저자는 골 연골 플러그의 과정을 포장에있는 그들의 도움을 UMC 위트레흐트에서 개발 역학 그룹에서 씨 제론 반 덴 베르그 씨 Matthijs Wassink 그들의 감사의 말씀을드립니다. 이 작품은 네덜란드 관절염 재단에서 부여에 의해 지원되었다.

참고 : 여기에 제시된 프로토콜은 최근의 연구 논문 6, 8, 9의 실험과 계산 과정에서 채택된다. 이 프로토콜은도 1에 도시되어있다. 사체 자료는 위트레흐트 대학의 수의학 교수의 허가를 수집 하였다. 1. 시료와 욕제 <ol><li> 차가운 인산 완충 식염수 (PBS)를 분사하면서 과열 이후 연골의 손상을 방지하기 위해 맞춤 제작 드릴 비트 (도 1)를 이용하여 사후 말 대퇴골로부터 원통 골 연골 플러그 (8.5 mm의 직경) 밖으로 드릴. </li><li> 오버레이 욕의 측면 확산을 최소화하기 위해 슬리브를 수축 플라스틱으로 골 연골 플러그 열 수축. <ol><li> 일에서 연골과 골 연골 플러그를 장착E는 플라스틱 슬리브 내부 수축 초기 상단과 열풍을 불어 따른다. 열 관련 손상을 방지하기 위해 연골의 표면에 젖은면 조각을 추가합니다. </li></ol></li><li> 별도 부과 (예를 들어, 650 μL, 420 mm의 ioxaglate, 분자량 (Mw) = 1,269 다 충전 = -1) 및 중성 (650 μL, 420 mm의 iodixanol, MW = 1,550 다) 용질 유한 화장실을 준비한다. </li><li> 주사기를 사용하여 연골의 표면에 제조 유한 화장실로드 및 RT (도 2A)에서 실험 동안에 증발을 방지하기 위해, 랩 샘플에 코르크 마개를 배치했다. 중립 용질 수송을 연구하기 위해, iodixanol 목욕을 배치하고 연골 표면에 음전하를 띤 용질 수송 장소 ioxaglate 목욕을 연구. </li></ol> 2. 이미징 및 이미지 처리 <ol><li> 마이크로 CT의 모터 구동 스테이지에 장착 주문품 홀더에 코르크 마개로 감싸 절연 샘플을 놓는다. 장소 t그 때문에 조영제 용액 위를 향하고 덮여 그 연골 표면을 샘플링. </li><li> 마이크로 CT (40 × 40 × 40 μm의 3 복셀의 크기가 2 분의 주사 시간을 90 kV로의 관전압 180 μA의 관전류) 연골 연골 판 구성된 시야를 사용하여 검색하고,의 유한 평형 상태 (48 시간)까지 여러 시점에서의 조를 (도 2A)에 도달한다. 농도 값이 시간에 따라 변화하지 않는 경우에 평형 상태가 달성된다. </li><li> 제조업체의 소프트웨어를 사용하여 관심 (ROI)의 지역의 위치를 용이하게하기 위해 초기 이미지에 따라 서로 다른 시점에서 3D 이미지를 등록합니다 (예를 들어, 분석). </li><li> 변환 3 차원 제조 업체의 소프트웨어를 사용하여 처리하기 전에 2D 태그가 지정된 이미지 파일 형식 (TIFF) 스택에 마이크로 CT 영상을 재구성. </li><li> 글로벌 세그먼트 (ImageJ에&gt; 조정&gt; 임계치) 연골 하골 이상에서 연골소프트웨어에서 목욕을 거짓말. </li><li> 상이한 시점에서 연골의 평균 그레이 값을 구하는 이전 단계에서 생성 된 연골 마스크를 사용하여 (은 ImageJ에&gt;&gt; 측정 분석). </li><li> 연골의 초기 욕 농도 및 초기 조영제 농도 주어 용질의 실제 농도 평균 그레이 값을 변환하는 선형 보정 곡선을 사용한다. 이전의 데이터는 그레이 값 조영제의 농도를 갖는 선형 관계를 유지한다는 사실을지지한다. </li><li> 실험 시점에 비해 용질 농도를 플롯. </li></ol> 3. 전산 모델링 주 :이 문제의 확산은 실험 경계 조건을 준수 (Z 축을 따라) (1D)에서 일어나는 것으로한다. 따라서, 형상 임의로 생성 할 수 있습니다. <ol><li> 유한 목욕을 기반으로 연골 멀티 존 모델을 구축 : 1) 연골 표면 영역으로 구성<html> (전체 연골 두께의 20 %), 중간 영역 (전체 연골 두께의 50 %)과 깊은 영역 (전체 연골 두께의 30 %) 12 FEBio 13, 14 (도 2b 2) 유한 욕). </li><li> FEBio에서 연골 및 목욕의 서로 다른 영역의 기계적 및 물리적 특성을 할당합니다. 영률 (MPa의 10)이 상부로 조 작용 삼투압 레지스트 따라서 과도한 변형에서 연골을 보호하기에 충분히 높은 것으로 가정 하였다. <ol><li> 10-3 mm의 유압 투과도를 사용 4 / Ns를 상기 시뮬레이션 8 욕 0 실제 사용 용질 확산 계수의 포아송 비, 9. </li></ol></li><li> 메쉬 (8 노드 삼선 육면체 요소)를 생성하고, 경계 근처를 세분화 (도 2B)클래스 = &quot;외부 참조&quot;&gt; 8, 9. </li><li> 바이 페이 - 용질 모델 <ol><li> 이것에 대응하는 욕조 유효 압력에서 용질의 초기 농도를 적용한다. 15 9에서 유효 압력의 설명을보십시오. </li><li> 다른 연골 영역에 소정의 확산 계수에 따른 시간 곡선 대 용질의 농도를 얻기 위해 과도 모드 모델을 실행. </li></ol></li><li> 다상 모델 참고 : 욕조와 조직 사이의 전기 변동은 욕조와 조직 모두에 두 가의 반대 이온을 추가하여 우회 할 수 있습니다. <ol><li> 정상 상태 모델 : 소기 FCD 값을 증가시키면서 연골 상부 욕 동일한 유효 유체 압력 및 농도를 사용한다. </li><li> 과도 모델 : 충분히 높은 욕 중에서의 용질 확산 계수를 유지하여 잘 교반 유한 조를 만든다. 주입목욕 욕조에 공기 인터페이스에서 용질의 농도 소정의 값에 도달한다. </li><li> 과도 : 이전 단계에서 소정의 용질 농도 경계 조건을 제거하고 그것의 실제 확산 계수로 유한 욕의 확산 계수를 되돌릴. </li><li> 연골 다른 영역에 적용 FCDs 및 확산 계수에 기초하여 용질 농도 - 시간 곡선을 획득하기 위해 모델을 실행. </li></ol></li><li> FEBio - MATLAB 인터페이스 <ol><li> 자동 FEBio 플롯 농도 - 시간 곡선 (FEBio-MATLAB 인터페이스) (8, 9)에서의 시뮬레이션을 수행하는 MATLAB 코드를 개발한다. </li><li> FEBio - MATLAB 인터페이스를 사용하여 연골 영역에서 확산 계수와 FCDs을 변경합니다. FEBio에서 모델을 실행하고, 용질의 농도 - 시간 곡선 9 8 압축을 풉니 다. </li><li> 실험적으로 얻어진 용질 농도 - 시간 곡선을 비교최소 루트에 따라 다른 연골 영역의 데이터 획득 확산 계수들의 세트 FCDs 제곱 오차 (RMSE) (8, 9)를 의미한다. </li></ol></li></ol><img alt="그림 1" class="xfigimg" src="/files/ftp_upload/54984/54984fig1.jpg" /> 그림 1 : 실험 설정. A) 사용자 정의 만든 드릴 비트를 사용하여 샘플 추출 과정. B) 마이크로 CT 이미징 절차 확산 프로세스를 모니터링한다. <a href="http://ecsource.jove.com/files/ftp_upload/54984/54984fig1large.jpg" target="_blank">이 그림의 더 큰 버전을 보려면 여기를 클릭하십시오.</a> <img alt="그림 2" class="xfigimg" src="/files/ftp_upload/54984/54984fig2.jpg" /> 그림 2 : 도식.A) 실험 설계. 유한 목욕, 연골 및 관련 메쉬의 표면, 중간과 깊은 영역으로 구성된 B) 다중 영역 계산 모델. <a href="http://ecsource.jove.com/files/ftp_upload/54984/54984fig2large.jpg" target="_blank">이 그림의 더 큰 버전을 보려면 여기를 클릭하십시오.</a>

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T., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Computed+tomography+detects+changes+in+contrast+agent+diffusion+after+collagen+cross-linking+typical+to+natural+aging+of+articular+cartilage.">Computed tomography detects changes in contrast agent diffusion after collagen cross-linking typical to natural aging of articular cartilage.</a> Osteoarthritis and Cartilage. 19 (10), 1190-1198 (2011).</li><li>Decker, S. G., Moeini, M., Chin, H. C., Rosenzweig, D. H., Quinn, T. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Adsorption+and+Distribution+of+Fluorescent+Solutes+near+the+Articular+Surface+of+Mechanically+Injured+Cartilage.">Adsorption and Distribution of Fluorescent Solutes near the Articular Surface of Mechanically Injured Cartilage.</a> Biophysical Journal. 105 (10), 2427-2436 (2013).</li><li>Silvast, T. S., Jurvelin, S. J., Tiitu, V., Quinn, T. M., Töyräs, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Bath+Concentration+of+Anionic+Contrast+Agents+Does+Not+Affect+Their+Diffusion+and+Distribution+in+Articular+cartilage+In+Vitro.">Bath Concentration of Anionic Contrast Agents Does Not Affect Their Diffusion and Distribution in Articular cartilage In Vitro.</a> Cartilage. 4 (1), 42-51 (2013).</li><li>Silvast, T. S., Jurvelin, J. S., Lammi, M. J., T&#246;yr&#228;s, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=pQCT+study+on+diffusion+and+equilibrium+distribution+of+iodinated+anionic+contrast+agent+in+human+articular+cartilage+-+associations+to+matrix+composition+and+integrity.">pQCT study on diffusion and equilibrium distribution of iodinated anionic contrast agent in human articular cartilage - associations to matrix composition and integrity.</a> Osteoarthritis and Cartilage. 17 (1), 26-32 (2009).</li><li>Pouran, B., Arbabi, V., Villamar, J., Weinans, H., Zadpoor, A. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Contrast+agent's+transport+across+healthy+articular+cartilage+under+various+bath+conditions.">Contrast agent's transport across healthy articular cartilage under various bath conditions.</a> , (2015).</li><li>Arbabi, V., Pouran, B., Weinans, H., Zadpoor, A. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Application+of+a+biphasic-solute+model+in+predicting+diffusive+properties+of+osteochondral+interface.">Application of a biphasic-solute model in predicting diffusive properties of osteochondral interface.</a> , (2016).</li></ol>

저자가 공개하는 게 없다.

우리는 관절 연골에서 중립 및 충전 용질의 확산을 연구하기 위해 유한 요소 모델링 절차와 결합 된 실험 프로토콜을 제시 하였다. 우리의 최근 연구에 따르면, 제안 된 모델은 정확하게 관절 연골 8, 9의 서로 다른 영역에서 중립 (이상성 - 용질) 모두의 전송 및 음전하 (다상) 용질을 설명 할 수있다. 널리 관절 연골이 기능적으로 같은 OA 진행 <sup class="xr...

<html> 분자 수송 관절 형 조인트의 항상성에 중요한 역할을하는 관절 치료제의 전달 연골 및 연골 조영 증강 영상 1, 2, 3. 이러한 연골 통합 intactness, 용질의 전하 및 크기뿐만 아니라 삼투압 및 전송 레이트 4, 5, 6에 영향을 미칠 수있는 연골 접촉 욕의 농도 요소. 각 영역은 다른 농도의 주요 세포 외 매트릭스 분자의 배향으로 구성되어 있기 때문에 중성 또는 충전 중 용질의 수송은, 관절 연골 영역과 상이 할 수있다, 즉, 프로테오글리칸 (PG를) 콜라겐 타입 II (1), 7, 8, 9,아가씨 = &quot;외부 참조&quot;&gt; 10, 11. 더 중요한 것은, 충전 용질의 수송은 관절 연골 (8, 9)에 걸쳐 증가 외 매트릭스 내에 고정 된 네거티브 전하를 포함 프로테오글리칸의 농도에 크게 의존 할 수있다. 골관절염 (OA)가되어 연골 걸쳐 확산 연구의 중요성을 나타내는, 진행됨에 따라 이러한 매개 변수, 특히 고정 된 전하 밀도 (FCD)는 콜라겐 원 섬유 연골 전체 수분 함량의 변화의 방향은 변화를 거칠 수있다. 현재의 연구에서, 프로토콜은 6, 8, 9, 정확하게 확산의 유한 목욕 모델에서 중립 및 충전 용질을 사용하여 다양한 경계 조건에서 확산을 조사하기 위해 제안 이전에 설정된 실험 및 전산 연구를 기반으로. 티그 방법은 연골 및 고급 이상성-용질 다상 유한 요소 모델에 의해지지되는 유한 욕조를 포함하는 시스템의 마이크로 컴퓨터 단층 촬영 영상 (마이크로 CT)으로 구성되어 제안. 이러한 모델은 관절 연골의 다양한 영역에 걸쳐 중성 및 하전 분자뿐만 아니라 FCDs의 확산 계수를 취득 할 수 있습니다. 이 모델을 사용하여, 하나는 연골 및 오버레이 유한 화장실 사이의 상호 작용을 조사하는 데 사용할 수 있습니다 확산 중립 및 충전 분자의 행동의 더 나은 이해를 얻을 수 있습니다.

<table border="0" cellpadding="0" cellspacing="0" width="688">
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Hexabrix</td>
			<td style="width:149px;">Guerbet</td>
			<td style="width:123px;">15HX005D</td>
			<td style="width:201px;">Negatively charged contrast agent</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Visipaque</td>
			<td style="width:149px;">GE healthcare</td>
			<td align="right" style="width:123px;">12570511</td>
			<td>Nuetral contrast agent</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">PBS</td>
			<td style="width:149px;">Life technologies</td>
			<td align="right" style="width:123px;">10010023</td>
			<td>Medium</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">micro-CT</td>
			<td style="width:149px;">Perkin Elmer</td>
			<td style="width:123px;"></td>
			<td>Monitoring diffusion</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Freezing-point osmometer</td>
			<td style="width:149px;">Advanced instruments</td>
			<td style="width:123px;"></td>
			<td>Measuring solution osmolality</td>
		</tr>
	</tbody>
</table>

an experimental and finite element protocol to investigate the transport of neutral and charged solutes across articular cartilage

<html> 골관절염 (OA)은 관절 연골과 연골 하골의 변성과 연관된 쇠약하게하는 질병이다. 그것은 상당한 화학적 열화, 즉 손실 프로테오글리칸 및 콜라겐 브릴 중단을 경험으로 관절 연골의 변성이 실질적으로 그 하중 기능을 손상시킨다. OA에서 화학적 손상 메커니즘을 조사하는 한 유망한 방법은 외부 용질에 연골 표본을 노출하고, 분자의 확산을 모니터하는 것입니다. 건강한 연골에 비해 상이한 확산 특성을 생성 관절 연골을 가로 질러 이동하는 동안 연골 손상 (즉, 농도 및 고분자 필수 구성)의 정도는 외부 용질 충돌하는 에너지 손실과 관련된다. 이 연구에서 우리는 몇 가지 단계로 구성되어 있으며 이전에 개발 된 실험 마이크로-C를 기반으로하는 프로토콜을 소개14px; &quot;&gt; omputed T omography (마이크로 CT) 및 유한 요소 모델링 제 이상성-용질 확산 계수를 얻기 다상 유한 요소 모델에 적용함으로써 뒤에 마이크로 CT를 사용하여 기록되어 충전 및 비 대전 요오드 분자의 수송. 연골 영역에 걸쳐 전하 밀도를 고정.

여기에 제공된 대표적인 결과는 이전의 연구 논문 6, 8, 9, 16에서 채택된다. OA에서 관절 연골은 큰 변화 가장 중요한 GAG 손실 및 콜라겐 원 섬유 손상 17, 18, 19을</su...

Watch this Scientific Journal Video about An Experimental and Finite Element Protocol to Investigate the Transport of Neutral and Charged Solutes across Articular Cartilage at JoVE.com

An Experimental and Finite Element Protocol to Investigate the Transport of Neutral and Charged Solutes across Articular Cartilage

우리는 최근에 개발 된 실험 및 수치 방법의 도움으로 관절 연골에서 충전 및 충전되지 않은 분자의 이동을 조사하기위한 프로토콜을 제안한다.

실험적 및 유한 요소 의정서는 관절 연골에서 중립 및 충전 용질의 운송을 조사하기 위해

Osteoarthritis (OA) is a debilitating disease that is associated with degeneration of articular cartilage and subchondral bone. Degeneration of articular ...

an-experimental-finite-element-protocol-to-investigate-transport

Universidad de Cartagena UDEC

Research

JoVE Journal

Bioengineering

6.2K 조회수.  Delft University of Technology (TU Delft).  우리는 최근에 개발 된 실험 및 수치 방법의 도움으로 관절 연골에서 충전 및 충전되지 않은 분자의 이동을 조사하기위한 프로토콜을 제안한다. 

비디오: An Experimental and Finite Element Protocol to Investigate the Transport of Neutral and Charged Solutes across Articular Cartilage

Patterned Photostimulation with Digital Micromirror Devices to Investigate Dendritic Integration Across Branch Points

Light is a versatile and precise means to control neuronal excitability. The recent introduction of light sensitive effectors such as channel-rhodopsin and caged neurotransmitters have led to interests in developing better means to control patterns of light in space and time that are useful for experimental neuroscience. One conventional strategy, employed in confocal and 2-photon microscopy, is to focus light to a diffraction limited spot and then scan that single spot sequentially over the region of interest. This approach becomes problematic if large areas have to be stimulated within a brief time window, a problem more applicable to photostimulation than for imaging. An alternate strategy is to project the complete spatial pattern on the target with the aid of a digital micromirror device (DMD). The DMD approach is appealing because the hardware components are relatively inexpensive and is supported by commercial interests. Because such a system is not available for upright microscopes, we will discuss the critical issues in the construction and operations of such a DMD system. Even though we will be primarily describing the construction of the system for UV photolysis, the modifications for building the much simpler visible light system for optogenetic experiments will also be provided. The UV photolysis system was used to carryout experiments to study a fundamental question in neuroscience, how are spatially distributed inputs integrated across distal dendritic branch points. The results suggest that integration can be non-linear across branch points and the supralinearity is largely mediated by NMDA receptors.

Digital micromirror devices (DMD) can generate complex patterns in time and space with which to control neuronal excitability. Issues relevant to the design, construction, and operation of DMD systems are discussed. Such a system enabled the demonstration of non-linear integration across distal dendritic branch points.

브랜치 지점 간 돌기 통합을 맡은 이후로 디지털 Micromirror 장치와 패턴 Photostimulation

Light is a versatile and precise means to control neuronal excitability. The recent introduction of light sensitive effectors such as channel-rhodopsin ...

연구

생체공학

Determination of the Transport Rate of Xenobiotics and Nanomaterials Across the Placenta using the ex vivo Human Placental Perfusion Model

Decades ago the human placenta was thought to be an impenetrable barrier between mother and unborn child. However, the discovery of thalidomide-induced birth defects and many later studies afterwards proved the opposite. Today several harmful xenobiotics like nicotine, heroin, methadone or drugs as well as environmental pollutants were described to overcome this barrier. With the growing use of nanotechnology, the placenta is likely to come into contact with novel nanoparticles either accidentally through exposure or intentionally in the case of potential nanomedical applications. Data from animal experiments cannot be extrapolated to humans because the placenta is the most species-specific mammalian organ 1. Therefore, the ex vivo dual recirculating human placental perfusion, developed by Panigel et al. in 1967 2 and continuously modified by Schneider et al. in 1972 3, can serve as an excellent model to study the transfer of xenobiotics or particles.
Here, we focus on the ex vivo dual recirculating human placental perfusion protocol and its further development to acquire reproducible results.
The placentae were obtained after informed consent of the mothers from uncomplicated term pregnancies undergoing caesarean delivery. The fetal and maternal vessels of an intact cotyledon were cannulated and perfused at least for five hours. As a model particle fluorescently labelled polystyrene particles with sizes of 80 and 500 nm in diameter were added to the maternal circuit. The 80 nm particles were able to cross the placental barrier and provide a perfect example for a substance which is transferred across the placenta to the fetus while the 500 nm particles were retained in the placental tissue or maternal circuit. The ex vivo human placental perfusion model is one of few models providing reliable information about the transport behavior of xenobiotics at an important tissue barrier which delivers predictive and clinical relevant data.

Determination of the Transport Rate of Xenobiotics and Nanomaterials Across the Placenta using the ex vivo Human Placental Perfusion Model

The ex vivo dual recirculating human placental perfusion model can be used to investigate the transfer of xenobiotics and nanoparticles across the human placenta. In this video protocol we describe the equipment and techniques required for a successful execution of a placenta perfusion.

태반의 맞은 편에 생체이 물과 나노 물질의 전송 속도의 결정을 사용하여<em&gt; 생체</em&gt; 인간 태반 관류 모델

Decades ago the human placenta was thought to be an impenetrable barrier between mother and unborn child. However, the discovery of thalidomide-induced ...

Models and Methods to Evaluate Transport of Drug Delivery Systems Across Cellular Barriers

Sub-micrometer carriers (nanocarriers; NCs) enhance efficacy of drugs by improving solubility, stability, circulation time, targeting, and release. Additionally, traversing cellular barriers in the body is crucial for both oral delivery of therapeutic NCs into the circulation and transport from the blood into tissues, where intervention is needed. NC transport across cellular barriers is achieved by: (i) the paracellular route, via transient disruption of the junctions that interlock adjacent cells, or (ii) the transcellular route, where materials are internalized by endocytosis, transported across the cell body, and secreted at the opposite cell surface (transyctosis). Delivery across cellular barriers can be facilitated by coupling therapeutics or their carriers with targeting agents that bind specifically to cell-surface markers involved in transport. Here, we provide methods to measure the extent and mechanism of NC transport across a model cell barrier, which consists of a monolayer of gastrointestinal (GI) epithelial cells grown on a porous membrane located in a transwell insert. Formation of a permeability barrier is confirmed by measuring transepithelial electrical resistance (TEER), transepithelial transport of a control substance, and immunostaining of tight junctions. As an example, ~200&#160;nm polymer NCs are used, which carry a therapeutic cargo and are coated with an antibody that targets a cell-surface determinant. The antibody or therapeutic cargo is labeled with 125I for radioisotope tracing and labeled NCs are added to the upper chamber over the cell monolayer for varying periods of time. NCs associated to the cells and/or transported to the underlying chamber can be detected. Measurement of free 125I allows subtraction of the degraded fraction. The paracellular route is assessed by determining potential changes caused by NC transport to the barrier parameters described above. Transcellular transport is determined by addressing the effect of modulating endocytosis and transcytosis pathways.

Many therapeutic applications require safe and efficient transport of drug carriers and their cargoes across cellular barriers in the body. This article describes an adaptation of established methods to evaluate the rate and mechanism of transport of drug nanocarriers (NCs) across cellular barriers, such as the gastrointestinal (GI) epithelium.

모델 및 세포 장벽에 걸쳐 약물 전달 시스템의 전송을 평가하는 방법

Sub-micrometer carriers (nanocarriers; NCs) enhance efficacy of drugs by improving solubility, stability, circulation time, targeting, and release. ...

Human Cartilage Tissue Fabrication Using Three-dimensional Inkjet Printing Technology

Bioprinting, which is based on thermal inkjet printing, is one of the most attractive enabling technologies in the field of tissue engineering and regenerative medicine. With digital control&#160;cells, scaffolds, and growth factors can be precisely deposited to the desired two-dimensional (2D) and three-dimensional (3D) locations rapidly. Therefore, this technology is an ideal approach to fabricate tissues mimicking their native anatomic structures. In order to engineer cartilage with native zonal organization, extracellular matrix composition (ECM), and mechanical properties, we developed a bioprinting platform using a commercial inkjet printer with simultaneous photopolymerization capable for 3D cartilage tissue engineering. Human chondrocytes suspended in poly(ethylene glycol) diacrylate (PEGDA) were printed for 3D neocartilage construction via layer-by-layer assembly. The printed cells were fixed at their original deposited positions, supported by the surrounding scaffold in simultaneous photopolymerization. The mechanical properties of the printed tissue were similar to the native cartilage. Compared to conventional tissue fabrication, which requires longer UV exposure, the viability of the printed cells with simultaneous photopolymerization was significantly higher. Printed neocartilage demonstrated excellent glycosaminoglycan (GAG) and collagen type II production, which was consistent with gene expression. Therefore, this platform is ideal for accurate cell distribution and arrangement for anatomic tissue engineering.

The methods described in this paper show how to convert a commercial inkjet printer into a bioprinter with simultaneous UV polymerization. The printer is capable of constructing 3D tissue structure with cells and biomaterials. The study demonstrated here constructed a 3D neocartilage.

입체 잉크젯 인쇄 기술을 사용하여 인간의 연골 조직 제작

Bioprinting, which is based on thermal inkjet printing, is one of the most attractive enabling technologies in the field of tissue engineering and ...

A Coupled Experiment-finite Element Modeling Methodology for Assessing High Strain Rate Mechanical Response of Soft Biomaterials

This study offers a combined experimental and finite element (FE) simulation approach for examining the mechanical behavior of soft biomaterials (e.g. brain, liver, tendon, fat, etc.) when exposed to high strain rates. This study utilized a Split-Hopkinson Pressure Bar (SHPB) to generate strain rates of 100-1,500 sec-1. The SHPB employed a striker bar consisting of a viscoelastic material (polycarbonate). A sample of the biomaterial was obtained shortly postmortem and prepared for SHPB testing. The specimen was interposed between the incident and transmitted bars, and the pneumatic components of the SHPB were activated to drive the striker bar toward the incident bar. The resulting impact generated a compressive stress wave (i.e. incident wave) that traveled through the incident bar. When the compressive stress wave reached the end of the incident bar, a portion continued forward through the sample and transmitted bar (i.e. transmitted wave) while another portion reversed through the incident bar as a tensile wave (i.e. reflected wave). These waves were measured using strain gages mounted on the incident and transmitted bars. The true stress-strain behavior of the sample was determined from equations based on wave propagation and dynamic force equilibrium. The experimental stress-strain response was three dimensional in nature because the specimen bulged. As such, the hydrostatic stress (first invariant) was used to generate the stress-strain response. In order to extract the uniaxial (one-dimensional) mechanical response of the tissue, an iterative coupled optimization was performed using experimental results and Finite Element Analysis (FEA), which contained an Internal State Variable (ISV) material model used for the tissue. The ISV material model used in the FE simulations of the experimental setup was iteratively calibrated (i.e. optimized) to the experimental data such that the experiment and FEA strain gage values and first invariant of stresses were in good agreement.

The current study prescribes a coupled experiment-finite element simulation methodology to obtain the uniaxial dynamic mechanical response of soft biomaterials (brain, liver, tendon, fat, etc.). The multiaxial experimental results that arose because of specimen bulging obtained from Split-Hopkinson Pressure Bar testing were rendered to a uniaxial true stress-strain behavior when simulated through iterative optimization of the finite element analysis of the biomaterial.

소프트 생체 재료의 높은 변형 속도 기계 응답을 평가하기위한 결합 실험 - 유한 요소 모델링 방법론

This study offers a combined experimental and finite element (FE) simulation approach for examining the mechanical behavior of soft biomaterials (e.g. ...

3D Hydrogel Scaffolds for Articular Chondrocyte Culture and Cartilage Generation

Human articular cartilage is highly susceptible to damage and has limited self-repair and regeneration potential. Cell-based strategies to engineer cartilage tissue offer a promising solution to repair articular cartilage. To select the optimal cell source for tissue repair, it is important to develop an appropriate culture platform to systematically examine the biological and biomechanical differences in the tissue-engineered cartilage by different cell sources. Here we applied a three-dimensional (3D) biomimetic hydrogel culture platform to systematically examine cartilage regeneration potential of juvenile, adult, and osteoarthritic (OA) chondrocytes. The 3D biomimetic hydrogel consisted of synthetic component poly(ethylene glycol) and bioactive component chondroitin sulfate, which provides a physiologically relevant microenvironment for in vitro culture of chondrocytes. In addition, the scaffold may be potentially used for cell delivery for cartilage repair in vivo. Cartilage tissue engineered in the scaffold can be evaluated using quantitative gene expression, immunofluorescence staining, biochemical assays, and mechanical testing. Utilizing these outcomes, we were able to characterize the differential regenerative potential of chondrocytes of varying age, both at the gene expression level and in the biochemical and biomechanical properties of the engineered cartilage tissue. The 3D culture model could be applied to investigate the molecular and functional differences among chondrocytes and progenitor cells from different stages of normal or aberrant development.

Cartilage repair represents an unmet medical challenge and cell-based approaches to engineer human articular cartilage are a promising solution. Here, we describe three-dimensional (3D) biomimetic hydrogels as an ideal tool for the expansion and maturation of human articular chondrocytes.

관절 연골 세포 문화와 연골 생성을위한 3D 하이드로 겔 공사장 공중 발판

Human articular cartilage is highly susceptible to damage and has limited self-repair and regeneration potential. Cell-based strategies to engineer ...

An Experimental Protocol for Assessing the Performance of New Ultrasound Probes Based on CMUT Technology in Application to Brain Imaging

The possibility to perform an early and repeatable assessment of imaging performance is fundamental in the design and development process of new ultrasound (US) probes. Particularly, a more realistic analysis with application-specific imaging targets can be extremely valuable to assess the expected performance of US probes in their potential clinical field of application.
The experimental protocol presented in this work was purposely designed to provide an application-specific assessment procedure for newly-developed US probe prototypes based on Capacitive Micromachined Ultrasonic Transducer (CMUT) technology in relation to brain imaging.
The protocol combines the use of a bovine brain fixed in formalin as the imaging target, which ensures both realism and repeatability of the described procedures, and of neuronavigation techniques borrowed from neurosurgery. The US probe is in fact connected to a motion tracking system which acquires position data and enables the superposition of US images to reference Magnetic Resonance (MR) images of the brain. This provides a means for human experts to perform a visual qualitative assessment of the US probe imaging performance and to compare acquisitions made with different probes. Moreover, the protocol relies on the use of a complete and open research and development system for US image acquisition, i.e. the Ultrasound Advanced Open Platform (ULA-OP) scanner.
The manuscript describes in detail the instruments and procedures involved in the protocol, in particular for&#160;the calibration, image acquisition and registration of US and MR images. The obtained results prove the effectiveness of the overall protocol presented, which is entirely open (within the limits of the instrumentation involved), repeatable, and covers the entire set of acquisition and processing activities for US images.

The development of new ultrasound (US) probes based on Capacitive Micromachined Ultrasonic Transducer (CMUT) technology requires an early realistic assessment of imaging capabilities. We describe a repeatable experimental protocol for US image acquisition and comparison with magnetic resonance images, using an ex vivo bovine brain as an imaging target.

뇌 이미징 응용 프로그램에서 CMUT 기술에 기반 하는 새로운 초음파 프로브의 성능을 평가 하기 위한 실험 프로토콜

The possibility to perform an early and repeatable assessment of imaging performance is fundamental in the design and development process of new ...

Creating a Structurally Realistic Finite Element Geometric Model of a Cardiomyocyte to Study the Role of Cellular Architecture in Cardiomyocyte Systems Biology

With the advent of three-dimensional (3D) imaging technologies such as electron tomography, serial-block-face scanning electron microscopy and confocal microscopy, the scientific community has unprecedented access to large datasets at sub-micrometer resolution that characterize the architectural remodeling that accompanies changes in cardiomyocyte function in health and disease. However, these datasets have been under-utilized for&#160;investigating the role of cellular architecture remodeling in cardiomyocyte function. The purpose of this protocol is to outline how to create an accurate finite element model of a cardiomyocyte using high resolution electron microscopy and confocal microscopy images. A detailed and accurate model of cellular architecture has significant potential to provide new insights into cardiomyocyte biology, more than experiments alone can garner. The power of this method lies in its ability to computationally fuse information from two disparate imaging modalities of cardiomyocyte ultrastructure to develop one unified and detailed model of the cardiomyocyte. This protocol outlines steps to integrate electron tomography and confocal microscopy images of adult male Wistar (name for a specific breed of albino rat) rat cardiomyocytes to develop a half-sarcomere finite element model of the cardiomyocyte. The procedure generates a 3D finite element model that contains an accurate, high-resolution depiction (on the order of ~35 nm) of the distribution of mitochondria, myofibrils and ryanodine receptor clusters that release the necessary calcium for cardiomyocyte contraction from the sarcoplasmic reticular network (SR) into the myofibril and cytosolic compartment. The model generated here as an illustration does not incorporate details of the transverse-tubule architecture or the sarcoplasmic reticular network and is therefore a minimal model of the cardiomyocyte. Nevertheless, the model can already be applied in simulation-based investigations into the role of cell structure in calcium signaling and mitochondrial bioenergetics, which is illustrated and discussed using two case studies that are presented following the detailed protocol.

This protocol outlines a novel method to create a spatially detailed finite element model of the intracellular architecture of cardiomyocytes from electron microscopy and confocal microscopy images. The power of this spatially detailed model is demonstrated using case studies in calcium signaling and bioenergetics.

Cardiomyocyte 시스템 생물학에서 세포 아키텍처의 역할을 연구 하기 Cardiomyocyte의 구조적으로 현실적인 유한 요소 형상 모델 생성

With the advent of three-dimensional (3D) imaging technologies such as electron tomography, serial-block-face scanning electron microscopy and confocal ...

A Novel Surgical Technique As a Foundation for In Vivo Partial Liver Engineering in Rat

Organ engineering is a novel strategy to generate liver organ substitutes that can potentially be used in transplantation. Recently, in vivo liver engineering, including in vivo organ decellularization followed by repopulation, has emerged as a promising approach over ex vivo liver engineering. However, postoperative survival was not achieved. The aim of this study is to develop a novel surgical technique of in vivo selective liver lobe perfusion in rats as a prerequisite for in vivo liver engineering. We generate a circuit bypass only through the left lateral lobe. Then, the left lateral lobe is perfused with heparinized saline. The experiment is performed with 4 groups (n = 3 rats per group) based on different perfusion times of 20 min, 2 h, 3 h, and 4 h. Survival, as well as the macroscopically visible change of color and the histologically determined absence of blood cells in the portal triad and the sinusoids, is taken as an indicator for a successful model establishment. After selective perfusion of the left lateral lobe, we observe that the left lateral lobe, indeed, turned from red to faint yellow. In a histological assessment, no blood cells are visible in the branch of the portal vein, the central vein, and the sinusoids. The left lateral lobe turns red after reopening the blocked vessels. 12/12 rats survived the procedure for more than one week. We are the first to report a surgical model for in vivo single liver lobe perfusion with a long survival period of more than one week. In contrast to the previously published report, the most important advantage of the technique presented here is that perfusion of 70% of the liver is maintained throughout the whole procedure. The establishment of this technique provides a foundation for in vivo partial liver engineering in rats, including decellularization and recellularization.

A Novel Surgical Technique As a Foundation for In Vivo Partial Liver Engineering in Rat

We establish a novel surgical technique for an in vivo single liver lobe perfusion model in rat as a prerequisite for further studying in vivo partial liver engineering in the future.

Vivo에서 부분 간 엔지니어링을에서 위한 쥐를 기초로 새로운 수술 기법

Organ engineering is a novel strategy to generate liver organ substitutes that can potentially be used in transplantation. Recently, in vivo liver ...

Fabrication of Decellularized Cartilage-derived Matrix Scaffolds

Osteochondral defects lack sufficient intrinsic repair capacity to regenerate functionally sound bone and cartilage tissue. To this extent, cartilage research has focused on the development of regenerative scaffolds. This article describes the development of scaffolds that are completely derived from natural cartilage extracellular matrix, coming from an equine donor. Potential applications of the scaffolds include producing allografts for cartilage repair, serving as a scaffold for osteochondral tissue engineering, and providing in vitro models to study tissue formation. By decellularizing the tissue, the donor cells are removed, but many of the natural bioactive cues are thought to be retained. The main advantage of using such a natural scaffold in comparison to a synthetically produced scaffold is that no further functionalization of polymers is required to drive osteochondral tissue regeneration. The cartilage-derived matrix scaffolds can be used for bone and cartilage tissue regeneration in both in vivo and in vitro settings.

Decellularized cartilage-derived scaffolds can be used as a scaffold to guide cartilage repair and as a means to regenerate osteochondral tissue. This paper describes the decellularization process in detail and provides suggestions to use these scaffolds in in vitro settings.

Decellularized 연골 파생 매트릭스 건설 기계 제조

Osteochondral defects lack sufficient intrinsic repair capacity to regenerate functionally sound bone and cartilage tissue. To this extent, cartilage ...