저자는 뉴욕 수도권 연구 얼라이언스 그랜트의 지원을 인정하고 싶습니다.

1. Bioprinting 플랫폼 구축 프린터 수정은 HP 데스크젯 500 열 잉크젯 프린터와 HP 51626a 검정 잉크 카트리지를 기반으로했다. <ol><li> 프린터 상단의 플라스틱 덮개를 분리하고 조심스럽게 덮개에서 제어판을 분리합니다. </li><li> 프린터 상단 부분과베이스 사이의 3 케이블 연결을 분리합니다. 기지에서 프린터 상단 부분을 제거합니다. </li><li> 프린터 상단 부분에서, 잉크 카트리지 아래 우측에있는 작은 플라스틱 및 고무 부속품 (프린트 헤드 세척 장치)를 제거한다. </li><li> 스프링 용지함 받침대를 제거합니다. </li><li> 플라스틱 급지 줄을 덮고있는 금속 플레이트를 제거합니다. </li><li> 손보고 또는 다른 절단 도구를 사용하여 중간 공급 휠 위치의 플라스틱 급지 줄을 잘라. </li><li> 이전 단계 이후에 노출 된 2 급지 바퀴를 제거합니다. 바퀴 플라스틱은 매우 단단하고 전자입니다톱 도움이 될 것입니다. </li><li> 통조림 공기와 에탄올 잎사귀를 사용하여 먼지와 이물질을 청소합니다. </li><li> 베이스에 프린터 상단 부분을 연결합니다. </li><li> 사용하기 전에 층류 후드에서 적어도 2 시간 동안 변경됨 프린터 살균 UV. </li><li> 손보고 또는 다른 절단 도구를 사용하여 HP의 51626a 잉크 카트리지의 뚜껑을 잘라. </li><li> 잉크를 비우고 연골의 아래쪽 아니라 저수지를 포함 필터를 제거합니다. </li><li> 철저하게 수돗물을 실행하여 카트리지를 씻어. </li><li> 초음파 처리는 10 분 동안 탈 이온화 (DI) 물에 카트리지 잔여 잉크를 제거합니다. </li><li> 반드시 모든 잉크가 제거 된 확인하기 위해 카트리지를 검사합니다. 멸균 DI 물 다음에 소독 용 70 % 에탄올로 철저하게 카트리지를, 린스 또는 스프레이. </li><li> 동시 광중합 능력을 제공하는 인쇄 플랫폼 위에 장파장 자외선 램프를 설치했다. </li><li> UV 광을 이용하여 인쇄 플랫폼에 UV 강도를 측정미터. 인쇄 제목의 강도가 4-8 mW의 / ㎠ (프린터 플랫폼 램프에서 약 25 CM) 그래서 UV 램프와 프린터 플랫폼 사이의 거리를 조정합니다. </li></ol> 2. Bioink 준비 <ol><li> 단층의 연골 세포의 확장 <ol><li> Dulbeccos의 세포 확장을위한 각 T175 조직 배양 플라스크에 넣고 판 5 백만 인간의 연골 세포는 10 %의 혈청 및 1X 페니실린 - 스트렙토 마이신-글루타민 (PSG)으로 보충 이글스 중간 (DMEM)을 수정. 5 % CO 2를 포함하는 가습 공기와 37 ° C에서 배양 세포. 플라스크는 85 %의 합류 때까지 3 일마다 배지를 변경합니다. 같은 구절에서 세포를 사용합니다. </li></ol></li><li> 10 % W / V의 최종 농도로 PBS에 PEGDA을 녹여 0.05 % W / V의 최종 농도에 광개시제 I-2959에 추가 솔루션을 살균 필터. </li><li> 5 × 10 6 세포의 준비 PEGDA 용액에서 배양 된 연골 세포를 일시 중지 /ML. </li></ol> 3. 연골 조직의 인쇄 <ol><li> 프린터와 노트북을 켭니다. </li><li> Microsoft Word 또는 어도비 포토샵을 사용하여 직경 4 mm의 고체 원의 인쇄 패턴을 만듭니다. <ol><li> 패턴의 위치를​​ 조정하고 플라스틱 금형에 정확하게 인쇄 할 수 있는지 확인하십시오. </li><li> 지지체의 원하는 두께에 도달하기 위해 필요한 인쇄 매수를 계산한다. 높이 4mm의 경우, (220) 인쇄가 원하는 발판을 만들어야합니다. </li></ol></li><li> 잉크 카트리지에 bioink를 넣습니다. 인쇄 중에 직접 UV의 노출로부터 보호하기 위해 알루미늄 호일로 카트리지를 커버. </li><li> 프린터로 인쇄 명령을 보냅니다. 프린터가 인쇄를 시작할 때 용지 센서를 당깁니다. 전체 인쇄 과정은 직경 4 ㎜, 높이 4 mm의 발판 미만 4 분 소요됩니다. </li><li> 전송은 24 웰 플레이트에 neocartilage을 인쇄하고 각 웰에 1.5 ml의 문화 매체를 추가 할 수 있습니다. </li></ol><ol><li> 어둠 속에서 15 분 동안 실온에서 LIVE / DEAD 생존 / 세포 독성 작업 솔루션에서 인쇄 neocartilage을 품어. </li><li> 반 세포 함유 하이드로 겔을 절단하고 절단 영역의 형광 이미지를 가지고. </li><li> 다섯 무작위로 촬영 한 이미지에 눈이 먼 관찰자 (녹색)과 죽음 (적색) 세포 라이브 계산합니다. 살아있는 및 죽은 세포의 총 수에 의해 생균 수를 분할함으로써 세포 생존도를 계산한다. </li></ol>

<ol>
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D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Direct+human+cartilage+repair+using+three-dimensional+bioprinting+technology.">Direct human cartilage repair using three-dimensional bioprinting technology.</a> Tissue Eng Part A. 18, 1304-1312 (2012).</li><li>Cui, X., Breitenkamp, K., Lotz, M., D'Lima, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Synergistic+action+of+fibroblast+growth+factor-2+and+transforming+growth+factor-beta1+enhances+bioprinted+human+neocartilage+formation.">Synergistic action of fibroblast growth factor-2 and transforming growth factor-beta1 enhances bioprinted human neocartilage formation.</a> Biotechnol. Bioeng. 109, 2357-2368 (2012).</li><li>Cui, X., Breitenkamp, K., Finn, M. G., Lotz, M., Colwell, C. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Direct+human+cartilage+repair+using+thermal+inkjet+printing+technology.">Direct human cartilage repair using thermal inkjet printing technology.</a> Osteoarthritis and Cartilage. 19, (2011).</li><li>Cui, X., Boland, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Simultaneous+deposition+of+human+microvascular+endothelial+cells+and+biomaterials+for+human+microvasculature+fabrication+using+inkjet+printing.">Simultaneous deposition of human microvascular endothelial cells and biomaterials for human microvasculature fabrication using inkjet printing.</a> NIP24/digital Fabrication 2008: 24th International Conference on Digital Printing Technologies, Technical Program and Proceedings. 24, 480-483 (2008).</li><li>Cui, X., Dean, D., Ruggeri, Z. M., Boland, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cell+damage+evaluation+of+thermal+inkjet+printed+Chinese+hamster+ovary+cells.">Cell damage evaluation of thermal inkjet printed Chinese hamster ovary cells.</a> Biotechnol. Bioeng. 106, 963-969 (2010).</li><li>Cui, X., Hasegawa, A., Lotz, M., D'Lima, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Structured+three-dimensional+co-culture+of+mesenchymal+stem+cells+with+meniscus+cells+promotes+meniscal+phenotype+without+hypertrophy.">Structured three-dimensional co-culture of mesenchymal stem cells with meniscus cells promotes meniscal phenotype without hypertrophy.</a> Biotechnol. Bioeng. 109, 2369-2380 (2012).</li><li>Cui, X., Gao, G., Qiu, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Accelerated+myotube+formation+using+bioprinting+technology+for+biosensor+applications.">Accelerated myotube formation using bioprinting technology for biosensor applications.</a> Biotechnol. Lett. 35, 315-321 (2013).</li><li>Cui, X., Boland, T., D'Lima, D. D., Lotz, M. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Thermal+inkjet+printing+in+tissue+engineering+and+regenerative+medicine.">Thermal inkjet printing in tissue engineering and regenerative medicine.</a> Recent Pat Drug Deliv Formul. 6, 149-155 (2012).</li><li>Bryant, S. J., Anseth, K. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Hydrogel+properties+influence+ECM+production+by+chondrocytes+photoencapsulated+in+poly(ethylene+glycol)+hydrogels.">Hydrogel properties influence ECM production by chondrocytes photoencapsulated in poly(ethylene glycol) hydrogels.</a> Journal of Biomedical Materials Research. 59, 63-72 (2002).</li><li>Elisseeff, J., 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=Photoencapsulation+of+chondrocytes+in+poly(ethylene+oxide)-based+semi-interpenetrating+networks.">Photoencapsulation of chondrocytes in poly(ethylene oxide)-based semi-interpenetrating networks.</a> Journal of Biomedical Materials Research. 51, 164-171 (2000).</li><li>Buskirk, W. A., 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=Development+of+A+High-Resolution+Thermal+Inkjet+Printhead.">Development of A High-Resolution Thermal Inkjet Printhead.</a> Hewlett-Packard Journal. 39, 55-61 (1988).</li><li>Harmon, J. P., Widder, J. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Integrating+the+Printhead+Into+the+HP+Deskjet+Printer.">Integrating the Printhead Into the HP Deskjet Printer.</a> Hewlett-Packard Journal. 39, 62-66 (1988).</li><li>Kim, T. K., 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=Experimental+model+for+cartilage+tissue+engineering+to+regenerate+the+zonal+organization+of+articular+cartilage.">Experimental model for cartilage tissue engineering to regenerate the zonal organization of articular cartilage.</a> Osteoarthritis and Cartilage. 11, 653-664 (2003).</li><li>Sharma, B., 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=Designing+zonal+organization+into+tissue-engineered+cartilage.">Designing zonal organization into tissue-engineered cartilage.</a> Tissue Engineering. 13, 405-414 (2007).</li><li>Cohen, D. L., Lipton, J. I., Bonassar, L. J., Lipson, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Additive+manufacturing+for+in+situ+repair+of+osteochondral+defects.">Additive manufacturing for in situ repair of osteochondral defects.</a> Biofabrication. 2, (2010).</li></ol>

저자는이 연구에 재정적 인 관심이 없다.

동시 광중합 용량이 3D bioprinting 시스템은 셀룰러 알지네이트 하이드로 겔 (16)을 압출 주사기를 사용하여 골 연골 결함 시츄 프린팅 소재의 최상의 이전에보고 된 방법보다 상당히 큰 인쇄 해상도를 제공한다. 높은 인쇄 해상도는 해부학 연골 띠 조직을 복원하는 연골 조직 공학에 특히 중요합니다. 계층 별 조립시 동시 광중합는 3D 건설을위한 세포 및 생체 재료 비계의 정확한 침착...

열 잉크젯 프린팅에 근거 Bioprinting은 조직 공학과 재생 의학 분야에서 가장 유망한 지원 기술의 하나이다. 디지털 제어 및 높은 처리량 프린트 헤드와 세포 골격 및 성장 인자는 정확히 3 차원 (3D) 위치를 빠르게 원하는 이차원 (2D)로 증착 될 수있다. 많은 성공적인 응용 프로그램은 조직 공학 및 재생 의학 1-9에서이 기술을 사용하여 달성되었습니다. 본 논문에서는 bioprinting 플랫폼은 수정 된 휴렛 팩커드 (HP) 데스크젯 500 열 잉크젯 프린터와 동시에 광중합 시스템과 함께 설립되었다. 폴리 (에틸렌 글리콜) (PEG)에서 제형 화 합성 하이드로 겔은 연골 세포 생존을 유지하고 연골 ECM 생산 10,11 홍보의 용량을 보여 주었다. 또한, photocrosslinkable PEG 동시 고분자 재료에 이상적 낮은 점도, 물에 매우 용해3D의 bioprinting 동안 rization. 본 논문에서는 폴리 (에틸렌) 글리콜 디 아크릴 레이트 (PEGDA, MW 3,400)에 현탁 인간의 연골 세포는 정확하게 3D 해상도 1,400 dpi로와 neocartilage 계층별로 레이어를 구성하는 인쇄되었다. 3 차원 지지체에 증착 된 세포 균질 분포는 우수한 기계적 특성 및 향상된 ECM 생산하여 연골 조직을 생성하는 관찰되었다. 대조적으로, 수동 조립에 의한 배양 2,3 불균일 연골 형성되었다 느린 비계 중합, 대신 그들의 초기 증착 위치의 겔의 바닥에 축적 된 세포.

<table border="0" cellpadding="0" cellspacing="0">
	<tbody>
		<tr height="22">
			<td height="22" style="height:22px;width:357px;">HP Deskjet 500 thermal inkjet printer</td>
			<td style="width:221px;">Hewlett-Packard</td>
			<td style="width:195px;">C2106a</td>
			<td style="width:433px;">Discontinued. Purchased refurbished from internet vendor.</td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:357px;">HP black ink cartridge</td>
			<td style="width:221px;">Hewlett-Packard</td>
			<td style="width:195px;">51626a</td>
			<td style="width:433px;"></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:357px;">Ultraviolet lamp</td>
			<td style="width:221px;">UVP</td>
			<td style="width:195px;">B-100AP</td>
			<td style="width:433px;"></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:357px;">UV light meter</td>
			<td style="width:221px;">General Tools</td>
			<td style="width:195px;">UV513AB</td>
			<td style="width:433px;"></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:357px;">Zeiss LSM 510 laser scanning microscope</td>
			<td style="width:221px;">Carl Zeiss</td>
			<td style="width:195px;">LSM 510</td>
			<td style="width:433px;"></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:357px;">Dulbeccos Modified Eagles Medium (DMEM)</td>
			<td style="width:221px;">Mediatech</td>
			<td style="width:195px;">10-013</td>
			<td style="width:433px;"></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:357px;">Penicillin-streptomycin-glutamine (PSG)</td>
			<td style="width:221px;">Invitrogen</td>
			<td style="width:195px;">10378-016</td>
			<td style="width:433px;"></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:357px;">Accutase cell dissociation reagent</td>
			<td style="width:221px;">Invitrogen</td>
			<td style="width:195px;">A11105-01</td>
			<td style="width:433px;"></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:357px;">Phosphate buffered saline (PBS)</td>
			<td style="width:221px;">Invitrogen</td>
			<td style="width:195px;">10010-023</td>
			<td style="width:433px;"></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:357px;">Live/Dead viability/cytotoxicity Kit</td>
			<td style="width:221px;">Invitrogen</td>
			<td style="width:195px;">L-3224</td>
			<td style="width:433px;"></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:357px;">Poly(ethylene glycol) diacrylate (PEGDA)</td>
			<td style="width:221px;">Glycosan Biosystems</td>
			<td style="width:195px;">GS700</td>
			<td style="width:433px;"></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:357px;">Irgacure 2959</td>
			<td style="width:221px;">Ciba Specialty Chemicals</td>
			<td style="width:195px;">I-2959</td>
			<td style="width:433px;"></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:357px;">Human articular chondrocytes</td>
			<td style="width:221px;">Lonza</td>
			<td style="width:195px;">CC-2550</td>
			<td style="width:433px;"></td>
		</tr>
	</tbody>
</table>

human cartilage tissue fabrication using three-dimensional inkjet printing technology

열 잉크젯 프린팅을 기반으로 Bioprinting는 조직 공학 및 재생 의학 분야에서 가장 매력적인 지원 기술 중 하나입니다. 디지털 제어 세포 골격 및 성장 인자는 정확히 3 차원 (3D) 위치 빠르게 원하는 이차원 (2D)로 증착 될 수로. 따라서,이 기술은 그 나라의 해부학 적 구조를 흉내 낸 조직을 조작 할 수있는 이상적인 방법입니다. 네이티브 존 조직, 세포 외 매트릭스 조성물 (ECM), 그리고 기계적 성질과 연골을 설계하기 위해, 우리는 3D 연골 조직 공학을위한 가능한 동시 광중합와 상용 잉크젯 프린터를 사용 bioprinting 플랫폼을 개발했다. 폴리 (에틸렌 글리콜) 디 아크릴 레이트 (PEGDA)에 현탁 인간의 연골 세포는 계층 별 어셈블리를 통해 3D neocartilage 건설에 인쇄되었다. 인쇄 균체 surro 지원 그들의 본래 증착 위치에 고정했다동시에 광중합에 발판을 unding. 인쇄 조직의 기계적 성질은 네이티브 연골 유사 하였다. 이상 자외선 노출을 필요로 기존의 조직 제조에 비해, 동시에 광중합 인쇄 된 세포의 생존 능력이 유의하게 높았다. 인쇄 neocartilage 우수한 글리코 사 미노 글리 칸 (GAG)과 유전자 발현과 일치했다 콜라겐 타입 II의 생산을 보여 주었다. 따라서이 플랫폼은 해부학 적 조직 공학에 대한 정확한 세포의 분포와 배치에 이상적입니다.

수정 된 열 잉크젯 프린터는 높은 처리량 및 우수한 세포 생존 능력의 세포와 비계 증착 할 수 있었다. 동시에 광중합 및 감광 바이오 물질과 결합,이 기술은 처음에 퇴적 위치로 세포와 다른 인쇄 물질을 고정 할 수 있습니다. 개질 열 잉크젯 프린터의 속성에 따라, 2D 인쇄 해상도는 130 (PL)의 하나의 잉크 방울 체적 300 dpi로 하였다. 3.6 kHz에서 발사 주파수 (12, 13)과 각각의 프린트 헤드 (50) ?...

Watch this Scientific Journal Video about Human Cartilage Tissue Fabrication Using Three-dimensional Inkjet Printing Technology at JoVE.com

Human Cartilage Tissue Fabrication Using Three-dimensional Inkjet Printing Technology

이 문서에 설명 된 방법은 동시 UV 중합과 bioprinter에 상용 잉크젯 프린터로 변환하는 방법을 보여줍니다. 프린터는 세포 및 생체 재료와 3 차원 조직 구조를 구축 할 수있다. 여기에 설명이 연구는 3D neocartilage을 건설했다.

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 ...

human-cartilage-tissue-fabrication-using-three-dimensional-inkjet

Research

JoVE Journal

Bioengineering

15.6K 조회수.  Rensselaer Polytechnic Institute.  이 문서에 설명 된 방법은 동시 UV 중합과 bioprinter에 상용 잉크젯 프린터로 변환하는 방법을 보여줍니다. 프린터는 세포 및 생체 재료와 3 차원 조직 구조를 구축 할 수있다. 여기에 설명이 연구는 3D neocartilage을 건설했다. 

비디오: Human Cartilage Tissue Fabrication Using Three-dimensional Inkjet Printing Technology

Planar and Three-Dimensional Printing of Conductive Inks

Printed electronics rely on low-cost, large-area fabrication routes to create flexible or multidimensional electronic, optoelectronic, and biomedical devices1-3. In this paper, we focus on one- (1D), two- (2D), and three-dimensional (3D) printing of conductive metallic inks in the form of flexible, stretchable, and spanning microelectrodes.
Direct-write assembly4,5 is a 1-to-3D printing technique that enables the fabrication of features ranging from simple lines to complex structures by the deposition of concentrated inks through fine nozzles (~0.1 - 250 &mu;m). This printing method consists of a computer-controlled 3-axis translation stage, an ink reservoir and nozzle, and 10x telescopic lens for visualization. Unlike inkjet printing, a droplet-based process, direct-write assembly involves the extrusion of ink filaments either in- or out-of-plane. The printed filaments typically conform to the nozzle size. Hence, microscale features (&lt; 1 &mu;m) can be patterned and assembled into larger arrays and multidimensional architectures.
In this paper, we first synthesize a highly concentrated silver nanoparticle ink for planar and 3D printing via direct-write assembly. Next, a standard protocol for printing microelectrodes in multidimensional motifs is demonstrated. Finally, applications of printed microelectrodes for electrically small antennas, solar cells, and light-emitting diodes are highlighted.

Planar and three-dimensional printing of conductive metallic inks is described. Our approach provides new avenues for fabricating printed electronic, optoelectronic, and biomedical devices in unusual layouts at the microscale.

전도성 잉크의 평면 및 3 차원 인쇄

Printed electronics rely on low-cost, large-area fabrication routes to create flexible or multidimensional electronic, optoelectronic, and biomedical ...

연구

생체공학

Generation of a Three-dimensional Full Thickness Skin Equivalent and Automated Wounding

In vitro models are a cost effective and ethical alternative to study cutaneous wound healing processes. Moreover, by using human cells, these models reflect the human wound situation better than animal models. Although two-dimensional models are widely used to investigate processes such as cellular migration and proliferation, models that are more complex are required to gain a deeper knowledge about wound healing. Besides a suitable model system, the generation of precise and reproducible wounds is crucial to ensure comparable results between different test runs. In this study, the generation of a three-dimensional full thickness skin equivalent to study wound healing is shown. The dermal part of the models is comprised of human dermal fibroblast embedded in a rat-tail collagen type I hydrogel. Following the inoculation with human epidermal keratinocytes and consequent culture at the air-liquid interface, a multilayered epidermis is formed on top of the models. To study the wound healing process, we additionally developed an automated wounding device, which generates standardized wounds in a sterile atmosphere.

The goal of this protocol is to build up a three-dimensional full thickness skin equivalent, which resembles natural skin. With a specifically constructed automated wounding device, precise and reproducible wounds can be generated under maintenance of sterility.

3 차원 전체 두께 피부의 세대 동등한 및 자동화 부상

In vitro models are a cost effective and ethical alternative to study cutaneous wound healing processes. Moreover, by using human cells, these models ...

Three-dimensional Biomimetic Technology: Novel Biorubber Creates Defined Micro- and Macro-scale Architectures in Collagen Hydrogels

Tissue scaffolds play a crucial role in the tissue regeneration process. The ideal scaffold must fulfill several requirements such as having proper composition, targeted modulus, and well-defined architectural features. Biomaterials that recapitulate the intrinsic architecture of in vivo tissue are vital for studying diseases as well as to facilitate the regeneration of lost and malformed soft tissue. A novel biofabrication technique was developed which combines state of the art imaging, three-dimensional (3D) printing, and selective enzymatic activity to create a new generation of biomaterials for research and clinical application. The developed material, Bovine Serum Albumin rubber, is reaction injected into a mold that upholds specific geometrical features. This sacrificial material allows the adequate transfer of architectural features to a natural scaffold material. The prototype consists of a 3D collagen scaffold with 4 and 3 mm channels that represent a branched architecture. This paper emphasizes the use of this biofabrication technique for the generation of natural constructs. This protocol utilizes a computer-aided software (CAD) to manufacture a solid mold which will be reaction injected with BSA rubber followed by the enzymatic digestion of the rubber, leaving its architectural features within the scaffold material.

An innovative biofabrication technique was developed to engineer three-dimensional constructs that resemble the architectural features, components, and mechanical properties of in vivo tissue. This technique features a newly developed sacrificial material, BSA rubber, which transfers detailed spatial features, reproducing the in vivo architectures of a wide variety of tissues.

세 가지 차원 생체 모방 기술 ​​: 소설 Biorubber는 콜라겐 하이드로 겔에 마이크로 및 매크로 스케일 아키텍처를 정의 작성합니다

Tissue scaffolds play a crucial role in the tissue regeneration process. The ideal scaffold must fulfill several requirements such as having proper ...

Fabrication of Inverted Colloidal Crystal Poly(ethylene glycol) Scaffold: A Three-dimensional Cell Culture Platform for Liver Tissue Engineering

The ability to maintain hepatocyte function in vitro, for the purpose of testing xenobiotics&#39; cytotoxicity, studying virus infection and developing drugs targeted at the liver, requires a platform in which cells receive proper biochemical and mechanical cues. Recent liver tissue engineering systems have employed three-dimensional (3D) scaffolds composed of synthetic or natural hydrogels, given their high water retention and their ability to provide the mechanical stimuli needed by the cells. There has been growing interest in the inverted colloidal crystal (ICC) scaffold, a recent development, which allows high spatial organization, homotypic and heterotypic cell interaction, as well as cell-extracellular matrix (ECM) interaction. Herein, we describe a protocol to fabricate the ICC scaffold using poly (ethylene glycol) diacrylate (PEGDA) and the particle leaching method. Briefly, a lattice is made from microsphere particles, after which a pre-polymer solution is added, properly polymerized, and the particles are then removed, or leached, using an organic solvent (e.g., tetrahydrofuran). The dissolution of the lattice results in a highly porous scaffold with controlled pore sizes and interconnectivities that allow media to reach cells more easily. This unique structure allows high surface area for the cells to adhere to as well as easy communication between pores, and the ability to coat the PEGDA ICC scaffold with proteins also shows a marked effect on cell performance. We analyze the morphology of the scaffold as well as the hepatocarcinoma cell (Huh-7.5) behavior in terms of viability and function to explore the effect of ICC structure and ECM coatings. Overall, this paper provides a detailed protocol of an emerging scaffold that has wide applications in tissue engineering, especially liver tissue engineering.

Fabrication of Inverted Colloidal Crystal Polyethylene glycol Scaffold: A Three-dimensional Cell Culture Platform for Liver Tissue Engineering

This manuscript presents a detailed protocol for the fabrication of an emerging three-dimensional hepatocyte culture platform, the inverted colloidal crystal scaffold, and the concomitant techniques to assess hepatocyte behavior. The size-controllable pores, interconnectivity and ability to conjugate extracellular matrix proteins to the poly(ethylene glycol) (PEG) scaffold enhance Huh-7.5 cell performance.

반전 콜로이드 결정체 폴리 (에틸렌 글리콜) 비계의 제작 : 간 조직 공학을위한 삼차원 세포 배양 플랫폼

The ability to maintain hepatocyte function in vitro, for the purpose of testing xenobiotics&#39; cytotoxicity, studying virus infection and developing ...

Fabrication of Three-dimensional Paper-based Microfluidic Devices for Immunoassays

Paper wicks fluids autonomously due to capillary action. By patterning paper with hydrophobic barriers, the transport of fluids can be controlled and directed within a layer of paper. Moreover, stacking multiple layers of patterned paper creates sophisticated three-dimensional microfluidic networks that can support the development of analytical and bioanalytical assays. Paper-based microfluidic devices are inexpensive, portable, easy to use, and require no external equipment to operate. As a result, they hold great promise as a platform for point-of-care diagnostics. In order to properly evaluate the utility and analytical performance of paper-based devices, suitable methods must be developed to ensure their manufacture is reproducible and at a scale that is appropriate for laboratory settings. In this manuscript, a method to fabricate a general device architecture that can be used for paper-based immunoassays is described. We use a form of additive manufacturing (multi-layer lamination) to prepare devices that comprise multiple layers of patterned paper and patterned adhesive. In addition to demonstrating the proper use of these three-dimensional paper-based microfluidic devices with an immunoassay for human chorionic gonadotropin (hCG), errors in the manufacturing process that may result in device failures are discussed. We expect this approach to manufacturing paper-based devices will find broad utility in the development of analytical applications designed specifically for limited-resource settings.

We detail a method to fabricate three-dimensional paper-based microfluidic devices for use in the development of immunoassays. Our approach to device assembly is a type of multilayer, additive manufacturing. We demonstrate a sandwich immunoassay to provide representative results for these types of paper-based devices.

면역 측정법에 대한 3 차원 종이 기반 미세 유체 장치의 제작

Paper wicks fluids autonomously due to capillary action. By patterning paper with hydrophobic barriers, the transport of fluids can be controlled and ...

Melt Electrospinning Writing of Three-dimensional Poly(&#949;-caprolactone) Scaffolds with Controllable Morphologies for Tissue Engineering Applications

This tutorial reflects on the fundamental principles and guidelines for electrospinning writing with polymer melts, an additive manufacturing technology with great potential for biomedical applications. The technique facilitates the direct deposition of biocompatible polymer fibers to fabricate well-ordered scaffolds in the sub-micron to micro scale range. The establishment of a stable, viscoelastic, polymer jet between a spinneret and a collector is achieved using an applied voltage and can be direct-written. A significant benefit of a typical porous scaffold is a high surface-to-volume ratio which provides increased effective adhesion sites for cell attachment and growth. Controlling the printing process by fine-tuning the system parameters enables high reproducibility in the quality of the printed scaffolds. It also provides a flexible manufacturing platform for users to tailor the morphological structures of the scaffolds to their specific requirements. For this purpose, we present a protocol to obtain different fiber diameters using melt electrospinning writing (MEW) with a guided amendment of the parameters, including flow rate, voltage and collection speed. Furthermore, we demonstrate how to optimize the jet, discuss often experienced technical challenges, explain troubleshooting techniques and showcase a wide range of printable scaffold architectures.

Melt Electrospinning Writing of Three-dimensional Poly(&amp;#949;-caprolactone) Scaffolds with Controllable Morphologies for Tissue Engineering Applications

This protocol serves as a comprehensive guideline to fabricate scaffolds via electrospinning with polymer melts in a direct writing mode. We systematically outline the process and define the appropriate parameter settings for achieving targeted scaffold architectures.

조직 공학 응용 프로그램에 대 한 제어 형태학과 3 차원 Poly(ε-caprolactone) 장비의 전기 글 녹아

This tutorial reflects on the fundamental principles and guidelines for electrospinning writing with polymer melts, an additive manufacturing technology ...

Culturing Mammalian Cells in Three-dimensional Peptide Scaffolds

A useful technique for culturing cells in a self-assembling nanofiber three-dimensional (3D) scaffold is described. This culture system recreates an environment that closely mimics the structural features of non-polarized tissue. Furthermore, the particular intrinsic nanofiber structure of the scaffold makes it transparent to visual light, which allows for easy visualization of the sample under microscopy. This advantage was largely used to study cell migration, organization, proliferation, and differentiation and thus any development of their particular cellular function by staining with specific dyes or probes. Furthermore, in this work, we describe the good performance of this system to easily study the redifferentiation of expanded human articular chondrocytes into cartilaginous tissue. Cells were encapsulated into self-assembling peptide scaffolds and cultured under specific conditions to promote chondrogenesis. Three-dimensional cultures showed good viability during the 4 weeks of the experiment. As expected, samples cultured with chondrogenic inducers (compared to non-induced controls) stained strongly positive for toluidine blue (which stains glycosaminoglycans (GAGs) that are highly present in cartilage extracellular matrix) and expressed specific molecular markers, including collagen type I, II and X, according to Western Blot analysis. This protocol is easy to perform and can be used at research laboratories, industries and for educational purposes in laboratory courses.

Here, we present a protocol to obtain 3D-culture systems in self-assembling peptide scaffolds to promote the differentiation of dedifferentiated human articular chondrocytes into cartilage-like tissue.

3 차원 펩 티 드 건설 기계에 포유류 세포 배양

A useful technique for culturing cells in a self-assembling nanofiber three-dimensional (3D) scaffold is described. This culture system recreates an ...

Three-dimensional Printing of Thermoplastic Materials to Create Automated Syringe Pumps with Feedback Control for Microfluidic Applications

Microfluidics has become a critical tool in research across the biological, chemical, and physical sciences. One important component of microfluidic experimentation is a stable fluid handling system capable of accurately providing an inlet flow rate or inlet pressure. Here, we have developed a syringe pump system capable of controlling and regulating the inlet fluid pressure delivered to a microfluidic device. This system was designed using low-cost materials and additive manufacturing principles, leveraging three-dimensional (3D) printing of thermoplastic materials and off-the-shelf components whenever possible. This system is composed of three main components: a syringe pump, a pressure transducer, and a programmable microcontroller. Within this paper, we detail a set of protocols for fabricating, assembling, and programming this syringe pump system. Furthermore, we have included representative results that demonstrate high-fidelity, feedback control of inlet pressure using this system. We expect this protocol will allow researchers to fabricate low-cost syringe pump systems, lowering the entry barrier for the use of microfluidics in biomedical, chemical, and materials research.

Here we present a protocol to construct a pressure-controlled syringe pump to be used in microfluidic applications. This syringe pump is made from an&#160;additively manufactured body, off-the-shelf hardware, and open-source electronics. The resulting system is low-cost, straightforward to build, and delivers well-regulated fluid flow to enable rapid microfluidic research.

인쇄를 만드는 열가 소성 재료의 3 차원 미세 응용 프로그램에 대 한 피드백 제어와 주사기 펌프를 자동

Microfluidics has become a critical tool in research across the biological, chemical, and physical sciences. One important component of microfluidic ...

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 ...

Pancreatic Tissue-Derived Extracellular Matrix Bioink for Printing 3D Cell-Laden Pancreatic Tissue Constructs

The transplantation of pancreatic islets is a promising treatment for patients who suffer from type 1 diabetes accompanied by hypoglycemia and secondary complications. However, islet transplantation still has several limitations such as the low viability of transplanted islets due to poor islet engraftment and hostile environments. In addition, the insulin-producing cells differentiated from human pluripotent stem cells have limited ability to secrete sufficient hormones that can regulate the blood glucose level; therefore, improving the maturation by culturing cells with proper microenvironmental cues is strongly required. In this article, we elucidate protocols for preparing a pancreatic tissue-derived decellularized extracellular matrix (pdECM) bioink to provide a beneficial microenvironment that can increase glucose sensitivity of pancreatic islets, followed by describing the processes for generating 3D pancreatic tissue constructs using a microextrusion-based bioprinting technique.

Decellularized extracellular matrix (dECM) can provide suitable microenvironmental cues to recapitulate the inherent functions of target tissues in an engineered construct. This article elucidates the protocols for the decellularization of pancreatic tissue, evaluation of pancreatic tissue-derived dECM bioink, and generation of 3D pancreatic tissue constructs using a bioprinting technique.

인쇄 3D 세포 -라덴 췌장 조직 구성을 인쇄를위한 췌장 조직 유래 세포 외 매트릭스 바이오 잉크

The transplantation of pancreatic islets is a promising treatment for patients who suffer from type 1 diabetes accompanied by hypoglycemia and secondary ...