作者承认国家健康与医学研究委员会澳大利亚（NHMRC＃1000674）的资助LJR福斯特。

1。制备SurgiLux解决方案<ol><li>准备2％（体积/体积）溶液中，在一个干净的玻璃烧杯中，使用去离子水的乙酸，使用在层流罩，以避免污染。 </li><li>称取0.02％（重量/体积）的发色团，吲哚花青绿，ICG，在无菌的Eppendorf管中，确保管被包裹在银箔，以防止任何光线穿透。 </li><li>使用干净的，一次性吸液管，约1毫升的稀乙酸溶液转移到试管中，以溶解染料，轻轻摇动，并保持在箔包裹。 </li><li>转移到烧杯中，并添加2％（重量/体积）的脱乙酰壳多糖粉末溶解ICG前加入无菌的磁力搅拌器。 </li><li>量的烧杯用Parafilm银箔，然后包裹在混合之前的72小时，在室温下在层流罩中的内容在每分钟约125转。 </li><li>的内容传送到干净的离心管中并离心，以15,000 xg离心15分钟，在4℃以除去任何particula德事。 </li><li>小心转移到一个干净的玻璃烧杯中，绿色SurgiLux溶液覆盖用石蜡膜然后包裹在银箔之前，贮存在冰箱中12小时，以增加粘度的溶液。 </li></ol> 2。铸造SurgiLux电影<ol><li>使用无菌注射器，8毫升冷SurgiLux溶液分配到一个干净的，直径为95毫米的陪替氏培养皿中，并轻轻地倾斜板，由该溶液以确保完全覆盖。溶液体积比变铸造区允许的薄膜厚度的控制，参见图1。 </li><li>删除任何明显的气泡，在使用消毒针头的尖端解决方案。银箔及地方的菜，在冰箱中以除去残留的微米级的气泡。 </li><li> 20分钟后，小心地取出培养皿中的冰箱，在层流罩，覆盖银箔，并制定解决方案蒸发，连续3周。 </li><li>在CO蒸发mplete，得分的外边缘的清晰的绿色SurgiLux膜的陪替氏培养皿中，并轻轻剥离&#39;远离餐具表面的膜。 </li><li> SurgiLux电影应该是灵活和容易操作，没有撕裂或断裂。 </li><li>存储银箔包裹在干燥条件下，在培养皿中，直到准备使用的的圆形SurgiLux薄膜。 </li></ol> 3。 SurgiLux粘结膜的激光激活<ol><li>为了证明激光激活的过程中，我们将使用一个牛的组织，如牛排切成15毫米宽，20毫米长度的大小。解剖的组织使用号码10手术刀片，以产生2个10毫米15在一条直线上。 </li><li>约在两片组织，使他们的边缘接触，但不重叠，并用棉签或纱布，轻轻地吸收任何多余的液体。 </li><li>接下来，切一块SurgiLux电影7×9毫米，并小心地将薄膜纵向跨越平分pIECE的组织，然后用干棉签轻轻按下。 </li><li> SurgiLux薄膜被激活使用红外线的二极管激光器在设定为120 mW。由于这是一类IIIB激光，采取适当的安全措施，包括使用适当的安全眼镜的所有人员。 </li><li>在角落里，开始照射SurgiLux用红外线激光，在120毫瓦和1毫米直径的束斑大小设置。通过束斑在绿膜在约1毫米每秒的速率。重复的照射过程两次以上。 </li></ol> 4。修复的力量<ol><li>小心地固定在拉伸测试仪的夹具的组织的端部。我们使用的是一个Instron Mini55系统用50牛顿的测力传感器。的最大载荷，拉伸强度，和断裂伸长率，计算使用的Bluehill计算机软件（美国）。装置从至少10个样品进行了测定组（n = 10）。 </li><li>以“松弛”然后分出组织片在1毫米每秒的速度，直到两块由SurgiLux带完全分离的组织保持在一起。 </li></ol>

<ol>
	<li>Kjaergard, H. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Suture+support:+is+it+advantageous.">Suture support: is it advantageous.</a> Am. J. Surg. 182, 15S-20S (2001).</li><li>Lauto, A., Mawad, D., Foster, L. J. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Adhesive+biomaterials+for+tissue+reconstruction.">Adhesive biomaterials for tissue reconstruction.</a> J. Chem. Tech. Biotech. 83, 464-472 (2008).</li><li>Fung, L. C., Mingin, G. C., Massicotte, M., Felsen, D., Poppas, D. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effects+of+temperature+on+tissue+thermal+injury+and+wound+strength+after+photochemical+wound+closure.">Effects of temperature on tissue thermal injury and wound strength after photochemical wound closure.</a> Lasers Surg. Med. 25, 285-290 (1999).</li><li>Piribo, <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Glues &amp; Sealants: Industry Background Report. , (2005).</li><li>Lauto, A., Hook, J., Doran, M., Camacho, F., Poole-Warren, L. A., Avolio, A., Foster, L. J. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Chitosan+adhesive+for+laser+tissue-welding:+in+vitro+characterisation.">Chitosan adhesive for laser tissue-welding: in vitro characterisation.</a> Lasers Surg. Med. 36, 193-201 (2005).</li><li>Lauto, A., Stoodley, M., Marcel, H., Avolio, A., Sarris, M., McKenzie, G., Sampson, D. D., Foster, L. J. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=In+vitro+and+in+vivo+tissue+repair+with+laser-activated+chitosan+adhesive.">In vitro and in vivo tissue repair with laser-activated chitosan adhesive.</a> Lasers Surg. Med. 39, 19-27 (2007).</li><li>Lauto, A., Foster, L. J. R., Avolio, A., Sampson, D., Raston, C., Sarris, M., McKenzie, G., Stoodley, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Sutureless+Nerve+Repair+with+Laser-Activated+Chitosan+Adhesive:+A+Pilot+in+vivo+Study.">Sutureless Nerve Repair with Laser-Activated Chitosan Adhesive: A Pilot in vivo Study.</a> J. Photomed. Laser. Surg. 26 (3), 227-234 (2008).</li><li>Mar&#231;al, H., Badylak, S. F., Sellaro, T. L., Lauto, A., Foster, L. J. R., Mahler, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+coalescence+of+decellularized+tissue+scaffolds,+laser-activated+chitosan+bioadhesive+and+olfactory+ensheathing+cells+for+tissue+repair+and+regeneration+of+the+spinal+cord.">The coalescence of decellularized tissue scaffolds, laser-activated chitosan bioadhesive and olfactory ensheathing cells for tissue repair and regeneration of the spinal cord.</a> Lasers Med. Sci. 23 (1), 96 (2008).</li><li>Foster, L. J. R., Thomson, K., Marcal, H., Butt, J., Watson, S., Wakefield, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+chitosan-vancomycin+composite+biomaterial+as+a+laser+activated+surgical+adhesive+with+regional+antimicrobial+activity.">A chitosan-vancomycin composite biomaterial as a laser activated surgical adhesive with regional antimicrobial activity.</a> Biomacromolecules. 11 (12), 3563-3570 (2010).</li><li>Shahbazi, J., Marcal, H., Watson, S., Wakefield, D., Sarris, M., Foster, L. J. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Sutureless+sealing+of+penetrating+corneal+wounds+using+a+laser-activated+thin+film+adhesive.">Sutureless sealing of penetrating corneal wounds using a laser-activated thin film adhesive.</a> Lasers Surg. Med. , .</li><li>Meyers, M. A., Chen, P. -. Y., Lin, A. Y. -. M., Seki, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Biological+materials:+Structure+and+mechanical+properties.">Biological materials: Structure and mechanical properties.</a> Prog. Mater. Sci. 53 (1), 1-206 (2008).</li></ol>

脱乙酰壳多糖可以在多种不同的分子量和不同程度的去乙醯化具有协同性作用（DDA）获得。脱乙酰壳多糖的纯度的变化可能会导致存在的微粒在SurgiLux溶液;离心是用来消除这些和应导致在一个透明的绿色溶液。但是，过滤还可以被用作一个附加的或替代的制造工序。至于与任何材料的加工，变化，，如壳聚糖DDA和分子量，有所得SurgiLux薄膜，生物和物质的理化性质，包括其粘合到组织的强度的影?...

<table border="1"><tbody><tr><td> 的试剂/设备名称 </td><td> 公司 </td><td> 目录编号 </td><td> 评论（可选） </td></tr><tr><td>壳聚糖</td><td> Sigma-Aldrich公司</td><td> 448877 </td><td></td></tr><tr><td>吲哚菁绿</td><td> Sigma-Aldrich公司</td><td> I2633 </td><td>也被称为Cardiogreen </td></tr><tr><td>醋酸</td><td> Sigma-Aldrich公司</td><td> 320099 </td><td></td></tr><tr><td>红外二极管激光器与光纤传输。 （808纳米，120毫瓦，梁芯200微米） </td><td> CNI激光器</td><td> FC-808 </td><td>可变高达5 W电源系统</td></tr><tr><td>激光防护眼镜</td><td> CNI激光器</td><td> LS-G </td><td></td></tr><tr><td>拉伸试验设备</td><td>英斯特朗私人有限公司</td><td> 5542 </td><td> 50 N称重传感器</td></tr></tbody></table>

a chitosan based, laser activated thin film surgical adhesive, 'surgilux': preparation and demonstration

缝合线是一个4000年的老技术，保持伤口愈合的“金标准”凭借其的修复强度（〜100千帕）。然而，缝合线可以作为感染的一个核心，在许多程序无法影响伤口修复，或功能性组织再生的干扰。1手术胶水和粘合剂，如那些基于纤维蛋白和氰基丙烯酸酯，已经研制出用作替代缝线这种伤口修复。然而，目前的商业粘合剂也有显着的缺点，从病毒和朊病毒的转让和与血纤维蛋白胶水的维修强度缺乏，组织的毒性和缺乏的生物相容性的氰基丙烯酸酯为基础的粘合剂。此外，目前可用的手术粘合剂往往是凝胶为基础的，并且可以延长固化时间，这限制了它们的应用程序2同样地，使用的UV激光器，以促进交联机制在蛋白质为基础的或白蛋白&#39;溶胶德尔斯的可导致DNA损伤，而激光组织焊接（LTW）易患热损伤的组织。3尽管他们的缺点，粘合剂和LTW已捕获的伤口闭合市场约30％报道超过美元5星十亿每年，一个重要的证明需要缝合的技术。 在无缝线技术的追求，我们利用壳聚糖作为一种生物材料发展的一个灵活的，薄膜，激光激活的手术粘合剂称为“SurgiLux”。这种新颖的的生物黏附使用FDA批准并成功应用于医学领域的应用和产品的各种生物材料和光子学相结合的独特。 SurgiLux克服与缝线和电流的手术粘合剂（ 见表1）相关联的所有的缺点。 在这份报告中，我们报告的制造的SurgiLux的相对简单的协议，并展示其激光激活和组织的熔接强度。 SurgiLux薄膜附着到胶原组织未经化学修饰，如交联和通过使用比较低的供电（120毫瓦）的红外激光，而不是的UV光的照射。壳聚糖膜胶原蛋白（约3千帕）有一个自然，而是弱的附着力，激光活化的壳聚糖基于SurgiLux电影强调了这种粘附强度，通过聚合物链相互作用的作为瞬态热膨胀的结果。没有这一“激活”过程，SurgiLux薄膜容易地除去。6-9 SurgiLux已经过测试，无论是在体外和体内的各种组织，包括神经，肠，硬脑膜和角膜。在所有情况下表现出良好的生物相容性，可以忽略不计热损伤照射的结果。6-10

离心分离导致的透明的绿色溶液，从而增加存储后的粘度在4-6℃下绿色溶液静置3周后，被转换成约20微米厚的透明的绿色SurgiLux膜，显示在视频中，很容易灵活。 一旦照射激光，SurgiLux膜债券到组织。在边缘处的膜组织出现收缩作为激光束通过的膜（ 图2），可以观察到这一点。无炭化或切除组织和薄膜应得到遵守。的接合强度SurgiLux到组织应该足以抬起平分的组?...

Watch this Scientific Journal Video about A Chitosan Based, Laser Activated Thin Film Surgical Adhesive, 'SurgiLux': Preparation and Demonstration at JoVE.com

A Chitosan Based, Laser Activated Thin Film Surgical Adhesive, &#039;SurgiLux&#039;: Preparation and Demonstration

一种新型的制造，柔性薄膜手术胶粘剂FDA批准的成分，壳聚糖和吲哚青绿。证明本胶原组织的粘接剂具有低功率的红外激光通过一个简单的激活过程的粘合。

基于几丁聚糖，激光激发薄膜手术胶粘剂，“SurgiLux的制备及示范

Sutures are a 4,000 year old technology that remain the 'gold-standard' for wound closure by virtue of their repair strength (~100 KPa). However, sutures ...

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Research

JoVE Journal

Bioengineering

A Chitosan Based, Laser Activated Thin Film Surgical Adhesive, 'SurgiLux': Preparation and Demonstration

13.1K 次观看.  University of New South Wales. 一种新型的制造，柔性薄膜手术胶粘剂FDA批准的成分，壳聚糖和吲哚青绿。证明本胶原组织的粘接剂具有低功率的红外激光通过一个简单的激活过程的粘合。

视频: A Chitosan Based, Laser Activated Thin Film Surgical Adhesive, 'SurgiLux': Preparation and Demonstration

Constructing a Collagen Hydrogel for the Delivery of Stem Cell-loaded Chitosan Microspheres

Multipotent stem cells have been shown to be extremely useful in the field of regenerative medicine1-3. However, in order to use these cells effectively for tissue regeneration, a number of variables must be taken into account. These variables include: the total volume and surface area of the implantation site, the mechanical properties of the tissue and the tissue microenvironment, which includes the amount of vascularization and the components of the extracellular matrix. Therefore, the materials being used to deliver these cells must be biocompatible with a defined chemical composition while maintaining a mechanical strength that mimics the host tissue. These materials must also be permeable to oxygen and nutrients to provide a favorable microenvironment for cells to attach and proliferate. Chitosan, a cationic polysaccharide with excellent biocompatibility, can be easily chemically modified and has a high affinity to bind with in vivo macromolecules4-5. Chitosan mimics the glycosaminoglycan portion of the extracellular matrix, enabling it to function as a substrate for cell adhesion, migration and proliferation. In this study we utilize chitosan in the form of microspheres to deliver adipose-derived stem cells (ASC) into a collagen based three-dimensional scaffold6. An ideal cell-to-microsphere ratio was determined with respect to incubation time and cell density to achieve maximum number of cells that could be loaded. Once ASC are seeded onto the chitosan microspheres (CSM), they are embedded in a collagen scaffold and can be maintained in culture for extended periods. In summary, this study provides a method to precisely deliver stem cells within a three dimensional biomaterial scaffold.

A major hurdle in current stem cell therapies is determining the most effective method to deliver these cells to host tissues. Here, we describe a chitosan-based delivery method that is efficient and simple in approach, while allowing adipose-derived stem cells to maintain their multipotency.

构建交付的干细胞壳聚糖微球的胶原凝胶

Multipotent stem cells have been shown to be extremely useful in the field of regenerative medicine1-3. However, in order to use these cells effectively ...

科研

生物工程

Fabrication and Application of Rose Bengal-chitosan Films in Laser Tissue Repair

Photochemical tissue bonding (PTB) is a sutureless technique for tissue repair, which is achieved by applying a solution of rose bengal (RB) between two tissue edges1,2. These are then irradiated by a laser that is selectively absorbed by the RB. The resulting photochemical reactions supposedly crosslink the collagen fibers in the tissue with minimal heat production3. In this report, RB has been incorporated in thin chitosan films to fabricate a novel tissue adhesive that is laser-activated. Adhesive films, based on chitosan and containing ~0.1 wt% RB, are fabricated and bonded to calf intestine and rat tibial nerves by a solid state laser (&lambda;=532 nm, Fluence~110 J/cm2, spot size~0.5 cm). A single-column tensiometer, interfaced with a personal computer, is used to test the bonding strength. The RB-chitosan adhesive bonds firmly to the intestine with a strength of 15 &plusmn; 6 kPa, (n=30). The adhesion strength drops to 2 &plusmn; 2 kPa (n=30) when the laser is not applied to the adhesive. The anastomosis of tibial nerves can be also completed without the use of sutures. A novel chitosan adhesive has been fabricated that bonds photochemically to tissue and does not require sutures.

Sutures are usually needed to repair tissue during surgical procedures. However, their application can be problematic as they are invasive and may damage tissue. The fabrication and application methods of a novel tissue adhesive are here reported. This adhesive film is laser-activated and does not require the use of sutures.

激光对生物组织的修复孟加拉玫瑰红壳聚糖膜的制备及应用

Photochemical tissue bonding (PTB) is a sutureless technique for tissue repair, which is achieved by applying a solution of rose bengal (RB) between two ...

Creating Adhesive and Soluble Gradients for Imaging Cell Migration with Fluorescence Microscopy

Cells can sense and migrate towards higher concentrations of adhesive cues such as the glycoproteins of the extracellular matrix and soluble cues such as growth factors. Here, we outline a method to create opposing gradients of adhesive and soluble cues in a microfluidic chamber, which is compatible with live cell imaging. A copolymer of poly-L-lysine and polyethylene glycol (PLL-PEG) is employed to passivate glass coverslips and prevent non-specific adsorption of biomolecules and cells. Next, microcontact printing or dip pen lithography are used to create tracks of streptavidin on the passivated surfaces to serve as anchoring points for the biotinylated peptide arginine-glycine-aspartic acid (RGD) as the adhesive cue. A microfluidic device is placed onto the modified surface and used to create the gradient of adhesive cues (100% RGD to 0% RGD) on the streptavidin tracks. Finally, the same microfluidic device is used to create a gradient of a chemoattractant such as fetal bovine serum (FBS), as the soluble cue in the opposite direction of the gradient of adhesive cues.

A method for the assembly of adhesive and soluble gradients in a microscopy chamber for live cell migration studies is described. The engineered environment combines antifouling surfaces and adhesive tracks with solution gradients and therefore allows one to determine the relative importance of guidance cues.

创建的胶粘剂和可溶性梯度荧光显微镜成像细胞迁移

Cells can sense and migrate towards higher concentrations of adhesive cues such as the glycoproteins of the extracellular matrix and soluble cues such as ...

Insertion of Flexible Neural Probes Using Rigid Stiffeners Attached with Biodissolvable Adhesive

Microelectrode arrays for neural interface devices that are made of biocompatible thin-film polymer are expected to have extended functional lifetime because the flexible material may minimize adverse tissue response caused by micromotion. However, their flexibility prevents them from being accurately inserted into neural tissue. This article demonstrates a method to temporarily attach a flexible microelectrode probe to a rigid stiffener using biodissolvable polyethylene glycol (PEG) to facilitate precise, surgical insertion of the probe. A unique stiffener design allows for uniform distribution of the PEG adhesive along the length of the probe. Flip-chip bonding, a common tool used in microelectronics packaging, enables accurate and repeatable alignment and attachment of the probe to the stiffener. The probe and stiffener are surgically implanted together, then the PEG is allowed to dissolve so that the stiffener can be extracted leaving the probe in place. Finally, an in vitro test method is used to evaluate stiffener extraction in an agarose gel model of brain tissue. This approach to implantation has proven particularly advantageous for longer flexible probes (&gt;3 mm). It also provides a feasible method to implant dual-sided flexible probes. To date, the technique has been used to obtain various in vivo recording data from the rat cortex.

Insertion of flexible neural microelectrode probes is enabled by attaching probes to rigid stiffeners with polyethylene glycol (PEG). A unique assembly process ensures uniform and repeatable attachment. After insertion into tissue, the PEG dissolves and the stiffener is extracted. An in vitro test method evaluates the technique in agarose gel.

柔性神经微探针的使用附加与Biodissolvable胶粘剂刚性加劲肋插入

Microelectrode  arrays for neural interface devices that are made of biocompatible thin-film  polymer are expected to have extended functional lifetime ...

Development of Amelogenin-chitosan Hydrogel for In Vitro Enamel Regrowth with a Dense Interface

Biomimetic enamel reconstruction is a significant topic in material science and dentistry as a novel approach for the treatment of dental caries or erosion. Amelogenin has been proven to be a critical protein for controlling the organized growth of apatite crystals. In this paper, we present a detailed protocol for superficial enamel reconstruction by using a novel amelogenin-chitosan hydrogel. Compared to other conventional treatments, such as topical fluoride and mouthwash, this method not only has the potential to prevent the development of dental caries but also promotes significant and durable enamel restoration. The organized enamel-like microstructure regulated by amelogenin assemblies can significantly improve the mechanical properties of etched enamel, while the dense enamel-restoration interface formed by an in situ regrowth of apatite crystals can improve the effectiveness and durability of restorations. Furthermore, chitosan hydrogel is easy to use and can suppress bacterial infection, which is the major risk factor for the occurrence of dental caries. Therefore, this biocompatible and biodegradable amelogenin-chitosan hydrogel shows promise as a biomaterial for the prevention, restoration, and treatment of defective enamel.

Development of Amelogenin-chitosan Hydrogel for In Vitro Enamel Regrowth with a Dense Interface

In this article, we describe a protocol for fabricating an amelogenin-chitosan hydrogel for superficial enamel reconstruction. Organized in situ growth of apatite crystals in the hydrogel formed a dense enamel-restoration interface, which will improve the effectiveness and durability of restorations.

釉原蛋白 - 壳聚糖水凝胶的发展<em&gt;体外</em&gt;搪瓷愈合了密集接口

Biomimetic enamel reconstruction is a significant topic in material science and dentistry as a novel approach for the treatment of dental caries or ...

Custom-designed Laser-based Heating Apparatus for Triggered Release of Cisplatin from Thermosensitive Liposomes with Magnetic Resonance Image Guidance

Liposomes have been employed as drug delivery systems to target solid tumors through exploitation of the enhanced permeability and retention (EPR) effect resulting in significant reductions in systemic toxicity. Nonetheless, insufficient release of encapsulated drug from liposomes has limited their clinical efficacy. Temperature-sensitive liposomes have been engineered to provide site-specific release of drug in order to overcome the problem of limited tumor drug bioavailability. Our lab has designed and developed a heat-activated thermosensitive liposome formulation of cisplatin (CDDP), known as HTLC, to provide triggered release of CDDP at solid tumors. Heat-activated delivery in vivo was achieved in murine models using a custom-built laser-based heating apparatus that provides a conformal heating pattern at the tumor site as confirmed by MR thermometry (MRT). A fiber optic temperature monitoring device was used to measure the temperature in real-time during the entire heating period with online adjustment of heat delivery by alternating the laser power. Drug delivery was optimized under magnetic resonance (MR) image guidance by co-encapsulation of an MR contrast agent (i.e.,&#160;gadoteridol) along with CDDP into the thermosensitive liposomes as a means to validate the heating protocol and to assess tumor accumulation. The heating protocol consisted of a preheating period of 5 min prior to administration of HTLC and 20 min heating post-injection. This heating protocol resulted in effective release of the encapsulated agents with the highest MR signal change observed in the heated tumor in comparison to the unheated tumor and muscle. This study demonstrated the successful application of the laser-based heating apparatus for preclinical thermosensitive liposome development and the importance of MR-guided validation of the heating protocol for optimization of drug delivery.

AN MRI-compatible&#160;custom-designed laser-based heating apparatus has been developed to provide local heating of subcutaneous tumors in order to activate release of agents from thermosensitive liposomes specifically at the tumor region.

定制设计的基于激光的加热装置顺铂触发释放的热敏脂质体与磁共振成像制导

Liposomes have been employed as drug delivery systems to target solid tumors through exploitation of the enhanced permeability and retention (EPR) effect ...

Automated Lipid Bilayer Membrane Formation Using a Polydimethylsiloxane Thin Film

An artificial lipid bilayer, or black lipid membrane (BLM), is a powerful tool for studying ion channels and protein interactions, as well as for biosensor applications. However, conventional BLM formation techniques have several drawbacks and they often require specific expertise and laborious processes. In particular, conventional BLMs suffer from low formation success rates and inconsistent membrane formation time. Here, we demonstrate a storable and transportable BLM formation system with controlled thinning-out time and enhanced BLM formation rate by replacing conventionally used films (polytetrafluoroethylene, polyoxymethylene, polystyrene) to polydimethylsiloxane (PDMS). In this experiment, a porous-structured polymer such as PDMS thin film is used. In addition, as opposed to conventionally used solvents with low viscosity, the use of squalene permitted a controlled thinning-out time via slow solvent absorption by PDMS, prolonging membrane lifetime. In addition, by using a mixture of squalene and hexadecane, the freezing point of the lipid solution was increased (~16 &#176;C), in addition, membrane precursors were produced that can be indefinitely stored and readily transported. These membrane precursors have reduced BLM formation time of &#60; 1 hr and achieved a BLM formation rate of ~80%. Moreover, ion channel experiments with gramicidin A demonstrated the feasibility of the membrane system.

We demonstrate a storable, transportable lipid bilayer formation system. A lipid bilayer membrane can be formed within 1 hr with over 80% success rate when a frozen membrane precursor is brought to ambient temperature. This system will reduce laborious processes and expertise associated with ion channels.

自动化脂双层膜的形成使用聚二甲基硅氧烷薄膜

An artificial lipid bilayer, or black lipid membrane (BLM), is a powerful tool for studying ion channels and protein interactions, as well as for ...

Image-guided, Laser-based Fabrication of Vascular-derived Microfluidic Networks

This detailed protocol outlines the implementation of image-guided, laser-based hydrogel degradation for the fabrication of vascular-derived microfluidic networks embedded in PEGDA hydrogels. Here, we describe the creation of virtual masks that allow for image-guided laser control; the photopolymerization of a micromolded PEGDA hydrogel, suitable for microfluidic network fabrication and pressure head-driven flow; the setup and use of a commercially available laser scanning confocal microscope paired with a femtosecond pulsed Ti:S laser to induce hydrogel degradation; and the imaging of fabricated microfluidic networks using fluorescent species and confocal microscopy. Much of the protocol is focused on the proper setup and implementation of the microscope software and microscope macro, as these are crucial steps in using a commercial microscope for microfluidic fabrication purposes that contain a number of intricacies. The image-guided component of this technique allows for the implementation of 3D image stacks or user-generated 3D models, thereby allowing for creative microfluidic design and for the fabrication of complex microfluidic systems of virtually any configuration. With an expected impact in tissue engineering, the methods outlined in this protocol could aid in the fabrication of advanced biomimetic microtissue constructs for organ- and human-on-a-chip devices. By mimicking the complex architecture, tortuosity, size, and density of in vivo vasculature, essential biological transport processes can be replicated in these constructs, leading to more accurate in vitro modeling of drug pharmacokinetics and disease.

This protocol outlines the implementation of image-guided, laser-based hydrogel degradation to fabricate vascular-derived, biomimetic microfluidic networks embedded in poly(ethylene glycol) diacrylate (PEGDA) hydrogels. These biomimetic microfluidic systems may be useful for tissue engineering applications, generation of in vitro disease models, and fabrication of advanced "on-a-chip" devices.

影像引导下，血管源性微流控网络的基于激光的研制

This detailed protocol outlines the implementation of image-guided, laser-based hydrogel degradation for the fabrication of vascular-derived microfluidic ...

Dry Film Photoresist-based Electrochemical Microfluidic Biosensor Platform: Device Fabrication, On-chip Assay Preparation, and System Operation

In recent years, biomarker diagnostics became an indispensable tool for the diagnosis of human disease, especially for the point-of-care diagnostics. An easy-to-use and low-cost sensor platform is highly desired to measure various types of analytes (e.g., biomarkers, hormones, and drugs) quantitatively and specifically. For this reason, dry film photoresist technology - enabling cheap, facile, and high-throughput fabrication - was used to manufacture the microfluidic biosensor presented here. Depending on the bioassay used afterwards, the versatile platform is capable of detecting various types of biomolecules. For the fabrication of the device, platinum electrodes are structured on a flexible polyimide (PI) foil in the only clean-room process step. The PI foil serves as a substrate for the electrodes, which are insulated with an epoxy-based photoresist. The microfluidic channel is subsequently generated by the development and lamination of dry film photoresist (DFR) foils onto the PI wafer. By using a hydrophobic stopping barrier in the channel, the channel is separated into two specific areas: an immobilization section for the enzyme-linked assay and an electrochemical measurement cell for the amperometric signal readout.
The on-chip bioassay immobilization is performed by the adsorption of the biomolecules to the channel surface. The glucose oxidase enzyme is used as a transducer for electrochemical signal generation. In the presence of the substrate, glucose, hydrogen peroxide is produced, which is detected at the platinum working electrode. The stop-flow technique is applied to obtain signal amplification along with rapid detection. Different biomolecules can quantitatively be measured by means of the introduced microfluidic system, giving an indication of different types of diseases, or, in regard to therapeutic drug monitoring, facilitating a personalized therapy.

A microfluidic biosensor platform was designed and fabricated using low-cost dry film photoresist technology for the rapid and sensitive quantification of various analytes. This single-use system allows for the electrochemical readout of on-chip-immobilized enzyme-linked assays by means of the stop-flow technique.

干膜 Photoresist-based 电化学微流控生物传感器平台: 器件制备、片上检测制备及系统运行

In recent years, biomarker diagnostics became an indispensable tool for the diagnosis of human disease, especially for the point-of-care diagnostics. An ...

Rat Model of Adhesive Capsulitis of the Shoulder

This proposal aims to create an in vivo rat model of adhesive capsulitis for researching potential treatment options for this condition and other etiologies of comparable arthrofibrosis. The model includes extra-articular fixation of the shoulder in rats via scapular to humeral suturing, resulting in a secondary contracture without invading the intra-articular space and resulting in decreased rotational ROM and increased joint stiffness.
We used 10 Sprague-Dawley rats for the purpose of this study. Baseline ROM measurements were taken before glenohumeral immobilization. The rats were subjected to 8 weeks of immobilization before the fixation sutures were removed and changes in ROM and joint stiffness were evaluated. To evaluate whether immobilization resulted in a significant reduction in ROM, changes in kinematics were calculated. ROM was measured at each time point in the follow-up period and was compared to the baseline internal and external ROM measurements. In order to evaluate the stiffness, joint kinetics were calculated by determining the differences in torque (text and tint ) needed to reach the initial external rotation of 60&#176; and initial internal rotation of 80&#176;.
After the removal of the extra-articular suture fixation on follow-up day 0, we found a 63% decrease in total ROM compared to baseline. We observed continuous improvement until week 5 of follow-up, with the progress slowing down around a 19% restriction. On week 8 of follow-up, there was still an 18% restriction of ROM. Additionally, on follow-up day 0, we found the torque increased by 13.3 Nmm when compared to baseline. On week 8, the total torque was measured to be 1.4 &#177; 0.2 Nmm higher than initial measurements. This work introduces a rat model of shoulder adhesive capsulitis with lasting reduced ROM and increased stiffness.

This protocol presents an in vivo rat model of adhesive capsulitis. The model includes an internal fixation of the glenohumeral joint with extra-articular suture fixation for an extended time, resulting in a decreased rotational range of motion (ROM) and increased joint stiffness.