저자는 LJR 포스터로 국민 건강과 호주의 의학 연구 협의회 (NHMRC # 1000674)에서 교부금을 인정합니다.

1. SurgiLux 솔루션의 준비 <ol><li> 깨끗한 유리 비커에 탈 이온수를 사용하여 아세트산의 2 % (V / V) 솔루션을 준비, 오염을 방지하기 위해 층류 흐름 후드를 사용합니다. </li><li> 발색단의 0.02 % (w / V)를 무게, 살균 Eppendorf 튜브에 ICG, 녹색 indocyanine, 튜브 등은 침투를 방지하기 위해 실버 호일에 포장되어 있는지 확인합니다. </li><li> 깨끗하고 일회용 피펫 사용, 염료를 분해 부드럽게 흔들어서 호일에 싸서 보관 튜브에 희석 아세트산 솔루션의 약 1 ML을 전송하기 만하면됩니다. </li><li> 비커에 solubilized ICG를 전송하고 살균 자기 교반기를 추가하기 전에 키토산 분말의 2 %를 (w / V)를 추가합니다. </li><li> Parafilm로 비커를 커버는 층류 후드에 상온 72 시간에 약 12​​5 rpm으로 내용을 혼합하기 전에 실버 호일에 포장. </li><li> 4에 15 분에 15,000 XG에서 깨끗한 원심 분리기 튜브와 원심 분리기로 내용을 전송 ° C 모든 particula를 제거하는테의 문제. </li><li> 조심스럽게 깨끗한 유리 비커에 녹색 SurgiLux 솔루션을 전송 솔루션의 점도를 높이기 위해 12 시간을 위해 냉장고에 저장하기 전에 실버 호일에 포장 한 후 Parafilm를 사용하여 다룹니다. </li></ol> 2. SurgiLux 영화의 주조 <ol><li> 멸균 주사기를 사용하여 95mm 직경의 깨끗하고 페트리 접시에 차가운 SurgiLux 솔루션의 8 ML을 투여하고, 부드럽게 솔루션 전체 범위를 보장하기 위해 판을 기울입니다. 주조 영역으로 솔루션 볼륨의 비율을 변화하면 막 두께의 제어를 허용, 그림 1을 참조하십시오. </li><li> 멸균 바늘의 끝을 사용하여 솔루션에 보이는 거품을 제거합니다. 잔여 미크론 크기의 거품을 제거하기 위해 냉장고에 실버 호일과 장소에서 요리를 다룹니다. </li><li> 20 분 후에 신중하게, 층류 후드에 냉장고에서 장소를 페트리 접시를 제거 실버 호일로 커버 3 주 동안 증발 할 수있는 솔루션을 둡니다. </li><li> 공동 후거리에 접시 표면에서 영화 &#39;껍질&#39;부드럽게 mplete 증발, 점수는 배양 접시에 투명한 녹색 SurgiLux 필름의 바깥 가장자리합니다. </li><li> SurgiLux 영화는 유연하고 쉽게 찢어 또는 한방울 흘리지 않고 조작해야합니다. </li><li> 사용 준비가 될 때까지 건조 조건은 호일에 싸서 배양 접시에있는 원형 SurgiLux 필름을 저장합니다. </li></ol> 3. SurgiLux 접착제 영화의 레이저 활성화 <ol><li> 레이저 활성화 과정을 설명하기 위해 우리는 다양한 15mm 및 20mm 길이의 크기로 절단 등 스테이크와 같은 소 조직의 조각을 사용합니다. 10mm로 15의 2 조각을 생산하는 10 번 수술 칼을 사용하여 직선의 조직을 해부. </li><li> 대략의 가장자리가 닿지 아니라 중복 및 면봉이나 거즈를 사용하므로 조직의 두 조각이 부드럽게 초과 액체를 흡수합니다. </li><li> 다음 SurgiLux 영화 7 X 9mm의 조각을 잘라 조심스럽게 bisected P에서 영화 길게 배치조직의 iece 한 후 마른 면봉으로 부드럽게 아래로 누르십시오. </li><li> SurgiLux 필름은 120 MW의 설정에 적외선 다이오드 레이저를 사용하여 활성화됩니다. 이 클래스 IIIB 레이저이기 때문에, 적절한 안전 조치는 모든 직원에 대한 적절한 안전 안경의 사용을 포함하여 이동해야합니다. </li><li> 구석에 시작, 120 MW로 설정 적외선 레이저와 1mm 직경의 빔 스폿 크기 SurgiLux을 비추다. 초당 약 1 mm의 속도로 녹색 필름을 통해 빔 스폿을 전달합니다. 조사 과정 두 번 이상 반복합니다. </li></ol> 4. 수리의 힘 <ol><li> 조심스럽게 인장 테스트 악기의 클램프에있는 조직의 끝을 고정하십시오. 우리는 50 뉴턴로드 셀과 Instron Mini55 시스템을 사용하고 있습니다. 휴식의 최대 하중, 인장 강도, 및 확장은 Bluehill 컴퓨터 소프트웨어 (USA)를 사용하여 계산되었다. 최소 10 샘플의 수단 (N = 10)으로 결정됩니다. </li><li> &#39;여유&#39;를 타고 위로 이동조직의 두 가지가 완전히 별도의 SurgiLux 스트립에 의해 함께 개최 될 때까지 다음, 초당 1 mm의 속도로 조직 조각을 분리합니다. </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>

관심 없음 충돌이 선언 없습니다.

키토산은 분자량 다양한과 deactylation의 다른도 (DDA)와 구할 수 있습니다. 키토산의 순도의 변화는 SurgiLux 솔루션의 미립자의 존재로 이어질 수 있으며, 원심 분리는 다음을 제거하고 투명한 녹색 솔루션을 초래한다 사용됩니다. 그러나, 여과는 추가 또는 대체 제조 단계로 사용할 수 있습니다. 와 같은 키토산 DDA와 분자 무게와 같은 자료 처리, 변화는 조직의 결합의 강도를 포함한 결과 SurgiLux 영화...

<table border="1"><tbody><tr><td> 시약 / 장비의 이름 </td><td> 회사 </td><td> 카탈로그 번호 </td><td> 코멘트 (선택 사항) </td></tr><tr><td> 키토산 </td><td> 시그마 - 알드리치 </td><td> 448877 </td><td></td></tr><tr><td> 그린 Indocyanine </td><td> 시그마 - 알드리치 </td><td> I2633 </td><td> 또한 Cardiogreen로 알려진 </td></tr><tr><td> 아세트산 </td><td> 시그마 - 알드리치 </td><td> 320099 </td><td></td></tr><tr><td> 섬유 배달 적외선 다이오드 레이저. (808 nm 정도, 120 MW, 빔 코어 200 μm) </td><td> CNI 레이저 </td><td> FC-808 </td><td> 가변 시스템을 최대 5 W 전원 </td></tr><tr><td> 레이저 안전 안경 </td><td> CNI 레이저 </td><td> LS-G </td><td></td></tr><tr><td> 인장 시​​험 장치 </td><td> Instron PTY (주) </td><td> 5542 </td><td> 50 N의로드 셀 </td></tr></tbody></table>

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

봉합은 수리 강도 (~ 100 KPa)의 미덕에 의해 상처 폐쇄에 대한 &#39;황금 표준&#39;남아 4,000년 오래된 기술입니다. 그러나, 봉합 감염의 근원 역할을 할 수 있으며 여러 절차에 상처 수리에 영향을 또는 기능 조직 재생을 방해 할 수 없습니다. 등 섬유소와 cyanoacrylates에 따라 그와 같은 일 외과 접착제와 접착제는 위해 봉합에 대한 대안으로 개발 된 이러한 상처의 수리. 그러나, 현재 상용 접착제는 조직 독성과 시아 노 아세틸렌 기반의 접착제에 대한 biocompatibility의 부족으로 섬유소의 접착제와 마찬가지로 바이러스와 prion 전송 및 수리 강도의 부족에 이르기까지 상당한 단점을 수 있습니다. 또한, 현재 수술 접착제는 젤을 기반으로하는 경향이 있으며 자신의 응용 프로그램을 제한 치료 시간을 연장 할 수 있습니다. 2 마찬가지로, UV 레이저의 사용이 단백질 기반이나 알부민 &#39;솔의 상호 연결 메커니즘을 촉진하기레이저 조직 용접 (LTW)가 자신의 단점, 접착제 및 LTW에도 불구하고 조직에 열 손상. 3 predisposes 동안 ders의 &#39;DNA 손상 될 수 있습니다 캡처 상처 폐쇄 시장의 약 30 %가 연간 미화 5 달러 억 초과로보고 sutureless 기술에 대한 필요성에 대한 중요한 증거 4. sutureless 기술의 추구에서 우리는 유연한 박막, &#39;SurgiLux&#39;이라고한다 레이저 활성 수술 접착제의 개발을위한 biomaterial로 키토산을 활용하고 있습니다. 이 소설의 bioadhesive는 FDA는 생물 의학 응용 프로그램 및 제품의 다양한 승인을 성공적으로 사용되는 biomaterials 및 포토닉스의 고유 한 조합을 사용합니다. SurgiLux는 봉합와 현재 수술 접착제 (표 1 참조)와 관련된 모든 단점을 극복. 이 프리젠 테이션에서 우리는 SurgiLux의 제조를위한 비교적 간단한 프로토콜을보고하고 설명의 레이저 활성화 및 조직 용접 강도. SurgiLux 영화는 교차 연결 및 조사를 통해 비교적 저전력 (120 MW) 적외선 레이저 대신 UV 라이트를 사용하는 화학 수정없이 collagenous 조직을 준수합니다. 키토산 영화가 콜라겐 (~ 3 KPa)에 자연 약한 접착제 매력을 가지고 키토산 기반 SurgiLux 영화의 레이저 활성화 과도 열 팽창의 결과로 폴리머 체인의 상호 작용을 통해이 부착의 힘을 강조하고 있습니다. 5이 &#39;정품&#39;절차없이 , SurgiLux 필름은 쉽게 제거됩니다. 6-9 SurgiLux은 신경, 내장, 경질 한 학교와 각막 등의 조직의 다양한 체외과 생체 모두에서 테스트되었습니다. 모든 경우에있어서 그 조사의 결과로 좋은 biocompatibility 및 무시할 열 손상을 보여 주었다. 6-10

원심 분리는 4-6로 저장 한 후 점도를 증가 투명한 녹색 솔루션 ° C.로 연결 3 주 동안 서 후, 녹색 솔루션은 동영상에서 보여준 것처럼, 약 20 미크론 두께 투명한 녹색 SurgiLux 영화로 변환되고, 쉽게 융통성이 있습니다. 조직에 레이저, SurgiLux 영화 채권과 방사선 주셔야합니다. 이것은 조직은 레이저 빔이 영화 (그림 2) 통과로 계약에 나타나는 영화의 가장자리에서...

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

소설의 제조, FDA 승인 재료의 유연한 박막 수술 접착제, 키토산, 녹색을 indocyanine이 설명되어 있습니다. 저전력 적외선 레이저로 간단한 인증 과정을 통해 collagenous 조직이 접착제의 결합이 입증되어 있습니다.

키토산 기반, 레이저 활성화 박막 외과 접착제, &#39;SurgiLux&#39;: 준비 및 데모

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

13.1K 조회수.  University of New South Wales. 소설의 제조, FDA 승인 재료의 유연한 박막 수술 접착제, 키토산, 녹색을 indocyanine이 설명되어 있습니다. 저전력 적외선 레이저로 간단한 인증 과정을 통해 collagenous 조직이 접착제의 결합이 입증되어 있습니다.

비디오: 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.

줄기 세포 - 로드된 키토산 Microspheres의 배달을 위해 콜라겐 히드로겔을 구축

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 ...

연구

생체공학

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.

제조 및 레이저 조직 복구에 장미 벵골 - 키토산 영화의 응용

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.

형광 현미경과 영상 셀 마이그레이션을위한 접착제 및 가용성식이 그라디언트를 만들기

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.

Biodissolvable 접착제가 부착 된 강성 보강재를 사용하여 유연한 신경 프로브의 삽입

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.

에 대한 성별 감정 - 키토산 하이드로 겔의 개발<em&gt; 체외</em고밀도 인터페이스&gt; 에나멜 재성장

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.

자기 공명 이미지 지침에 열성 리포좀에서 시스플라틴의 트리거 된 릴리스를위한 맞춤 설계 레이저 기반의 가열 장치

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.

폴리 디메틸 실록산 박막을 사용하여 자동화 지질 이중층 제막

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.

혈관 유래 미세 유체 네트워크의 이미지 유도, 레이저 기반의 제조

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.

드라이 필름 포토 레지스트 기반 전기 화학 미세 바이오 센서 플랫폼: 장치 제작, 온 칩 시험 준비 및 시스템 운영

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.