属性强，可用于阿归养血糖浆中阿胶及牛皮源的检测。     

Timing of Uterine Artery Embolization for Leiomyoma during the Menstrual Cycle

PurposeTo investigate differences in outcomes of uterine artery embolization (UAE) for leiomyoma when performed during different phases of the menstrual cycle.Materials and MethodsIn this single-institution retrospective analysis, 111 premenopausal patients (median [range] age, 44 [33–52] years) undergoing UAE for symptomatic leiomyoma between June 2014 and February 2020 were included. Twenty-one patients underwent UAE in the menstrual phase (the early follicular phase), 27 in the late follicular phase, and 63 in the luteal phase. Baseline characteristics and technical and peri-procedural outcomes were compared among groups. Leiomyoma infarction on contrast-enhanced magnetic resonance imaging 1 week after UAE and 4-month outcomes, including changes in the Uterine Fibroid Symptom and Quality of Life questionnaire scores, the volume reduction rates of the uterus and largest leiomyoma, follicle stimulating hormone values, adverse events, and amenorrhea, were compared among groups.ResultsA 4-month follow-up was completed for all patients. No significant differences were observed among groups in baseline characteristics or technical and peri-procedural outcomes. There were no significant differences in the multivariate-adjusted 1-week infarction rates of all leiomyoma volumes (P = .161) or multivariate-adjusted 4-month outcomes, including changes in the Uterine Fibroid Symptom and Quality of Life questionnaire symptoms and total scores (P = .864 and P = .798, respectively), the volume reduction rates of the uterus and the largest leiomyoma (P = .865 and P = .965, respectively), and follicle stimulating hormone values (P = .186) among the groups. No significant differences were noted in the 4-month adverse events (P = .260) or amenorrhea (P = .793) among the groups.ConclusionsThe present study demonstrated no significant differences in the outcomes of UAE for leiomyoma when performed during different phases of the menstrual cycle.     

A 2,5-Dihydroxybenzoic Acid–Gelatin Conjugate: The Synthesis,Antiviral Activity and Mechanism of Antiviral Action Against Two Alphaherpesviruses

Various natural and synthetic polyanionic polymers with different chemical structures are known to exhibit potent antiviral activity in vitro toward a variety of enveloped viruses and may be considered as promising therapeutic agents. A water-soluble conjugate of 2,5-dihydroxybezoic acid (2,5-DHBA) with gelatin was synthesized by laccase-catalyzed oxidation of 2,5-DHBA in the presence of gelatin, and its antiviral activity against pseudorabies virus (PRV) and bovine herpesvirus type 1 (BoHV-1), two members of the Alphaherpesvirinae subfamily, was studied. The conjugate produced no direct cytotoxic effect on cells, and did not inhibit cell growth at concentrations up to 1000 µg/mL. It exhibited potent antiviral activity against PRV (IC₅₀, 1.5–15 µg/mL for different virus strains) and BoHV-1 (IC₅₀, 0.5–0.7 µg/mL). When present during virus adsorption, the conjugate strongly inhibited the attachment of PRV and BoHV-1 to cells. The 2,5-DHBA–gelatin conjugate had no direct virucidal effect on the viruses and did not influence their penetration into cells, cell-to-cell spread, production of infectious virus particles in cells, and expression of PRV glycoproteins E and B. The results indicated that the 2,5-DHBA–gelatin conjugate strongly inhibits the adsorption of alphaherpesviruses to cells and can be a promising synthetic polymer for the development of antiviral formulations against alphaherpesvirus infections.     

Evaluation of Arterial Impairment after Experimental Gelatin Sponge Embolization in a Rabbit Renal Model

ObjectiveArterial stenosis is a major obstacle for subsequent interventional procedures. We hypothesized that the stenosis is caused by gelatin sponge embolization and performed an experimental study in a rabbit renal model.ResultsGelatin sponge particles were mainly observed in the segmental and interlobar arteries. Transmural inflammation of the embolized arterial wall and mild thickening of the media were observed 1 week after embolization. Resorption of the gelatin sponge and organization of thrombus accompanied by foreign body reactions, were observed from 2 to 4 weeks after embolization. Microscopic images of the 3 weeks group showed vessel lumens filled mostly with organized thrombi, resulting in severe stenosis. Additionally, vessels showed a thickened intima that contained migrating smooth muscle cells and accompanying interruption of the internal elastic lamina. The migrating smooth muscle cells were distributed around the recanalized arterial lumen.ConclusionGelatin sponge embolization may induce arterial stenosis by causing organized thrombus and intimal hyperplasia, which consists of migrating smooth muscle cells and intimal collagen deposits.     

可吸收止血流体明胶预防术后脑脊液漏

目的:探索可吸收止血流体明胶预防术后脑脊液漏的有效性。方法:对2013年3月至9月有硬脊膜撕裂脑脊液漏的脊柱损伤17例患者进行回顾性分析,男16例,女1例;年龄16～67岁,平均(39.6±15.4)岁;颈椎1例,胸椎9例,胸腰段4例,腰椎3例;爆裂骨折4例,骨折脱位13例。神经损伤ASIA分级:A级12例,B级2例,D级2例,E级1例。车祸伤2例,高处坠落伤10例,重物砸伤4例,地震时从楼梯滚落1例。术中应用可吸收止血流体明胶封堵以预防术后脑脊液漏。记录术后每日的引流量。结果:17例患者有15例术后无脑脊液漏出现。2例术后出现脑脊液漏者引流管保留至术后第6～7天。术后未出现头痛、头晕、发热、颈项抵抗、皮疹、切口不愈合、切口感染、血肿、神经症状加重等并发症。随访9个月均未见到切口周围有异常现象。结论:术中应用可吸收止血流体明胶封堵硬脊膜撕裂预防术后脑脊液漏是有效的方法。