The authors have no acknowledgements.

이러한 프로토콜은 우리 기관의 인간 연구 윤리위원회의 지침을 따르십시오. 1. 기존의 각막 콜라겐 CXL (C-CXL) 환자 (1)의 제조 <ol><li> 오일 수술 전, 1 % 필로 카핀 두 번 처리 된 눈의 날을 삭제했습니다. </li><li> 수술실에서 무균 조건에서, 그 / 그녀를 다시 환자 거짓말. </li><li> 국소 마취에게 같은 oxybuprocaine 0.4 %를 관리. </li><li> 눈과 두 번 살균 요오드와 눈 주위의 피부를 청소합니다. </li><li> 열린 눈을 유지하기 위해 뚜껑 검경을 사용합니다. </li></ol> 2. 상피 제거 <ol><li> 원 각막 마커 각막의 중앙 9.0 mm로 표시합니다. </li><li> 무딘 주걱을 사용하여 기계적인 변연 절제술에 의한 각막 상피​​의 중심 7.0-9.0 mm를 제거합니다. </li></ol> 3. 리보플라빈 응용 프로그램 <ol><li> 0.1 %가 일에 20 % 덱스 트란과 리보플라빈 적용<html> 전자 각막 20 분마다 분. </li></ol> 4. UVA 조사 <ol><li> 3 mW의 / cm 2 (5.4 J / cm 2 표면 도즈)의 조도로 30 분간 작동 거리 5cm에서 370 nm 파장의 빛으로 UVA 각막을 조사. </li></ol><img alt="그림 1" src="/files/ftp_upload/53119/53119fig1.jpg" /> 그림 1. C-CXL의 UVA 조사 각막 3 mW의 / cm 2 (5.4 J / cm 2 표면 도즈)의 조도로 30 분간 5cm 작동 거리에서 370 nm 파장의 UVA 광 조사한다. <a href="https://www.jove.com/files/ftp_upload/53119/53119fig1large.jpg" target="_blank">이 그림의 더 큰 버전을 보려면 여기를 클릭하십시오.</a> <ol start="2"><li> 조사 기간 동안, 각막에 매 5 분 리보플라빈 방울을 적용합니다. </li><li> 조사 기간 동안, 국소 마취 (oxybuprocaine 0.4 %) 필요한 경우를 추가합니다. </li></ol>외과 &quot;&gt; 5. 끝 <ol><li> 넣어 항생제 방울 (토 브라 마이신 0.3 %) 및 ​​운영의 눈에 인공 눈물 (히알루 론산은 0.18 % 방울). </li><li> 재 상피화가 완료 될 때까지 수술 끝에 붕대 소프트 콘택트 렌즈를 배치했다. 재 상피화는 일반적으로 삼일 소요됩니다. </li><li> 같은 해열 (500 mg)을 더한 코데인 (30 mg)을, 6 약을 하루에 같은 진통제를 처방. </li><li> 각막의 재 상피화 후, 스테로이드와 국소 치료 (국소 덱사메타손 1 ㎎ / ㎖)을 시작 3-4 주 동안 계속합니다. 또한, 1 개월 인공 눈물을 하루에 4 번을 사용합니다. </li></ol> 2. 가속 각막 콜라겐 CXL (A-CXL) 환자 (1)의 제조 <ol><li> 오일 수술 전, 1 % 필로 카핀 두 번 처리 된 눈의 날을 삭제했습니다. </li><li> 수술실에서 무균 조건에서, 그 / 그녀를 다시 환자 거짓말. </li><li> 국소 마취에게 같은 oxybuprocaine 0.4 %를 관리. </li><li> 전자를 청소너희 두 번 살균 요오드와 눈 주위의 피부. </li><li> 열린 눈을 유지하기 위해 뚜껑 검경을 사용합니다. </li></ol> 2. 상피 제거 <ol><li> 각막 원 마커로 각막의 중앙 9.0 mm 마크 </li><li> 무딘 주걱을 사용하여 기계적인 변연 절제술에 의한 각막 상피​​의 중심 7.0-9.0 mm를 제거합니다. </li></ol> 3. 리보플라빈 응용 프로그램 <ol><li> 10 분 동안 각막에 덱스 트란없이 매 2 분 0.1 % 리보플라빈을 적용합니다. </li></ol> 4. UVA 조사 <ol><li> 30 mW의 / cm 2 (5.4 J / cm 2 표면 선량)와 3 분간 5cm 작동 거리에서의 방사 조도에서 370 nm 파장의 빛으로 UVA 각막을 조사. </li><li> (oxybuproca &amp; # 239; 0.4 %를 북동) 조사하는 동안, 국소 마취를 추가 필요. </li></ol> 수술 5. 끝 <ol><li> 항생제 방울 (토 브라 마이신 0.3 %), 인공 눈물을 (히알루 론산은 0을 삭제 배치<html> 작동 눈에 0.18 %). </li><li> 재 상피화가 완료 될 때까지 수술 끝에 붕대 소프트 콘택트 렌즈를 배치했다. 재 상피화는 일반적으로 삼일 소요됩니다. </li><li> 같은 해열 (500 mg)을 더한 코데인 (30 mg)을, 6 약을 하루에 같은 진통제를 처방. </li><li> 각막의 재 상피화 후, 스테로이드와 국소 치료 (국소 덱사메타손 1 ㎎ / ㎖)을 시작 3-4 주 동안 계속합니다. 또한, 1 개월 인공 눈물을 하루에 4 번을 사용합니다. </li></ol> 3. 이온 토 포레 시스 (I-CXL) 환자 (1)의 제조 <ol><li> 오일 수술 전, 1 % 필로 카핀 두 번 처리 된 눈의 날을 삭제했습니다. </li><li> 수술실에서 무균 조건에서, 그 / 그녀를 다시 환자 거짓말. </li><li> 국소 마취에게 같은 oxybuprocaine 0.4 %를 관리. </li><li> 눈과 두 번 살균 요오드와 눈 주위의 피부를 청소합니다. </li><li> 열린 눈을 유지하기 위해 뚜껑 검경을 사용합니다. </li></ol><html> _step &quot;&gt; 2. 이온 토 포레 시스 장치를 놓습니다. <ol><li> 수술 분야에서 이마에 스티커 수동 전극을 적용합니다. </li><li> 열린 눈, 활성 전극, 흡입 링을 적용합니다. 흡인을 해제하기 전에 각막의 주변에 흡입 링의 중심. </li></ol><img alt="그림 2" src="/files/ftp_upload/53119/53119fig2.jpg" /> 그림 2. 이온 토 포레 시스 장치. 수동 전극은 수술 분야에서 이마와 활성 전극, 흡입 링에 적용되는, 열린 눈에 적용된다. <a href="https://www.jove.com/files/ftp_upload/53119/53119fig2large.jpg" target="_blank">이 그림의 더 큰 버전을 보려면 여기를 클릭하십시오.</a> 3. 리보플라빈 응용 프로그램 <ol><li> 0.1 hypoosmolar %는 덱스 트란없이 리보플라빈과 흡입 반지를 입력합니다. </li></ol><html> JPG &quot;/&gt; I-CXL 그림 3. 리보플라빈 응용 프로그램입니다. 흡입 링은 덱스 트란없이 hypoosmolar 0.1 % riboflavine으로 가득합니다. <a href="https://www.jove.com/files/ftp_upload/53119/53119fig3large.jpg" target="_blank">이 그림의 더 큰 버전을 보려면 여기를 클릭하십시오.</a> <ol start="2"><li> 0.2 mA의 전류를 시작하고 점차적으로 5 분 (그림 4)의 총 이온 토 포레 시스 시간을 1.0 mA로 증가. </li></ol><img alt="그림 4" src="/files/ftp_upload/53119/53119fig4.jpg" /> 침투 리보플라빈 그림 4. 이온 삼투압 장치. 전류가 처음에 0.2 mA이며, 점차적으로 1.0 mA로 증가. 총 이온 토 포레 시스 시간은 5 분입니다. <a href="https://www.jove.com/files/ftp_upload/53119/53119fig4large.jpg" target="_blank">이 그림의 더 큰 버전을 보려면 여기를 클릭하십시오.</a> EP &quot;&gt; 4. UVA 조사 <ol><li> 경우 10mW / ㎠ (5.4 J / cm 2 표면 도즈), 9 분 동안 5cm 작동 거리에서의 방사 조도에서 370 nm 파장의 빛으로 UVA 각막을 조사. </li><li> (oxybuproca &amp; # 239; 0.4 %를 북동) 조사하는 동안, 국소 마취를 추가 필요. </li></ol> 수술 5. 끝 <ol><li> 항생제 방울 (토 브라 마이신 0.3 %) 및 ​​운영의 눈에 인공 눈물을 (히알루 론산은 0.18 % 방울) 놓습니다. </li><li> 같은 해열 (500 mg)을 더한 코데인 (30 mg)을, 6 약을 하루에 같은 진통제를 처방. </li><li> 수술 후 스테로이드와 국소 치료 (국소 덱사메타손 1 ㎎ / ㎖)을 시작 3-4 주 동안 계속합니다. 또한, 1 개월 인공 눈물을 하루에 4 번을 사용합니다. </li></ol>

<ol>
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N. .. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=In+vivo+confocal+microscopy+analyses+of+corneal+microstructural+changes+in+a+prospective+study+of+collagen+cross-linking+in+keratoconus.">In vivo confocal microscopy analyses of corneal microstructural changes in a prospective study of collagen cross-linking in keratoconus.</a> Ophthalmology. 121 (2), 469-474 (2014).</li><li>Touboul, D., 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=Corneal+confocal+microscopy+following+conventional,+transepithelial,+and+accelerated+corneal+collagen+cross-linking+procedures+for+keratoconus.">Corneal confocal microscopy following conventional, transepithelial, and accelerated corneal collagen cross-linking procedures for keratoconus.</a> J Refract Surg. 28 (11), 769-776 (2012).</li><li>Bouheraoua, 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=Optical+coherence+tomography+and+confocal+microscopy+following+three+different+protocols+of+corneal+collagen-crosslinking+in+keratoconus.">Optical coherence tomography and confocal microscopy following three different protocols of corneal collagen-crosslinking in keratoconus.</a> Invest Ophthalmol Vis Sci. 55 (11), 7601-7609 (2014).</li><li>Hafezi, F., Mrochen, M., Iseli, H. P., Seiler, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Collagen+crosslinking+with+ultraviolet-A+and+hypoosmolar+riboflavin+solution+in+thin+corneas.">Collagen crosslinking with ultraviolet-A and hypoosmolar riboflavin solution in thin corneas.</a> J Cataract Refract Surg. 35 (4), 621-624 (2009).</li><li>C&#305;nar, Y., 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=Comparison+of+accelerated+and+conventional+corneal+collagen+cross-linking+for+progressive+keratoconus.">Comparison of accelerated and conventional corneal collagen cross-linking for progressive keratoconus.</a> Cutan Ocul Toxicol. 33 (3), 218-222 (2013).</li><li>Cing&#252;, A. K., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Transient+corneal+endothelial+changes+following+accelerated+collagen+cross-linking+for+the+treatment+of+progressive+keratoconus.">Transient corneal endothelial changes following accelerated collagen cross-linking for the treatment of progressive keratoconus.</a> Cutan Ocul Toxicol. 33 (2), 127-131 (2013).</li><li>Spoerl, E., Mrochen, M., Sliney, D., Trokel, S., Seiler, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Safety+of+UVA-riboflavin+cross-linking+of+the+cornea.">Safety of UVA-riboflavin cross-linking of the cornea.</a> Cornea. 26 (4), 385-389 (2007).</li><li>Gokhale, N. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Corneal+endothelial+damage+after+collagen+cross-linking+treatment.">Corneal endothelial damage after collagen cross-linking treatment.</a> Cornea. 30 (12), 1495-1498 (2011).</li><li>Rootman, D. 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=Pharmacokinetics+and+safety+of+transcorneal+iontophoresis+of+tobramycin+in+the+rabbit.">Pharmacokinetics and safety of transcorneal iontophoresis of tobramycin in the rabbit.</a> Invest Ophthalmol Vis Sci. 29 (9), 1397-1401 (1998).</li><li>Vinciguerra, 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=Transepithelial+iontophoresis+corneal+collagen+cross-linking+for+progressive+keratoconus:+initial+clinical+outcomes.">Transepithelial iontophoresis corneal collagen cross-linking for progressive keratoconus: initial clinical outcomes.</a> J Refract Surg. 30 (11), 746-753 (2014).</li><li>Caporossi, A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Riboflavin-UVA-induced+corneal+collagen+cross-linking+in+pediatric+patients.">Riboflavin-UVA-induced corneal collagen cross-linking in pediatric patients.</a> Cornea. 31 (3), 227-231 (2012).</li><li>Buzzonetti, L., Petrocelli, G., Valente, P., Larossi, G., Ardia, R., Petroni, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Iontophoretic+transepithelial+corneal+cross-linking+to+halt+keratoconus+in+pediatric+cases:+15-month+follow-up.">Iontophoretic transepithelial corneal cross-linking to halt keratoconus in pediatric cases: 15-month follow-up.</a> Cornea. 34 (5), 512-515 (2015).</li><li>Baiocchi, S., Mazzotta, C., Cerretani, D., Caporossi, T., Caporossi, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Corneal+crosslinking:+riboflavin+concentration+in+corneal+stroma+exposed+with+and+without+epithelium.">Corneal crosslinking: riboflavin concentration in corneal stroma exposed with and without epithelium.</a> J Cataract Refract Surg. 35 (5), 893-899 (2009).</li><li>Wollensak, G., Iomdina, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Biomechanical+and+histological+changes+after+corneal+crosslinking+with+and+without+epithelial+debridement.">Biomechanical and histological changes after corneal crosslinking with and without epithelial debridement.</a> J Cataract Refract Surg. 35 (3), 540-546 (2009).</li><li>Soeters, N., Wisse, R. P., Godefrooij, D. A., Imhof, S. M., Tahzib, N. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Transepithelial+versus+epithelium-off+corneal+cross-linking+for+the+treatment+of+progressive+keratoconus:+a+randomized+controlled+trial.">Transepithelial versus epithelium-off corneal cross-linking for the treatment of progressive keratoconus: a randomized controlled trial.</a> Am J Ophthalmol. 159 (5), 821-828 (2015).</li></ol>

The authors have nothing to disclose.

UVA 조사 및 리보플라빈을 사용하여 CXL은 원추 각막의 진행을 체포에 대한 표준 치료입니다. 리보플라빈 3 UVA 조사에 화학적 공유 결합 (가교)을 유도 감광제이다. 각막,이 현상은 각막 강성을 증가 콜라겐 섬유 사이의 상호 링크를 작성합니다. 이 현상을 잘 설명 하였지만, 지금까지 intracorneal 가교의 직접적인 증거는 없었다. 그럼에도 불구하고, 여러 연구 CXL 따라서 C-3-8의 효과를 ...

원추 각막은 일반적으로 각막 모양의 변형의 결과로 일반 인구 1 천 1에보고 된 양자 및 진보적 인 각막 확장증이며, 따라서 시력이 감소했다. 각막 조기 사춘기에 보통 존재하는 질병은 일반적으로 안정 될 때 진행하는 경향이 환자의 일생 동안 가변 될 수 있지만, 삶의 제 십년에 셋째까지 진행한다. 원추 각막의 진행을 중단함으로써, 가교는 연기 또는 회피 각막 목표로하고있다. 현재까지 임상 연구에서 입증 된 진보적 인 원추 각막의 유일한 안전하고 효과적으로 치료 강성을 증가시키고 따라서 원추 각막의 진행 3-8을 중단하는 것을 목표로 기존의 각막 콜라겐 교차 결합 (C-CXL) 프로토콜입니다. 수술 시간 및 감염성 각막염이나 각막 안개 9로 C-CXL의 다른 가능한 위험 요인을 줄이기 위해 몇 가지 개선 된 프로토콜이에설명. 첫째, 가속 CXL (A-CXL)에, UVA의 높은 조도는 시간을 단축 (10)를 통해 각막에 전달된다. 둘째, 상피 괴사에 대한 필요성을 피하기 위해, transepithelial 접근법이 이용되어왔다. 종래의 프로토콜 (11)에 비교할 때 불행히도, 이들은 제한된 성공. CXL 동안 각막 리보플라빈 배달 최근 transepithelial 방법은 이온 토 포레 시스 (I-CXL)이지만,이 처리의 엄격한 평가는 아직 12 실시되지 않았다. 이온 토 포레 시스는 작은 전류를 조직을 통하여 이온화 약물의 침투를 향상시키기 위해 적용되는 비 침습 기술이다. 이온 삼투압에 의한 CXL에서, 리보플라빈은 각막 상피​​를 통해 침투하는 이온화된다. 생체 공 초점 현미경 (IVCM)는 같은 원추 각막 (13) 등의 질환에서 비정상적인 각막의 세포 변화를 강조 할 수있는 각막을 영상화하는 방법이다. 사실, IVCM서브 기저 신경 얼기의 밀도에서 특정 감소 원추 각막에서 각막의 모든 레이어에 변경을 증명하고 기질은 13-15 각막 간질했다. 또한, IVCM는 C-CXL (16) 후 각막의 미세 구조 분석을위한 매우 편리한 것으로 입증되었습니다. 각막 분계선은 300 μm의 (17, 18)의 깊이에서 C-CXL 후 hyperreflective OCT (AS) 전 안부 빛 간섭 단층 촬영에서 볼 줄 1개월으로 설명되어 있습니다. IVCM 다음의 C-CXL 300 ㎛의 깊이로 각막 실질 세포의 부재를 포함하여 각막의 구조 변경에 대한 정보를 제공합니다. 이 무 세포 영역의 깊이뿐만 아니라 각막 간질 내에서 경계선의 깊이가 간섭 단층 1과의 CXL 처리 (19), 및 각막 분계선 깊이 측정의 유효 깊이와 관련이있는 것으로 보인다 10월로 밝혀 CXL 개월 후에는 효과적인 임상 적으로 제안되어왔다CXL 효과 (18)의 평가 방법. 본 연구에서 우리는 10월 및 공 초점 현미경 AS에 의해 각막 간질 분계선의 측정을 사용하여 세 가지 (기존, 가속 및 이온 토 포레 시스) 각막 콜라겐 가교 프로토콜의 효율성을 조사합니다. 우리는 또한 정량적으로 세 가지 치료 후 각막 미세 구조 변화를 분석하기 위해 IVCM을 사용했다.

<table>
	<tbody>
		<tr>
			<td></td>
			<td></td>
			<td>Riboflavin&#160;&#160;&#160;&#160;&#160;&#160;&#160; Product number</td>
			<td></td>
		</tr>
		<tr>
			<td>C-CXL</td>
			<td>Sooft SPA, Montegiorgio, Italy</td>
			<td>Ricrolin&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160; 468465-6</td>
			<td></td>
		</tr>
		<tr>
			<td>A-CXL</td>
			<td>Avedro Inc, Waltham, Massachusetts</td>
			<td>VibeX&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160; 520-01863-006</td>
			<td></td>
		</tr>
		<tr>
			<td>I-CXL</td>
			<td>Sooft SPA, Montegiorgio, Italy</td>
			<td>Ricrolin+&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160; 975481-6</td>
			<td>Passive electrode: PROTENS ELITE 4848LE/ Active electrode: IONTOFOR CXL</td>
		</tr>
		<tr>
			<td></td>
			<td></td>
			<td>UVA Machine</td>
			<td></td>
		</tr>
		<tr>
			<td>X-Vega</td>
			<td></td>
			<td></td>
			<td>UVA: 3 mW/cm2 30 min</td>
		</tr>
		<tr>
			<td>KXL System</td>
			<td></td>
			<td></td>
			<td>UVA: 30 mW/cm2 10 min</td>
		</tr>
		<tr>
			<td>X-Vega</td>
			<td></td>
			<td></td>
			<td>UVA: 10 mW/cm2 9 min</td>
		</tr>
	</tbody>
</table>

three different protocols of corneal collagen crosslinking in keratoconus: conventional, accelerated and iontophoresis

원추 각막은 양자 및 진보적 인 각막 확장증입니다. 그 진행을 느리게하기 위하여, 각막 콜라겐 가교 (CXL)은 최근 효율적인 치료 방법으로 도입되었다. 생물학적 및 화학적 과학, 가교 반응 분자 사이에 형성되는 새로운 화학 결합을 의미한다. 따라서, 각막 콜라겐 CXL의 목적은 합성 각막 기질에 콜라겐 섬유 사이의 가교의 형성을 증가시키는 것이다. 종래 CXL (C-CXL) 프로토콜의 효율은 이미 여러 임상 연구에 도시되었다는 사실에도 불구하고, 절차 및 각막 상피​​의 제거 기간에도 향상 것이다. 따라서, 두 개의 새로운 최적화 CXL 프로토콜 응집성 평가를 제공하기 위해, 우리는 세 CXL 치료의 하나받은 원추 각막 환자 연구 : 이온 도입 (I-CXL)를 CXL (A-CXL), 및 종래 CXL을 (가속 C-CXL). - CXL는 6 시간 빠른 CXL 절차 U입니다여전히 상피 제거하는 등 10 시간 더 높은 UVA의 조도를 노래하지만. 이온 삼투 요법은 작은 전류가 각막에 걸쳐 리보플라빈 침투를 개선하기 위해인가되는 transepithelial 비 침습 기술이다. OCT (AS) 및 생체 공 초점 현미경 (IVCM)에서 전 안부 빛 간섭 단층 촬영을 사용하여, 우리는 치료 침투의 깊이에 대해 기존의 CXL 프로토콜은 진보적 인 원추 각막 치료를위한 표준 유지 결론 지었다. 가속 CXL은 얇은 각막을 치료하는, 빠르고 효과적이고 안전한 대안이 될 것으로 보인다. 이온 삼투 요법의 사용은 여전히​​ 연구되고 있으며 더욱 신중 고려되어야한다.

<html> 각막 경계 라인 (73.6 SD) 301.6 ㎛의 평균 깊이의 경우 92 %에 10월로 볼 수 있었다 <img alt="그림 5" src="/files/ftp_upload/53119/53119fig5.jpg" /> C-CXL 후 그림 5. 분계선. OCT (AS) 고해상도 각막 전 안부 빛 간섭 단층 촬영 검사 기존의 각막 콜라겐 교차 후 358 μm의 (흰색 화살표), 1 개월 평균 깊이에서 각막 기질 분계선을 시각화 연결 (C-CXL). 스케...

Watch this Scientific Journal Video about Three Different Protocols of Corneal Collagen Crosslinking in Keratoconus: Conventional, Accelerated and Iontophoresis at JoVE.com

Three Different Protocols of Corneal Collagen Crosslinking in Keratoconus: Conventional, Accelerated and Iontophoresis

Corneal collagen cross-linking (CXL) is the only conservative treatment currently available to halt keratoconus progression by improving the biomechanical rigidity of the corneal stroma. The aim of this manuscript is to highlight the methods of three different protocols of CXL: conventional CXL (C-CXL), accelerated CXL (A-CXL), and iontophoresis CXL (I-CXL).

원추 각막의 각막 콜라겐 가교의 세 가지 서로 다른 프로토콜 : 기존의가, 가속 및 이온 토 포레 시스

Keratoconus is a bilateral and progressive corneal ectasia. In order to slow down its progression, corneal collagen cross-linking (CXL) has recently been ...

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19.8K 조회수.  Quinze-Vingts National Ophthalmology Hospital. Corneal collagen cross-linking (CXL) is the only conservative treatment currently available to halt keratoconus progression by improving the biomechanical rigidity of the corneal stroma. The aim of this manuscript is to highlight the methods of three different protocols of CXL: conventional CXL (C-CXL), accelerated CXL (A-CXL), and iontophoresis CXL (I-CXL).

비디오: Three Different Protocols of Corneal Collagen Crosslinking in Keratoconus: Conventional, Accelerated and Iontophoresis

Staining Protocols for Human Pancreatic Islets

Estimates of islet area and numbers and endocrine cell composition in the adult human pancreas vary from several hundred thousand to several million and beta mass ranges from 500 to 1500 mg 1-3. With this known heterogeneity, a standard processing and staining procedure was developed so that pancreatic regions were clearly defined and islets characterized using rigorous histopathology and immunolocalization examinations. 
Standardized procedures for processing human pancreas recovered from organ donors are described in part 1 of this series. The pancreas is processed into 3 main regions (head, body, tail) followed by transverse sections. Transverse sections from the pancreas head are further divided, as indicated based on size, and numbered alphabetically to denote subsections. This standardization allows for a complete cross sectional analysis of the head region including the uncinate region which contains islets composed primarily of pancreatic polypeptide cells to the tail region.
The current report comprises part 2 of this series and describes the procedures used for serial sectioning and histopathological characterization of the pancreatic paraffin sections with an emphasis on islet endocrine cells, replication, and T-cell infiltrates. Pathology of pancreatic sections is intended to characterize both exocrine, ductular, and endocrine components. The exocrine compartment is evaluated for the presence of pancreatitis (active or chronic), atrophy, fibrosis, and fat, as well as the duct system, particularly in relationship to the presence of pancreatic intraductal neoplasia4. Islets are evaluated for morphology, size, and density, endocrine cells, inflammation, fibrosis, amyloid, and the presence of replicating or apoptotic cells using H&amp;E and IHC stains. 
The final component described in part 2 is the provision of the stained slides as digitized whole slide images. The digitized slides are organized by case and pancreas region in an online pathology database creating a virtual biobank. Access to this online collection is currently provided to over 200 clinicians and scientists involved in type 1 diabetes research. The online database provides a means for rapid and complete data sharing and for investigators to select blocks for paraffin or frozen serial sections.

This video demonstrates procedures for characterization of human pancreatic islets using hematoxylin and eosin (H&E) and immunohistochemistry (IHC). Pancreatic sections from head, body, and tail regions are stained by both H&amp;E and IHC to determine islet endocrine composition (insulin, glucagon, and pancreatic polypeptide), cell replication (Ki67), and inflammatory infiltrates (H&E, CD3). The uncinate region is localized using IHC for pancreatic polypeptide.

인간의 췌장 Islets위한 염색법 프로토콜

Estimates of islet area and numbers and endocrine cell composition in the adult human pancreas vary from several hundred thousand to several million and ...

연구

의학

An Alkali-burn Injury Model of Corneal Neovascularization in the Mouse

Under normal conditions, the cornea is avascular, and this transparency is essential for maintaining good visual acuity. Neovascularization (NV) of the cornea, which can be caused by trauma, keratoplasty or infectious disease, breaks down the so called &lsquo;angiogenic privilege' of the cornea and forms the basis of multiple visual pathologies that may even lead to blindness. Although there are several treatment options available, the fundamental medical need presented by corneal neovascular pathologies remains unmet. In order to develop safe, effective, and targeted therapies, a reliable model of corneal NV and pharmacological intervention is required. Here, we describe an alkali-burn injury corneal neovascularization model in the mouse. This protocol provides a method for the application of a controlled alkali-burn injury to the cornea, administration of a pharmacological compound of interest, and visualization of the result. This method could prove instrumental for studying the mechanisms and opportunities for intervention in corneal NV and other neovascular disorders.

Neovascularization (NV) of the cornea can complicate multiple visual pathologies. Utilizing a controlled, alkali-burn injury model, a quantifiable level of corneal NV can be produced for mechanistic study of corneal NV and evaluation of potential therapies for neovascular disorders.

마우스의 각막 신생 혈관의 알칼리 화상 상해 모델

Under normal conditions, the cornea is avascular, and this transparency is essential for maintaining good visual acuity. Neovascularization (NV) of the ...

A Rat Model of Ventricular Fibrillation and Resuscitation by Conventional Closed-chest Technique

A rat model of electrically-induced ventricular fibrillation followed by cardiac resuscitation using a closed chest technique that incorporates the basic components of cardiopulmonary resuscitation in humans is herein described. The model was developed in 1988 and has been used in approximately 70 peer-reviewed publications examining a myriad of resuscitation aspects including its physiology and pathophysiology, determinants of resuscitability, pharmacologic interventions, and even the effects of cell therapies. The model featured in this presentation includes: (1) vascular catheterization to measure aortic and right atrial pressures, to measure cardiac output by thermodilution, and to electrically induce ventricular fibrillation; and (2) tracheal intubation for positive pressure ventilation with oxygen enriched gas and assessment of the end-tidal CO2. A typical sequence of intervention entails: (1) electrical induction of ventricular fibrillation, (2) chest compression using a mechanical piston device concomitantly with positive pressure ventilation delivering oxygen-enriched gas, (3) electrical shocks to terminate ventricular fibrillation and reestablish cardiac activity, (4) assessment of post-resuscitation hemodynamic and metabolic function, and (5) assessment of survival and recovery of organ function. A robust inventory of measurements is available that includes &#8211; but is not limited to &#8211; hemodynamic, metabolic, and tissue measurements. The model has been highly effective in developing new resuscitation concepts and examining novel therapeutic interventions before their testing in larger and translationally more relevant animal models of cardiac arrest and resuscitation.

This article describes a rat model of electrically-induced ventricular fibrillation and resuscitation by chest compression, ventilation, and delivery of electrical shocks that simulates an episode of sudden cardiac arrest and conventional cardiopulmonary resuscitation. The model enables gathering insights on the pathophysiology of cardiac arrest and exploration of new resuscitation strategies.

기존의 폐쇄 가슴 기술에 의해 쥐의 심실 세동의 모델 및 소생

A rat model of electrically-induced ventricular fibrillation followed by cardiac resuscitation using a closed chest technique that incorporates the basic ...

Echocardiographic Approaches and Protocols for Comprehensive Phenotypic Characterization of Valvular Heart Disease in Mice

The aim of this manuscript and accompanying video is to provide an overview of the methods and approaches used for imaging heart valve function in rodents, with detailed descriptions of the appropriate methods for anesthesia, the echocardiographic windows used, the imaging planes and probe orientations for image acquisition, the methods for data analysis, and the limitations of emerging technologies for the evaluation of cardiac and valvular function. Importantly, we also highlight several future areas of research in cardiac and heart valve imaging that may be leveraged to gain insights into the pathogenesis of valve disease in preclinical animal models. We propose that using a systematic approach to evaluating cardiac and heart valve function in mice can result in more robust and reproducible data, as well as facilitate the discovery of previously underappreciated phenotypes in genetically-altered and/or physiologically-stressed mice.

This protocol provides a detailed description of the echocardiographic approach for comprehensive phenotyping of heart and heart valve function in mice.

마우스의 심장 판막 질환의 종합 표현형 특성에 대한 심 초음파 접근 및 프로토콜

The aim of this manuscript and accompanying video is to provide an overview of the methods and approaches used for imaging heart valve function in ...

Induction of Accelerated Atherosclerosis in Mice: The "Wire-Injury" Model

Atherosclerosis is a proliferative fibro-inflammatory disease developing in the arterial wall, inducing a deficient blood flow or a lack of blood flow. Moreover, by rupture of the defective vascular wall, atherosclerosis induces occlusive thrombus formation, which represents the main cause of myocardial infarction or stroke and the most frequent cause of death. Despite the advances in the cardiovascular field, many questions remain unanswered, and additional basic research is essential to improve our understanding of the molecular mechanisms during atherosclerosis and its effects. Due to limited clinical studies, there is a need for representative animal models recreating atherosclerotic conditions such as neointima formation after stent implantation, balloon angioplasty, or endarterectomy. Since the mouse presents many advantages and is the most frequently used model for studying molecular processes, the current study proposes an invasive procedure of endothelial denudation, also known as the wire-injury model, which is representative of the human condition of neointima formation in arteries after revascularization procedures.

Induction of Accelerated Atherosclerosis in Mice: The &quot;Wire-Injury&quot; Model

This study describes an invasive procedure for the induction of accelerated atherosclerosis in mice. In comparison to other methods using electric- or cryo-induced injury, mechanical-induced injury mimics the human condition of restenosis after revascularization therapies and is ideal for the study of the molecular mechanisms involved.

마우스에서 가속 죽상 경화증의 유도: "와이어 부상" 모델

Atherosclerosis is a proliferative fibro-inflammatory disease developing in the arterial wall, inducing a deficient blood flow or a lack of blood flow. ...

Corneal Epithelial Abrasion with Ocular Burr As a Model for Cornea Wound Healing

The murine cornea provides an excellent model to study wound healing. The cornea is the outermost layer of the eye, and thus is the first defense to injury. In fact, the most common type of eye injury found in clinic is a corneal abrasion. Here, we utilize an ocular burr to induce an abrasion resulting in removal of the corneal epithelium in vivo on anesthetized mice. This method allows for targeted and reproducible epithelial disruption, leaving other areas intact. In addition, we describe the visualization of the abraded epithelium with fluorescein staining and provide concrete advice on how to visualize the abraded cornea. Then, we follow the timeline of wound healing 0, 18, and 72 h after abrasion, until the wound is re-epithelialized. The epithelial abrasion model of corneal injury is ideal for studies on epithelial cell proliferation, migration and re-epithelialization of the corneal layers. However, this method is not optimal to study stromal activation during wound healing, because the ocular burr does not penetrate to the stromal cell layers. This method is also suitable for clinical applications, for example, pre-clinical test of drug effectiveness.

This protocol describes a method to inflict an abrasion to the ocular surface of the mouse, and to follow the wound healing process thereafter. The protocol takes advantage of an ocular burr to partially remove the surface epithelium of the eye in anaesthetized mice.

각 막에 대 한 모델로 눈 버와 각 막 상피 마모 상처 치유

The murine cornea provides an excellent model to study wound healing. The cornea is the outermost layer of the eye, and thus is the first defense to ...

Simultaneous Laryngopharyngeal and Conventional Esophageal pH Monitoring

In addition to typical reflux symptoms, many patients with gastroesophageal reflux disease (GERD) present with extraesophageal symptoms such as cough, hoarseness or asthma, which can be caused by laryngopharyngeal reflux (LPR). Due to their multifactorial origin, those symptoms can be a great diagnostic and therapeutic challenge. Esophageal pH-monitoring is commonly used to determine abnormal esophageal acid exposure and confirm the diagnosis of GERD. However, for better evaluation of acid exposure above the upper esophageal sphincter, a new laryngopharyngeal pH measurement system is now available and may lead to more reliable results in patients with predominantly extraesophageal symptoms. This article aims to present a standardized protocol for simultaneous pH measurement using esophageal and laryngopharyngeal pH probes in order to obtain acid exposure scores from both measurements.

Laryngopharyngeal pH monitoring has been specifically designed to measure acid exposure above the upper esophageal sphincter and complements diagnostic evaluation in patients that present with mainly extraesophageal reflux symptoms. Patients with suspected laryngopharyngeal reflux (LPR) were evaluated using distal esophageal and laryngopharyngeal pH testing simultaneously.

In addition to typical reflux symptoms, many patients with gastroesophageal reflux disease (GERD) present with extraesophageal symptoms such as cough, ...

Three-Dimensional Collagen Matrix Scaffold Implantation as a Liver Regeneration Strategy

Liver diseases are the leading cause of death worldwide. Excessive alcohol consumption, a high-fat diet, and hepatitis C virus infection promote fibrosis, cirrhosis, and/or hepatocellular carcinoma. Liver transplantation is the clinically recommended procedure to improve and extend the life span of patients in advanced disease stages. However, only 10% of transplants are successful, with organ availability, presurgical and postsurgical procedures, and elevated costs directly correlated with that result. Extracellular matrix (ECM) scaffolds have emerged as an alternative for tissue restoration. Biocompatibility and graft acceptance are the main beneficial characteristics of those biomaterials. Although the capacity to restore the size and correct function of the liver has been evaluated in liver hepatectomy models, the use of scaffolds or some kind of support to replace the volume of the extirpated liver mass has not been assessed.
Partial hepatectomy was performed in a rat liver with the xenoimplantation of a collagen matrix scaffold (CMS) from a bovine condyle. Left liver lobe tissue was removed (approximately 40%), and an equal proportion of CMS was surgically implanted. Liver function tests were evaluated before and after the surgical procedure. After days 3, 14, and 21, the animals were euthanized, and macroscopic and histologic evaluations were performed. On days 3 and 14, adipose tissue was observed surrounding the CMS, with no clinical evidence of rejection or infection, as was vessel neoformation and CMS reabsorption at day 21. There was histologic evidence of an insignificant inflammation process and migration of adjacent cells to the CMS, observed with the hematoxylin and eosin (H&#38;E) and Masson&#39;s trichrome staining. The CMS was shown to perform well in liver tissue and could be a useful alternative for studying tissue regeneration and repair in chronic liver diseases.

Liver diseases are induced by many causes that promote fibrosis or cirrhosis. Transplantation is the only option for recovering health. However, given the scarcity of transplantable organs, alternatives must be explored. Our research proposes the implantation of collagen scaffolds in liver tissue from an animal model.

간 재생 전략으로 3차원 콜라겐 매트릭스 스캐폴드 이식

Liver diseases are the leading cause of death worldwide. Excessive alcohol consumption, a high-fat diet, and hepatitis C virus infection promote fibrosis, ...

Ex Vivo and In Vivo Animal Models for Mechanical and Chemical Injuries of Corneal Epithelium

Corneal injury to the ocular surface, including chemical burn and trauma, may cause severe scarring, symblepharon, corneal limbal stem cells deficiency, and result in a large, persistent corneal epithelial defect. Epithelial defect with the following corneal opacity and peripheral neovascularization result in irreversible visual impairment and hinder future management, especially keratoplasty. Since the animal model can be used as an effective drug development platform, models of corneal injury to the mouse and alkali burn to rabbit corneal epithelium are developed here. New Zealand white rabbit is used in the alkali burn model. Different concentrations of sodium hydroxide can be applied onto the central circular area of the cornea for 30 s under intramuscular and topical anesthesia. After copious isotonic normal saline irrigation, residual loose corneal epithelium was removed with corneal burr deep down to the Bowman's layer within this circular area. Wound healing was documented by fluorescein staining under Cobalt blue light. C57BL/6 mice were used in the traumatic model of murine corneal epithelium. The murine central cornea was marked using a skin punch, 2 mm in diameter, and then debrided by a corneal rust ring remover with a 0.5 mm burr under a stereomicroscope. These models can be prospectively used to validate the therapeutic effect of eye drops or mixed agents such as stem cells, which potentially facilitate corneal epithelial regeneration. By observing corneal opacity, peripheral neovascularization, and conjunctival congestion with stereomicroscope and imaging software, therapeutic effects in these animal models can be monitored.

Ex Vivo and In Vivo Animal Models for Mechanical and Chemical Injuries of Corneal Epithelium

Here, animal models based on mouse and rabbit are developed for mechanical and chemical injury of corneal epithelium to screen new therapeutics and the underlying mechanism.

각막 상피의 기계적 및 화학적 손상에 대한 Ex Vivo 및 In Vivo 동물 모델

Corneal injury to the ocular surface, including chemical burn and trauma, may cause severe scarring, symblepharon, corneal limbal stem cells deficiency, ...

A Human Corneal Organ Culture Model of Descemet's Stripping Only with Accelerated Healing Stimulated by Engineered Fibroblast Growth Factor 1

Fuchs Endothelial Corneal Dystrophy (FECD) results from dysfunctional corneal endothelial cells (CECs) and is currently treated by transplantation of the whole cornea or Descemet's membrane. Recent developments in ocular surgery have established Descemet's Stripping Only (DSO), a surgical technique in which a central circle of guttae-dense Descemet's membrane is removed to allow for the migration of CECs onto the smooth stroma, restoring function and vision to the cornea. While this potential treatment option is of high interest in the field of ophthalmic research, no successful ex vivo models of DSO have been established and clinical data is limited. This work presents a novel wound-healing model simulating DSO in human donor corneas. Using this approach to evaluate the efficacy of the human engineered FGF1 (NM141), we found that treatment accelerated healing via stimulation of migration and proliferation of CECs. This finding was confirmed in 11 pairs of human corneas with signs of dystrophy reported by the eye banks in order to verify that these results can be replicated in patients with Fuchs' Dystrophy, as the target population of the DSO procedure.

Descemet's Stripping Only is an experimental procedure wherein patients with central corneal guttae resulting from Fuchs Endothelial Corneal Dystrophy have Descemet's membrane stripped for peripheral cells to regenerate the endothelial layer. We present novel methodology simulating DSO in dystrophic human corneas ex vivo with accelerated healing stimulated by eFGF1 (NM141).

조작된 섬유아세포 성장 인자 1에 의해 자극된 가속화된 치유를 통해서만 Descemet의 스트리핑의 인간 각막 배양 모델

Fuchs Endothelial Corneal Dystrophy (FECD) results from dysfunctional corneal endothelial cells (CECs) and is currently treated by transplantation of the ...