이 작품 재정적으로 협동 연구 센터 CRC에 의해 세포 치료 제조, 첨가제 Biomanufacturing와 뮌헨의 기술적인 대학의 고급 연구를 위한 연구소에서 호주 연구 협의회 아크 센터를 지원 하고있다. 이 연구는 호주 연구 협의회 산업 변환 훈련 센터 첨가제 Biomanufacturing http://www.additivebiomanufacturing.org (IC160100026)에 의해 실시 됐다. 기사, 책, 텔레비전 이나 라디오 프로그램, 전자 매체, 또는 프로젝트에 관련 된 다른 문학 작품에 대 한 사이트를 방문 하시기 바랍니다. 또한, 저자 기꺼이 인정 마리아 Flandes Iparraguirre 촬영, 필립 허바드는 음성에 대 한에 Luise Grossmann 지원에 대 한 촬영과 편집에 대 한.

1. 재료 준비
<ol>
	<li>깔때기와 3 mL 플라스틱 주사기에 PCL의 2 세대를 오픈 엔드에 피스톤을 삽입.</li>
	<li>8 헤 포인트 팁 위쪽으로 개통 집계 가까이에 공기 방울 수 있도록 65 ° C에서 preheated 오븐에 주사기를 놓습니다.</li>
	<li>녹은 물자 안에서 덫을 놓은 공기를 분리 하는 얇은 개체와 피스톤을 밀어.</li>
	<li>그것은 10 분 후에 더 이상 고분자 투명 하면 달성 된다 실내 온도 아래로 멋진 보자.</li>
	<li>그것은 사용 될 때까지 건조 하 고 어두운 환경에서 실 온에서 PCL 사전 로드 된 주사기를 저장 합니다.</li>
</ol>
2. 하드웨어 및 소프트웨어 설치
<ol>
	<li>주사기와 공기 압력 시스템에 주사기를 연결 하는 다른 끝에 배럴 어댑터를 23 G 플랫 흘린된 바늘 (spinneret)를 연결 합니다.</li>
	<li>인쇄 머리에 주사기를 놓고 머리의 하단 측면에 황동 부분에서 1mm 띄는 spinneret 팁 때까지 아래로 누릅니다.</li>
	<li>무대에서 수집을 탑재 하 고 표면 먼지 또는 잔류 폴리머 제거를 70% (vol/vol) 에탄올과 인쇄 헤드 청소.</li>
	<li>spinneret와 컬렉터 사이 12 m m 높은 개체를 배치 하 여 작동 거리를 설정 하 고 spinneret 팁은 단지 그것을 접촉 될 때까지 프린트 헤드를 내립니다.</li>
	<li>각각 spinneret 지역 및 주사기, 65 ° C 82 ° C와 전기 상자에 온도 조절기를 조정 하 고 용융은 PCL에 전원.</li>
	<li>폴리머 용 해 될 때까지 적어도 10 분 기다립니다 고 1.8 바 하는 레 귤 레이 터를 설정 하 여 공기 압력을 시작.</li>
	<li>크기와 모양, 간 멘 거리 및 발판의 레이어 및 프로세스의 컬렉션 속도의 수를 정의 하는 G-코드를 준비 합니다. 참고: 평평 하 고 관 건설 기계 조작에 대 한 자세한 템플릿은 토론 장 (표 2)에 제공 됩니다.</li>
	<li>수동으로 모든 접지 케이블 인클로저 및 벽 플러그에 안전 하 게 연결 되어 있는지 다시 확인 합니다.</li>
	<li>컴퓨터에 소프트웨어를 (예를 들어, 마 하 3)를 시작 하 고 준비 된 G-코드를 업로드.</li>
</ol>
3. 비 계 제조
<ol>
	<li>인클로저, 안전 내부 고정기를 연결 하 고 트리거는 spinneret에 높은 전압 공급의 전면 도어를 닫습니다. 참고: 일단 문을 열면 예를 들어 때 인쇄 완료 또는 비상시, 높은 전압 강하 및 발판 제거할 수 있습니다 안전 하 게.</li>
	<li>테일러 콘 형성 되 고 섬유 컬렉터 쪽으로 배출 될 때까지 0.2 k v 단계에서 점차 높은 전압을 증가 ( 그림 1D에 참조 모범적인 테일러 콘).</li>
	<li>폴리머 용융을 5 분 동안 운동 없이 제트를 안정화 하기 위해 여전히 수집기 접시에 돌출 될 수 있습니다. 새로운 인쇄를 시작 하기 전에 재료의 더미를 제거 합니다.</li>
	<li>G-코드 시작 됩니다 지점 위에 인쇄 헤드를 이동 하려면 키보드에서 커서를 사용 합니다.</li>
	<li>컴퓨터에 소프트웨어에서 G-코드를 시작 합니다.</li>
</ol>
4. 섬유 직경 조정
<ol>
	<li>작동 거리 (12mm) 및 온도 레 귤 레이 터 (82 ° C와 spinneret 지역 및 주사기, 65 ° C 각각) 일정 수준에 전에 설명한 대로에 단계 2.4 및 2.5. 참고: 다른 직경을 조정의 요약 표 3에서 제공 됩니다.</li>
	<li>(3-10 µ m) 작은 크기의 직경을 가진 섬유를 인쇄 합니다. 0.8 바 공기 압력 수준을 낮추기, 8에 적용 된 전압 kV 및 수집기 1700 m m/분 속도.</li>
	<li>매체와 인쇄 섬유 직경 크기의 (10-20 µ m). 공기 압력 조절 레벨 바, 11 설정된 전압 1.5 kV와 낮은 컬렉션-1200 m m/분 속도.</li>
	<li>큰 직경 (20-30 μ m)와 인쇄 섬유. 2.6 바 공기 압력 레벨 증가, 변경 12 적용된 전압 kV 및 감소 컬렉션 700 m m/분 속도.</li>
</ol>
5. 제트 최적화
<ol>
	<li>제트 강한 led 향상 된 가시성에 대 한 인클로저 외부에서 빛을 밝히는.</li>
	<li>1 분에 대 한 섬유의 동작을 관찰 하 고 작은 단계, 즉 0.1에서에서 프로세스를 최적화 하는 시스템 매개 변수를 조정 위해 적용된 전압, 100 mm/min 컬렉션 속도 공기 압력에 대 한 0.1 kV. 참고: 요약 표 4에서 주어진 다.</li>
	<li>정기적으로 공기 압력 감소, 속도 증가 및 섬유의 비행 경로 3 분 이상 안정적인 전차 곡선을 닮을 때까지 전압을 최소화 하 여 동작을 왜곡 안정화 합니다.</li>
	<li>전압을 증가, 공기 압력을 감소 시키고 수집기의 속도 줄여 제트 뒤에 후행의 비행 경로 수정 합니다. 섬유의 비행 경로 전차 곡선 모양으로 다시 이동 될 때까지 이러한 조치를 적용 합니다.</li>
	<li>섬유가 적용된 전압 감소, 수집기의 속도 증가 및 제트기의 비행 경로 유지 전차 곡선의 모양을 다시 때까지 공기 압력을 증가 하 여 컬렉터를 향해 수직으로 여행 하지 마십시오.</li>
</ol>
6. 비 계 컬렉션
<ol>
	<li>인쇄 완료 되 면 문을 열고 커서를 사용 하 여 더 나은 접근성에 대 한 문 쪽으로 수집가 격판덮개를 이동 합니다.</li>
	<li>에탄올 70% (vol/vol) 믹스와 발판을 스프레이 하 고 가시는 수집기에서 분리 될 때까지 10 초를 기다립니다.</li>
	<li>족집게와 인클로저에서 해제 잡아 한쪽 가장자리에 의해 완성 된 비 계를 수집 합니다.</li>
</ol>
7. 문제 해결
<ol>
	<li>적용된 전압이 감소 또는 청취 표시 spinneret 또는 크래킹 잡음 사이 불꽃 경우 즉시 문을 열어.</li>
	<li>잠재적인 점화 시 불 점화 수 있습니다으로 인클로저 내부에서 모든 유해 물질 및 에탄올 70% (vol/vol) 등 액체를 제거 합니다.</li>
	<li>프로그램 G-코드는 spinneret 모든 레이어 완료 후 발판을 인쇄 하는 지역에서 이동. 이 spinneret가 마지막으로 중지 하는 위치 지점 위에 소재 축적을 방지 합니다.</li>
	<li>확대경 아래 spinneret을 확인 하 고이 크게 테일러 콘의 균질 성에 영향을 미칠 것으로 spinneret 손상 인지 확인 합니다.</li>
</ol>

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W., 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=Additive+manufacturing+of+tissues+and+organs.">Additive manufacturing of tissues and organs.</a> Prog. in pol Sci. 37 (8), 1079-1104 (2012).</li><li>Hochleitner, G., 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=Fibre+pulsing+during+melt+electrospinning+writing.">Fibre pulsing during melt electrospinning writing.</a> BioNanoMaterials. 17, 159 (2016).</li><li>Persano, L., Camposeo, A., Tekmen, C., Pisignano, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Industrial+Upscaling+of+Electrospinning+and+Applications+of+Polymer+Nanofibers:+A+Review.">Industrial Upscaling of Electrospinning and Applications of Polymer Nanofibers: A Review.</a> Macromol. Mater. Eng. 298 (5), 504-520 (2013).</li><li>Wunner, F. M., 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=.">.</a> Comprehensive Biomaterials II. , 217-235 (2017).</li><li>Loh, Q. L., Choong, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Three-Dimensional+Scaffolds+for+Tissue+Engineering+Applications:+Role+of+Porosity+and+Pore+Size.">Three-Dimensional Scaffolds for Tissue Engineering Applications: Role of Porosity and Pore Size.</a> Tissue Eng Part B Rev. 19 (6), 485-502 (2013).</li><li>Ristovski, N., 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=Improved+fabrication+of+melt+electrospun+tissue+engineering+scaffolds+using+direct+writing+and+advanced+electric+field+control.">Improved fabrication of melt electrospun tissue engineering scaffolds using direct writing and advanced electric field control.</a> Biointerphases. 10 (1), 011006 (2015).</li><li>Wei, C., Dong, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Direct+fabrication+of+high-resolution+three-dimensional+polymeric+scaffolds+using+electrohydrodynamic+hot+jet+plotting.">Direct fabrication of high-resolution three-dimensional polymeric scaffolds using electrohydrodynamic hot jet plotting.</a> J Micromech Microeng. 23 (2), 025017 (2013).</li><li>Hacker, C., 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=Electrospinning+of+polymer+melt+:+steps+towards+an+upscaled+multi-jet+process.">Electrospinning of polymer melt : steps towards an upscaled multi-jet process.</a> , 71-76 (2009).</li><li>Nayak, R., Padhye, R., Kyratzis, I. L., Truong, Y. B., Arnold, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Recent+advances+in+nanofibre+fabrication+techniques.">Recent advances in nanofibre fabrication techniques.</a> Text. Res. J. 82 (2), 129-147 (2012).</li><li>Li, H., 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=Interjet+distance+in+needleless+melt+differential+electrospinning+with+umbellate+nozzles.">Interjet distance in needleless melt differential electrospinning with umbellate nozzles.</a> J. Appl. Polym. Sci. 131 (15), (2014).</li><li>Liao, S., 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=Effect+of+humidity+on+melt+electrospun+polycaprolactone+scaffolds.">Effect of humidity on melt electrospun polycaprolactone scaffolds.</a> BioNanoMaterials. 17, 173 (2016).</li><li>Detta, N., 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=Melt+electrospinning+of+polycaprolactone+and+its+blends+with+poly(ethylene+glycol).">Melt electrospinning of polycaprolactone and its blends with poly(ethylene glycol).</a> Polym Int. 59 (11), 1558-1562 (2010).</li><li>Hochleitner, G., H&#252;mmer, J. F., Luxenhofer, R., Groll, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=High+definition+fibrous+poly(2-ethyl-2-oxazoline)+scaffolds+through+melt+electrospinning+writing.">High definition fibrous poly(2-ethyl-2-oxazoline) scaffolds through melt electrospinning writing.</a> Polymer. 55 (20), 5017-5023 (2014).</li><li>Chen, F., 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=Additive+Manufacturing+of+a+Photo-Cross-Linkable+Polymer+via+Direct+Melt+Electrospinning+Writing+for+Producing+High+Strength+Structures.">Additive Manufacturing of a Photo-Cross-Linkable Polymer via Direct Melt Electrospinning Writing for Producing High Strength Structures.</a> Biomacromolecules. 17 (1), 208-214 (2016).</li><li>Hochleitner, G., 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=Additive+manufacturing+of+scaffolds+with+sub-micron+filaments+via+melt+electrospinning+writing.">Additive manufacturing of scaffolds with sub-micron filaments via melt electrospinning writing.</a> Biofabrication. 7 (3), 035002 (2015).</li></ol>

저자는 공개 없다.

의료 분야에서도 전에 대 한 혁신적인 솔루션을 찾기 위해 오전 통합 21st 세기에 대 한 새로운 패러다임을 제공 합니다. "바이오-제조"의 소위 필드 상승세 이며 제조 기술에서 혁신 테 응용 프로그램에 대 한 매우 정교한 아키텍처의 생산을 활성화. 직접 쓰기 모드 (여기 뮤)에 폴리머 용융 전기 볼 가장 유망한 제조 후보 중 하나로 테 커뮤니티의 요구를 준수 nanoscale에 미크론에 생체 재료?...

셀에 대 한 3 차원 (3D) 생체 구조 제조 조직에 첨가제 biomanufacturing의 주요 기여 중 하나입니다 공학 (테), 사용자 지정 된 생체, 세포, 생 화 확 적인 요인, 적용 하 여 조직 복원을 목표로 또는 그들의 조합입니다. 따라서, 건설 기계 테 응용 프로그램에 대 한 주요 요구 사항을 포함: 생체 재료, 타겟된 셀 침공에 대 한 제어 형태학 속성에 대 한 최적화 된 표면 속성에서 제조 셀 상호 작용 향상 1.
뮤 높게 주문한 ultrathin 섬유 형태학2첨가제 제조 (자주 라는 3D 인쇄) 및 폴리머 메시의 생산을 위한 전기의 원리를 결합 하는 용 매 없는 제조 기술입니다. 직접 쓰기 접근이 고 정확 하 게 프로그램 된 코드3, G-코드 라고 하는 섬유를 예금. 용 해 electrospun 구조는 현재 각각 다공성 평평 하 고 관 건설 기계를 조작 하는 플랫4,5 또는 굴대6,7 수집기를 사용 하 여 준비.
이 기술은 의료 학년 폴리머 poly(ε-caprolactone) (PCL), 우수한 생체 적합성8선물 등을 직접 인쇄 테 및 재생 의학 (RM) 커뮤니티 가능성 때문에 상당한 혜택을 제공 합니다. 다른 장점 높은 조직 방식으로 높은 표면 볼륨 비율의 건설 기계를 조작 하는 섬유를 입금 하 여 크기와 다공성의 분포를 사용자 지정할 수 있습니다. 뮤를 수행 하기 전에 폴리머는 먼저 열9의 응용 프로그램이 필요 합니다. 일단 액체 상태에서 적용 된 공기 압력 높은 전압 소스에 연결 된 금속 spinneret를 통해 밖으로 흘러 그것 강요 합니다. 표면 장력 및 접지 수집기에 정전 된 물방울의 매력 사이 힘 균형 테일러 콘 제트10의 방출 다음의 형성에 지도 한다.
이미지와이 프로토콜에 사용 되는 내부 빌드 뮤 장치의 그리기 회로도 그림 1에 표시 됩니다. 그것은 또한 단 열 테이프를 사용 하 여 난방 요소와는 spinneret 주변 전기 충전된 황동 부분 사이의 전기 방전을 방지 하기의 원리를 보여 줍니다. 부족 한 단열재는 구현 된 하드웨어의 내부 손상으로 이어질 것 이다.
3 시스템 매개 변수 (온도, 수집 속도 공기 압력)의 조정에 따라 뮤 토론 섹션에서 설명 하는 다른 직경을 가진 섬유의 제조 수 있습니다. 그러나 대부분의 경우에, 미세 조정 및 제트의 최적화가 됩니다 필요한 안정적인 제트 방출 될 것입니다 전에. 전기 여행 제트기의 시각화 과정의 일관성을 확인 하는 효과적인 방법입니다. 이상적인 경우, 비행 경로 시스템 매개 변수11에 의해 제어 되는 힘 균형 결과로 획득 전차 곡선을 닮았다. 또한, 마이크로-와 매크로-구조는 건설 기계 폴리머 제트12의 비행 경로에 따라 달라 집니다. 다른 편향 동작 및 최적화에 대 한 조치의 상세한 테이블 토론 섹션에서 제공 됩니다.
현재 연구에서 선물이 뮤 기술을 사용 하 여 높은 제어 섬유 장비의 제조에 대 한 제조 단계를 설명 하는 프로토콜. 이 작품, 의료 학년 PCL (분자량 95-140 kg/mol)이 의료 학년 PCL 기술 학년 이상의 순도 개선 하 고 그것의 기계 및 처리 속성은 뮤로 사용 되었다. PCL의 범위를 처리 하는 광범위 한 용융 그것의 낮은 융해 점 (60 ° C) 및 높은 열 안정성에서 유래. 또한, PCL 느린 속도 생 분해성 폴리머, 그것에 게 많은 조직 공학 응용 프로그램13에 대 한 훌륭한 소재 이다.
이 연구에 대 한 온도 및 수집기 거리 유지 됩니다 상수 (65 ° C와 주사기 및 spinneret의 온도 대 한 82 ° C (각각) 및 컬렉터 거리 12 mm); 그러나 적용 전압, 수집 속도 공기 압력,, 타겟된 직경을 가진 섬유를 조작 하 변화 될 것입니다. 뮤 건설 기계를 사용 하 여 게시 된 연구의 상세한 목록 결과 섹션에서 제공 되 고 테와 RM (표 1)의 분야에 대 한 다른 응용 프로그램을 보여.

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>Plastic syringe</td>
			<td>Nordson Australia Pty Ltd</td>
			<td>7012072</td>
			<td>EFD BARREL O 3mL Clear 50</td>
		</tr>
		<tr>
			<td>Medical grade Poly (&#949;-caprolactone) (mPCL)</td>
			<td>Corbion Purac, The Netherlands</td>
			<td>PURASORB PC12</td>
			<td></td>
		</tr>
		<tr>
			<td>23 GA needle</td>
			<td>Nordson Australia Pty Ltd</td>
			<td>7018302</td>
			<td>#23GP .013 X .25 ORANGE 50 PC</td>
		</tr>
		<tr>
			<td>Plunger</td>
			<td>Nordson Australia Pty Ltd</td>
			<td>7012166</td>
			<td>PISTON O 3mL WH WIPER 50</td>
		</tr>
		<tr>
			<td>Pressure adapter</td>
			<td>Nordson Australia Pty Ltd</td>
			<td>7012059</td>
			<td>ADAPTER ASM O 3mL BL 1.8M</td>
		</tr>
		<tr>
			<td>Aluminium collector</td>
			<td>Action Aluminium, Australia</td>
			<td>SHP2</td>
			<td>Sheet 5005 H34</td>
		</tr>
		<tr>
			<td>Acrylic glass</td>
			<td>Mulford Plastics Pty Ltd</td>
			<td>ACC6-13094</td>
			<td></td>
		</tr>
		<tr>
			<td>Mach 3 software</td>
			<td>Art Soft</td>
			<td></td>
			<td>Purchased online</td>
		</tr>
		<tr>
			<td>Safety switch interlock</td>
			<td>RS components Pty Ltd</td>
			<td>12621330</td>
			<td></td>
		</tr>
		<tr>
			<td>High voltage generator</td>
			<td>EMCO High Voltage Co.</td>
			<td>DX250R</td>
			<td></td>
		</tr>
		<tr>
			<td>Temperature controller</td>
			<td>WATLOW</td>
			<td>PM9R1FJ</td>
			<td></td>
		</tr>
		<tr>
			<td>X and Y positioning slide</td>
			<td>VELMEX Inc.</td>
			<td>XN-10-0020-M011</td>
			<td></td>
		</tr>
	</tbody>
</table>

melt electrospinning writing of three-dimensional poly(&#949;-caprolactone) scaffolds with controllable morphologies for tissue engineering applications

이 자습서의 기본 원리에 반영 하 고 전기 폴리머와 쓰기에 대 한 지침 녹는, 생명 의학 어플리케이션에 대 한 좋은 잠재력을 가진 첨가제 제조 기술. 기술은 마이크로 눈금 범위를 서브 미크론에서 정돈된 건설 기계를 조작 하는 생체 고분자 섬유의 직접 증 착을 촉진 한다. 안정, 점 탄성의 설립, 폴리머 jet는 spinneret와 컬렉터 사이 적용 되는 전압을 사용 하 여 이루어집니다 그리고 직접 작성 될 수 있습니다. 전형적인 다공성 발판의 중요 한 이점은 셀 첨부 파일 및 성장에 대 한 증가 효과적인 접착 사이트를 제공 하는 높은 표면 볼륨 비율입니다. 시스템 매개 변수를 조정 하 여 인쇄 프로세스를 제어 인쇄 장비의 품질에서 높은 재현성을 수 있습니다. 그것은 또한 사용자가 그들의 특정 요구 사항에 건설 기계 형태학 상 구조에 맞게 유연한 제조 플랫폼을 제공 합니다. 이 목적을 위해 우리 가이드 개정 매개 변수, 흐름 속도, 전압 및 수집 속도 등의 다른 섬유 직경 하 여 용융 전기 쓰기 (뮤)를 얻기 위해 프로토콜 제시. 또한, 제트 최적화 자주 경험된 기술 과제 토론, 문제 해결 기법을 설명 하 고 인쇄 비 계 아키텍처의 넓은 범위를 보여주는 하는 방법을 보여 줍니다.

컬렉션의 두 가지 방법은 일반적으로 평면 고 고 곡 괭이는 뮤에 사용 됩니다. 결과 아키텍처의 G-코드 (표 2), 소프트웨어에 의해 실행 되는 프로그램에 따라 달라 집니다.
플랫 컬렉션 편평한 수집가 적용 하는 가장 일반적인 방법을 참조 하며 미리 프로그램 된 G-코드를 참조 하는 자료의 직접 증 착...

Watch this Scientific Journal Video about Melt Electrospinning Writing of Three-dimensional PolyÃŽÂµ-caprolactone Scaffolds with Controllable Morphologies for Tissue Engineering Applications at JoVE.c

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

이 프로토콜은 폴리머와 전기를 통해 건설 기계를 조작 하는 포괄적인 지침 직접 쓰기 모드에 녹아 있습니다. 우리는 체계적으로 과정을 약술 하 고 타겟된 비 계 구조를 달성 하기 위한 적절 한 매개 변수 설정 정의.

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 ...

melt-electrospinning-writing-three-dimensional-poly-caprolactone

Research

JoVE Journal

Bioengineering

15.0K 조회수.  Queensland University of Technology (QUT). 이 프로토콜은 폴리머와 전기를 통해 건설 기계를 조작 하는 포괄적인 지침 직접 쓰기 모드에 녹아 있습니다. 우리는 체계적으로 과정을 약술 하 고 타겟된 비 계 구조를 달성 하기 위한 적절 한 매개 변수 설정 정의.

비디오: Melt Electrospinning Writing of Three-dimensional Poly(ε-caprolactone) Scaffolds with Controllable Morphologies for Tissue Engineering Applications

Elastomeric PGS Scaffolds in Arterial Tissue Engineering

Cardiovascular disease is one of the leading cause of mortality in the US and especially, coronary artery disease increases with an aging population and increasing obesity1. Currently, bypass surgery using autologous vessels, allografts, and synthetic grafts are known as a commonly used for arterial substitutes2. However, these grafts have limited applications when an inner diameter of arteries is less than 6 mm due to low availability, thrombotic complications, compliance mismatch, and late intimal hyperplasia3,4. To overcome these limitations, tissue engineering has been successfully applied as a promising alternative to develop small-diameter arterial constructs that are nonthrombogenic, robust, and compliant. Several previous studies have developed small-diameter arterial constructs with tri-lamellar structure, excellent mechanical properties and burst pressure comparable to native arteries5,6. While high tensile strength and burst pressure by increasing collagen production from a rigid material or cell sheet scaffold, these constructs still had low elastin production and compliance, which is a major problem to cause graft failure after implantation. Considering these issues, we hypothesized that an elastometric biomaterial combined with mechanical conditioning would provide elasticity and conduct mechanical signals more efficiently to vascular cells, which increase extracellular matrix production and support cellular orientation.
The objective of this report is to introduce a fabrication technique of porous tubular scaffolds and a dynamic mechanical conditioning for applying them to arterial tissue engineering. We used a biodegradable elastomer, poly (glycerol sebacate) (PGS)7 for fabricating porous tubular scaffolds from the salt fusion method. Adult primary baboon smooth muscle cells (SMCs) were seeded on the lumen of scaffolds, which cultured in our designed pulsatile flow bioreactor for 3 weeks. PGS scaffolds had consistent thickness and randomly distributed macro- and micro-pores. Mechanical conditioning from pulsatile flow bioreactor supported SMC orientation and enhanced ECM production in scaffolds. These results suggest that elastomeric scaffolds and mechanical conditioning of bioreactor culture may be a promising method for arterial tissue engineering.

Elastomeric PGS scaffolds with vascular smooth muscle cells cultured in a pulsatile flow bioreactor may lead to promising small-diameter arterial constructs with native ECM production in a relatively short culture period.

탄성 PGS는 아티 조직 공학 공사장 공중 발판

Cardiovascular disease is one of the leading cause of mortality in the US and especially, coronary artery disease increases with an aging population and ...

연구

생체공학

Preparation of 3D Fibrin Scaffolds for Stem Cell Culture Applications

Stem cells are found in naturally occurring 3D microenvironments in vivo, which are often referred to as the stem cell niche 1. Culturing stem cells inside of 3D biomaterial scaffolds provides a way to accurately mimic these microenvironments, providing an advantage over traditional 2D culture methods using polystyrene as well as a method for engineering replacement tissues 2. While 2D tissue culture polystrene has been used for the majority of cell culture experiments, 3D biomaterial scaffolds can more closely replicate the microenvironments found in vivo by enabling more accurate establishment of cell polarity in the environment and possessing biochemical and mechanical properties similar to soft tissue.3 A variety of naturally derived and synthetic biomaterial scaffolds have been investigated as 3D environments for supporting stem cell growth. While synthetic scaffolds can be synthesized to have a greater range of mechanical and chemical properties and often have greater reproducibility, natural biomaterials are often composed of proteins and polysaccharides found in the extracelluar matrix and as a result contain binding sites for cell adhesion and readily support cell culture. Fibrin scaffolds, produced by polymerizing the protein fibrinogen obtained from plasma, have been widely investigated for a variety of tissue engineering applications both in vitro and in vivo 4. Such scaffolds can be modified using a variety of methods to incorporate controlled release systems for delivering therapeutic factors 5. Previous work has shown that such scaffolds can be used to successfully culture embryonic stem cells and this scaffold-based culture system can be used to screen the effects of various growth factors on the differentiation of the stem cells seeded inside 6,7.
 This protocol details the process of polymerizing fibrin scaffolds from fibrinogen solutions using the enzymatic activity of thrombin. The process takes 2 days to complete, including an overnight dialysis step for the fibrinogen solution to remove citrates that inhibit polymerization. These detailed methods rely on fibrinogen concentrations determined to be optimal for embryonic and induced pluripotent stem cell culture. Other groups have further investigated fibrin scaffolds for a wide range of cell types and applications - demonstrating the versatility of this approach 8-12.

This work details the preparation of 3D fibrin scaffolds for culturing and differentiating plutipotent stem cells. Such scaffolds can be used to screen the effects of various biological compounds on stem cell behavior as well as modified to contain drug delivery systems.

3D 섬유소의 준비는 줄기 세포 배양 용도 공사장 공중 발판

Stem cells are found in naturally occurring 3D microenvironments in vivo, which are often referred to as the stem cell niche 1. Culturing stem cells ...

Self-reporting Scaffolds for 3-Dimensional Cell Culture

Culturing cells in 3D on appropriate scaffolds is thought to better mimic the in vivo microenvironment and increase cell-cell interactions. The resulting 3D cellular construct can often be more relevant to studying the molecular events and cell-cell interactions than similar experiments studied in 2D. To create effective 3D cultures with high cell viability throughout the scaffold the culture conditions such as oxygen and pH need to be carefully controlled as gradients in analyte concentration can exist throughout the 3D construct. Here we describe the methods of preparing biocompatible pH responsive sol-gel nanosensors and their incorporation into poly(lactic-co-glycolic acid) (PLGA) electrospun scaffolds along with their subsequent preparation for the culture of mammalian cells. The pH responsive scaffolds can be used as tools to determine microenvironmental pH within a 3D cellular construct. Furthermore, we detail the delivery of pH responsive nanosensors to the intracellular environment of mammalian cells whose growth was supported by electrospun PLGA scaffolds. The cytoplasmic location of the pH responsive nanosensors can be utilized to monitor intracellular pH (pHi) during ongoing experimentation.

Biocompatible pH responsive sol-gel nanosensors can be incorporated into poly(lactic-co-glycolic acid) (PLGA) electrospun scaffolds. The produced self-reporting scaffolds can be used for in&#160;situ monitoring of microenvironmental conditions whilst culturing cells upon the scaffold. This is beneficial as the 3D cellular construct can be monitored in real-time without disturbing the experiment.

3 차원 세포 배양을위한 자기보고 발판

Culturing cells in 3D on appropriate scaffolds is thought to better mimic the in vivo microenvironment and increase cell-cell interactions. The resulting ...

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 ...

Electrospinning Growth Factor Releasing Microspheres into Fibrous Scaffolds

This procedure describes a method to fabricate a multifaceted substrate to direct nerve cell growth. This system incorporates mechanical, topographical, adhesive and chemical signals. Mechanical properties are controlled by the type of material used to fabricate the electrospun fibers. In this protocol we use 30% methacrylated Hyaluronic Acid (HA), which has a tensile modulus of ~500 Pa, to produce a soft fibrous scaffold. Electrospinning on to a rotating mandrel produces aligned fibers to create a topographical cue. Adhesion is achieved by coating the scaffold with fibronectin. The primary challenge addressed herein is providing a chemical signal throughout the depth of the scaffold for extended periods. This procedure describes fabricating&#160;poly(lactic-co-glycolic acid) (PLGA) microspheres that contain Nerve Growth Factor (NGF) and directly impregnating the scaffold with these microspheres during the electrospinning process. Due to the harsh production environment, including high sheer forces and electrical charges, protein viability is measured after production. The system provides protein release for over 60 days and has been shown to promote primary nerve cell growth.

This protocol combines electrospinning and microspheres to develop tissue engineered scaffolds to direct neurons. Nerve growth factor was encapsulated within PLGA microspheres and electrospun into Hyaluronic Acid (HA) fibrous scaffolds. The protein bioactivity was tested by seeding the scaffolds with primary chick Dorsal Root Ganglia and culturing for 4-6 days.

섬유 공사장 공중 발판으로 마이크로 스피어를 해제 전기 방사의 성장 인자

This procedure describes a method to fabricate a multifaceted substrate to direct nerve cell growth.  This system incorporates mechanical, topographical, ...

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 ...

Fabrication of Mechanically Tunable and Bioactive Metal Scaffolds for Biomedical Applications

Biometal systems have been widely used for biomedical applications, in particular, as load-bearing materials. However, major challenges are high stiffness and low bioactivity of metals. In this study, we have developed a new method towards fabricating a new type of bioactive and mechanically reliable porous metal scaffolds-densified porous Ti scaffolds. The method consists of two fabrication processes, 1) the fabrication of porous Ti scaffolds by dynamic freeze casting, and 2) coating and densification of the porous scaffolds. The dynamic freeze casting method to fabricate porous Ti scaffolds allowed the densification of porous scaffolds by minimizing the chemical contamination and structural defects. The densification process is distinctive for three reasons. First, the densification process is simple, because it requires a control of only one parameter (degree of densification). Second, it is effective, as it achieves mechanical enhancement and sustainable release of biomolecules from porous scaffolds. Third, it has broad applications, as it is also applicable to the fabrication of functionally graded porous scaffolds by spatially varied strain during densification.

Bioactive and mechanically reliable metal scaffolds have been fabricated through a method which consists of two processes, dynamic freeze casting for the fabrication of porous Ti, and coating and densification of the Ti scaffolds. The densification process is simple, effective and applicable to the fabrication of functionally graded scaffolds.

Biometal systems have been widely used for biomedical applications, in particular, as load-bearing materials. However, major challenges are high stiffness ...

Engineering Three-dimensional Epithelial Tissues Embedded within Extracellular Matrix

The architecture of branched organs such as the lungs, kidneys, and mammary glands arises through the developmental process of branching morphogenesis, which is regulated by a variety of soluble and physical signals in the microenvironment. Described here is a method created to study the process of branching morphogenesis by forming engineered three-dimensional (3D) epithelial tissues of defined shape and size that are completely embedded within an extracellular matrix (ECM). This method enables the formation of arrays of identical tissues and enables the control of a variety of environmental factors, including tissue geometry, spacing, and ECM composition. This method can also be combined with widely used techniques such as traction force microscopy (TFM) to gain more information about the interactions between cells and their surrounding ECM. The protocol can be used to investigate a variety of cell and tissue processes beyond branching morphogenesis, including cancer invasion.

This manuscript describes a soft lithography-based technique to engineer uniform arrays of three-dimensional (3D) epithelial tissues of defined geometry surrounded by extracellular matrix. This method is amenable to a wide variety of cell types and experimental contexts and allows for high-throughput screening of identical replicates.

세포 외 매트릭스 내에서 임베디드 공학 3 차원 상피 조직

The architecture of branched organs such as the lungs, kidneys, and mammary glands arises through the developmental process of branching morphogenesis, ...

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 ...

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 ...