Name: three different protocols of corneal collagen crosslinking in keratoconus: conventional, accelerated and iontophoresis
Uploaded: 2015-11-12T13:00:00.000+00:00
Duration: 7 min 29 s
Description: Watch this Scientific Journal Video about Three Different Protocols of Corneal Collagen Crosslinking in Keratoconus: Conventional, Accelerated and Iontophoresis at JoVE.com

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://jove.unicartagenaproxy.elogim.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://jove.unicartagenaproxy.elogim.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://jove.unicartagenaproxy.elogim.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://jove.unicartagenaproxy.elogim.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>

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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&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp; Product number</td><td></td></tr><tr><td>C-CXL</td><td>Sooft SPA, Montegiorgio, Italy</td><td>Ricrolin&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp; 468465-6</td><td></td></tr><tr><td>A-CXL</td><td>Avedro Inc, Waltham, Massachusetts</td><td>VibeX&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp; 520-01863-006</td><td></td></tr><tr><td>I-CXL</td><td>Sooft SPA, Montegiorgio, Italy</td><td>Ricrolin+&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp; 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/cm&lt;sup&gt;2&lt;/sup&gt; 30 min</td></tr><tr><td>KXL System</td><td></td><td></td><td>UVA: 30 mW/cm&lt;sup&gt;2&lt;/sup&gt; 10 min</td></tr><tr><td>X-Vega</td><td></td><td></td><td>UVA: 10 mW/cm&lt;sup&gt;2&lt;/sup&gt; 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 ...

three-different-protocols-corneal-collagen-crosslinking-keratoconus

Universidad de Cartagena UDEC

Research

JoVE Journal

Medicine

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

Adenoviral Gene Therapy for Diabetic Keratopathy: Effects on Wound Healing and Stem Cell Marker Expression in Human Organ-cultured Corneas and Limbal Epithelial Cells

The goal of this protocol is to describe molecular alterations in human diabetic corneas and demonstrate how they can be alleviated by adenoviral gene therapy in organ-cultured corneas. The diabetic corneal disease is a complication of diabetes with frequent abnormalities of corneal nerves and epithelial wound healing. We have also documented significantly altered expression of several putative epithelial stem cell markers in human diabetic corneas. To alleviate these changes, adenoviral gene therapy was successfully implemented using the upregulation of c-met proto-oncogene expression and/or the downregulation of proteinases matrix metalloproteinase-10 (MMP-10) and cathepsin F. This therapy accelerated wound healing in diabetic corneas even when only the limbal stem cell compartment was transduced. The best results were obtained with combined treatment. For possible patient transplantation of normalized stem cells, an example is also presented of the optimization of gene transduction in stem cell-enriched cultures using polycationic enhancers. This approach may be useful not only for the selected genes but also for the other mediators of corneal epithelial wound healing and stem cell function.

An example of adenoviral gene therapy in the human diabetic organ-cultured corneas is presented towards the normalization of delayed wound healing and markedly reduced epithelial stem cell marker expression in these corneas. It also describes the optimization of this process in stem cell-enriched limbal epithelial cultures.

당뇨병 각막에 대한 아데노 바이러스 유전자 치료 : 인간 장기 배양 각막 및 윤부 상피 세포에 상처 치유에 효과 및 세포 마커의 발현 줄기

The goal of this protocol is to describe molecular alterations in human diabetic corneas and demonstrate how they can be alleviated by adenoviral gene ...

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발생학

Corneal Donor Tissue Preparation for Endothelial Keratoplasty

Over the past ten years, corneal transplantation surgical techniques have undergone revolutionary changes1,2. Since its inception, traditional full thickness corneal transplantation has been the treatment to restore sight in those limited by corneal disease. Some disadvantages to this approach include a high degree of post-operative astigmatism, lack of predictable refractive outcome, and disturbance to the ocular surface. The development of Descemet's stripping endothelial keratoplasty (DSEK), transplanting only the posterior corneal stroma, Descemet's membrane, and endothelium, has dramatically changed treatment of corneal endothelial disease. DSEK is performed through a smaller incision; this technique avoids 'open sky' surgery with its risk of hemorrhage or expulsion, decreases the incidence of postoperative wound dehiscence, reduces unpredictable refractive outcomes, and may decrease the rate of transplant rejection3-6.
Initially, cornea donor posterior lamellar dissection for DSEK was performed manually1 resulting in variable graft thickness and damage to the delicate corneal endothelial tissue during tissue processing. Automated lamellar dissection (Descemet's stripping automated endothelial keratoplasty, DSAEK) was developed to address these issues. Automated dissection utilizes the same technology as LASIK corneal flap creation with a mechanical microkeratome blade that helps to create uniform and thin tissue grafts for DSAEK surgery with minimal corneal endothelial cell loss in tissue processing.
Eye banks have been providing full thickness corneas for surgical transplantation for many years. In 2006, eye banks began to develop methodologies for supplying precut corneal tissue for endothelial keratoplasty. With the input of corneal surgeons, eye banks have developed thorough protocols to safely and effectively prepare posterior lamellar tissue for DSAEK surgery. This can be performed preoperatively at the eye bank. Research shows no significant difference in terms of the quality of the tissue7 or patient outcomes8,9 using eye bank precut tissue versus surgeon-prepared tissue for DSAEK surgery. For most corneal surgeons, the availability of precut DSAEK corneal tissue saves time and money10, and reduces the stress of performing the donor corneal dissection in the operating room. In part because of the ability of the eye banks to provide high quality posterior lamellar corneal in a timely manner, DSAEK has become the standard of care for surgical management of corneal endothelial disease.
The procedure that we are describing is the preparation of the posterior lamellar cornea at the eye bank for transplantation in DSAEK surgery (Figure 1).

Endothelial corneal transplantation is a surgical technique for treatment of posterior corneal diseases. Mechanical microkeratome dissection to prepare tissue results in thinner, more symmetric grafts with less endothelial cell loss and improved outcomes. Dissections can be performed at the eye bank prior to corneal transplantation surgery.

내피 Keratoplasty을위한 각막 기증자 조직 준비

Over the past ten years, corneal transplantation surgical techniques have undergone revolutionary changes1,2. Since its inception, traditional full ...

의학

Combination of Microstereolithography and Electrospinning to Produce Membranes Equipped with Niches for Corneal Regeneration

Corneal problems affect millions of people worldwide reducing their quality of life significantly. Corneal disease can be caused by illnesses such as Aniridia or Steven Johnson Syndrome as well as by external factors such as chemical burns or radiation. Current treatments are (i) the use of corneal grafts and (ii) the use of stem cell expanded in the laboratory and delivered on carriers (e.g., amniotic membrane); these treatments are relatively successful but unfortunately they can fail after 3-5 years. There is a need to design and manufacture new corneal biomaterial devices able to mimic in detail the physiological environment where stem cells reside in the cornea. Limbal stem cells are located in the limbus (circular area between cornea and sclera) in specific niches known as the Palisades of Vogt. In this work we have developed a new platform technology which combines two cutting-edge manufacturing techniques (microstereolithography and electrospinning) for the fabrication of corneal membranes that mimic to a certain extent the limbus. Our membranes contain artificial micropockets which aim to provide cells with protection as the Palisades of Vogt do in the eye.

We report a technique for the fabrication of micropockets within electrospun membranes in which to study cell behavior. Specifically, we describe a combination of microstereolithography and electrospinning for the production of PLGA (Poly(lactide-co-glycolide)) corneal biomaterial devices equipped with microfeatures.

마이크로 광 및 전기 방사의 조합은 각막 재생을위한 틈새를 장착 멤브레인을 생산하는

Corneal problems affect millions of people worldwide reducing their quality of life significantly. Corneal disease can be caused by illnesses such as ...

생체공학

An &#8220;All-laser&#8221; Endothelial Transplant

The &#8220;all laser&#8221; assisted endothelial keratoplasty is a procedure that is performed with a femtosecond laser used to cut the donor tissue at an intended depth, and a near infrared diode laser to weld the corneal tissue. The proposed technique enables to reach the three main goals in endothelial keratoplasty: a precise control in the thickness of the donor tissue; its easy insertion in the recipient bed and a reduced risk of donor lenticule dislocation. The donor cornea thickness is measured in the surgery room with optical coherence tomography (OCT), in order to correctly design the donor tissue dimensions. A femtosecond laser is used to cut the donor cornea. The recipient eye is prepared by manual stripping of the descemetic membrane. The donor endothelium is inserted into a Busin-injector, the peripheral inner side is stained with a proper chromophore (a water solution of Indocyanine Green) and then it is pulled in the anterior chamber. The transplanted tissue is placed in the final and correct location and then diode laser welding is induced from outside the eyeball. The procedure has been performed on more than 15 patients evidencing an improvement in surgery performances, with a good recovery of visual acuity and a reduced donor lenticule dislocation event.

An all laser procedure is proposed in the endothelial transplant. The surgical technique is based on the use of a femtosecond laser to prepare the donor tissue. Laser welding technique is then used to secure the donor endothelium in the correct position.

&quot;모든 레이저&quot;내피 세포 이식

The &#8220;all laser&#8221; assisted endothelial keratoplasty is a procedure that is performed with a femtosecond laser used to cut the donor tissue at an ...

Scleral Cross-linking Using Riboflavin and Ultraviolet-A Radiation for Prevention of Axial Myopia in a Rabbit Model

Myopic individuals, especially those with severe myopia, are at higher-than-normal risk of cataract, glaucoma, retinal detachment and chorioretinal abnormalities. In addition, pathological myopia is a common irreversible cause of visual impairment and blindness1-3. Our study demonstrates the effect of scleral crosslinking using riboflavin and ultraviolet-A radiation on the development of axial myopia in a rabbit model. The axial length of the eyeball was measured by A-scan ultrasound in New Zealand white rabbits aged 13 days (male and female). The eye then underwent 360&#176; conjunctival peritomy with scleral crosslinking, followed by tarsorrhaphy. Axial elongation was induced in 13 day-old New Zealand rabbits by suturing their right eye eyelids (tarsorrhaphy). The eyes were divided into quadrants, and every quadrant had two scleral irradiation zones, each with an area of 0.2 cm&#178; and a radius of 4 mm. Crosslinking was performed by dropping 0.1% dextran-free riboflavin-5-phosphate onto the irradiation zones 20 sec before ultraviolet-A irradiation and every 20 sec during the 200 sec irradiation time. UVA radiation (370 nm) was applied perpendicular to the sclera at 57 mW/cm&#178; (total UVA light dose, 57 J/cm&#178;). Tarsorrhaphies were removed on day 55, followed by repeated axial length measurements. This study demonstrates that scleral crosslinking with riboflavin and ultraviolet-A radiation effectively prevents occlusion-induced axial elongation in a rabbit model.

We demonstrate the effect of scleral crosslinking with riboflavin and UVA on an axial elongation rabbit eye. Axial elongation was induced in 13 day-old New Zealand rabbits (male and female) by suturing their right eye eyelids (tarsorrhaphy).

공막 래빗 모델에 축 방향 근시의 예방 리보플라빈 및 자외선-A 방사선을 사용하여 교차 결합

Myopic individuals, especially those with severe myopia, are at higher-than-normal risk of cataract, glaucoma, retinal detachment and chorioretinal ...

Second Harmonic Generation Signals in Rabbit Sclera As a Tool for Evaluation of Therapeutic Tissue Cross-linking (TXL) for Myopia

Methods to strengthen tissue by introducing chemical bonds (non-enzymatic cross-linking) into structural proteins (fibrillar collagens) for therapy include photochemical cross-linking and tissue cross-linking (TXL) methods. Such methods for inducing mechanical tissue property changes are being employed to the cornea in corneal thinning (mechanically weakened) disorders such as keratoconus as well as the sclera in progressive myopia, where thinning and weakening of the posterior sclera occurs and likely contributes to axial elongation. The primary target proteins for such tissue strengthening are fibrillar collagens which constitute the great majority of dry weight proteins in the cornea and sclera. Fortuitously, fibrillar collagens are the main source of second harmonic generation signals in the tissue extracellular space. Therefore, modifications of the collagen proteins, such as those induced through cross-linking therapies, could potentially be detected and quantitated through the use of second harmonic generation microscopy (SHGM). Monitoring SHGM signals through the use of a laser scanning microscopy system coupled with an infrared excitation light source is an exciting modern imaging method that is enjoying widespread usage in the biomedical sciences. Thus, the present study was undertaken in order to evaluate the use of SHGM microscopy as a means to measure induced cross-linking effects in ex vivo rabbit sclera, following an injection of a chemical cross-linking agent into the sub-Tenon&#39;s space (sT), an injection approach that is standard practice for causing ocular anesthesia during ophthalmologic clinical procedures. The chemical cross-linking agent, sodium hydroxymethylglycinate (SMG), is from a class of cosmetic preservatives known as formaldehyde releasing agents (FARs). Scleral changes following reaction with SMG resulted in increases in SHG signals and correlated with shifts in thermal denaturation temperature, a standard method for evaluating induced tissue cross-linking effects.

Second Harmonic Generation Signals in Rabbit Sclera As a Tool for Evaluation of Therapeutic Tissue Cross-linking TXL for Myopia

This protocol describes techniques for evaluating chemical cross-linking of the rabbit sclera using&#160;second harmonic generation imaging and differential scanning calorimetry.

치료 조직 (TXL) 근 시에 대 한 교차 연결의 평가 위한 도구로 토끼 Sclera에 제 2 고조파 발생 신호

Methods to strengthen tissue by introducing chemical bonds (non-enzymatic cross-linking) into structural proteins (fibrillar collagens) for therapy ...

Ex Vivo Corneal Organ Culture Model for Wound Healing Studies

The cornea has been used extensively as a model system to study wound healing. The ability to generate and utilize primary mammalian cells in two dimensional (2D) and three dimensional (3D) culture has generated a wealth of information not only about corneal biology but also about wound healing, myofibroblast biology, and scarring in general. The goal of the protocol is an assay system for quantifying myofibroblast development, which characterizes scarring. We demonstrate a corneal organ culture ex vivo model using pig eyes. In this anterior keratectomy wound, corneas still in the globe are wounded with a circular blade called a trephine. A plug of approximately 1/3 of the anterior cornea is removed including the epithelium, the basement membrane, and the anterior part of the stroma. After wounding, corneas are cut from the globe, mounted on a collagen/agar base, and cultured for two weeks in supplemented-serum free medium with stabilized vitamin C to augment cell proliferation and extracellular matrix secretion by resident fibroblasts. Activation of myofibroblasts in the anterior stroma is evident in the healed cornea. This model can be used to assay wound closure, the development of myofibroblasts and fibrotic markers, and for toxicology studies. In addition, the effects of small molecule inhibitors as well as lipid-mediated siRNA transfection for gene knockdown can be tested in this system.

A protocol for an ex vivo corneal organ culture model useful for wound healing studies is described. This model system can be used to assess the effects of agents to promote regenerative healing or drug toxicity in an organized 3D multicellular environment.

Ex Vivo 막 기관 문화 모델에 대 한 상처 치유 연구

The cornea has been used extensively as a model system to study wound healing. The ability to generate and utilize primary mammalian cells in two ...

Full-Field Optical Coherence Microscopy for Histology-Like Analysis of Stromal Features in Corneal Grafts

The quality of donor corneal stroma, which makes up about 90% of total corneal thickness, is likely to be one of the main, if not the major, limiting factor(s) for success of deep anterior lamellar and penetrating keratoplasty. These are surgical procedures that involve replacing part or all of the diseased corneal layers, respectively, by donated tissue, the graft, taken from a recently deceased individual. However, means to evaluate stromal quality of corneal grafts in eye banks are limited and lack the capability of high-resolution quantitative assessment of disease indicators. Full-field optical coherence microscopy (FF-OCM), permitting high-resolution 3D imaging of fresh or fixed ex vivo biological tissue samples, is a non-invasive technique well suited for donor cornea assessment. Here we describe a method for the qualitative and quantitative analysis of corneal stroma using FF-OCM. The protocol has been successfully applied to normal donor corneas and pathological corneal buttons, and can be used to identify healthy and pathologic features on both the macroscopic and microscopic level, thereby facilitating the detection of stromal disorders that could compromise the outcome of keratoplasty. By improving the graft quality control, this protocol has the potential to result in better selection (and rejection) of donor tissues and hence decreased graft failure.

We describe use of full-field optical coherence microscopy as a method for high quality assessment of corneal donor stroma. This protocol can be used to identify features indicative of health or disease, and is aimed at improving the screening and selection of donor tissues, and hence the results of keratoplasty.

각막 이식편의 기질 특징에 대한 조직학과 유사한 분석을 위한 전체 필드 광학 간섭 현미경

The quality of donor corneal stroma, which makes up about 90% of total corneal thickness, is likely to be one of the main, if not the major, limiting ...

면역학 및 감염병학

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 ...

축삭 재생 연구를 위한 각막 감각 섬유 손상 프로토콜

Three Strategies to Induce Neurotrophic Keratitis and Nerve Regeneration in Murine Cornea

The cornea is a transparent tissue that covers the eye and is crucial for clear vision. It is the most innervated tissue in the body. This innervation provides sensation and trophic function to the eye and contributes to preserving corneal integrity. The pathological disruption of this innervation is termed neurotrophic keratitis. This can be triggered by injury to the eye, surgery, or disease. In this study, we propose three different protocols for inflicting damage on the innervation in ways that recapitulate the three types of cases generally encountered in the clinic.
The first method consists in making an abrasion of the epithelium with an ophthalmic burr. This involves the removal of the epithelial layer, the free nerve endings, and the subbasal plexus in a manner similar to the photorefractive keratectomy surgery performed in the clinic. The second method&#160;only targets the innervation by sectioning it at the periphery with a biopsy punch, maintaining the integrity of the epithelium. This method is similar to the first steps of lamellar keratoplasty and leads to a degeneration of the innervation followed by regrowth of the axons in the central cornea. The last method damages the innervation of a transgenic mouse model using a multiphoton microscope, which specifically localizes the site of cauterization of the fluorescent nerve fibers. This method inflicts the same damage as photokeratitis, an overexposure to UV light.
This study describes different options for investigating the physiopathology of corneal innervation, particularly the degeneration and regeneration of the axons. Promoting regeneration is crucial for avoiding such complications as epithelium defects or even perforation of the cornea. The proposed models can help test new pharmacological molecules or gene therapy that enhance nerve regeneration and limit disease progression.

저자 스포트라이트: 혁신적인 모델을 통한 각막 신경 분포 연구의 발전 

Here, we propose three different methods of damaging the sensory fibres innervating the cornea. These methods facilitate the study of axon regeneration in mice. These three methods, which are adaptable to other animal models, are ideal for the study of corneal innervation physiology and regeneration.

쥐 각막의 신경 영양성 각막염 및 신경 재생을 유도하는 세 가지 전략

The cornea is a transparent tissue that covers the eye and is crucial for clear vision. It is the most innervated tissue in the body. This innervation ...