저자는 실험실에서 자신의 전문적이고 우수한 기술 작업 양 달리아 셀라 씨와 에미 샤론에 감사합니다.

동물 연구에서 동물들의 사용에 ARVO 해상도에 따라 처리 하였다. 연구 프로토콜은 실험 동물 연구를위한 제도적위원회 (.; 021211 승인없이 022-4598-2)에 의해 승인되었다. 수술 # 1 1. 준비 <ol><li> 무게와 케타민 염산염 100 ㎎ / ㎖ (100 ㎎ / ㎏)과 자일 라진 하이드로 클로라이드 25 ㎎ / ㎖ (12.5 ㎎ / ㎏)의 근육 내 주사로 13 일 된 뉴질랜드 흰 토끼를 마취. 이러한 투여 량으로 인해 수술의 길이로 사용된다. 고통 반사의 부족에 의해 마취의 적절한 수준을 보장. </li><li> 멸균 면봉을 사용하여,이 건조 각막을 방지하기 위해 비 작동 눈 안연고 소량을 적용한다. 건조에서 각막을 방지하기 위해 작업 중에 생리 식염수가 운영하는 눈의 각막에 떨어 0.9 %를 적용합니다. </li><li> oxybuprocain (국소 마취 후 각각의 눈에 세 개의 축 방향 길이 측정을 수행다음 예 염산염 0.4 %), 초음파 A-스캐너를 사용하고 측정을 평균. 중앙 각막에 수직으로 프로브를 적용합니다. </li></ol> 2. 수술 # 1 - 사전 가교 단계 <ol><li> 안과 운영 현미경 수술 안과 집게와 가위를 사용하여, 수술 안과 집게와 가위로 360 ° 결막 절개를합니다. </li><li> 각진 집게 또는 근육 후크 네 개의 안구 직근 근육을 확인하고 2-0 꼰 실크 비 바늘로 봉합으로 그들을 분리를 사용하여. 주 : 실크 봉합사가 원하는 방향으로 세계를 움직이는 데 도움이됩니다. 안구는 네 개의 직근 근육 (사 사분면) 사이의 사분면으로 나누어 져 있습니다. <ol><li> 피부 마커 각 사분면에서 두 영역, 적도 공막 하나와 후방 공막 하나와 마크. 이러한 조사 영역이다. </li></ol></li><li> 0.1 % 덱스 트란없는 리보플라빈 -5- phosphat을 함유하는 5 ㎖ 주사기로 3 ㎖를 준비이자형. 26 G 눈물 정맥 또는 25 G 테이퍼 hydrodelineator에 주사기를 연결합니다. </li><li> 하향 경사 주문품 광섬유에 연결된 UV A (370 ㎚) 광원을 포함하는 조사 장치를 준비한다. 에너지 전력 및 교정의 측정에 따라 (아래 참고 참조), 57 mW의 / cm²에 장치를 설정합니다. 주 : 교정 제조업체에서 상업적으로 실시 하였다. 이 방법은 57 J / cm²의 총 UVA 광 용량을 제공합니다. (0.2 cm (2)에 57 mW의 / cm 2 2.2 J 0.2 cm 당 2 스폿의 누적로드 200 초 동안 11.4 mW의 / 0.2 cm (2)를 사용하여 11.4 mW의 / 0.2 cm 2이다) </li></ol> 3. 수술 # 1 - 가교 <ol><li> 선택한 사분면의 반대 방향으로 실크 봉합사를 당겨 안구를 움직여 처리 수 (하부 시간 사분면을 처리 할 때 예를 위로 당기고 코). 2-0 꼰 실크 비 니들 봉합사를 사용하여 안구 운동을 수행하는 전각 직근을 solates. </li><li> 조사가 시작되기 전에 조사 영역 20 초에 0.1 % 덱스 트란없는 리보플라빈 -5- 인산을 포함하는 감광제 솔루션을 적용합니다. </li><li> 200 초 조사 기간 동안 하향 경사 주문품 광섬유를 사용하는 조사 영역을 조사한다. 4mm의 반경 0.2 cm² 각 영역의 면적을 측정한다. </li><li> 200 초 조사 기간 동안 조사 영역에 20 초마다 0.1 % 덱스 트란없는 리보플라빈 -5- 인산염을 함유하는 감광제 용액을 적용한다. 주 : 조사 및이 외과 의사에 의해 동시에 리보플라빈의 낙하를 수행합니다. </li><li> 반복 네 개의 안구 사분면의 각 3.1-3.4 단계를 반복합니다. </li></ol> 4. 수술 # 1 - 포스트 크로스 연결 단계 <ol><li> 눈 주위의 털을 면도. 외과 안과 가위를 사용하여 뚜껑 마진을 손질하고 부드럽게 상부와 4-0 꼰 실크 아이보리 색상을 사용하여 아래 눈꺼풀 봉합 - 3/8 원 재절 커팅 - 13mm 길이 C-3 바늘 (안검 봉합). 절차 (클로람페니콜 5 %)의 단부에 뚜껑 가장자리에 안연고 소량을 적용한다. </li></ol> 5. 수술 # 1 - 포스트 수술 케어 <ol><li> 복구하는 동안 가열 램프 아래에있는 동물을 유지합니다. 이 흉골 드러 누움을 유지하기 위해 충분한 의식을 회복 할 때까지 무인 동물을 두지 마십시오. </li><li> 완전히 복구 될 때까지 새로운 침대와 깨끗한 새장에 동물을 배치합니다. 복구 후, 동물은 동물의 방으로 돌아갑니다. 경우에만 깨어 자신의 어머니에게 동물을 돌려줍니다. </li><li> 처음 48 시간 동안 일반 조건 및 질병이나 감염의 징후에 관한 동물을 검사합니다. 토끼의 행동에 변화가 관찰되는 경우 필요에 따라 섹션 1. 관리]의 진통에 따라 진통제를 관리 할 수​​ 있습니다. (설피 린은 50 ㎎ / kg마다 6 시간 방울). </li></ol> 6. 수술 # 2 <ol><li> 오십 오일의 후urgery # 1, 무게와 케타민 히드로 클로라이드 100 ㎎ / ㎖ (100 ㎎ / ㎏)와 자일 라진 하이드로 클로라이드 25 ㎎ / ㎖ (12.5 밀리그램 / kg)을 근육 주사하여 마취 된 토끼. </li><li> 안과 운영 현미경 수술 안과 가위를 사용하여 안검 봉합을 제거합니다. 국소 마취 (oxybuprocaine 염산염 0.4 %), 초음파 A-스캐너를 사용한 후 각각의 눈에 세 축 방향 길이 측정을 수행하고 측정 값을 평균. 중앙 각막에 수직으로 프로브를 적용합니다. </li><li> 복강 페노바비탈 나트륨 하였다 (200 mg / 15 kg 체중)로 토끼를 안락사. </li></ol>

<ol>
	<li>McBrien, N. A., Gentle, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Role+of+the+sclera+in+the+development+and+pathological+complications+of+myopia.">Role of the sclera in the development and pathological complications of myopia.</a> Prog. Retin. Eye Res. 22, 307-338 (2003).</li><li>Saw, S. M., Gazzard, G., Au Eong, K. -. G., Tan, D. T. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Myopia:+attempts+to+arrest+progression.">Myopia: attempts to arrest progression.</a> Br. J. Ophthalmol. 86, 1306-1311 (2002).</li><li>Bullimore, M. A., Jones, L. A., Moeschberger, M. L., Zadnik, K., Payor, R. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+retrospective+study+of+myopia+progression+in+adult+contact+lens+wearers.">A retrospective study of myopia progression in adult contact lens wearers.</a> Invest Ophthalmol Vis Sci. 43, 2110-2113 (2002).</li><li>Rose, K., Smith, W. E., Morgan, I., Mitchell, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+increasing+prevalence+of+myopia:+implications+for+Australia.">The increasing prevalence of myopia: implications for Australia.</a> Clin. Exp. Ophthalmol. 29, 116-120 (2001).</li><li>Saw, S. M., Gazzard, G., Shih-Yen, E. C., Chua, W. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Myopia+and+associated+pathological+complications.">Myopia and associated pathological complications.</a> Ophthal. Physiol. Opt. 25, 381-391 (2005).</li><li>Tano, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Pathologic+myopia+-+where+are+we+now.">Pathologic myopia - where are we now.</a> Am. J. Ophthalmol. 134, 645-660 (2002).</li><li>Greene, P. R., Guyton, D. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Time+course+of+rhesus+lid-suture+myopia.">Time course of rhesus lid-suture myopia.</a> Exp. Eye Res. 42, 529-534 (1986).</li><li>McBrien, N. A., Norton, T. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+development+of+experimental+myopia+and+ocular+component+dimensions+in+monocularly+lid-sutured+tree+shrews+(Tupaia+belangeri).">The development of experimental myopia and ocular component dimensions in monocularly lid-sutured tree shrews (Tupaia belangeri).</a> Vision Res. 32, 843-852 (1992).</li><li>McKanna, J. A., Casagrande, V. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Reduced+lens+development+in+lid-suture+myopia.">Reduced lens development in lid-suture myopia.</a> Exp. Eye Res. 26, 715-723 (1978).</li><li>Shapiro, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Experimental+visual+deprivation+and+myopia.">Experimental visual deprivation and myopia.</a> Doc. Ophthalmol. Proc. Ser. 28, 193-195 (1981).</li><li>Wiesel, T. N., Raviola, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Increase+in+axial+length+of+the+macaque+monkey+eye+after+corneal+opacification.">Increase in axial length of the macaque monkey eye after corneal opacification.</a> Invest. Ophthalmol. Vis. Sci. 18, 1232-1236 (1979).</li><li>Weiss, A. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Unilateral+high+myopia:+optical+components,+associated+factors+and+visual+outcomes.">Unilateral high myopia: optical components, associated factors and visual outcomes.</a> Br J Ophthalmol. 87, 1025-1031 (2003).</li><li>Mechanic, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Crosslinking+of+collagen+in+a+heritable+disorder+of+connective+tissue:+Ehlers-Danlos+syndrome.">Crosslinking of collagen in a heritable disorder of connective tissue: Ehlers-Danlos syndrome.</a> Biochem. Biophys. Res. Com. 47, 267-272 (1972).</li><li>Curtin, B. J., Iwamoto, T., Renaldo, D. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Normal+and+staphylomatous+sclera+of+high+myopia.+An+electron+microscopic+study.">Normal and staphylomatous sclera of high myopia. An electron microscopic study.</a> Arch. Ophthalmol. 97, 912-915 (1979).</li><li>Liu, K. R., Chen, M. S., Ko, L. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Electron+microscopic+studies+of+the+scleral+collagen+fibre+in+excessively+high+myopia.">Electron microscopic studies of the scleral collagen fibre in excessively high myopia.</a> J. Formosan Med. Assoc. 85, 1032-1038 (1986).</li><li>Funata, M., Tokoro, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Scleral+change+in+experimentally+myopic+monkeys.">Scleral change in experimentally myopic monkeys.</a> Graefes Arch. Clin. Exp. Ophthalmol. 228, 174-179 (1990).</li><li>McBrien, N. A., Norton, T. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Prevention+of+collagen+cross-linking+increases+form+deprivation+myopia+in+tree+shrew.">Prevention of collagen cross-linking increases form deprivation myopia in tree shrew.</a> Exp. Eye Res. 59, 475-486 (1994).</li><li>Iomdina, E. N., Daragan, V. A., Ilyina, E. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Certain+biomechanical+properties+and+cross+linking+of+the+scleral+shell+of+the+eye+in+progressive+myopia.">Certain biomechanical properties and cross linking of the scleral shell of the eye in progressive myopia.</a> Proceedings of XIVth Congress on Biomechanics. Paris: International Society of Biomechanics. , 616-617 (1993).</li><li>Wollensak, G., Sp&#246;rl, E., 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=Riboflavin+/+ultraviolet-A-induced+collagen+crosslinking+for+the+treatment+of+keratoconus.">Riboflavin / ultraviolet-A-induced collagen crosslinking for the treatment of keratoconus.</a> Am. J. Ophthalmol. 135, 620-627 (2003).</li><li>Wollensak, G., Sp&#246;rl, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Collagencrosslinking+of+human+and+porcine+sclera.">Collagencrosslinking of human and porcine sclera.</a> J. Cataract Refract. Surg. 30, 689-695 (2004).</li><li>Wollensak, G., Iomdina, E., Dittert, D. -. D., Salamatina, O., Stoltenburg, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cross-linking+of+scleral+collagen+in+the+rabbit+using+riboflavin+and+UVA.">Cross-linking of scleral collagen in the rabbit using riboflavin and UVA.</a> Acta Ophthalmol. Scand. 83, 477-482 (2005).</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=Long-term+biomechanical+properties+of+rabbit+sclera+after+collagen+crosslinking+using+riboflavin+and+ultraviolet+(UVA).">Long-term biomechanical properties of rabbit sclera after collagen crosslinking using riboflavin and ultraviolet (UVA).</a> Acta Ophthalmol. 87, 193-198 (2009).</li><li>Wildsoet, C. F., Norton, T. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Toward+controlling+myopia+progression.">Toward controlling myopia progression.</a> Optom. Vis. Sci. 76, 341-342 (1999).</li><li>Avetisov, E. S., Tarutta, E. P., Iomdina, E. N., Vinetskaya, M. I., Andreyeva, M. I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Nonsurgical+and+surgical+methods+of+sclera+reinforcement+in+progressive+myopia.">Nonsurgical and surgical methods of sclera reinforcement in progressive myopia.</a> Acta Ophthalmol. Scand. 75, 618-623 (1997).</li><li>Thompson, F. B. <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+sclera+reinforcement+technique.">A simplified sclera reinforcement technique.</a> Am. J. Ophthalmol. 86, 782-790 (1978).</li><li>Whitmore, W. G., Curtin, B. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Scleral+reinforcement:+two+case+reports.">Scleral reinforcement: two case reports.</a> Ophthalmic Surg. 18, 503-505 (1987).</li><li>Dotan, A., Kremer, I., Livnat, T., Zigler, A., Weinberger, D., Bourla, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Scleral+cross-linking+using+riboflavin+and+ultraviolet-A+radiation+for+prevention+of+progressive+myopia+in+a+rabbit+model.">Scleral cross-linking using riboflavin and ultraviolet-A radiation for prevention of progressive myopia in a rabbit model.</a> Exp Eye Res. 127, 190-195 (2014).</li></ol>

저자는 더 경쟁 재정적 이해 관계가 없음을 선언합니다.

우리는 리보플라빈 및 UVA 조사로 가교 결합 기술을 사용하여 토끼 모델에서 축 근시 예방 생체 내 연구에서 최초로 제시한다. 다른 실험 동물 연구는 이러한 유형에 사용될 수 있지만, 우리는 주로 인해 눈의 크기와 공막 표면 상에 가교 결합을 수행 할 필요로 토끼를 선택 하였다. 우리는 토끼 공막을 노출하고 윗 눈꺼풀과 아랫 눈꺼풀을 봉합하는 까다로운 절차로 것으로 나타났습니다. 우리는 뚜껑 마진 트리밍과 눈꺼풀이 제대로 정정에 대한 필요없이 주어진 기간 동안 닫혀 ​​있는지 확인하기 위해 우리의 프로토콜에 언급 된 봉합사를 사용하는 것이 좋습니다. 고려해야 할 중요한 것은 조사 장치 및 필요한 전력 에너지 광섬유를 보정 할 필요가있다. H &amp; E 조직 학적 슬라이드는 가교 절차를 시행 눈에 망막에 독성 변화를 보이지 않았다. 또한, 조사 에너지 및 노출 시간의 위치에 포커스 양, 망막 맥락막과 공막에 리보플라빈 및 UVA 조사의 잠재적 독성을 해결하기 위해 수행되어야한다. 이 연구 영장 고려의 몇 가지 제한. 만 수술 전 및 수술 후 축 길이를 측정 하였다. 안구 축 길이는 스캔을 초음파에 의해 측정 하였다. 우리는 생체 특성, 굴절 분석이나 공막의 생체 역학적 특성을 평가하지 않았다. 함께, 우리는 토끼 눈의 축 신장에 리보플라빈과 UVA와 공막 가교의 효과가 근시 연구에 긍정적 인 영향을 미칠 것이라고 생각합니다.

근시는 굴절 장애의 가장 보편적이다. 미국과 유럽에서 근시의 유병률은 약 30 %로보고하고, 아시아 국가에서 일반 인구의 1, 2의 60 %까지 영향을줍니다. 근시 진행은 보통 2 년 간격으로 3 위에 약 -0.5 디옵터의 속도로, 근시의 최대 50 %에서 발생합니다. 근시에 의해 부과 된 건강 비용은 안경에 대한 비용, 콘택트 렌즈 및 굴절 수술과 녹내장, 백내장, 망막 박리 및 시각 장애 4 ~ 6의 증가 된 건강 위험과 관련된 비용을 포함, 상당한 수 있습니다. 근시의 동물 연구에서 시력 감소는 눈과 각막 문신 (11)에서 짧은 거리에서 눈꺼풀 봉합 7-10, occluder에의 배치에 의해 유도된다. 인공 근시는 이들 연구에서 발생하도록하지만, 폐색 처리가없는 시력 상실 실험 카 같이 매우 어린 동물에서 수행되어야성인 표본에 밖으로 폭발물 공격이 성​​공 증명했다. 심한 근시의 중요한 기능 중 하나는 Ehlers- 같이 인해 공막의 일부 대사 장애로 인해 시각적 박탈 12 후 emmetropization의 방해 피드백 메커니즘에 아마 공막의 진보적 인 숱과 공막의 병리학 적 변화 Danlos 증후군 13. 궁극적으로, 두 메커니즘으로 인해 이러한 fibrillogenesis (16)의 감소 콜라겐 섬유 직경 14, 15 및 장애와 근시 공막의 구조적 이상에 스트레칭과 공막, 망막 및 맥락막의 숱로 이어집니다. 몇몇 연구는 손상된 콜라겐 가교 근시 공막 17-18 약화 과정에서 중요한 요소가 있음을 보여 주었다. WOLLENSAK 외. 19-21 리보플라빈 및 자외선 A (UVA) 조사에 감광제를 도포하여 콜라겐 가교를 유도 (370 ㎚)체외 (19)의 돼지와 인간의 공막의 강성을 크게, 157 % 증가 및 생체 내에서 토끼 공막 강성의 465% 증가 (탄성 계수) (20)를 언급했다. 강성 4 개월 3 일 후, 320.4 %, 277.6 %로 증가, 8 개월 (영률) 후 22 5백2퍼센트 : 가교 결합은 생체 내 토끼 공막의 장기적 효과가 있었다. 근시 진행을 체포 ​​치료 시도가 23-26 발표되었다 그러나이 방법의 성공은 논쟁의 여지가있다. 진행성 근시 방지의 어떠한 효과적인 수단은 지금까지 발견되지 않았다. 근시의 원인은 아직 논란, 그 처리는 도전을 포즈. 이러한 발견에 기초하여, 그 공막 가교 근시 진행 공막 기반 치료를위한 수단으로서 역할을 할 수 있다고 가정한다. 이 연구의 목적은 공막 콜라겐 crosslinki 검사하는시각적 축 폐색에 의해 유도 축 근시의 개발에 영향을 ng를.

<table border="0" cellpadding="0" cellspacing="0" width="667">
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:244px;">2-0 braided silk non-needled sutures&#160;</td>
			<td style="width:137px;">ETHICON</td>
			<td style="width:119px;">W193</td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="25">
			<td height="25" style="height:25px;">4-0 braided silk ivory color&#160;</td>
			<td style="width:137px;">ETHICON</td>
			<td style="width:119px;">W816</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:244px;">0.1% dextran-free riboflavin-5-phosphate 1mg:1ml</td>
			<td style="width:137px;">Concept for Pharmacy Ltd</td>
			<td style="width:119px;">D2-5025</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">UV A (370nm) light source&#160;</td>
			<td>O/E LAND Inc</td>
			<td style="width:119px;">NCSU033B</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Beveled down custom made fiber optic&#160;</td>
			<td>Prizmatix Ltd</td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="49">
			<td height="49" style="height:49px;width:244px;">26G lacrimal cannula&#160;</td>
			<td style="width:137px;">Beaver-visitec International Ltd.&#160;</td>
			<td style="width:119px;">REF581276</td>
			<td></td>
		</tr>
		<tr height="44">
			<td height="44" style="height:44px;width:244px;">25G tapered hydrodelineator [Blumenthal]&#160;</td>
			<td style="width:137px;">Beaver-visitec International Ltd.&#160;</td>
			<td style="width:119px;">REF585107</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:244px;">13 days old rabbits</td>
			<td style="width:137px;">Harlan&#160;</td>
			<td style="width:119px;">1NZWR40</td>
			<td></td>
		</tr>
		<tr height="23">
			<td height="23" style="height:23px;width:244px;">Ultrasonic biometer</td>
			<td>Allergan-Humphrey</td>
			<td style="width:119px;">820-519</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:244px;">Skin marker</td>
			<td style="width:137px;">Devon</td>
			<td style="width:119px;">4237101664X</td>
			<td></td>
		</tr>
	</tbody>
</table>

scleral cross-linking using riboflavin and ultraviolet-a radiation for prevention of axial myopia in a rabbit model

근시 개인, 심한 근시 특히 사람들은, 백내장, 녹내장, 망막 박리와 맥락 망막 이상 높은 정상보다 위험이 있습니다. 또한, 병적 근시는 시각 장애인 1-3의 일반적인 원인은 비가 역적이다. 본 연구는 토끼 모델에서 축 근시의 개발 리보플라빈 및 자외선 방사선을 사용하여 가교 공막의 효과를 보여준다. 안구 축 길이는 십삼일 (암수) 세 뉴질랜드 흰 토끼에 주사 초음파에 의해 측정 하였다. 눈은 안검 봉합을 다음 공막 가교와 360 ° 결막 절개를을 시행 하였다. 축 방향 신장은 자신의 오른쪽 눈의 눈꺼풀 (안검 봉합을) 봉합에 의해 십삼일 된 뉴질랜드 토끼 유도 하였다. 눈은 사분면으로 분할하고, 사분면마다 0.2 cm²의 면적 4 mm의 반경이 공막 조사 영역 각각 있었다. 가교는 0.1 %를 놓는 방법으로 수행 하였다덱스 트란없는-A 자외선 조사 전, 조사 영역이 20 초 및 200 초의 조사 시간 동안 매 20 초 상 리보플라빈 -5- 인산염. UVA 방사선 (370 나노 미터)은 57 mW의 / cm² (총 UVA 광 용량, 57 J / cm²)에서 공막에 수직으로 적용되었다. Tarsorrhaphies 반복 축 길이를 측정 한 후, 하루 55에 제거 하였다. 이 연구는 리보플라빈 자외선-A 방사선 공막 가교 효과적으로 토끼 모델에서 폐색에 의한 축 방향 신장을 방지 함을 보여줍니다.

도 1 및도 2에 그래픽으로 두 ​​군의 축 길이 측정 값을 보여준다. 왼쪽 눈은 (그림 1)에서 운영하지 않는 동안 그룹 1 토끼는 오른쪽 눈에 공막 가교 및 안검 봉합을 시행 하였다. 왼쪽 눈은 (그림 2)에서 운영하지 않는 동안 2 군 토끼는 오른쪽 눈에만 절개를하고 안검 봉합을 시행 하였다. 공막 가교 및 안검 봉합을 시행 한 1 군으로, 오른쪽 눈 축 길이를 평균 3.61 ± 0.76 mm의 평균 차이를 나중에 눈꺼풀 봉합 전 10.68 ± 0.74 mm와 14.29 ± 0.3 mm 오십오일를 측정 하였다. unsutured / 작동이되지 않는 왼쪽 눈에 해당하는 값은 4.44 ± 0.81 mm (그림 1)의 평균 차이, 10.70 ± 0.79 mm와 15.14 ± 0.32 mm이었다. 봉합의 축 길이의 비교및 폐쇄 단계의 끝에서 unsutured 눈은 봉합 눈에 덜 순증 한 것으로 밝혀졌습니다. 만 절개를하고 tarsorrhaphies 시행 2 군에서, 오른쪽 눈 축 길이를 평균은 5.19 ± 0.85 mm의 평균 차이를 나중에 눈꺼풀 봉합 전 10.50 ± 0.67 mm와 15.69 ± 0.39 mm 오십오일를 측정 하였다. unsutured / 작동이되지 않는 왼쪽 눈에 해당하는 값은 4.20 ± 0.67 mm (그림 2)의 평균 차이, 10.54 ± 0.71 mm와 14.74 ± 0.38 mm이었다. 교합 단계의 끝에서 봉합 및 unsutured 눈의 축 길이의 비교는 봉합 눈에 더 순증 한 것으로 밝혀졌습니다. 2 군 (5.19 ± 0.85 mm) 및 1 군 (3.61 ± 0.76 mm)의 오른쪽 눈의 축 방향 길이의 평균 변화의 비교 폐색 (p)의 말에 상당히 낮은 값을 산출하세 가교 절차 (P &lt;0.001, 비모수 맨 - 휘트니 시험) 시행 눈 (오십오일). 왼쪽 눈에 평균 축 방향 길이의 집단 간 차이 (4.20 ± 0.67 mm 대 4.44 ± 0.81 mm)는 (P = 0.39, 맨 - 휘트니 비 파라 메트릭 테스트) 통계적으로 유의하지 않았다. <img alt="그림 1" src="/files/ftp_upload/53201/53201fig1.jpg" /> 그림 1. 이전과 공막 가교과 왼쪽 눈 축 측정 대 안검 봉합 후 오른쪽 눈 축 측정. 공막 가교 및 안검 봉합 (RE 시작) 55 일 후 안검 봉합을 제거 (RE 종료) 후 전에 오른쪽 눈의 축 방향 길이를 의미. 베이스 라인에서 왼쪽 눈의 평균 축 방향 길이 (LE 시작) 55 일 후 (LE 끝). 왼쪽 눈은 수술하지 않은 및 개방했다. 오류 막대는 평균의 표준 오차를 나타냅니다. (permissio로 재 인쇄N 기준 27)에서. <a href="https://www-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/53201/53201fig1large.jpg" target="_blank">이 그림의 더 큰 버전을 보려면 여기를 클릭하십시오.</a> <img alt="그림 2" src="/files/ftp_upload/53201/53201fig2.jpg" /> 그림 2. 이전과 왼쪽 눈 축 측정 대 안검 봉합 후 오른쪽 눈 축 측정. 축 안검 봉합 (RE 시작) 전에 오른쪽 눈의 길이, 55 일 후 안검 봉합을 제거 (RE 종료) 후 평균. (가)베이스 라인에서 왼쪽 눈의 축 방향 길이를 의미 국지적 인 왼쪽 눈에 작동되지 않았고 개방 된 (LE 시작) 55 일 후 (LE 끝). 오류 막대는 평균의 표준 오차를 나타냅니다. (참조 (27)로부터 허가를 재 인쇄). <a href="https://www-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/53201/53201fig2large.jpg" target="_blank">이 그림의 더 큰 버전을 보려면 여기를 클릭하십시오</a>.

Watch this Scientific Journal Video about Scleral Cross-linking Using Riboflavin and Ultraviolet-A Radiation for Prevention of Axial Myopia in a Rabbit Model at JoVE.com

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

우리는 축 신장 토끼 눈에 리보플라빈과 UVA와 공막 가교의 효과를 보여줍니다. 축 방향 신장은 오른쪽 눈의 눈꺼풀 (안검 봉합을) 봉합하여 (남성과 여성) 십삼일 된 뉴질랜드 토끼 유도 하였다.

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 ...

scleral-cross-linking-using-riboflavin-ultraviolet-radiation-for

Nanyang Technological University

Research

JoVE Journal

Medicine

8.8K 조회수.  Rabin Medical Center, Beilinson Campus.  우리는 축 신장 토끼 눈에 리보플라빈과 UVA와 공막 가교의 효과를 보여줍니다. 축 방향 신장은 오른쪽 눈의 눈꺼풀 (안검 봉합을) 봉합하여 (남성과 여성) 십삼일 된 뉴질랜드 토끼 유도 하였다. 

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

In vivo Bioluminescence Imaging of Tumor Hypoxia Dynamics of Breast Cancer Brain Metastasis in a Mouse Model

It is well recognized that tumor hypoxia plays an important role in promoting malignant progression and affecting therapeutic response negatively. There is little knowledge about in situ, in vivo, tumor hypoxia during intracranial development of malignant brain tumors because of lack of efficient means to monitor it in these deep-seated orthotopic tumors. Bioluminescence imaging (BLI), based on the detection of light emitted by living cells expressing a luciferase gene, has been rapidly adopted for cancer research, in particular, to evaluate tumor growth or tumor size changes in response to treatment in preclinical animal studies. Moreover, by expressing a reporter gene under the control of a promoter sequence, the specific gene expression can be monitored non-invasively by BLI. Under hypoxic stress, signaling responses are mediated mainly via the hypoxia inducible factor-1&alpha; (HIF-1&alpha;) to drive transcription of various genes. Therefore, we have used a HIF-1&alpha; reporter construct, 5HRE-ODD-luc, stably transfected into human breast cancer MDA-MB231 cells (MDA-MB231/5HRE-ODD-luc). In vitro HIF-1&alpha; bioluminescence assay is performed by incubating the transfected cells in a hypoxic chamber (0.1% O2) for 24 hr before BLI, while the cells in normoxia (21% O2) serve as a control. Significantly higher photon flux observed for the cells under hypoxia suggests an increased HIF-1&alpha; binding to its promoter (HRE elements), as compared to those in normoxia. Cells are injected directly into the mouse brain to establish a breast cancer brain metastasis model. In vivo bioluminescence imaging of tumor hypoxia dynamics is initiated 2 wks after implantation and repeated once a week. BLI reveals increasing light signals from the brain as the tumor progresses, indicating increased intracranial tumor hypoxia. Histological and immunohistochemical studies are used to confirm the in vivo imaging results. Here, we will introduce approaches of in vitro HIF-1&alpha; bioluminescence assay, surgical establishment of a breast cancer brain metastasis in a nude mouse and application of in vivo bioluminescence imaging to monitor intracranial tumor hypoxia.

In vivo Bioluminescence Imaging of Tumor Hypoxia Dynamics of Breast Cancer Brain Metastasis in a Mouse Model

Bioluminescence imaging of hypoxia inducible factor-1&alpha; activity is applied to monitor intracranial tumor hypoxia development in a breast cancer brain metastasis mouse model.

유방암 뇌 전이의 종양 Hypoxia 역학 생체내의 Bioluminescence 이미징에서는 마우스 모델에서

It is well recognized that tumor hypoxia plays an important role in promoting malignant progression and affecting therapeutic response negatively. There ...

연구

의학

Stereotactic Intracranial Implantation and In vivo Bioluminescent Imaging of Tumor Xenografts in a Mouse Model System of Glioblastoma Multiforme

Glioblastoma multiforme (GBM) is a high-grade primary brain cancer with a median survival of only 14.6 months in humans despite standard tri-modality treatment consisting of surgical resection, post-operative radiation therapy and temozolomide chemotherapy 1. New therapeutic approaches are clearly needed to improve patient survival and quality of life. The development of more effective treatment strategies would be aided by animal models of GBM that recapitulate human disease yet allow serial imaging to monitor tumor growth and treatment response. In this paper, we describe our technique for the precise stereotactic implantation of bio-imageable GBM cancer cells into the brains of nude mice resulting in tumor xenografts that recapitulate key clinical features of GBM 2. This method yields tumors that are reproducible and are located in precise anatomic locations while allowing in vivo bioluminescent imaging to serially monitor intracranial xenograft growth and response to treatments 3-5. This method is also well-tolerated by the animals with low perioperative morbidity and mortality.

Stereotactic Intracranial Implantation and In vivo Bioluminescent Imaging of Tumor Xenografts in a Mouse Model System of Glioblastoma Multiforme

We describe an integrated method for the precise, stereotactic implantation of human glioblastoma multiforme cells into the brains of nude mice and subsequent serial in vivo imaging to monitor growth and response to treatment of the resultant xenografts.

Stereotactic Intracranial 주입 및<em&gt; 생체에</emGlioblastoma의 Multiforme의 마우스 모델 시스템에서 종양 Xenografts의&gt; 발광 생물 이미징

Glioblastoma multiforme (GBM) is a high-grade primary brain cancer with a median survival of only 14.6 months in humans despite standard tri-modality ...

Matrix-assisted Autologous Chondrocyte Transplantation for Remodeling and Repair of Chondral Defects in a Rabbit Model

Articular cartilage defects are considered a major health problem because articular cartilage has a limited capacity for self-regeneration 1. Untreated cartilage lesions lead to ongoing pain, negatively affect the quality of life and predispose for osteoarthritis. During the last decades, several surgical techniques have been developed to treat such lesions. However, until now it was not possible to achieve a full repair in terms of covering the defect with hyaline articular cartilage or of providing satisfactory long-term recovery 2-4. Therefore, articular cartilage injuries remain a prime target for regenerative techniques such as Tissue Engineering. In contrast to other surgical techniques, which often lead to the formation of fibrous or fibrocartilaginous tissue, Tissue Engineering aims at fully restoring the complex structure and properties of the original articular cartilage by using the chondrogenic potential of transplanted cells. Recent developments opened up promising possibilities for regenerative cartilage therapies.
The first cell based approach for the treatment of full-thickness cartilage or osteochondral lesions was performed in 1994 by Lars Peterson and Mats Brittberg who pioneered clinical autologous chondrocyte implantation (ACI) 5. Today, the technique is clinically well-established for the treatment of large hyaline cartilage defects of the knee, maintaining good clinical results even 10 to 20 years after implantation 6. In recent years, the implantation of autologous chondrocytes underwent a rapid progression. The use of an artificial three-dimensional collagen-matrix on which cells are subsequently replanted became more and more popular 7-9.
MACT comprises of two surgical procedures: First, in order to collect chondrocytes, a cartilage biopsy needs to be performed from a non weight-bearing cartilage area of the knee joint. Then, chondrocytes are being extracted, purified and expanded to a sufficient cell number in vitro. Chondrocytes are then seeded onto a three-dimensional matrix and can subsequently be re-implanted. When preparing a tissue-engineered implant, proliferation rate and differentiation capacity are crucial for a successful tissue regeneration 10. The use of a three-dimensional matrix as a cell carrier is thought to support these cellular characteristics 11.
The following protocol will summarize and demonstrate a technique for the isolation of chondrocytes from cartilage biopsies, their proliferation in vitro and their seeding onto a 3D-matrix (Chondro-Gide, Geistlich Biomaterials, Wollhusen, Switzerland). Finally, the implantation of the cell-matrix-constructs into artificially created chondral defects of a rabbit's knee joint will be described. This technique can be used as an experimental setting for further experiments of cartilage repair.

An experimental technique for the treatment of chondral defects in the rabbit's knee joint is described. The implantation of autologous chondrocytes seeded on a matrix is a well-accepted method for the remodeling and repair of articular cartilage lesions providing satisfying long-term results. Matrix-assisted autologous chondrocyte transplantation (MACT) offers a standardized and clinically established implantation method.

토끼 모델에서 Chondral 결함의 리모델링 및 수리를위한 매트릭스를 이용한자가 연골 세포 이식

Articular cartilage defects are considered a major health problem because articular cartilage has a limited capacity for self-regeneration 1. Untreated ...

Screening for Melanoma Modifiers using a Zebrafish Autochthonous Tumor Model

Genomic studies of human cancers have yielded a wealth of information about genes that are altered in tumors1,2,3. A challenge arising from these studies is that many genes are altered, and it can be difficult to distinguish genetic alterations that drove tumorigenesis from that those arose incidentally during transformation. To draw this distinction it is beneficial to have an assay that can quantitatively measure the effect of an altered gene on tumor initiation and other processes that enable tumors to persist and disseminate. Here we present a rapid means to screen large numbers of candidate melanoma modifiers in zebrafish using an autochthonous tumor model4 that encompasses steps required for melanoma initiation and maintenance. A key reagent in this assay is the miniCoopR vector, which couples a wild-type copy of the mitfa melanocyte specification factor to a Gateway recombination cassette into which candidate melanoma genes can be recombined5. The miniCoopR vector has a mitfa rescuing minigene which contains the promoter, open reading frame and 3'-untranslated region of the wild-type mitfa gene. It allows us to make constructs using full-length open reading frames of candidate melanoma modifiers. These individual clones can then be injected into single cell Tg(mitfa:BRAFV600E);p53(lf);mitfa(lf)zebrafish embryos. The miniCoopR vector gets integrated by Tol2-mediated transgenesis6 and rescues melanocytes. Because they are physically coupled to the mitfa rescuing minigene, candidate genes are expressed in rescued melanocytes, some of which will transform and develop into tumors. The effect of a candidate gene on melanoma initiation and melanoma cell properties can be measured using melanoma-free survival curves, invasion assays, antibody staining and transplantation assays.

A rapid way to screen for melanoma modifiers using a zebrafish autochthonous tumor model is presented. It takes advantage of the miniCoopR vector which allows for expression of candidate melanoma genes in melanocytes. A method to obtain melanoma-free survival curves, an invasion assay, a protocol for antibody staining of scale melanocytes and a melanoma transplantation assay are described.

Zebrafish Autochthonous 종양 모델을 사용하여 흑종 조절을위한 검사

Genomic studies of  human cancers have yielded a wealth of information about genes that are altered  in tumors1,2,3. A challenge arising from these ...

The In ovo CAM-assay as a Xenograft Model for Sarcoma

Sarcoma is a very rare disease that is heterogeneous in nature, all hampering the development of new therapies. Sarcoma patients are ideal candidates for personalized medicine after stratification, explaining the current interest in developing a reproducible and low-cost xenotransplant model for this disease. The chick chorioallantoic membrane is a natural immunodeficient host capable of sustaining grafted tissues and cells without species-specific restrictions. In addition, it is easily accessed, manipulated and imaged using optical and fluorescence stereomicroscopy. Histology further allows detailed analysis of heterotypic cellular interactions.
This protocol describes in detail the in ovo grafting of the chorioallantoic membrane with fresh sarcoma-derived tumor tissues, their single cell suspensions, and permanent and transient fluorescently labeled established sarcoma cell lines (Saos-2 and SW1353). The chick survival rates are up to 75%. The model is used to study graft- (viability, Ki67 proliferation index, necrosis, infiltration) and host (fibroblast infiltration, vascular ingrowth) behavior. For localized grafting of single cell suspensions, ECM gel provides significant advantages over inert containment materials. The Ki67 proliferation index is related to the distance of the cells from the surface of the CAM and the duration of application on the CAM, the latter determining a time frame for the addition of therapeutic products.

The In ovo CAM-assay as a Xenograft Model for Sarcoma

The in ovo chorioallantoic membrane (CAM) is grafted with fresh sarcoma-derived tumor tissues, their single cell suspensions, and permanent and transient fluorescently labeled established sarcoma cell lines. The model is used to study graft- (viability, Ki67 proliferation index, necrosis, infiltration) and host (fibroblast infiltration, vascular ingrowth) behavior.

<em&gt; OVO에서</em육종을위한 Xenograftmodel으로&gt; CAM-분석

Sarcoma is a very rare disease that is heterogeneous in nature, all hampering the development of new therapies. Sarcoma patients are ideal candidates for ...

The Rabbit Blood-shunt Model for the Study of Acute and Late Sequelae of Subarachnoid Hemorrhage: Technical Aspects

Early brain injury and delayed cerebral vasospasm both contribute to unfavorable outcomes after subarachnoid hemorrhage (SAH). Reproducible and controllable animal models that simulate both conditions are presently uncommon. Therefore, new models are needed in order to mimic human pathophysiological conditions resulting from SAH.
This report describes the technical nuances of a rabbit blood-shunt SAH model that enables control of intracerebral pressure (ICP). An extracorporeal shunt is placed between the arterial system and the subarachnoid space, which enables examiner-independent SAH in a closed cranium. Step-by-step procedural instructions and necessary equipment are described, as well as technical considerations to produce the model with minimal mortality and morbidity. Important details required for successful surgical creation of this robust, simple and consistent ICP-controlled SAH rabbit model are described.

The experimental intracranial pressure-controlled blood shunt subarachnoid hemorrhage (SAH) model in the rabbit combines the standard procedures &#8212; subclavian artery cannulation and transcutaneous cisterna magna puncture, which enables close mimicking of human pathophysiological conditions after SAH. We present step-by-step instructions and discuss key surgical points for successful experimental SAH creation.

지주막 하 출혈의 급성과 늦은 후유증의 연구에 대한 토끼 피 션트 모델 : 기술적 측면

Early brain injury and delayed cerebral vasospasm both contribute to unfavorable outcomes after subarachnoid hemorrhage (SAH). Reproducible and ...

Diffuse Optical Spectroscopy for the Quantitative Assessment of Acute Ionizing Radiation Induced Skin Toxicity Using a Mouse Model

Acute skin toxicities from ionizing radiation (IR) are a common side effect from therapeutic courses of external beam radiation therapy (RT) and negatively impact patient quality of life and long term survival. Advances in the understanding of the biological pathways associated with normal tissue toxicities have allowed for the development of interventional drugs, however, current response studies are limited by a lack of quantitative metrics for assessing the severity of skin reactions. Here we present a diffuse optical spectroscopic (DOS) approach that provides quantitative optical biomarkers of skin response to radiation. We describe the instrumentation design of the DOS system as well as the inversion algorithm for extracting the optical parameters. Finally, to demonstrate clinical utility, we present representative data from a pre-clinical mouse model of radiation induced erythema and compare the results with a commonly employed visual scoring. The described DOS method offers an objective, high through-put evaluation of skin toxicity via functional response that is translatable to the clinical setting.

We present a diffuse optical spectroscopic (DOS) approach that provides quantitative optical biomarkers of skin response to radiation. We describe DOS instrumentation design, optical parameters extraction algorithms and the animal handling procedures required to yield representative data from a pre-clinical mouse model of radiation induced erythema.

마우스 모델을 사용하여 급성 이온화 방사선 유발 피부 독성의 정량적 평가를위한 광학 분광학 확산

Acute skin toxicities from ionizing radiation (IR) are a common side effect from therapeutic courses of external beam radiation therapy (RT) and ...

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 ...

Isometric Contractility Measurement of the Mouse Mesenteric Artery Using Wire Myography

The wire myograph technique is used to assess the contractility of vascular smooth muscles in response to depolarization, GPCR agonists/inhibitors and drugs. It is widely used in many studies on the physiological functions of vascular smooth muscle, the pathogenesis of vascular diseases such as hypertension, and the development of smooth muscle relaxant drugs. The mouse is a widely used model animal with a large pool of disease models and genetically modified strains. We introduced this method to measure the isometric contraction of mouse mesenteric artery in detail. A 1.4-mm segment of mouse mesenteric resistance artery was isolated and mounted on a myograph chamber by passing two steel wires through its lumen. After equilibration and normalization steps, the vessel segment was potentiated by a high-K+ solution twice prior to the contraction assay. As an example of the application of this method in drug development, we measured the relaxant effect of a novel natural substance, neoliensinine, isolated from a Chinese herb, embryos of the lotus seed (Nelumbo nucifera Gaertn.) on mouse mesenteric arteries. The vessel segments mounted on the myograph chamber were stimulated with a high-K+ solution. When the force tension reached a stable sustained phase, cumulative doses of neoliensinine were added to the chamber. We found that neoliensinine had a dose-dependent relaxant effect on smooth muscle contraction, thus suggesting that it bears potential activity against hypertension. In addition, as the vessel segment can survive at least 4 hours after mounting and maintain contractility induced by the high-K+ solution for many times, we suggest that the wire myograph system may be used for the time-consuming process of drug screening.

The wire myograph technique is used to investigate vascular smooth muscle functions and screen new drugs. We report a detailed protocol for measuring the isometric contractility of the mouse mesenteric artery and for screening new relaxants of vascular smooth muscle.

Myography 와이어를 사용 하 여 마우스 Mesenteric 동맥의 isometric 수축 측정

The wire myograph technique is used to assess the contractility of vascular smooth muscles in response to depolarization, GPCR agonists/inhibitors and ...

Targeted and Selective Treatment of Pluripotent Stem Cell-derived Teratomas Using External Beam Radiation in a Small-animal Model

The growing number of victims of &#34;stem cell tourism,&#34; the unregulated transplantation of stem cells worldwide, has raised concerns about the safety of stem cell transplantation. Although the transplantation of differentiated rather than undifferentiated cells is common practice, teratomas can still arise from the presence of residual undifferentiated stem cells at the time of transplant or from spontaneous mutations in differentiated cells. Because stem cell therapies are often delivered into anatomically sensitive sites, even small tumors can be clinically devastating, resulting in blindness, paralysis, cognitive abnormalities, and cardiovascular dysfunction. Surgical access to these sites may also be limited, leaving patients with few therapeutic options. Controlling stem cell misbehavior is, therefore, critical for the clinical translation of stem cell therapy.
External beam radiation offers an effective means of delivering targeted therapy to decrease the teratoma burden while minimizing injury to surrounding organs. Additionally, this method avoids genetic manipulation or viral transduction of stem cells-which are associated with additional clinical safety and efficacy concerns. Here, we describe a protocol to create pluripotent stem cell-derived teratomas in mice and to apply external beam radiation therapy to selectively ablate these tumors in vivo.

Research on treatment strategies for pluripotent stem cell-derived teratomas is important for the clinical translation of stem cell therapy. Here, we describe a protocol to, first, generate stem cell-derived teratomas in mice and, then, to selectively target and treat these tumors in vivo using a small-animal irradiator.

만능 줄기 세포 유래 Teratomas 외부 빔 방사선을 사용 하 여 작은 동물 모델에서의 타겟 및 선택적 치료

The growing number of victims of &#34;stem cell tourism,&#34; the unregulated transplantation of stem cells worldwide, has raised concerns about the ...