저자는 고급 안과 재료의 개발에 대한 우리의 결제 수단 NSERC 20/20 네트워크를 인정하고 싶습니다.

모든 실험은 대학의 워털루의 동물 연구 윤리위원회에 의해 설명 된 모든 관련 지침에 따라 준수에 완료되었다. 소 눈이 넉넉한 지역 도살장에서 기증된다. 1. 아이 모델 <ol><li> 설계 및 금형 (13)의 제조 <ol><li> 인간의 성인 눈의 평균 생리 학적 차원에 따라 눈 모델을 디자인합니다. (13) </li><li> 안구와 눈 모델의 눈꺼풀 조각 사이에 250 ㎛의 간격을 둡니다. 컴퓨터 지원 설계 (CAD) 소프트웨어를 사용하여 각 금형을 설계한다. </li><li> 의 AutoCAD 또는 솔리드 웍스의 새로운 .cad 파일 또는 .sldprt 파일을 만듭니다. 인간의 안구 / 눈꺼풀의 3D 모델을 생성합니다. 모델의 금형을 만들고 .STL 파일로 금형을 저장합니다. </li><li> 3D 프린터 소프트웨어로 가져 오기 .STL 파일 (replicator2에 대한 예를 들어, makeware). 인쇄의 매개 변수 (위치, 희소성, 크기, 방향, 부드러움 등을 지정 </EM&gt;) 13. </li><li> 3D 프린터는 읽기위한 G 코드 파일로 파일을 저장합니다. 이러한 PLA (폴리 락트산), ABS (아크릴로 니트릴 부타디엔 스티렌), PC (폴리 카보네이트), 또는 이들의 조합으로서 선택 물질은 몰드 (13)를 인쇄한다. </li><li> 선택의 재료의 원하는 필라멘트를 설치합니다. 읽을 수있는 3D 프린터에 G 코드 파일을 가져옵니다. 금형을 인쇄합니다. 주 : 눈 모델에 매끄러운 표면이 요구되는 경우 또는, (CNC) 머신을 제어하는​​ 컴퓨터의 수치를 이용하여 눈 금형을 생산한다. CNC 몰드 생산 용 금형 재료가 더 이상 열적 플라스틱에 한정되지 않지만, 금속, 세라믹, 폴리 테트라 플루오로 에틸렌과 같은 화학적 저항성 중합체로 연장된다. </li><li> 절단 드릴에 연결되어있는 CNC 소프트웨어 인터페이스를 엽니 다. 전면, 상단, 측면 및 제어 소프트웨어 인터페이스에서 이전에 건설 안구 / 눈꺼풀 모델 금형의 사시도에 따라 3D 금형을 구축합니다. 에 적합한 매개 변수를 선택가공 (비트 크기, 기판 소재, 재료 두께) 금형을 잘라 진행합니다. </li></ol></li><li> PDMS를 사용하여 접안 렌즈의 합성 <ol><li> 주사기를 사용하여, PDMS (폴리 디메틸 실록산) 염기 10 ml의 부피를 측정하고, 15 내지 50 mL의 원심 분리 튜브로 채운다. PDMS의 총 중량 엘라스토머 솔루션의 V / w는 10 %를 추가합니다. 교반 막대를 사용하여 용액 잘 섞는다. </li><li> 안구 및 눈꺼풀 금형에 PDMS 솔루션을 따르십시오. PDMS의 RT는 O / N에 정착 할 수 있도록 (또는 적어도 12 시간)하여 중합을 시작하고, 기포가 중합체로부터 분해 할 수 있도록. 참고 : 상승 또는 확장 할 수있는 PDMS에 남아있는 기포가 없는지 확인합니다. </li><li> 그 후, 1 시간, 5 분 동안 150 ° C (302 ° F)의 75 ° C (167 ° F) 오븐에 금형을 넣어. 부드러운 젤의 경우, PDMS이 완전히 중합 적어도 48 시간 동안 실온에서 앉아 보자. </li><li> 몇 분 동안 냉동실에 샘플을 넣어; 이는 PDMS를 축소하고 단순화몰드로부터 표본을 제거. 얇은 주걱을 사용하여 금형의 접안 렌즈의 압축을 풉니 다. </li><li> 안구 및 눈꺼풀 편 사이의 공간에 용액을 전달하기 위해, 1/16 &quot;동일 다리 결합기 튜브 커넥터 1/16&quot;X 1/8 &quot;폴리 테트라 플루오로 에틸렌 튜브를 연결하고 튜브 구멍의 눈꺼풀 피스에 부착 . </li></ol></li><li> 아가로 오스를 사용하여 안구 조각의 합성 주 : 안구 조각을 아가 로스 등의 다른 중합체를 사용하여 합성 할 수있다. 다음 절차는 또한 PDA (포테이​​토 덱 스트로스 배지) 또는 SDA (사부로 포도당 한천)로서 한천 유형의 다양한 눈 조각을 생산하도록 변형시킬 수있다. <ol><li> 2 % (2 ㎍ / 100 ㎖) 겔 측정 아가 2 g의 생성 및 초순수 100 ㎖와 혼합한다. 아가로 오스가 완전히 용해되도록 종기 (100 ° C)에 대한 해결책을 가져옵니다. 용액을 5 분 동안 식히십시오. </li><li> 안구 금형에 솔루션을 붓고 솔루션은 3 냉각 할 수 있도록RT에서 0 분. 주걱으로 안구 조각을 제거합니다. A의 안구 한천 보관 -20 나중에 사용하기 위해 ° C 냉장고. 미생물학 연구의 경우, 고압 증기 멸균 및 / 또는 자외선 조사에 의한 안구 금형을 소독. </li></ol></li><li> PDMS 안구에 소 각막의 설립 참고 :.이 프로토콜은 Parekh 씨 등에서 적응하고있다 (14) <ol><li> 층류 후드 무균 조건에서 소 각막 절개 및 혼합을 수행한다. 눈을 취득하고 당일을 해부. </li><li> 사용하기 전에 10 분 동안의 흐름 후드를 켜고 70 % 에탄올 알코올로 소독. 모든 자료 및 악기는 273 ° F / 45 분 동안 133 ° C에서 고압 증기 멸균에 의해 멸균하고, 흐름 후드 입구에서 더 이상 4 인치 위치에 있는지 확인합니다. </li><li> 2 분 동안 희석 된 포비돈 요오드 용액을 넣은 비커에 소 눈 담근다. 인산염 완충 식염수를 넣은 비커에 눈 린스 (PBS) pH 7.4. 사용 포셉 부드럽게 유리 페트리 접시, 각막면이 위를 향에 눈을 넣습니다. </li><li> 무딘 끝 해부 가위로 공막 부착 지점에서 절단하여 과도한 근육과 지방 조직을 제거합니다. 동물 폐기물 지정된 멸균 비이커에 초과 조직을 폐기하십시오. </li><li> 마이크로 가위를 사용하여 눈의 결막을 제거합니다. 윤부로부터 최소 1 cm의 거리를 유지, 멸균 거즈로 눈을 감 쌉니다. </li><li> 기본 맥락막 및 유리체의 침투를 방지하며 표면으로되도록 메스를 사용하여 약 2mm 윤부 영역으로부터 공막 절개. 조심스럽게 자연 곡률에서 각막을 변형없이 메스 또는 해부 가위를 사용하여 360 °로 절개를 확장합니다. </li><li> 미세 집게로, 눈의 각막을 제거합니다. 집게를 사용하여 조심스럽게 부착 포도막 조직을 제거하고 PBS로 각막을 씻어. </li><li> 문화 멸균 용기에 31ºC에서 각막을 저장매체 (매체 (199)와 같은) 조직의 수분과 영양 세포를 유지하기 위해 3 % 소 태아 혈청을 함유. </li><li> 실험에 앞서, PDMS의 안구에 절제 각막 휴식, 그리고 전문 클립 - 온과 함께 두 조각을 고정. </li></ol></li></ol> 2. 깜박임 플랫폼 <ol><li> 깜박임 플랫폼의 설계 및 생산 주 : 깜박 플랫폼이 세 가지 기능 부분으로 구성되어있다 : 눈 모델 (섹션 1에서 설명), 기어 시스템, 전자 시스템. <ol><li> 디자인은 눈 모델 (섹션 1.1)에 기술 된 것과 유사한 CAD 및 3D 인쇄를 사용하여 점멸 플랫폼 제조. 는 접안 렌즈의 횡 방향 및 회전 동작에 모터의 회전 간단한 변환되도록 기어 시스템을 설계한다. 15 </li><li> 피니언과 기어기구를 이용하여 눈꺼풀 편에 연결된 피니언의 횡 방향 운동에 스테핑 모터의 회전 운동을 변환. </li><li> 사용공액 기어에있어서, 세 개의 다른 안구 피스 세 개 (또는 그 이상)의 회전 운동에 스테퍼 모터의 하나의 회전 운동을 증폭한다. </li><li> 둘 사이의 거리가 일정하게되도록, 두 기어 시스템 눈꺼풀 용 및 안구에 대한 하나의 정렬. 마이크로 모터 차폐, 2 개의 모터를 전자 시스템을 조립한다. 참고 : 깜박이는 운동으로 기어 시스템에 의해 번역됩니다 회전 모터를 제공하는 두 개의 스테퍼 모터를 사용합니다. </li><li> 마이크로 컨트롤러에 적층 모터 방패로 구성된 시스템에 두 개의 스테퍼 모터를 연결합니다. 연결 및 오픈 소스 소프트웨어 제품과 함께 작동하는 전자 부품을 구성합니다. </li><li> 분당 발사 속도 (RPM), 전방 발사 수, 후방 발사 수 및 회전 스타일 같은 모터 파라미터를 제어하도록 시스템을 프로그램. 참고 : 자세한 내용은 보충 &quot;아두 이노 코드 파일&quot;을 참조하십시오. </li><li> 석사에서 시스템 소프트웨어를 다운로드제조 업체들이 적시 한 &#39;웹 사이트. </li><li> 소프트웨어를 설치하고 엽니 다. 원하는 구성에 스텝 모터를 제어하는​​ 코드를 작성한다. 연구자에 의해 정의 된 모터가 원하는 방식으로 이동할 수 있도록 전자 시스템에 전원을 공급하는 공급원과 시스템을 연결한다. 참고 : 추가 &quot;아두 이노 코드 파일&quot;을 참조하십시오. </li></ol></li><li> 미세 유체와 어셈블리 (인공 눈물 솔루션) <ol><li> 합성 안구와 눈꺼풀 조각을 타고 눈 모델에 대한 해당 클립 기능에 그들을 미끄러. 눈꺼풀 조각 (섹션 1.2.5)과 미세 유체 펌프에 주사기와 결합하고 위치 튜브를 연결합니다. 테스트 플랫폼을 실행하고 일관된 움직임을 확인합니다. </li><li> 프라임 튜브와 인공 눈물 액 (ATS)의 꾸준한 흐름을 확인합니다. ATS에 대한 레시피는 이전에보고 된 바있다. (16) </li><li> 수동으로 평평한 평면 함께 눈 모델 부품을 이동하도록 상기 안구와 시선뚜껑 접촉하고있다. 원하는 값으로 미세 유체 펌프의 유량을 설정한다. 1.5 μL / 분에 생리 흐름 속도를 설정합니다. (17) </li><li> 실험을 시작하기 위해 펌프와 액추에이터를 시작합니다. 약물 전달 실험을 위해, 안구 부분에 약물 함유 콘택트 렌즈를 배치했다. </li><li> 플로우를 통해 유체가 12 웰 플레이트에 똑 수 있습니다. 원하는 설정된 시간 간격, 예컨대 UV-비스 분광법이나 형광 검출 등의 일반적인 방법을 사용하여 분석 물 또는 약물 농도를 정량화. 1,4,18을 </li><li> 콘택트 렌즈에 눈물 성분의 침착을 평가하는 연구를 들면, &quot;안구&quot;조각에 콘택트 렌즈를 배치합니다. 버려 질 수있는 관류 액을 모은다. </li><li> 원하는 시간 간격 후에 안구 부분에서 콘택트 렌즈를 제거하고 같은 초점 현미경으로 추가 분석을 위해 렌즈를 준비한다. </li></ol></li></ol>

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E., Plummer, C., Chauhan, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Extended+drug+delivery+by+contact+lenses+for+glaucoma+therapy.">Extended drug delivery by contact lenses for glaucoma therapy.</a> J Control Release. 162, 152-158 (2012).</li><li>Faschinger, C., Mossbock, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Continuous+24+h+monitoring+of+changes+in+intraocular+pressure+with+the+wireless+contact+lens+sensor+Triggerfish.+First+results+in+patients.">Continuous 24 h monitoring of changes in intraocular pressure with the wireless contact lens sensor Triggerfish. First results in patients.</a> Der Ophthalmologe : Zeitschrift der Deutschen Ophthalmologischen Gesellschaft. 107, 918-922 (2010).</li><li>Shaw, A. J., Davis, B. A., Collins, M. J., Carney, L. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+technique+to+measure+eyelid+pressure+using+piezoresistive+sensors.">A technique to measure eyelid pressure using piezoresistive sensors.</a> IEEE transactions on bio-medical engineering. 56, 2512-2517 (2009).</li><li>Liao, Y. T., Yao, H. F., Lingley, A., Parviz, B., Otis, B. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+3-mu+W+CMOS+glucose+sensor+for+wireless+contact-lens+tear+glucose+monitoring.">A 3-mu W CMOS glucose sensor for wireless contact-lens tear glucose monitoring.</a> Ieee J Solid-St Circ. 47, 335-344 (2012).</li><li>Coster, D. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Cornea. , (2002).</li><li>Parekh, 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+simplified+technique+for+in+situ+excision+of+cornea+and+evisceration+of+retinal+tissue+from+human+ocular+globe.">A simplified technique for in situ excision of cornea and evisceration of retinal tissue from human ocular globe.</a> Journal of visualized experiments : JoVE. , e3765 (2012).</li><li>Way, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Gear+and+pinion.">Gear and pinion.</a> US patent. , (1942).</li><li>Lorentz, 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=Contact+lens+physical+properties+and+lipid+deposition+in+a+novel+characterized+artificial+tear+solution.">Contact lens physical properties and lipid deposition in a novel characterized artificial tear solution.</a> Molecular vision. 17, 3392-3405 (2011).</li><li>Furukawa, R. E., Polse, K. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Changes+in+tear+flow+accompanying+aging.">Changes in tear flow accompanying aging.</a> American journal of optometry and physiological optics. 55, 69-74 (1978).</li><li>Bajgrowicz, M., Phan, C. M., Subbaraman, L., Jones, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Release+of+ciprofloxacin+and+moxifloxacin+from+daily+disposable+contact+lenses+from+an+in+vitro+eye+model.">Release of ciprofloxacin and moxifloxacin from daily disposable contact lenses from an in vitro eye model.</a> Invest Ophthalmol Vis Sci. , (2015).</li><li>Luensmann, D., Zhang, F., Subbaraman, L., Sheardown, H., Jones, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Localization+of+lysozyme+sorption+to+conventional+and+silicone+hydrogel+contact+lenses+using+confocal+microscopy.">Localization of lysozyme sorption to conventional and silicone hydrogel contact lenses using confocal microscopy.</a> Current eye research. 34, 683-697 (2009).</li><li>Tieppo, A., Pate, K. M., Byrne, M. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=In+vitro+controlled+release+of+an+anti-inflammatory+from+daily+disposable+therapeutic+contact+lenses+under+physiological+ocular+tear+flow.">In vitro controlled release of an anti-inflammatory from daily disposable therapeutic contact lenses under physiological ocular tear flow.</a> Eur J Pharm Biopharm. 81, 170-177 (2012).</li><li>Ali, 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=Zero-order+therapeutic+release+from+imprinted+hydrogel+contact+lenses+within+in+vitro+physiological+ocular+tear+flow.">Zero-order therapeutic release from imprinted hydrogel contact lenses within in vitro physiological ocular tear flow.</a> J Control Release. 124, 154-162 (2007).</li><li>White, C. J., McBride, M. K., Pate, K. M., Tieppo, A., Byrne, M. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Extended+release+of+high+molecular+weight+hydroxypropyl+methylcellulose+from+molecularly+imprinted,+extended+wear+silicone+hydrogel+contact+lenses.">Extended release of high molecular weight hydroxypropyl methylcellulose from molecularly imprinted, extended wear silicone hydrogel contact lenses.</a> Biomaterials. 32, 5698-5705 (2011).</li><li>Kaczmarek, J. C., Tieppo, A., White, C. J., Byrne, M. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Adjusting+biomaterial+composition+to+achieve+controlled+multiple-day+release+of+dexamethasone+from+an+extended-wear+silicone+hydrogel+contact+lens.">Adjusting biomaterial composition to achieve controlled multiple-day release of dexamethasone from an extended-wear silicone hydrogel contact lens.</a> J Biomater Sci Polym Ed. 25, 88-100 (2014).</li><li>Mohammadi, S., Postnikoff, C., Wright, A. M., Gorbet, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Design+and+development+of+an+in+vitro+tear+replenishment+system.">Design and development of an in vitro tear replenishment system.</a> Ann Biomed Eng. 42, 1923-1931 (2014).</li><li>Lorentz, H., Heynen, M., Khan, W., Trieu, D., Jones, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+impact+of+intermittent+air+exposure+on+lipid+deposition.">The impact of intermittent air exposure on lipid deposition.</a> Optom Vis Sci. 89, 1574-1581 (2012).</li><li>Peng, C. C., Fajardo, N. P., Razunguzwa, T., Radke, C. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=In+vitro+spoilation+of+silicone-hydrogel+soft+contact+lenses+in+a+model-blink+cell.">In vitro spoilation of silicone-hydrogel soft contact lenses in a model-blink cell.</a> Optom Vis Sci. 92, 768-780 (2015).</li><li>Liu, P., 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=Dissolution+studies+of+poorly+soluble+drug+nanosuspensions+in+non-sink+conditions.">Dissolution studies of poorly soluble drug nanosuspensions in non-sink conditions.</a> AAPS PharmSciTech. 14, 748-756 (2013).</li></ol>

저자는 공개 아무것도 없어.

거기에 특별한주의를 필요로하는 프로토콜 내에서 세 가지 중요한 단계는 다음과 같습니다 설계 및 금형 생산 (섹션 1.1), 플랫폼 어셈블리 (제 2.2.1-2.2.3)을하고, 실험 실행을 감시 (제 2.2.4-2.2.7 ). 설계 및 금형 (1.1 절)의 제조의 측면에서, 안구 부분은 인간 각막의 크기에 따라 설계되어야한다. 그러나, 완벽한 시판 콘택트 렌즈 (CL)에 맞는 생성 될 수있는 안구 편 전에 몰드의 복수의 원형을 요구할 ?...

콘택트 렌즈 (CL) 내의 관심 분야의 두 가지 중요한 부분이 불편 CL 및 신규 애플리케이션의 개발을 포함한다. CL의 불편 함을 기본 메커니즘을 해명하는 것은 수십 년 동안 필드를 회피 한 문제입니다. 8 소설의 개발은 이러한 약물 전달 장치 1,3,9 및 바이오 센서 등의 기능 CL에서, 10-12 성장 관심의 영역입니다, 상당한 잠재 시장과. 두 경우에, 시험 관내 모델에서 정교한는 개발 단계에서 렌즈 재료 및 디자인 특성 적절한 선택을 지원하기 위해 적절한 정보를 제공 할 것이다. 불행하게도, CL에서 다른 눈 관련 응용 프로그램을 평가하기위한 시험 관내 모델에 전류가 상대적으로 원유와 단순한입니다. 전통적으로, 눈물 성막 또는 약물 전달을 평가하는 시험 관내 연구 CL은 고정 된 부피의 유체를 포함하는 정적 대량 튜브에서 수행되는 GREATLY 생리 양을 초과합니다. 또한,이 단순한 모델은 천연 눈물 흐름 성분과 안구 환경 인자를 정의하는 둘 점멸 반사를 없다. 정교한, 생리 학적으로 중요한 눈 &quot;모델&quot;의 개발은 멀티 - 징계 접근 방식을 필요로하고 생체 검증에 상당한 필요합니다. 이러한 이유로, 우리의 생체 외 눈 모델에 대한 기본 골격 모델이 차후의 업그레이드 및 변조를 통해 개선 될 수있는, 범용성이 높은 것이다. 최신 모델은 눈물 양, 눈물 흐름의 기계적 마모 및 대기에의 노출을 시뮬레이션 할 수있다. 목표는 생체 내 및 생체 관측에 예측하고 무료로 의미있는 결과를 제공 할 것 체외 모델을 만드는 것입니다.

<table border="0" cellpadding="0" cellspacing="0" width="1129">
	<tbody>
		<tr height="42">
			<td height="42" style="height:42px;width:281px;">Arduino Uno R3 (Atmega328 - assembled)</td>
			<td style="width:125px;">Adafruit</td>
			<td style="width:451px;">50</td>
			<td style="width:273px;">Board</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:281px;">Stepper motor</td>
			<td style="width:125px;">Adafruit</td>
			<td style="width:451px;">324</td>
			<td>Motor and Motor shield</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:281px;">Equal Leg Coupler 1.6mm 1/16&#34;</td>
			<td style="width:125px;">VWR</td>
			<td style="width:451px;">CA11009-280</td>
			<td>50 pcs of tube connector</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:281px;">Tubing PT/SIL 1/16&#34;x1/8&#34;</td>
			<td style="width:125px;">VWR</td>
			<td style="width:451px;">16211-316</td>
			<td>Case of 50feet</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:281px;">PDMS</td>
			<td style="width:125px;">Dow Corning</td>
			<td style="width:451px;">Sylgard 184 Solar Cell Encapsulation</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:281px;">Agarose, Type 1-A, low EEO</td>
			<td style="width:125px;">Sigma-Aldrich</td>
			<td style="width:451px;">A0169-25G</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:281px;">PHD UltraTM</td>
			<td style="width:125px;">Harvard Apparatus</td>
			<td style="width:451px;">703006</td>
			<td>MicroFluidic Pump</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:281px;">Bovine cornea</td>
			<td style="width:125px;">Cargill, Guelph/ON</td>
			<td style="width:451px;"></td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:281px;">Soldidworks</td>
			<td style="width:125px;">Dassault Systemes</td>
			<td style="width:451px;"></td>
			<td>Software</td>
		</tr>
		<tr height="63">
			<td height="63" style="height:63px;width:281px;">3-D printing</td>
			<td style="width:125px;">University of Waterloo - 3D Print Centre</td>
			<td style="width:451px;"></td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:281px;">Dissection tools</td>
			<td style="width:125px;">Fine Science Tools</td>
			<td style="width:451px;"></td>
			<td>General dissection tools</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:281px;">Medium 199</td>
			<td style="width:125px;">Sigma-Aldrich</td>
			<td style="width:451px;"></td>
			<td>Culture medium storage for cornea</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:281px;">Fetal bovine serum</td>
			<td style="width:125px;">Thermo Fisher</td>
			<td style="width:451px;"></td>
			<td>Add to culture medium, 3% total volume</td>
		</tr>
	</tbody>
</table>

development of an in vitro ocular platform to test contact lenses

Currently, in vitro evaluations of contact lenses (CLs) for drug delivery are typically performed in large volume vials,1-6 which fail to mimic physiological tear volumes.7 The traditional model also lacks the natural tear flow component and the blinking reflex, both of which are defining factors of the ocular environment. The development of a novel model is described in this study, which consists of a unique 2-piece design, eyeball and eyelid piece, capable of mimicking physiological tear volume. The models are created from 3-D printed molds (Polytetrafluoroethylene or Teflon molds), which can be used to generate eye models from various polymers, such as polydimethylsiloxane (PDMS) and agar. Further modifications to the eye pieces, such as the integration of an explanted human or animal cornea or human corneal construct, will permit for more complex in vitro ocular studies. A commercial microfluidic syringe pump is integrated with the platform to emulate physiological tear secretion. Air exposure and mechanical wear are achieved using two mechanical actuators, of which one moves the eyelid piece laterally, and the other moves the eyeballeyepiece circularly. The model has been used to evaluate CLs for drug delivery and deposition of tear components on CLs.

기계 숍에서 3-D 인쇄에서 얻어진 합성 눈 금형은도 1에 나타내었다. 이들 주형은 원하는 특성으로 접안 렌즈를 생산하기 위해, 아가 로스 PDMS와 같은 중합체의 다양한 이용 될 수있다. 미세 유체 주사기 펌프로 눈 모델 플랫폼의 손짓 조립체는도 2에 도시된다.이 플랫폼과 눈꺼풀 피스의 움직임 아웃 횡 통해 안구 부재의 회전, 대기에의 노출을 통?...

Watch this Scientific Journal Video about Development of an In Vitro Ocular Platform to Test Contact Lenses at JoVE.com

Development of an In Vitro Ocular Platform to Test Contact Lenses

콘택트 렌즈 CLS () 및 기타 눈 관련 응용 프로그램을 평가하는 현재 시험관 모델은 심각하게 제한됩니다. 제시된 안 플랫폼은 생리적 눈물 흐름, 눈물 양, 공기 노출과 기계적 마모를 시뮬레이션합니다. 이 시스템은 매우 다목적이고 CL에서 분석하여 시험 관내에서 다양한 적용 할 수있다.

개발<em&gt; 체외</em&gt; 안구 플랫폼은 콘택트 렌즈를 테스트하는

Currently, in vitro evaluations of contact lenses (CLs) for drug delivery are typically performed in large volume vials,1-6 which fail to mimic ...

development-of-an-in-vitro-ocular-platform-to-test-contact-lenses

Research

JoVE Journal

Bioengineering

Development of an In Vitro Ocular Platform to Test Contact Lenses

10.5K 조회수.  University of Waterloo.  콘택트 렌즈 CLS () 및 기타 눈 관련 응용 프로그램을 평가하는 현재 시험관 모델은 심각하게 제한됩니다. 제시된 안 플랫폼은 생리적 눈물 흐름, 눈물 양, 공기 노출과 기계적 마모를 시뮬레이션합니다. 이 시스템은 매우 다목적이고 CL에서 분석하여 시험 관내에서 다양한 적용 할 수있다. 

비디오: Development of an In Vitro Ocular Platform to Test Contact Lenses

Tissue Engineering of a Human 3D in vitro Tumor Test System

Cancer is one of the leading causes of death worldwide. Current therapeutic strategies are predominantly developed in 2D culture systems, which inadequately reflect physiological conditions in vivo. Biological 3D matrices provide cells an environment in which cells can self-organize, allowing the study of tissue organization and cell differentiation. Such scaffolds can be seeded with a mixture of different cell types to study direct 3D cell-cell-interactions. To mimic the 3D complexity of cancer tumors, our group has developed a 3D in vitro tumor test system.	
	Our 3D tissue test system models the in vivo situation of malignant peripheral nerve sheath tumors (MPNSTs), which we established with our decellularized porcine jejunal segment derived biological vascularized scaffold (BioVaSc). In our model, we reseeded a modified BioVaSc matrix with primary fibroblasts, microvascular endothelial cells (mvECs) and the S462 tumor cell line. For static culture, the vascular structure of the BioVaSc is removed and the remaining scaffold is cut open on one side (Small Intestinal Submucosa SIS-Muc). The resulting matrix is then fixed between two metal rings (cell crowns).	
	Another option is to culture the cell-seeded SIS-Muc in a flow bioreactor system that exposes the cells to shear stress. Here, the bioreactor is connected to a peristaltic pump in a self-constructed incubator. A computer regulates the arterial oxygen and nutrient supply via parameters such as blood pressure, temperature, and flow rate. This setup allows for a dynamic culture with either pressure-regulated pulsatile or constant flow.	
	In this study, we could successfully establish both a static and dynamic 3D culture system for MPNSTs. The ability to model cancer tumors in a more natural 3D environment will enable the discovery, testing, and validation of future pharmaceuticals in a human-like model.

Tissue Engineering of a Human 3D in vitro Tumor Test System

Methods to create human 3D tumor tissues as test systems are described. These technologies are based on a decellularized Biological Vascularized Scaffold (BioVaSc), primary human cells and a tumor cell line, which can be cultured under static as well as under dynamic conditions in a flow bioreactor.

인간의 3 차원의 조직 공학<em&gt; 시험관</em&gt; 종양 테스트 시스템

Cancer is one of the leading causes of death worldwide.  Current therapeutic strategies are predominantly developed in 2D culture  systems, which ...

연구

생체공학

Development of Amelogenin-chitosan Hydrogel for In Vitro Enamel Regrowth with a Dense Interface

Biomimetic enamel reconstruction is a significant topic in material science and dentistry as a novel approach for the treatment of dental caries or erosion. Amelogenin has been proven to be a critical protein for controlling the organized growth of apatite crystals. In this paper, we present a detailed protocol for superficial enamel reconstruction by using a novel amelogenin-chitosan hydrogel. Compared to other conventional treatments, such as topical fluoride and mouthwash, this method not only has the potential to prevent the development of dental caries but also promotes significant and durable enamel restoration. The organized enamel-like microstructure regulated by amelogenin assemblies can significantly improve the mechanical properties of etched enamel, while the dense enamel-restoration interface formed by an in situ regrowth of apatite crystals can improve the effectiveness and durability of restorations. Furthermore, chitosan hydrogel is easy to use and can suppress bacterial infection, which is the major risk factor for the occurrence of dental caries. Therefore, this biocompatible and biodegradable amelogenin-chitosan hydrogel shows promise as a biomaterial for the prevention, restoration, and treatment of defective enamel.

Development of Amelogenin-chitosan Hydrogel for In Vitro Enamel Regrowth with a Dense Interface

In this article, we describe a protocol for fabricating an amelogenin-chitosan hydrogel for superficial enamel reconstruction. Organized in situ growth of apatite crystals in the hydrogel formed a dense enamel-restoration interface, which will improve the effectiveness and durability of restorations.

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

Biomimetic enamel reconstruction is a significant topic in material science and dentistry as a novel approach for the treatment of dental caries or ...

Eye Irritation Test (EIT) for Hazard Identification of Eye Irritating Chemicals using Reconstructed Human Cornea-like Epithelial (RhCE) Tissue Model

To comply with the Seventh Amendment to the EU Cosmetics Directive and EU REACH legislation, validated non-animal alternative methods for reliable and accurate assessment of ocular toxicity in man are needed. To address this need, we have developed an eye irritation test (EIT) which utilizes a three dimensional reconstructed human cornea-like epithelial (RhCE) tissue model that is based on normal human cells. The EIT is able to separate ocular&#160;irritants and corrosives (GHS Categories 1 and 2 combined) and those that do not require labeling (GHS No Category). The test utilizes two separate protocols, one designed for liquid chemicals and a second, similar protocol for solid test articles. The EIT prediction model uses a single exposure period (30 min for liquids, 6 hr for solids) and a single tissue viability cut-off (60.0% as determined by the MTT assay). Based on the results for 83 chemicals (44 liquids and 39 solids) EIT achieved 95.5/68.2/ and 81.8% sensitivity/specificity and accuracy (SS&#38;A) for liquids, 100.0/68.4/ and 84.6% SS&#38;A for solids, and 97.6/68.3/ and 83.1% for overall SS&#38;A. The EIT will contribute significantly to classifying the ocular irritation potential of a wide range of liquid and solid chemicals without the use of animals to meet regulatory testing requirements. The EpiOcular EIT method was implemented in 2015 into the OECD Test Guidelines as TG 492.

Eye Irritation Test EIT for Hazard Identification of Eye Irritating Chemicals using Reconstructed Human Cornea-like Epithelial RhCE Tissue Model

We have developed an eye irritation test which utilizes a three dimensional reconstructed human cornea-like epithelial (RhCE) tissue model. The test is able to discriminate between ocular irritant and corrosive materials (GHS Categories 1 and 2 combined) and those that do not require labeling (GHS No Category).

복원 된 인간의 각막 모양의 상피 (RHCE) 조직 모델을 사용하여 눈에 자극성 화학 물질의 위험 식별을위한 눈 자극 시험 (EIT)

To comply with the Seventh Amendment to the EU Cosmetics Directive and EU REACH legislation, validated non-animal alternative methods for reliable and ...

In Vitro Model of Physiological and Pathological Blood Flow with Application to Investigations of Vascular Cell Remodeling

Vascular disease is a common cause of death within the United States. Herein, we present a method to examine the contribution of flow dynamics towards vascular disease pathologies. Unhealthy arteries often present with wall stiffening, scarring, or partial stenosis which may all affect fluid flow rates, and the magnitude of pulsatile flow, or pulsatility index. Replication of various flow conditions is the result of tuning a flow pressure damping chamber downstream of a blood pump. Introduction of air within a closed flow system allows for a compressible medium to absorb pulsatile pressure from the pump, and therefore vary the pulsatility index. The method described herein is simply reproduced, with highly controllable input, and easily measurable results. Some limitations are recreation of the complex physiological pulse waveform, which is only approximated by the system. Endothelial cells, smooth muscle cells, and fibroblasts are affected by the blood flow through the artery. The dynamic component of blood flow is determined by the cardiac output and arterial wall compliance. Vascular cell mechano-transduction of flow dynamics may trigger cytokine release and cross-talk between cell types within the artery. Co-culture of vascular cells is a more accurate picture reflecting cell-cell interaction on the blood vessel wall and vascular response to mechanical signaling. Contribution of flow dynamics, including the cell response to the dynamic and mean (or steady) components of flow, is therefore an important metric in determining disease pathology and treatment efficacy. Through introducing an in vitro co-culture model and pressure damping downstream of blood pump which produces simulated cardiac output, various arterial disease pathologies may be investigated.

In Vitro Model of Physiological and Pathological Blood Flow with Application to Investigations of Vascular Cell Remodeling

This protocol replicates physiological or pathological blood flow in vitro to aid in determining cell response in disease pathologies. By introducing a pressure damping chamber downstream of a blood pump, blood flow across the vasculature can be recapitulated and imposed on a monolayer of vascular endothelium or a mimetic co-culture.

혈관 세포 리모델링의 조사에 생리의 체외 모델 및 응용 프로그램과 함께 병리학 혈액 흐름

Vascular disease is a common cause of death within the United States. Herein, we present a method to examine the contribution of flow dynamics towards ...

An In Vitro Organ Culture Model of the Murine Intervertebral Disc

Intervertebral disc (IVD) degeneration is a significant contributor to low back pain. The IVD is a fibrocartilaginous joint that serves to transmit and dampen loads in the spine. The IVD consists of a proteoglycan-rich nucleus pulposus (NP) and a collagen-rich annulus fibrosis (AF) sandwiched by cartilaginous end-plates. Together with the adjacent vertebrae, the vertebrae-IVD structure forms a functional spine unit (FSU). These microstructures contain unique cell types as well as unique extracellular matrices. Whole organ culture of the FSU preserves the native extracellular matrix, cell differentiation phenotypes, and cellular-matrix interactions. Thus, organ culture techniques are particularly useful for investigating the complex biological mechanisms of the IVD. Here, we describe a high-throughput approach for culturing whole lumbar mouse FSUs that provides an ideal platform for studying disease mechanisms and therapies for the IVD. Furthermore, we describe several applications that utilize this organ culture method to conduct further studies including contrast-enhanced microCT imaging and three-dimensional high-resolution finite element modeling of the IVD.

An In Vitro Organ Culture Model of the Murine Intervertebral Disc

Whole organ culture of the intervertebral disc (IVD) preserves the native extracellular matrix, cell phenotypes, and cellular-matrix interactions. Here we describe an IVD culture system using mouse lumbar and caudal IVDs in their functional spinal units and several applications utilizing this system.

<em&gt; 체외</em쥐의 추간판의&gt; 장기 문화 모델

Intervertebral disc (IVD) degeneration is a significant contributor to low back pain. The IVD is a fibrocartilaginous joint that serves to transmit and ...

Assembly and Tracking of Microbial Community Development within a Microwell Array Platform

The development of microbial communities depends on a combination of complex deterministic and stochastic factors that can dramatically alter the spatial distribution and activities of community members. We have developed a microwell array platform that can be used to rapidly assemble and track thousands of bacterial communities in parallel. This protocol highlights the utility of the platform and describes its use for optically monitoring the development of simple, two-member communities within an ensemble of arrays within the platform. This demonstration uses two mutants of Pseudomonas aeruginosa, part of a series of mutants developed to study Type VI secretion pathogenicity. Chromosomal inserts of either mCherry or GFP genes facilitate the constitutive expression of fluorescent proteins with distinct emission wavelengths that can be used to monitor community member abundance and location within each microwell. This protocol describes a detailed method for assembling mixtures of bacteria into the wells of the array and using time-lapse fluorescence imaging and quantitative image analysis to measure the relative growth of each member population over time. The seeding and assembly of the microwell platform, the imaging procedures necessary for the quantitative analysis of microbial communities within the array, and the methods that can be used to reveal interactions between microbial species area all discussed.

The development of microbial communities depends on a combination of factors, including environmental architecture, member abundance, traits, and interactions. This protocol describes a synthetic, microfabricated environment for the simultaneous tracking of thousands of communities contained in femtoliter wells, where key factors such as niche size and confinement can be approximated.

마이크로 웰 어레이 플랫폼 내 미생물 공동체 개발의 구성 및 추적

The development of microbial communities depends on a combination of complex deterministic and stochastic factors that can dramatically alter the spatial ...

Development of an Electrochemical DNA Biosensor to Detect a Foodborne Pathogen

Vibrio parahaemolyticus (V. parahaemolyticus) is a common foodborne pathogen that contributes to a large proportion of public health problems globally, significantly affecting the rate of human mortality and morbidity. Conventional methods for the detection of V. parahaemolyticus such as culture-based methods, immunological assays, and molecular-based methods require complicated sample handling and are time-consuming, tedious, and costly. Recently, biosensors have proven to be a promising and comprehensive detection method with the advantages of fast detection, cost-effectiveness, and practicality. This research focuses on developing a rapid method of detecting V. parahaemolyticus with high selectivity and sensitivity using the principles of DNA hybridization. In the work, characterization of synthesized polylactic acid-stabilized gold nanoparticles (PLA-AuNPs) was achieved using X-ray Diffraction (XRD), Ultraviolet-visible Spectroscopy (UV-Vis), Transmission Electron Microscopy (TEM), Field-emission Scanning Electron Microscopy (FESEM), and Cyclic Voltammetry (CV). We also carried out further testing of stability, sensitivity, and reproducibility of the PLA-AuNPs. We found that the PLA-AuNPs formed a sound structure of stabilized nanoparticles in aqueous solution. We also observed that the sensitivity improved as a result of the smaller charge transfer resistance (Rct) value and an increase of active surface area (0.41 cm2). The development of our DNA biosensor was based on modification of a screen-printed carbon electrode (SPCE) with PLA-AuNPs and using methylene blue (MB) as the redox indicator. We assessed the immobilization and hybridization events by differential pulse voltammetry (DPV). We found that complementary, non-complementary, and mismatched oligonucleotides were specifically distinguished by the fabricated biosensor. It also showed reliably sensitive detection in cross-reactivity studies against various food-borne pathogens and in the identification of V. parahaemolyticus in fresh cockles.

A protocol for the development of an electrochemical DNA biosensor comprising a polylactic acid-stabilized, gold nanoparticles-modified, screen-printed carbon electrode to detect Vibrio parahaemolyticus is presented.

Foodborne 병원 체를 검출 하는 전기 화학 DNA 바이오 센서의 개발

Vibrio parahaemolyticus (V. parahaemolyticus) is a common foodborne pathogen that contributes to a large proportion of public health problems globally, ...

An Ultrahigh-throughput Microfluidic Platform for Single-cell Genome Sequencing

Sequencing technologies have undergone a paradigm shift from bulk to single-cell resolution in response to an evolving understanding of the role of cellular heterogeneity in biological systems. However, single-cell sequencing of large populations has been hampered by limitations in processing genomes for sequencing. In this paper, we describe a method for single-cell genome sequencing (SiC-seq) which uses droplet microfluidics to isolate, amplify, and barcode the genomes of single cells. Cell encapsulation in microgels allows the compartmentalized purification and tagmentation of DNA, while a microfluidic merger efficiently pairs each genome with a unique single-cell oligonucleotide barcode, allowing &gt;50,000 single cells to be sequenced per run. The sequencing data is demultiplexed by barcode, generating groups of reads originating from single cells. As a high-throughput and low-bias method of single-cell sequencing, SiC-seq will enable a broader range of genomic studies targeted at diverse cell populations.

Single-cell sequencing reveals genotypic heterogeneity in biological systems, but current technologies lack the throughput necessary for the deep profiling of community composition and function. Here, we describe a microfluidic workflow for sequencing &gt;50,000 single-cell genomes from diverse cell populations.

단일 셀 게놈 시퀀싱 초고 처리량 미세 플랫폼

Sequencing technologies have undergone a paradigm shift from bulk to single-cell resolution in response to an evolving understanding of the role of ...

Rodent Behavioral Testing to Assess Functional Deficits Caused by Microelectrode Implantation in the Rat Motor Cortex

Medical devices implanted in the brain hold tremendous potential. As part of a Brain Machine Interface (BMI) system, intracortical microelectrodes demonstrate the ability to record action potentials from individual or small groups of neurons. Such recorded signals have successfully been used to allow patients to interface with or control&#160;computers, robotic limbs, and their own limbs. However, previous animal studies have shown that a microelectrode implantation in the brain not only damages the surrounding tissue but can also result in functional deficits. Here, we discuss a series of behavioral tests to quantify potential motor impairments following the implantation of intracortical microelectrodes into the motor cortex of a rat. The methods for open field grid, ladder crossing, and grip strength testing provide valuable information regarding the potential complications resulting from a microelectrode implantation. The results of the behavioral testing are correlated with endpoint histology, providing additional information on the pathological outcomes and impacts of this procedure on the adjacent tissue.

We have shown that a microelectrode implantation in the motor cortex of rats causes immediate and lasting motor deficits. The methods proposed herein outline a microelectrode implantation surgery and three rodent behavioral tasks to elucidate potential changes in the fine or gross motor function due to implantation-caused damage to the motor cortex.

설치류 쥐 모터 피 질에서 Microelectrode 주입으로 인 한 기능적 적자를 평가 하기 위해 테스트 행동

Medical devices implanted in the brain hold tremendous potential. As part of a Brain Machine Interface (BMI) system, intracortical microelectrodes ...

Advanced 3D Liver Models for In vitro Genotoxicity Testing Following Long-Term Nanomaterial Exposure

Due to the rapid development and implementation of a diverse array of engineered nanomaterials (ENM), exposure to ENM is inevitable and the development of robust, predictive in vitro test systems is essential. Hepatic toxicology is key when considering ENM exposure, as the liver serves a vital role in metabolic homeostasis and detoxification as well as being a major site of ENM accumulation post exposure. Based upon this and the accepted understanding that 2D hepatocyte models do not accurately mimic the complexities of intricate multi-cellular interactions and metabolic activity observed in vivo, there is a greater focus on the development of physiologically relevant 3D liver models tailored for ENM hazard assessment purposes in vitro. In line with the principles of the 3Rs to replace, reduce and refine animal experimentation, a 3D HepG2 cell-line based liver model has been developed, which is a user friendly, cost effective system that can support both extended and repeated ENM exposure regimes (&#8804;14 days). These spheroid models (&#8805;500 &#181;m in diameter) retain their proliferative capacity (i.e., dividing cell models) allowing them to be coupled with the &#8216;gold standard&#8217; micronucleus assay to effectively assess genotoxicity in vitro. Their ability to report on a range of toxicological endpoints (e.g., liver function, (pro-)inflammatory response, cytotoxicity and genotoxicity) has been characterized using several ENMs across both acute (24 h) and long-term (120 h) exposure regimes. This 3D in vitro hepatic model has the capacity to be utilized for evaluating more realistic ENM exposures, thereby providing a future in vitro approach to better support ENM hazard assessment in a routine and easily accessible manner.

This procedure was established to be used for developing advanced 3D hepatic cultures in vitro, which can provide a more physiologically relevant assessment of the genotoxic hazards associated with nanomaterial exposures over both an acute or long-term, repeated dose regimes.

장기 나노 물질 노출 다음 체외 물질 독성 테스트를 위한 고급 3D 간 모델

Due to the rapid development and implementation of a diverse array of engineered nanomaterials (ENM), exposure to ENM is inevitable and the development of ...

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