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Preparation of myocardial patches from DiI-labeled rat bone marrow mesenchymal stem cells and neonatal rat cardiomyocytes contact co-cultured on polycaprolactone film DiI标记大鼠骨髓间充质干细胞和乳鼠心肌细胞在聚己内酯膜上接触共培养制备心肌贴片
IF 4 3区 医学 Q2 Engineering Pub Date : 2022-05-12 DOI: 10.1088/1748-605X/ac6f38
Zichang Zhang, Fan Zhou, Jianwei Zheng, J. Mu, P. Bo, Bin You
To provide better treatment of myocardial infarction, DiI-labeled bone marrow mesenchymal stem cells (BMSCs) were contact co-cultured with cardiomyocytes (CMs) on polycaprolactone (PCL) film to prepare myocardial patches. BMSCs from Sprague Dawley rats were isolated, cultured, and characterized for expression of surface markers by flow cytometry. CMs were isolated from suckling rats. After BMSCs were cultured for three generations, they were labeled with DiI dye. DiI-labeled BMSCs were co-cultured with CMs on PCL film in the experimental group, while CMs were replaced with the same amount of unlabeled BMSCs in the control group. After 24 h, cell growth was observed by light microscopy and cells were fixed for scanning electron microscopy (SEM). After 7 d of co-culture, cells were stained for immunofluorescence detection of myocardial markers cardiac troponin T (cTnT) and α-actin. Differentiation of BMSCs on PCL was observed by fluorescence microscopy. The efficiency of BMSC differentiation into CMs was analyzed by flow cytometry on the first and seventh days of co-culture. CMs were stained with calcein alone and contact co-cultured with DiI-labeled BMSCs on PCL film to observe intercellular dye transfer. Finally, cells were stained for immunofluorescence detection of connexin 43 (Cx43) expression and to observe the relationship between gap junctions and contact co-culture. BMSCs were identified by flow cytometry as strongly positive for CD90 and CD44H, and negative for CD11b/c and CD45. After co-culture for 24 h, cells were observed to have attached to PCL by light microscopy. Upon appropriate excitation, DiI-labeled BMSCs exhibited red fluorescence, while unlabeled CMs did not. SEM revealed a large number of cells on the PCL membrane and their cell state appeared normal. On the seventh day, some DiI-labeled BMSCs expressed cTnT and α-actin. Flow cytometry showed that the rate of stem cell differentiation in the experimental group was significantly higher than the control group on the seventh day (20.12% > 3.49%, P < 0.05). From the second day of co-culture, immunofluorescence staining for Cx43 revealed green fluorescent puncta in some BMSCs; from the third day of co-culture, a portion of BMSCs exhibited green fluorescence in dye transfer tests. Contact co-culture of DiI-labeled BMSCs and CMs on PCL film generated primary myocardial patches. The mechanism by which contact co-culture promoted differentiation of the myocardial patch may be related to gap junctions and gap junction-mediated intercellular signaling pathways.
为了更好地治疗心肌梗死,DiI标记的骨髓间充质干细胞(BMSC)与心肌细胞(CM)在聚己内酯(PCL)膜上接触共培养,制备心肌贴片。分离、培养来自Sprague-Dawley大鼠的BMSC,并通过流式细胞术表征表面标记物的表达。CM是从乳鼠中分离出来的。BMSCs培养三代后,用DiI染料进行标记。在实验组中,DiI标记的BMSC与PCL膜上的CM共培养,而在对照组中,用相同量的未标记的BMSCs代替CM。24小时后,通过光学显微镜观察细胞生长,并将细胞固定用于扫描电子显微镜(SEM)。共培养7天后,对细胞进行染色,以进行心肌标记物肌钙蛋白T(cTnT)和α-肌动蛋白的免疫荧光检测。通过荧光显微镜观察骨髓基质干细胞在PCL上的分化。在共培养的第一天和第七天通过流式细胞术分析BMSC分化为CMs的效率。CM单独用钙黄绿素染色,并在PCL膜上与DiI标记的BMSC接触共培养,以观察细胞间染料转移。最后,对细胞进行染色,以免疫荧光检测连接蛋白43(Cx43)的表达,并观察间隙连接和接触共培养之间的关系。通过流式细胞术鉴定BMSCs为CD90和CD44H强阳性,CD11b/c和CD45阴性。共培养24小时后,通过光学显微镜观察到细胞已经附着在PCL上。在适当的激发下,DiI标记的BMSC表现出红色荧光,而未标记的CM则没有。扫描电镜显示PCL膜上有大量细胞,细胞状态正常。第7天,一些DiI标记的骨髓基质干细胞表达cTnT和α-肌动蛋白。流式细胞术显示,实验组干细胞分化率在第7天显著高于对照组(20.12%>3.49%,P<0.05);从共培养的第三天起,部分BMSC在染料转移测试中表现出绿色荧光。DiI标记的BMSCs和CMs在PCL膜上的接触共培养产生原发性心肌斑块。接触共培养促进心肌片分化的机制可能与间隙连接和间隙连接介导的细胞间信号通路有关。
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引用次数: 2
A methacrylated hyaluronic acid network reinforced Pluronic F-127 gel for treatment of bacterial keratitis 甲基丙烯酸透明质酸网络增强Pluronic F-127凝胶治疗细菌性角膜炎
IF 4 3区 医学 Q2 Engineering Pub Date : 2022-05-11 DOI: 10.1088/1748-605X/ac6ea9
Anyang Wang, Lina Dong, Zhongwei Guo, Wei Sun, S. Mi
In this study, we developed a novel in situ thermoresponsive gel by introducing crosslinked methacrylated hyaluronic acid (HA-MA) networks into Pluronic F-127 (PF-127) gel (HP gel) to achieve sustained levofloxacin (LFX) delivery in bacterial keratitis treatment. The interactions between PF-127 molecules and HA-MA networks were studied by scanning electron microscopy, rheology, dynamic light scattering, differential scanning calorimetry, and small angle x-ray scattering. The results showed that the HP gel exhibited a higher critical gelling temperature and lower viscosity than the PF-127 gel (P gel), and could form a uniform thin layer on the ocular surface. Moreover, the drug release profile and gel dissolution rate revealed that the HA-MA network could retard the diffusion and dissolution of drug molecules and prolong the drug release time, which corresponded to an enhanced antibacterial ability of the HP-LFX gel. Furthermore, the HP gel exhibited low cytotoxicity to human corneal epithelial cells. Finally, an in vivo pharmacodynamic study was conducted with rabbit keratitis models. An improved treatment efficacy was observed after application of the HP-LFX gels. This study highlights the potential of HP gels in ophthalmic drug delivery.
在这项研究中,我们开发了一种新的原位热响应凝胶,通过将交联甲基丙烯酸透明质酸(HA-MA)网络引入Pluronic F-127 (PF-127)凝胶(HP凝胶)中,以实现治疗细菌性角膜炎的左氧氟沙星(LFX)的持续递送。通过扫描电镜、流变学、动态光散射、差示扫描量热法和小角x射线散射等方法研究了PF-127分子与HA-MA网络的相互作用。结果表明,HP凝胶比PF-127凝胶(P凝胶)具有更高的临界胶凝温度和更低的粘度,并能在眼表面形成均匀的薄层。此外,药物释放谱和凝胶溶出率显示HA-MA网络可以延缓药物分子的扩散和溶解,延长药物释放时间,这与HP-LFX凝胶的抗菌能力增强相对应。此外,HP凝胶对人角膜上皮细胞具有较低的细胞毒性。最后,对兔角膜炎模型进行了体内药效学研究。应用HP-LFX凝胶后,观察到治疗效果的改善。这项研究强调了HP凝胶在眼科给药方面的潜力。
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引用次数: 2
Environmentally friendly fabrication of electrospun nanofibers made of polycaprolactone, chitosan and κ-carrageenan (PCL/CS/κ-C) 聚己内酯、壳聚糖和κ-卡拉胶(PCL/CS/κ-C)静电纺纳米纤维的环保制备
IF 4 3区 医学 Q2 Engineering Pub Date : 2022-05-11 DOI: 10.1088/1748-605X/ac6eaa
Z. Vargas-Osorio, Florian Ruther, Si Chen, S. Sengupta, L. Liverani, M. Michálek, D. Galusek, A. Boccaccini
Electrospun fibers based on biodegradable polyanionic or polycationic biopolymers are highly beneficial for biomedical applications. In this work, electrospun nanofibers made from poly(epsilon caprolactone) (PCL), chitosan (CS) and κ-carrageenan (κ-C) were successfully fabricated using several mixtures of benign solvents containing formic acid and acetic acid. The addition of κ-C improved the preparation procedure for the production of PCL/CS fibers by electrospinning. Moreover, a polymer mixture was selected to be stored at −20 °C for one month with the purpose to study the properties of the resulting fiber mat. The results indicated that fiber characteristics were not seriously compromised compared to the ones of those fabricated with the original solution, which represents an important reduction in produced waste. Thus, the interactions that occur between positively and negatively charged hydrophilic polysaccharides might induce higher stability to the linear aliphatic polyester in the polymer mixture. All fiber mats were morphologically, physico-chemically and mechanically characterized, showing average fiber diameters in the nano scale. A direct cell viability assay using ST-2 cells demonstrated cell proliferation after seven days of incubation for all prepared fiber mats, confirming their suitability as potential candidates for bone tissue engineering and wound healing applications.
以可生物降解的聚阴离子或聚阳离子生物聚合物为基础的静电纺丝纤维在生物医学上有着广泛的应用。本研究以聚己内酯(PCL)、壳聚糖(CS)和κ-卡拉胶(κ-C)为原料,采用甲酸和乙酸混合溶剂制备了电纺丝纳米纤维。κ c的加入改善了静电纺丝生产PCL/CS纤维的制备工艺。此外,选择一种聚合物混合物在- 20°C下储存一个月,目的是研究所得纤维垫的性能。结果表明,与使用原始溶液制造的纤维相比,纤维特性没有受到严重损害,这代表了产生废物的重要减少。因此,带正电荷和负电荷的亲水性多糖之间的相互作用可能会导致聚合物混合物中线性脂肪族聚酯具有更高的稳定性。所有纤维垫都进行了形态、物理化学和机械表征,显示出纳米尺度的平均纤维直径。使用ST-2细胞进行的直接细胞活力测定显示,所有制备的纤维垫在孵育7天后都有细胞增殖,证实了它们作为骨组织工程和伤口愈合应用的潜在候选者的适用性。
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引用次数: 8
The construction of a self-assembled coating with chitosan-grafted reduced graphene oxide on porous calcium polyphosphate scaffolds for bone tissue engineering 壳聚糖接枝还原氧化石墨烯自组装涂层在多孔聚磷酸钙支架上的构建
IF 4 3区 医学 Q2 Engineering Pub Date : 2022-05-11 DOI: 10.1088/1748-605X/ac6eab
Hong-Tao Ding, Xu Peng, Xiaoshuang Yu, Mengyue Hu, C. Wan, Ningning Lei, Yihao Luo, Xixun Yu
Bone regeneration in large bone defects remains one of the major challenges in orthopedic surgery. Calcium polyphosphate (CPP) scaffolds possess excellent biocompatibility and exhibits good bone ingrowth. However, the present CPP scaffolds lack enough osteoinductive activity to facilitate bone regeneration at bone defects that exceed the critical size threshold. To endow CPP scaffolds with improved osteoinductive activity for better bone regeneration, in this study, a self-assembled coating with chitosan-grafted reduced graphene oxide (CS-rGO) sheets was successfully constructed onto the surface of CPP scaffolds through strong electrostatic interaction and hydrogen bonds. Our results showed that the obtained CPP/CS-rGO composite scaffolds exhibited highly improved biomineralization and considerable antibacterial activity. More importantly, CPP/CS-rGO composite scaffolds could drive osteogenic differentiation of BMSCs and significantly up-regulate the expression of osteogenesis-related proteins in vitro. Meanwhile, the CS-rGO coating could inhibit aseptic loosening and improve interfacial osseointegration through stimulating bone marrow mesenchymal stem cells (BMSCs) to secrete more osteoprotegerin (OPG) and lesser receptor activator of nuclear factor-κB ligand (RANKL). Overall, the CS-rGO coating adjusts CPP scaffolds’ biological environment interface and endows CPP scaffolds with more bioactivity.
大骨缺损的骨再生仍然是骨科手术的主要挑战之一。聚磷酸钙(CPP)支架具有良好的生物相容性和良好的骨内生长。然而,目前的CPP支架缺乏足够的骨诱导活性来促进超过临界尺寸阈值的骨缺损处的骨再生。为了增强CPP支架的骨诱导活性,实现更好的骨再生,本研究通过强静电相互作用和氢键,成功地在CPP支架表面构建了壳聚糖接枝还原氧化石墨烯(CS-rGO)片的自组装涂层。我们的结果表明,所获得的CPP/CS-rGO复合支架表现出高度改善的生物矿化和相当大的抗菌活性。更重要的是,CPP/CS-rGO复合支架可以在体外驱动BMSCs的成骨分化,并显著上调成骨相关蛋白的表达。同时,CS-rGO涂层可以通过刺激骨髓间充质干细胞分泌更多的骨保护素(OPG)和更少的核因子-κB配体受体激活剂(RANKL)来抑制无菌性松动并改善界面骨整合。总之,CS-rGO涂层调节了CPP支架的生物环境界面,赋予CPP支架更大的生物活性。
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引用次数: 2
Innovative electrospun PCL/fibroin/l-dopa scaffolds scaffolds supporting bone tissue regeneration 新型电纺丝PCL/丝素/左旋多巴支架:支持骨组织再生的支架
IF 4 3区 医学 Q2 Engineering Pub Date : 2022-05-03 DOI: 10.1088/1748-605X/ac6c68
E. Marin, Orion Yoshikawa, F. Boschetto, T. Honma, T. Adachi, Wenliang Zhu, Huaizhong Xu, N. Kanamura, Toshiro Yamamoto, G. Pezzotti
Poly-caprolactone is one of the most promising biocompatible polymers on the market, in particular for temporary devices that are not subjected to high physiological loads. Even if completely resorbable in various biological environments, poly-caprolactione does not play any specific biological role in supporting tissue regeneration and for this reason has a limited range of possible applications. In this preliminary work, for the first time l-dopa and fibroin have been combined with electrospun poly-caprolactone fibers in order to induce bioactive effects and, in particular, stimulate the proliferation, adhesion and osteoconduction of the polymeric fibers. Results showed that addition of low-molecular weight fibroin reduces the mechanical strength of the fibers while promoting the formation of mineralized deposits, when tested in vitro with KUSA-A1 mesenchymal cells. l-dopa, on the other hand, improved the mechanical properties and stimulated the formation of agglomerates of mineralized deposits containing calcium and phosphorous with high specific volume. The combination of the two substances resulted in good mechanical properties and higher amounts of mineralized deposits formed in vitro.
聚己内酯是市场上最有前途的生物相容性聚合物之一,特别是用于不受高生理负荷的临时装置。即使在各种生物环境中完全可吸收,聚己内旋酮在支持组织再生方面也没有任何特定的生物学作用,因此可能的应用范围有限。本初步研究首次将左旋多巴和丝素与电纺丝聚己内酯纤维结合,以诱导生物活性,特别是刺激聚合纤维的增殖、粘附和骨传导。结果表明,低分子量纤维蛋白的加入降低了纤维的机械强度,同时促进了矿化沉积物的形成,并与kasa - a1间充质细胞进行了体外实验。另一方面,左旋多巴改善了机械性能,刺激了高比体积含钙磷矿化沉积团块的形成。这两种物质的结合导致了良好的机械性能和体外形成的较高数量的矿化沉积物。
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引用次数: 1
Production and characterization of a coconut oil incorporated gelatin-based film and its potential biomedical application 椰子油明胶基薄膜的生产和表征及其潜在的生物医学应用
IF 4 3区 医学 Q2 Engineering Pub Date : 2022-05-03 DOI: 10.1088/1748-605X/ac6c67
Mehlika Karamanlioglu, Serap Yesilkir-Baydar
The influence of coconut oil (CO) on a gelatin-based film was investigated when used as a potential wound dressing material. There is limited study on CO in protein-based wound dressing materials. Therefore, in this study a self-supporting, continuous and homogenous CO incorporated gelatin-based film was formulated and obtained by solution casting method. The influence of CO on physicochemical and thermal properties of gelatin-based film was also determined. Moreover, the effect CO in gelatin films on cell viability and cell migration was analysed with a preliminary cell culture study. Homogenous dispersion of 10% (w/w) CO was obtained in films when 3% (v/w) Tween 80, a surfactant, was incorporated to 20% (w/w) plasticized gelatin film forming solution. Effect of CO on gelatin-based film was observed via phase separation by scanning electron microscopy analysis. Water uptake of gelatin film with no CO, GE film; and 10% (w/w) CO incorporated GE film, GE-CO, were 320% and 210%, respectively, after 3 h in water. Fourier transform infrared spectroscopy analysis showed triglyceride component of CO and increased hydrogen bonding between NH groups of gelatin in GE-CO films. Differential scanning calorimetry results suggested a more ordered structure of GE-CO film due to an increase in melt-like transition temperature and melting enthalpy of GE-CO film. CO content also increased cell viability, assessed by XTT assay since cell viability was approximately 100% when L929 cell culture was incubated with GE-CO of 5–100 μg ml−1. Moreover, GE-CO samples within 5–25 μg ml−1 concentration range, increased proliferation of L929 cells since cell viability was significantly higher than the 100% viable cell culture control (P < 0.05) which is also an indication of efficient healing. However, GE decreased viability of L929 cells significantly at 100–10 μg ml−1 concentration range (P < 0.05) and were toxic at concentrations of 100, 75 and 50 μg ml−1 which decreased ∼50% of the viability of the cells. Scratch Assay to assess in vitro wound healing showed cell migration towards scratch after 24 h as an indication of wound healing only in GE-CO samples. This study showed that, CO could efficiently be added to gelatin-based films for preparation of a primary wound dressing biomaterial which is also demonstrated to have a promising wound healing effect for minor wounds.
当用作潜在的伤口敷料材料时,研究了椰子油(CO)对明胶基薄膜的影响。关于蛋白质类伤口敷料材料中CO的研究有限。因此,在本研究中,通过溶液浇铸法配制并获得了一种自支撑、连续且均匀的CO掺入明胶基薄膜。还测定了CO对明胶基薄膜理化性能和热性能的影响。此外,通过初步的细胞培养研究,分析了明胶膜中CO对细胞活力和细胞迁移的影响。当将表面活性剂3%(v/w)吐温80掺入20%(w/w)增塑明胶成膜溶液中时,在膜中获得10%(w/w)CO的均匀分散。通过扫描电子显微镜分析,通过相分离观察了CO对明胶基薄膜的影响。不含CO、GE膜的明胶膜的吸水性;和10%(w/w)CO掺入的GE膜GE-CO在水中3小时后分别为320%和210%。傅立叶变换红外光谱分析表明,在GE-CO薄膜中,CO的甘油三酯成分和明胶NH基团之间的氢键增加。差示扫描量热法结果表明,由于GE-CO薄膜的类熔体转变温度和熔融焓的增加,GE-CO膜的结构更加有序。通过XTT测定评估,CO含量也增加了细胞活力,因为当L929细胞培养物与5–100μg ml−1的GE-CO孵育时,细胞活力约为100%。此外,在5–25μg ml−1浓度范围内的GE-CO样品增加了L929细胞的增殖,因为细胞活力显著高于100%活力的细胞培养对照(P<0.05),这也是有效愈合的指标。然而,GE在100–10μg ml−1浓度范围内显著降低L929细胞的活力(P<0.05),在100、75和50μg ml–1浓度下具有毒性,降低了细胞活力的50%。评估体外伤口愈合的划痕试验显示,细胞在24小时后向划痕迁移,这仅在GE-CO样品中表明伤口愈合。这项研究表明,CO可以有效地添加到明胶基薄膜中,用于制备初级伤口敷料生物材料,该生物材料也被证明对轻微伤口具有良好的伤口愈合效果。
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引用次数: 1
Hybrid electrospun scaffolds based on polylactic acid/ PAMAM dendrimer/gemini surfactant for enhancement of synergistic antibacterial ability for biomedical application 聚乳酸/PAMAM树状大分子/双子表面活性剂复合电纺支架增强协同抗菌能力的生物医学应用
IF 4 3区 医学 Q2 Engineering Pub Date : 2022-04-29 DOI: 10.1088/1748-605X/ac6bd7
Rasool Shabanloo, S. Akbari, M. Mirsalehi
Hybrid electrospun scaffolds based on poly (L-lactic acid) (PLLA)/poly (amidoamine) (PAMAM-G2) dendrimer/gemini surfactant were fabricated for the enhancement of synergistic antibacterial activities. The second generation of poly (amidoamine) (PAMAM-G2) and cationic gemini surfactant were utilized to functionalize the optimum electrospun scaffolds. The gelatination process was utilized to improve the wettability of PLLA scaffolds to extend cell attachment and cell proliferation. PLLA nanofibrous scaffolds were characterized by energy dispersion x-ray, scanning electron microscopy images, mechanical properties, water contact angle, Fourier transform infrared spectroscopy, zeta potential and antibacterial assessment. In vitro cell biocompatibility was evaluated by 3-(4, 5- dimethylthiazoyl-2-yl)-2, 5-diphenyltetrazolium bromide assay and morphology of PC-12 cells cultured on hybrid nanofibrous scaffolds and gelatinized ones. The results indicated that the optimum scaffolds could successfully modify the characteristics of PLLA scaffolds leading to much more appropriate physical and chemical properties. In addition, gelatinized nanofibrous scaffolds reveal more wettability enhancing cell attachment and proliferation. Furthermore, using poly (amidoamine) (PAMAM-G2) and gemini surfactant reveals synergetic antibacterial activity due to the competition between both cationic groups of PAMAM and gemini surfactant. Finally, improved cell adhesion and cell viability on modified scaffolds were confirmed. These favorable properties give a chance for these scaffolds to be used in a wide variety of biomedical applications.
制备了基于聚L-乳酸(PLLA)/聚氨基胺(PAMAM-G2)树状大分子/双子表面活性剂的混合电纺支架,以增强协同抗菌活性。利用第二代聚氨基胺(PAMAM-G2)和阳离子双子表面活性剂对最佳的电纺支架进行了功能化。凝胶化过程用于改善PLLA支架的润湿性,以延长细胞附着和细胞增殖。通过能量分散x射线、扫描电子显微镜图像、力学性能、水接触角、傅立叶变换红外光谱、ζ电位和抗菌评价对PLLA纳米纤维支架进行了表征。通过3-(4,5-二甲基噻唑基-2-基)-2,5-二苯基四唑溴化测定法和在混合纳米纤维支架和凝胶支架上培养的PC-12细胞的形态来评估体外细胞生物相容性。结果表明,优化的支架可以成功地改变PLLA支架的特性,从而获得更合适的物理和化学性能。此外,凝胶化的纳米纤维支架显示出更多的润湿性,增强了细胞的附着和增殖。此外,由于PAMAM和双子表面活性剂的两个阳离子基团之间的竞争,使用聚氨基胺(PAMAM-G2)和双子表面表面活性剂显示出协同抗菌活性。最后,证实了在修饰的支架上细胞粘附性和细胞活力的改善。这些有利的性质为这些支架在各种生物医学应用中的应用提供了机会。
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引用次数: 1
Tailored alginate/PCL-gelatin-β-TCP membrane for guided bone regeneration 定制藻酸盐/PCL明胶-β-TCP膜引导骨再生
IF 4 3区 医学 Q2 Engineering Pub Date : 2022-04-29 DOI: 10.1088/1748-605X/ac6bd8
Gyeongjin Joo, Myeongki Park, Seong-su Park, G. Tripathi, Byong-Taek Lee
Membranes prepared for guided bone regeneration (GBR) signify valued resources, inhibiting fibrosis and assisting bone regenration. However, existing membranes lack bone regenerative capacity or adequate degradation profile. An alginate-casted polycaprolactone-gelatin-β-tricalcium phosphate dual membrane was fabricated by electrospinning and casting processes to enhance new bone formation under a GBR process. Porous membranes were synthesized with suitable hydrophilicity, swelling, and degradation behavior to confirm the compatibility of the product in the body. Furthermore, osteoblast-type cell toxicity and cell adhesion results showed that the electrospun membrane offered compatible environment to cells while the alginate sheet was found capable enough to supress the cellular attachment, but was a non-toxic material. Post-implantation, the in-vivo outcomes of the dual-layered membrane, showed appreciable bone formation. Significantly, osteoid islands had fused in the membrane group by eight weeks. The infiltration of fibrous tissues was blocked by the alginate membrane, and the ingrowth of new bone was enhanced. Immunocytochemical analysis indicated that the dual membrane could direct more proteins which control mineralization and convene osteoconductive properties of tissue-engineered bone grafts.
为引导骨再生(GBR)制备的膜意味着有价值的资源,可以抑制纤维化并帮助骨再生。然而,现有的膜缺乏骨再生能力或足够的降解特性。通过静电纺丝和浇铸工艺制备了藻酸盐浇铸聚己内酯明胶-β-磷酸三钙双膜,以增强GBR工艺下的新骨形成。合成了具有合适亲水性、溶胀性和降解行为的多孔膜,以确认产品在体内的兼容性。此外,成骨细胞型细胞毒性和细胞粘附结果表明,电纺膜为细胞提供了相容的环境,而海藻酸盐片足以抑制细胞附着,但是一种无毒材料。植入后,双层膜的体内结果显示有明显的骨形成。值得注意的是,到8周时,膜组的类骨岛已经融合。藻酸盐膜阻断了纤维组织的浸润,促进了新骨的向内生长。免疫细胞化学分析表明,双膜可以引导更多的蛋白质控制矿化,并召集组织工程骨移植物的骨传导特性。
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引用次数: 4
Preparation and pharmacokinetics of glycyrrhetinic acid and cell transmembrane peptides modified with liposomes for liver targeted-delivery 肝靶向给药脂质体修饰的甘次酸和细胞跨膜肽的制备及其药代动力学
IF 4 3区 医学 Q2 Engineering Pub Date : 2022-04-28 DOI: 10.1088/1748-605X/ac6b73
Li Li, Anqi Chen, Bingmi Liu, Hao Pan, Yanjie Yu, Yu Liu
The article presents a hepatocellular carcinoma cell surface-specific ligand glycyrrhetinic acid (GA) and cell-penetrating peptide (TAT) with good cell membrane penetration to modify the anti-tumor drug pingyangmycin (PYM) liver delivery system, which achieve targeted delivery of drugs and improve anti-tumor efficiency. In this study, we synthesized the pingyangmycin liposome modified by glycyrrhetinic acid and cell penetrating peptide(GA-TAT-PYM-L) and evaluated the anti-tumor effect of GA-TAT-PYM-L in vitro. Using the 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenylte-trazolium bromidecell proliferation method, GA-TAT-PYM-L had a stronger inhibitory effect on HepG2 cells than the free drug PYM at the same concentration. Acridine orange-ethidium bromide staining assays showed that GA-TAT-PYM-L had stronger apoptosis promotion effects on HepG2 cells in comparison to PYM. Pharmacokinetic studies indicated that, compared with PYM, GA-TAT-PYM-L enhanced mean residence time (MRT0–∞) and area under curve (AUC0–∞) by about 2.79-fold and 2.45-fold. The T 1/2 was prolonged to 140.23 ± 14.13 min. Tissue distribution results showed that the PYM concentrations in livers from the GA-TAT-PYM-L group were always higher than other tissues at each monitoring period after 5 min, indicating that GA-TAT-PYM-L can achieve liver targeting.
本文提出一种具有良好细胞膜穿透性的肝癌细胞表面特异性配体甘草酸(GA)和细胞穿透肽(TAT)修饰抗肿瘤药物平阳霉素(PYM)肝脏递送系统,实现药物靶向递送,提高抗肿瘤效率。本研究合成了甘次酸和细胞穿透肽修饰的平阳霉素脂质体(GA-TAT-PYM-L),并对GA-TAT-PYM-L体外抗肿瘤作用进行了评价。采用3-(4,5 -二甲基噻唑-2-基)- 2,5 -二苯基曲唑溴化细胞增殖法,GA-TAT-PYM-L对HepG2细胞的抑制作用强于相同浓度的游离药PYM。吖啶橙-溴化乙啶染色实验表明,GA-TAT-PYM-L对HepG2细胞的凋亡促进作用强于PYM。药代动力学研究表明,与PYM相比,GA-TAT-PYM-L的平均停留时间(MRT0 -∞)和曲线下面积(AUC0 -∞)分别提高了约2.79倍和2.45倍。t1 /2延长至140.23±14.13 min。组织分布结果显示,在5 min后的每个监测周期,GA-TAT-PYM-L组肝脏中PYM浓度始终高于其他组织,表明GA-TAT-PYM-L能够实现肝脏靶向。
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引用次数: 1
Dental resin composites with improved antibacterial and mineralization properties via incorporating zinc/strontium-doped hydroxyapatite as functional fillers 锌/锶掺杂羟基磷灰石作为功能填料提高牙科树脂复合材料的抗菌和矿化性能
IF 4 3区 医学 Q2 Engineering Pub Date : 2022-04-28 DOI: 10.1088/1748-605X/ac6b72
Yechen Li, Daixing Zhang, Zhuo Wan, Xiaoping Yang, Q. Cai
This study intends to improve the antibacterial and mineralization performance of photocurable dental resin composites (DRCs) to reduce the possibility of repair failure caused by secondary caries. To the end, functionalized hydroxyapatite (HAp), including Zn-doped (Zn/HAp) and Sr-doped HAp (Sr/HAp), were added into the bisphenol A glycidyl methacrylate and triethylene glycol dimethacrylate mixture, providing the DRCs with antibacterial and mineralization capacity, respectively. By controlling the total amount of inorganic filler at 70 wt%, these HAp powders were introduced into the resin matrix with barium glass powder (BaGP), while the ratios of HAp to aGP varied from 0:70 to 8:62. And the 8 wt% of HAp could be pure HAp, Zn/HAp, Sr/HAp, or Zn/HAp +Sr/HAp in different ratios (i.e. 2:6, 4:4, 6:2). Though the fillers varied, the obtained DRCs displayed similar micro-morphology, flexural strength (∼110 MPa) and modulus (∼7 GPa), and Vickers hardness (∼65). When the doping amounts of Sr2+/Zn2+ reached 15 mol% of Ca2+ in the Sr/HAp and Zn/HAp, the DRCs displayed a high antibacterial activity by killing ∼95% Staphylococcus aureus, and induced rich mineral deposition on surface in simulated body fluid. The incorporation of the Zn/HAp and Sr/HAp into the DRCs did not cause significant cytotoxicity, with L929 fibroblasts remaining >99% viability as cultured in extracts made from the DRCs. Therein, the DRC preparations containing both Zn/HAp and Sr/HAp have achieved improvements in both the biomineralization and antibacterial performance, as well as, having sufficient mechanical properties and excellent biocompatibility for dental restoration.
本研究旨在提高光固化牙科树脂复合材料(DRCs)的抗菌和矿化性能,以降低继发龋导致修复失败的可能性。最后,将功能化羟基磷灰石(HAp),包括Zn掺杂的(Zn/HAp)和Sr掺杂的HAp(Sr/HAp),加入到双酚A甲基丙烯酸缩水甘油酯和二甲基丙烯酸三乙二醇酯的混合物中,分别为DRCs提供抗菌和矿化能力。通过将无机填料的总量控制在70wt%,将这些HAp粉末引入具有钡玻璃粉末(BaGP)的树脂基体中,同时HAp与aGP的比率在0:70至8:62之间变化。并且8wt%的HAp可以是不同比例(即2:6、4:4、6:2)的纯HAp、Zn/HAp、Sr/HAp或Zn/HAp+Sr/HAp。尽管填料各不相同,但所获得的DRCs显示出相似的微观形态、弯曲强度(~110 MPa)和模量(~7 GPa)以及维氏硬度(~65)。当Sr2+/Zn2+的掺杂量达到Sr/HAp和Zn/HAp中Ca2+的15mol%时,DRCs通过杀死~95%的金黄色葡萄球菌表现出高抗菌活性,并在模拟体液中诱导表面富矿物质沉积。Zn/HAp和Sr/HAp掺入DRCs不会引起显著的细胞毒性,在DRCs提取物中培养的L929成纤维细胞保持>99%的活力。其中,同时含有Zn/HAp和Sr/HAp的DRC制剂在生物矿化和抗菌性能方面都有所提高,并且具有足够的力学性能和优异的生物相容性,可用于牙齿修复。
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引用次数: 9
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Biomedical materials
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