우리는 자금에 대한 미국 심장 협회 미드 - 애틀랜틱 제휴를 (0555407U, GLB) 감사드립니다.

<ol><li> 이전 electrospinning로 콜라겐은 육개월 소 진피에서 초산 기반의 프로세스를 사용하여 추출해야합니다. 이 추출된 콜라겐은 다음 dissolutions, precipitations 및 dialyses의 후속 시리즈를 통해 정화됩니다. 일단 콜라겐이 동결 건조된이며, electrospinning 과정을 시작할 수 있습니다. </li><li> polycaprolactone (PCL), 엘라스틴 (ELAS) 및 콜라겐 (콜) : 세 가지 자료 massed 있습니다. 이 자료는 별도로 용해됩니다 1,1​​,1,3,3,3 MG / ML, 각각 100 MG / ML, 200 MG / ML, 그리고 70 농도에 hexafluoro - 2 - 프로파놀 (HFP, TCI 미국). </li><li> 자료 98-2-0의 체적 비율에 다섯 가지의 섬광 튜브에 혼합 아르 PCL - ELAS - 콜 (intima), 45-45-10, 55-35-10 및 65-25-10 PCL - ELAS - 콜 (미디어), 그리고 70-0-30 PCL - ELAS - 콜 (adventitia). </li><li> 버스트, 봉합 보존 및 컴플 라 이언스 테스트를 위해 다층 혈관 이식은 3-1 구성에서 electrospun 동안 uni​​axial 인장 시험 및 개별 레이어 컴플 라 이언스 테스트를 위해 자료의 각 개별적으로 electrospun되었습니다. Uniaxial 인장 시험 공사장 공중 발판이 평평하고 직사각형했다 (2.5 cm x 세로 10.2 cm 두께 X 0.3 cm 길이) 개별 레이어와 다층 이식을 electrospin하는 데 사용되는 2mm 직경의 mandrels 이후 테스트 샘플을 펀칭 너무 작은되었습니다. </li><li> 모두 개인 및 다층 electrospinning 들어, 폴리머 솔루션은 18 게이지 야기한 팁 바늘로 비닐 3 ML 백톤 디킨슨 주사기에 로드된되었고, 세트 디스 펜 서리 속도 주사기 펌프에 위치. 다층 electrospinning 속도가 전환과 계층에 따라 다양한 동안 개인 electrospinning 요금, ML / HR 4로 설정되었습니다. 흐름 속도와 볼륨과 같은되었습니다 : intima 4 ML / 시간의 속도로 electrospun했고 0.5 ML의 볼륨 2 ML / 시간 각각 0.2 ML에 대한 내막과 중간 주사기를 모두 결합 변화가옵니다. 내막 주사기는 다음 차단되었으며 중간 계층은 ML / HR 2 ML / 시간 각 고분자 용액의 0.2 ML에 대한 언론과 adventitia 사이의 전이에 의해 다음 4 0.6 ML에 대한 회전을 허용했다. 마지막으로, 미디어가 중지되었으며 a​​dventitia 4 ML / 시간에서 0.4 ML에 대한 회전을 허용했다. </li><li> 모든 샘플은 이전에 모든 시험 1-18시간 50 MM EDC의 상호 연결되었습니다. </li><li> Uniaxial 인장 시험은 24 시간 동안 PBS에 수산화 한 electrospun 시트 6 샘플에서 수행되었다. &quot;개 - 뼈&quot;모양의 샘플 electrospun 매트에서 펀치와 100 N 부하 셀 및 10.0 mm / 분 확장 속도 MTS Bionix 200 테스트 시스템에서 테스트되었습니다. 피크 응력, 계수, 그리고 휴식의 변형은 TestWorks 버전 4를 사용하여 계산되었다. </li><li> 봉합사 보존 시험은 37 24 시간 동안 PBS에 담가 여섯 가지 electrospun의 이식 6 2mm 내경 관형 표본, ° C, MTS Bionix 200 테스트 시스템에서 50 N로드 셀 (MTS 시스템 주식 회사)와 함께에서 수행되었습니다 미국 국립 표준 연구소 2에서 설명한 절차에 걸쳐 직선에 따라 150.0 mm / 분의 확장 속도. 5-0 상용 PDS II 보라색 monofilament의 봉합은 샘플의 끝에서 2mm를 배치하고 봉합이 이식을 통해 뽑아했다 때까지 연장되었습니다. 피크 부하는 TestWorks 버전 4를 사용하여 g - 무력에 기록되었다. </li><li> 버스트 강도 테스트는 여섯 가지 electrospun의 이식 6 샘플에서 수행되었다. 버스트 튜브, 길이 2~3cm은 장치에 연결된, 그리고 2-0 실크 봉합사와 보안 1.5 mm 직경의 유두를 통해 장착되어, PBS에 수산화되었습니다. 공기 튜브가 2 버스트 때까지 5 mmHg / s의 속도로 압력을 증가, 시스템에 도입되었다. 결과는 튜브가 파열되는 mmHg의 압력으로보고됩니다. </li><li> 동적 준수는 가상의 생리 조건에서 3cm의 길이에서 여섯 가지 electrospun의 이식에서 가져온 여섯 2mm 내경 관형 이식에 대한 결정했습니다. 개별 레이어와 다층 구조 관형 모두 동일한 조건에서 electrospun 및 테스트되었습니다. 표본은 37 PBS로 가득 조직 성장 기술에 의해 개발된 기계 자극 (항목) 생물 반응기 ° C. 통해 지능형 조직 공학 테스트되었습니다 생물 반응기는 50분의 90 세 가지 압력 수준 80분의 120, 그리고 110분의 150 mmHg 수축기 / diastolic가 2 조사 있던 이식의 내부에 1Hz 순환 압력 변화, 3을 제공했습니다. </li></ol> 대표 결과 electrospinning 프로토콜이 올바르게 수행하는 경우, 최종 제품은 부드러운 원활한 튜브 레이어 사이의 박리없이 초기 징후와 함께해야합니다. uniaxial 인장 시험, 파열 강도 시험, 봉합 보존 테스트 및 컴플 라 이언스 테스트가 수행되는 경우, 결과는 중간 계층 강성이 증가로, 엘라스틴의 감소 금액과 관련된 기계적 성질이 stiffer 튜브를 입증해야한다는 의미합니다.

<ol>
	<li>McClure, M. J., Sell, S. A., Barnes, C. P., Bowen, W. C., Bowlin, G. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cross-linking+Electrospun+Polydioxanone-Soluble+Elastin+Blends:+Material+Characterization.">Cross-linking Electrospun Polydioxanone-Soluble Elastin Blends: Material Characterization.</a> Journal of Engineered Fibers and Fabrics. 3, 1-10 (2008).</li><li>. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Cardiovascular implants - Vascular graft protheses. VP20, (1994).</li><li>McClure, M. J., Sell, S. A., Simpson, D. G., Walpoth, B. H., Bowlin, G. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+three-layered+electrospun+matrix+to+mimic+native+arterial+architecture+using+polycaprolactone,+elastin,+and+collagen:+A+preliminary+study.">A three-layered electrospun matrix to mimic native arterial architecture using polycaprolactone, elastin, and collagen: A preliminary study.</a> Acta Biomater. , (2010).</li></ol>

몇몇 저자는 미국이 논문에서 제시에 관한 기술을 출원 국제 특허를 가지고 있고,이 기술은 Organogenesis 주식 허가되었습니다

본 연구의 가장 중요한 부분은 electrospinning 과정입니다. 3-1 입력 출력 노즐을 사용하는 경우, arching 및 충전 손실이 발생할 수 있습니다. 이 발생하면, 회전하는 폴리머와 관련된 전압 용접, &quot;젖은&quot;섬유가 발생하고 세 가지 레이어의 각 사이의 박리를 만들 줄어 듭니다. 따라서, 지속적인 전기 잠재력은 이상적인 다층 튜브를 얻기 위해 필수적입니다. 준수 불일치는 작?...

<table><tbody><tr><td>Polycaprolactone</td><td>Sigma-Aldrich</td><td></td><td></td></tr><tr><td>Elastin</td><td>Elastin Products Company</td><td></td><td></td></tr><tr><td>Collagen</td><td></td><td></td><td></td></tr><tr><td>Acetic Acid</td><td>Fisher Scientific</td><td></td><td></td></tr><tr><td>Sodium Chloride</td><td>Fisher Scientific</td><td></td><td></td></tr><tr><td>TRIS</td><td>Fisher Scientific</td><td></td><td></td></tr><tr><td>6 L Avanti J-HC Centrifuge</td><td>Beckman Coulter Inc.</td><td></td><td></td></tr><tr><td>1,1,1,3,3,3 hexafluoro-2-propanol</td><td>TCI America</td><td></td><td></td></tr><tr><td>3 ml Becton Dickinson Syringe</td><td></td><td></td><td></td></tr><tr><td>CZE1000R Rack Mount Power Supply</td><td>Spellman</td><td></td><td></td></tr><tr><td>High-Pressure Syringe Pump</td><td>Cole-Parmer</td><td></td><td></td></tr><tr><td>1-ethyl-3-(3-dimethylaminopropyl)-carbodiimide</td><td>Pierce, Thermo Scientific</td><td></td><td></td></tr><tr><td>70 % ethanol</td><td>Sigma-Aldrich</td><td></td><td></td></tr><tr><td>ImageTool 3.0 software</td><td>Shareware provided by UTHSCSA</td><td></td><td></td></tr><tr><td>Scanning Electron Microscope</td><td>Carl Zeiss, Inc.</td><td></td><td></td></tr><tr><td>MTS Bionix 200 testing system</td><td>MTS Systems Corp.</td><td></td><td></td></tr><tr><td>TestWorks version 4</td><td></td><td></td><td></td></tr><tr><td>Intelligent Tissue Engineering via Mechanical Stimulation (ITEMS) Bioreactor</td><td>Tissue Growth Technologies</td><td></td><td></td></tr></tbody></table>

tri-layered electrospinning to mimic native arterial architecture using polycaprolactone, elastin, and collagen: a preliminary study

기본 동맥 통해 콜라겐과 엘라스틴은 기계 백본을 제공하는 선박의 파열을 방지하고, 타악기 deformations에 따라 복구를 추진, 중요한 역할을한다. 본 연구의 목표는 엘라스틴의 추가와 45-45-10의 결합, 55-35로 콜라겐과 폴리 (caprolactone) (PCL)으로 구성된 다층 electrospun 도관을 fabricating하여 기본 동맥의 구조를 모방하는 것이었다 - 조직 재생에 도움이되는 유지하면서 10, 65-25-10는 PCL - ELAS - 콜, 기본 동맥 조직을 나타내는 기계적 성질을 설명합니다. 전체 이식 및 개별 레이어는 uniaxial 인장 시험, 동적 준수, 봉합 보존 및 버스트 강도를 사용하여 분석했다. 11.8 MPA - uniaxial 결과는 2.0의 평균 계수 범위를 시연하는 동안 2.8 % / 100 mmHg, - 준수 결과는 중간 / 중간 계층에 대한 변경 사항이 0.8에 이르기까지 전체 그래프트 준수 값 전체 그래프트 동작을 변경하는 것으로 나타났습니다. 계수 및 규정 준수 데이터를 모두 원래 동맥의 범위 내에서 값을 표시합니다. 수학 모델링은 레이어 강성의 변화는 전체 원주 벽 스트레스에 미치는 영향 및 디자인 지원은 재료의 최고의 기계적 조합을 달성할 수있을만큼 보여주 구현되었습니다. 전반적으로, 결과는 그래프트는 레이어 속성을 변경하여 트라이 - 계층 구조를 모방하기위한 수 지적했다.

Watch this Scientific Journal Video about Tri-layered Electrospinning to Mimic Native Arterial Architecture using Polycaprolactone, Elastin, and Collagen: A Preliminary Study at JoVE.com

Tri-layered Electrospinning to Mimic Native Arterial Architecture using Polycaprolactone, Elastin, and Collagen: A Preliminary Study

본 연구의 목적은 동맥 벽 원래 세 계층 아키텍처를 모방하는 것이었다. 이것을 달성하기 위해, electrospinning은 3-1 (입출력) 노즐의 사용과 고용 및 polycaprolactone, 엘라스틴, 콜라겐의 결합되었다.

Polycaprolactone, 엘라스틴, 콜라겐을 사용하여 네이티브 아티 건축을 모방하기 위해 트라이 - 계층 Electrospinning : 예비 연구

Throughout native artery, collagen and elastin play an important role, providing a mechanical backbone, preventing vessel rupture, and promoting recovery ...

tri-layered-electrospinning-to-mimic-native-arterial-architecture

Research

JoVE Journal

Bioengineering

12.8K 조회수.  Virginia Commonwealth University. 본 연구의 목적은 동맥 벽 원래 세 계층 아키텍처를 모방하는 것이었다. 이것을 달성하기 위해, electrospinning은 3-1 (입출력) 노즐의 사용과 고용 및 polycaprolactone, 엘라스틴, 콜라겐의 결합되었다.

비디오: Tri-layered Electrospinning to Mimic Native Arterial Architecture using Polycaprolactone, Elastin, and Collagen: A Preliminary Study

Postproduction Processing of Electrospun Fibres for Tissue Engineering

Electrospinning is a commonly used and versatile method to produce scaffolds (often biodegradable) for 3D tissue engineering.1, 2, 3 Many tissues in vivo undergo biaxial distension to varying extents such as skin, bladder, pelvic floor and even the hard palate as children grow. In producing scaffolds for these purposes there is a need to develop scaffolds of appropriate biomechanical properties (whether achieved without or with cells) and which are sterile for clinical use. The focus of this paper is not how to establish basic electrospinning parameters (as there is extensive literature on electrospinning) but on how to modify spun scaffolds post production to make them fit for tissue engineering purposes - here thickness, mechanical properties and sterilisation (required for clinical use) are considered and we also describe how cells can be cultured on scaffolds and subjected to biaxial strain to condition them for specific applications.
Electrospinning tends to produce thin sheets; as the electrospinning collector becomes coated with insulating fibres it becomes a poor conductor such that fibres no longer deposit on it. Hence we describe approaches to produce thicker structures by heat or vapour annealing increasing the strength of scaffolds but not necessarily the elasticity. Sequential spinning of scaffolds of different polymers to achieve complex scaffolds is also described. Sterilisation methodologies can adversely affect strength and elasticity of scaffolds. We compare three methods for their effects on the biomechanical properties on electrospun scaffolds of poly lactic-co-glycolic acid (PLGA).
Imaging of cells on scaffolds and assessment of production of extracellular matrix (ECM) proteins by cells on scaffolds is described. Culturing cells on scaffolds in vitro can improve scaffold strength and elasticity but the tissue engineering literature shows that cells often fail to produce appropriate ECM when cultured under static conditions. There are few commercial systems available that allow one to culture cells on scaffolds under dynamic conditioning regimes - one example is the Bose Electroforce 3100 which can be used to exert a conditioning programme on cells in scaffolds held using mechanical grips within a media filled chamber.4 An approach to a budget cell culture bioreactor for controlled distortion in 2 dimensions is described. We show that cells can be induced to produce elastin under these conditions. Finally assessment of the biomechanical properties of processed scaffolds cultured with or without cells is described.

Electrospun scaffolds can be processed post production for tissue engineering applications. Here we describe methods for spinning complex scaffolds (by consecutive spinning), for making thicker scaffolds (by multi-layering using heat or vapour annealing), for achieving sterility (aseptic production or sterilisation post production) and for achieving appropriate biomechanical properties.

조직 공학을위한 Electrospun의 Fibres의 Postproduction 처리

Electrospinning is a commonly used and versatile method to produce  scaffolds (often biodegradable) for 3D tissue engineering.1, 2, 3 Many tissues in vivo ...

연구

생체공학

Electrospun Fibrous Scaffolds of Poly(glycerol-dodecanedioate) for Engineering Neural Tissues From Mouse Embryonic Stem Cells

For tissue engineering applications, the preparation of biodegradable and biocompatible scaffolds is the most desirable but challenging task. &#160;Among the various fabrication methods, electrospinning is the most attractive one due to its simplicity and versatility. Additionally, electrospun nanofibers mimic the size of natural extracellular matrix ensuring additional support for cell survival and growth. This study showed the viability of the fabrication of long fibers spanning a larger deposit area for a novel biodegradable and biocompatible polymer named&#160;poly(glycerol-dodecanoate) (PGD)1 by using a newly designed collector for electrospinning. PGD exhibits unique elastic properties with similar mechanical properties to nerve tissues, thus it is suitable for neural tissue engineering applications. The synthesis and fabrication set-up for making fibrous scaffolding materials was simple, highly reproducible, and inexpensive. In biocompatibility testing, cells derived from mouse embryonic stem cells could adhere to and grow on the electrospun PGD fibers. In summary, this protocol provided a versatile fabrication method for making PGD electrospun fibers to support the growth of mouse embryonic stem cell derived neural lineage cells.

Electrospun Fibrous Scaffolds of Polyglycerol-dodecanedioate for Engineering Neural Tissues From Mouse Embryonic Stem Cells

Synthesis and fabrication of electrospun long fibers spanning a larger deposit area via a newly designed collector from a novel biodegradable polymer named poly(glycerol-dodecanoate) (PGD) was reported. The fibers were able to support the growth of cells derived from mouse pluripotent stem cells.

마우스 배아 줄기 세포에서 신경 조직 공학을위한 폴리의 전기 방사 섬유 발판 (글리세롤 dodecanedioate)

For tissue engineering applications, the preparation of biodegradable and biocompatible scaffolds is the most desirable but challenging task. &#160;Among ...

Electrospun Nanofiber Scaffolds with Gradations in Fiber Organization

The goal of this protocol is to report a simple method for generating nanofiber scaffolds with gradations in fiber organization and test their possible applications in controlling cell morphology/orientation. Nanofiber organization is controlled with a new fabrication apparatus that enables the gradual decrease of fiber organization in a scaffold. Changing the alignment of fibers is achieved through decreasing deposition time of random electrospun fibers on a uniaxially aligned fiber mat. By covering the collector with a moving barrier/mask, along the same axis as fiber deposition, the organizational structure is easily controlled. For tissue engineering purposes, adipose-derived stem cells can be seeded to these scaffolds. Stem cells undergo morphological changes as a result of their position on the varied organizational structure, and can potentially differentiate into different cell types depending on their locations. Additionally, the graded organization of fibers enhances the biomimicry of nanofiber scaffolds so they more closely resemble the natural orientations of collagen nanofibers at tendon-to-bone insertion site compared to traditional scaffolds. Through nanoencapsulation, the gradated fibers also afford the possibility to construct chemical gradients in fiber scaffolds, and thereby further strengthen their potential applications in fast screening of cell-materials interaction and interfacial tissue regeneration. This technique enables the production of continuous gradient scaffolds, but it also can potentially produce fibers in discrete steps by controlling the movement of the moving barrier/mask in a discrete fashion.

Here, we present a protocol to fabricate electrospun nanofiber scaffolds with gradated organization of fibers and explore their applications in regulating cell morphology/orientation. Gradients with regard to physical and chemical properties of the nanofiber scaffolds offer a wide variety of applications in the biomedical field.

섬유 조직의 그라데이션으로 전기 방사 나노 섬유 공사장 공중 발판

The goal of this protocol is to report a simple method for generating nanofiber scaffolds with gradations in fiber organization and test their possible ...

Adapting the Electrospinning Process to Provide Three Unique Environments for a Tri-layered In Vitro Model of the Airway Wall

Electrospinning is a highly adaptable method producing porous 3D fibrous scaffolds that can be exploited in in vitro cell culture. Alterations to intrinsic parameters within the process allow a high degree of control over scaffold characteristics including fiber diameter, alignment and porosity. By developing scaffolds with similar dimensions and topographies to organ- or tissue-specific extracellular matrices (ECM), micro-environments representative to those that cells are exposed to in situ can be created. 
The airway bronchiole wall, comprised of three main micro-environments, was selected as a model tissue. Using decellularized airway ECM as a guide, we electrospun the non-degradable polymer, polyethylene terephthalate (PET), by three different protocols to produce three individual electrospun scaffolds optimized for epithelial, fibroblast or smooth muscle cell-culture. Using a commercially available bioreactor system, we stably co-cultured the three cell-types to provide an in vitro model of the airway wall over an extended time period.
This model highlights the potential for such methods being employed in in vitro diagnostic studies investigating important inter-cellular cross-talk mechanisms or assessing novel pharmaceutical targets, by providing a relevant platform to allow the culture of fully differentiated adult cells within 3D, tissue-specific environments.

Adapting the Electrospinning Process to Provide Three Unique Environments for a Tri-layered In Vitro Model of the Airway Wall

Advancements in biomaterial technologies enable the development of three-dimensional multi-cell-type constructs. We have developed electrospinning protocols to produce three individual scaffolds to culture the main structural cells of the airway to provide a 3D in vitro model of the airway bronchiole wall.

전기 방사를 적응하는 것은 트라이 계층에 대한 세 가지 독특한 환경을 제공하기 위해<em&gt; 체외</em&gt;기도 벽의 모델

Electrospinning is a highly adaptable method producing porous 3D fibrous scaffolds that can be exploited in in vitro cell culture. Alterations to ...

Fabricating Superhydrophobic Polymeric Materials for Biomedical Applications

Superhydrophobic materials, with surfaces possessing permanent or metastable non-wetted states, are of interest for a number of biomedical and industrial applications. Here we describe how electrospinning or electrospraying a polymer mixture containing a biodegradable, biocompatible aliphatic polyester (e.g., polycaprolactone and poly(lactide-co-glycolide)), as the major component, doped with a hydrophobic copolymer composed of the polyester and a stearate-modified poly(glycerol carbonate) affords a superhydrophobic biomaterial. The fabrication techniques of electrospinning or electrospraying provide the enhanced surface roughness and porosity on and within the fibers or the particles, respectively. The use of a low surface energy copolymer dopant that blends with the polyester and can be stably electrospun or electrosprayed affords these superhydrophobic materials. Important parameters such as fiber size, copolymer dopant composition and/or concentration, and their effects on wettability are discussed. This combination of polymer chemistry and process engineering affords a versatile approach to develop application-specific materials using scalable techniques, which are likely generalizable to a wider class of polymers for a variety of applications.

Two- and three-dimensional superhydrophobic polymeric materials are prepared by electrospinning or electrospraying biodegradable polymers blended with a lower surface energy polymer of similar composition.

바이오 메디컬 응용 프로그램에 대한 초 소수성 고분자 재료를 제조

Superhydrophobic materials, with surfaces possessing permanent or metastable non-wetted states, are of interest for a number of biomedical and industrial ...

Electrospinning Fibrous Polymer Scaffolds for Tissue Engineering and Cell Culture

As the field of tissue engineering evolves, there is a tremendous demand to produce more suitable materials and processing techniques in order to address the requirements (e.g., mechanics and vascularity) of more intricate organs and tissues. Electrospinning is a popular technique to create fibrous scaffolds that mimic the architecture and size scale of the native extracellular matrix. These fibrous scaffolds are also useful as cell culture substrates since the fibers can be used to direct cellular behavior, including stem cell differentiation (see extensive reviews by Mauck et al. and Sill et al. for more information). In this article, we describe the general process of electrospinning polymers and as an example, electrospin a reactive hyaluronic acid capable of crosslinking with light exposure (see Ifkovits et al. for a review on photocrosslinkable materials). We also introduce further processing capabilities such as photopatterning and multi-polymer scaffold formation. Photopatterning can be used to create scaffolds with channels and multi-scale porosity to increase cellular infiltration and tissue distribution. Multi-polymer scaffolds are useful to better tune the properties (mechanics and degradation) of a scaffold, including tailored porosity for cellular infiltration. Furthermore, these techniques can be extended to include a wide array of polymers and reactive macromers to create complex scaffolds that provide the cues necessary for the development of successful tissue engineered constructs.

The process of electrospinning polymers for tissue engineering and cell culture is addressed in this article. Specifically, the electrospinning of photoreactive macromers with additional processing capabilities of photopatterning and multi-polymer electrospinning is described.

조직 공학 및 세포 배양을위한 섬유 고분자의 공사장 공중 발판을 Electrospinning

As the field of tissue engineering evolves, there is a tremendous demand to produce more suitable materials and processing techniques in order to address ...

생물학

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.

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 ...

Controlling Electrospun Polymer Morphology for Tissue Engineering Demonstrated Using hepG2 Cell Line

Electrospinning affords researchers the opportunity to fabricate reproducible micro to nanoscale polymer fibers. The 3D fibrous architecture of electrospun polymers is regarded as a structural imitation of the extracellular matrix (ECM). Hence, electrospun fibers fabricated from biocompatible polymers have been widely investigated by tissue engineering researchers for their potential role as an artificial ECM for guiding tissue growth both in vitro and in vivo. All cells are acutely sensitive to their mechanical environment. This has been demonstrated by the discovery of multiple mechanotransduction pathways intrinsically linked to the cytoskeletal actin filaments. The cytoskeleton acts as a mechanical sensor that can direct the functionality and differentiation of the host cell depending on the stiffness and morphology of its substrate. Electrospun fibers can be tuned both in terms of fiber size and morphology to easily modulate the mechanical environment within a fibrous polymer scaffold. Here, methods for electrospinning polycaprolactone (PCL) for three distinct morphologies at two different fiber diameters are described. The morphological fiber categories consist of randomly oriented fibers, aligned fibers, and porous cryogenically spun fibers, with 1 &#181;m and 5 &#181;m diameters. The methods detailed within this study are proposed as a platform for investigating the effect of electrospun fiber architecture on tissue generation. Understanding these effects will allow researchers to optimize the mechanical properties of electrospun fibers and demonstrate the potential of this technology more thoroughly.

This method provides the means to test different polycaprolactone fiber morphologies and topographies for the purpose of tissue engineering. Small and large fibers are fabricated with random orientations, aligned orientations, and also porous cryogenically electrospun structures and used as platforms for cell culture.

Electrospinning affords researchers the opportunity to fabricate reproducible micro to nanoscale polymer fibers. The 3D fibrous architecture of ...

3D Human Myocardial Tissue Generation Using Melt Electrospinning Writing of Polycaprolactone Scaffolds and hiPSC-Derived Cardiac Cells

The development of functional human cardiac tissues holds significant promise for advancing applications in drug screening, disease modeling, and regenerative medicine. This protocol describes the stepwise fabrication of 3D myocardial tissues with advanced mimicry of native cardiac structure by combining melt electrospinning writing (MEW) polycaprolactone (PCL) scaffolds with fibrin hydrogels and human induced pluripotent stem cell (hiPSC)-derived cardiac cells. The process involves embedding a mixture of cardiomyocytes (hiPSC-CMs) and cardiac fibroblasts (hiPSC-CFs) within a fibrin matrix to create mini-tissues, with structural support provided by MEW-generated scaffolds. These fibrillar scaffolds are fabricated at the micro- to nanoscale, allowing for precise control over fiber architecture, which plays a key role in organizing cell distribution and alignment. Meanwhile, the fibrin matrix promotes cell viability and mimics the extracellular environment. Characterization of the generated tissues reveals well-organized sarcomeres within hiPSC-CMs, along with stable contractile activity. The tissues demonstrate consistent spontaneous beating as early as two days post-seeding, with sustained functionality over time. The combination of hiPSC-CFs with hiPSC-CMs enhances the structural integrity of the tissues while supporting long-term cell viability. This approach offers a reproducible, adaptable, and scalable method for creating biomimetic cardiac tissue models, providing a versatile platform for preclinical drug testing, mechanistic studies of cardiac disease, and potential regenerative therapies.

A reproducible method is presented to generate 3D myocardial tissues combining melt electrospinning writing (MEW) polycaprolactone (PCL) scaffolds and fibrin hydrogels with hiPSC-derived cardiomyocytes and fibroblasts. This technique offers precise control over scaffold architecture and can be applied in preclinical drug testing and cardiac disease modeling.

폴리카프로락톤 골격 및 hiPSC 유래 심장 세포의 용융 전기방사 쓰기를 사용한 3D 인간 심근 조직 생성

The development of functional human cardiac tissues holds significant promise for advancing applications in drug screening, disease modeling, and ...

Combining 3D-Printing and Electrospinning to Manufacture Biomimetic Heart Valve Leaflets

Electrospinning has become a widely used technique in cardiovascular tissue engineering as it offers the possibility to create (micro-)fibrous scaffolds with adjustable properties. The aim of this study was to create multilayered scaffolds mimicking the architectural fiber characteristics of human heart valve leaflets using conductive 3D-printed collectors.
Models of aortic valve cusps were created using commercial computer-aided design (CAD) software. Conductive polylactic acid was used to fabricate 3D-printed leaflet templates. These cusp negatives were integrated into a specifically designed, rotating electrospinning mandrel. Three layers of polyurethane were spun onto the collector, mimicking the fiber orientation of human heart valves. Surface and fiber structure was assessed with a scanning electron microscope (SEM). The application of fluorescent dye additionally permitted the microscopic visualization of the multilayered fiber structure. Tensile testing was performed to assess the biomechanical properties of the scaffolds.
3D-printing of essential parts for the electrospinning rig was possible in a short time for a low budget. The aortic valve cusps created following this protocol were three-layered, with a fiber diameter of 4.1 &#177; 1.6 &#181;m. SEM imaging revealed an even distribution of fibers. Fluorescence microscopy revealed individual layers with differently aligned fibers, with each layer precisely reaching the desired fiber configuration. The produced scaffolds showed high tensile strength, especially along the direction of alignment. The printing files for the different collectors are available as Supplemental File 1, Supplemental File 2, Supplemental File 3, Supplemental File 4, and Supplemental File 5.
With a highly specialized setup and workflow protocol, it is possible to mimic tissues with complex fiber structures over multiple layers. Spinning directly on 3D-printed collectors creates considerable flexibility in manufacturing 3D shapes at low production costs.

The presented method offers an innovative way for engineering biomimetic fiber structures in three-dimensional (3D) scaffolds (e.g., heart valve leaflets). 3D-printed, conductive geometries were used to determine shape and dimensions. Fiber orientation and characteristics were individually adjustable for each layer. Multiple samples could be manufactured in one setup.

3D 프린팅과 전기방사를 결합하여 생체모방 심장 판막 전단지 제조

Electrospinning has become a widely used technique in cardiovascular tissue engineering as it offers the possibility to create (micro-)fibrous scaffolds ...