This study was supported and funded by the Avenir Foundation (Fondation de l'Avenir, 255 rue de Vaugirard, 75719 Paris cedex 15, Paris, France). This study would never have been conducted without the precious help of the veterinary team of the Laboratory of Biosurgical Research from the Alain Carpentier Foundation.

모든 동물은 동물 연구 윤리위원회 (농업의 프랑스 교육부의 지침)에 따라 처리 하였다. 이 프로토콜은 파리 데카르트 대학에서 동물 실험과 승인 지역 윤리위원회의 승인을받은 (등록 번호 MESR 2035.02을, 학부 의학 파리 데카르트, 파리, 프랑스의). 1. ADSC 문화 및 라벨링 <ol><li> 민간 기관에서 확인 ADSCs를 얻습니다. 37 ° C, 5 %, 10 % 태아 송아지 혈청과 1 % 항생제 (페니실린과 스트렙토 마이신)를 포함한 알파 최소 필수 매체와 CO 2의 문화 동종 ADSCs. </li><li> 표준 셀 확장 절차 (37 ° C와 5 % CO 2)를 사용하여 동물 당 약 24 × 10 6 ADSCs를 얻습니다. 표준 트립신 용액 (a T150 접시 10 ㎖)를 사용하여 수확 ADSCs. 혈구와 셀 계산을 수행합니다. </li><li> 전지이다 PKH67 염색 홍보를 사용하여 이식 전날 라벨을 수행ocedure 후술. 주 : 염색 과정의 목적은 이식 후 세포의 추적을 허용하는 것이다. <ol><li> 2 × 10 6 ADSCs 레이블을하기 위해, 용액 A를 준비하고 해결 B (희석제 C 1 mL를 PKH67의 4 μL) (100 희석제 C의 μL 2 × 10 6 세포). </li><li> 2 분 솔루션 A와 B를 품어. 그런 다음, 1 분 동안 태아 송아지 혈청 100 μL를 추가합니다. </li><li> 5 분 동안 1,500 XG에서 구연산 RPMI 300 μL와 솔루션 (RPMI의 298.5 μL와 구연산의 1.5 μL) 원심 분리기를 씻어. 이 작업을 2 회 반복합니다. 표지 된 세포를 유지하고 시트 건설 될 때까지 빛으로부터 보호. </li></ol></li></ol> 두 번 ADSC 매 구축 <ol><li> 날은 이식 전에 PKH 라벨링되기 전에, 상술 한 세포 배양 배지의 웰 당 4 ㎖, 함께 12 웰 온도 - 응답 세포 배양 접시를 준비한다. 1.5 × 10 6 PKH-labele와 함께 요리를 시드웰 당 D 세포는 12 시간 (37 ℃, 5 % CO 2) 배양한다. </li><li> 이식 당일에, 30 분 동안 실온에서 배양 접시에 의해 합류 세포 시트를 분리. </li><li> 조심스럽게 피펫 한 ADSC 시트를 대기음과 소수성 종이 (1.5 cm 직경)에 레이어. 지원 막으로이 용지를 사용하십시오. </li><li> 다른 ADSC 시트를 잡고 부드럽게 이중층 구조를 획득하기 위해 다른 하나의 상부에 증착. </li><li> (동물 당 4) 필요한만큼 ADSC 시트 구조를 얻기 위해이 절차를 반복합니다. </li></ol> 3. 식도 내시경 점막하 절개 (ESD) <ol><li> 이식 당일, 근육 내 케타민 10 ㎎ / kg과 동물을 미리 의료를 8 밀리그램 / kg의 정맥 내 프로포폴로 유도. 그런 다음, 기관 내 삽관을 수행하고 이소 플루 란 2.5 %의 흡입으로 마취를 유지한다. </li><li> 동물은 전신 마취하에되면, gastrosc으로 ESD를 수행OPE하는 비디오 스코프 및 전기 수술 장치. <ol><li> 40cm의 치열 궁에서 45cm까지 헤미 - 원주 등의 마크를 획득하기 위해 내시경 나이프와 소프트 응고를 사용합니다. </li><li> 근육 층으로부터 점막층의 분리 점막하 층으로 인디고 카민 염료를 함유하는 글리세롤의 용액을 주입한다. </li><li> 원주 절개를 얻을 수 endocut의 I 모드 내시경 칼을 사용합니다. </li><li> 말단 절개에 근위 절개에서 이동, 점막하 절개를 수행하기 위해 강제 응고 모드 내시경 칼을 사용합니다. </li></ol></li><li> ESD 피 프로 시저의 끝에서, 근육 층을 노출 헤미 원주 길이 5cm 식도 흉터를 관찰한다. </li></ol> 4. 내시경 이식 <ol><li> 즉시 ESD 후, 내시경 4 번 ADSC 매 구조로 동물을 이식. <ol><li> 내시경 집게와 가진 ADSC 시트 구조를 파악큰 투명 캡 내시경을 이용하여 상처 부위로 수송 중에 보호. </li><li> 궤양 침대에 ADSC 매 구조의 전체 표면을 적용합니다. </li><li> 부드럽게 완전하고 안정된 생착을 얻기 위해 10 분 동안 ADSC 시트 구조가 적용된다. 애플리케이션의 내시경 겸자 내시경 캡을 사용하여 ADSC의 전면은 식도의 점막하 층에 구성 적용된다. </li></ol></li><li> 네 이식 세포 접착 이중 시트 구조를 얻기 위하여이 과정을 각각의 동물에 대한 네 번 반복한다. 참고 : 목표는 부상 영역의 approximatively 절반을 커버하는 것입니다. </li></ol> 5. 수술 후 평가 및 후속 <ol><li> 각 동물의 경우, 0.2 mg의 모르핀 / kg 세 번 첫날과에 소메 프라 졸의 28 일 안티 - 산 처리 (40 밀리그램 / 일)과 하루의 근육 내 주사와 후 ESD의 진통을 보장합니다. amoxicill와 예방 적 항생제를 수행7 일간 (1g / 일)이다. 다음 날 하루 1 액체 및 고체 권한을 부여합니다. </li><li> 이십팔일에 대한 동물과 함께하십시오. 감미로운 Pinkas의 연하 곤란 점수 (16)와 무게 변화 측정을 사용하여 매일 임상 평가를 얻습니다. </li><li> 복합 협착 평가 : 예약 된 날짜 3, 14, 28에 전신 마취 하에서, 협착 발생의 복합 평가를 수행합니다. <ol><li> 흉터 설명 협착 측정 및 용지지지 막 검출하여 내시경 평가를 수행한다. 협착을 통해 교차하는 위 내시경을 사용하여 협착을 측정합니다 (위 내시경의 직경이 8mm)입니다. 염증 측면에주의를 기울이고, 흉터를 관찰 (점막 발적, 점막 부종 및 자연 출혈 점막)과 흉터 내의 부재 또는 용지지지 막의 존재를. </li><li> 흉터 침대에 직접 내시경 연산자 운하를 통해 pCLE 녹색 프로브를 적용합니다. 자발적 검색및 PKH67 표지 ADSC 시트 구조의 생착과 호환 녹색 신호를 조직했다. </li><li> 방사능 후프 (앞 왼쪽 편 빈도)와 Baryte 용의 식도의 2 직교 발생률을 수행합니다. 협착 평가 협소 입사 계속) 17, 18 (협착 (% 정도) = 협착 하에서 단축 1- (길이 / 협착 하에서 정상 축)의 길이는 100 X)]한다. </li></ol></li><li> 동물 희생. <ol><li> 28 일째에, 관찰 기간이 종료되면 동물을 희생을 수행한다. 동물은 전신 마취하에있는 동안, 정맥 페노바르비탈 100 ㎎ / ㎏을 주입. 하트 비트의 부재과 움직임을 호흡의 임상 시험에 사용하는 동물의 죽음을 확인합니다. </li><li> 각 (10 % 포르말린 고정, 파라핀 삽입, 4 μm의 두께 섹션과) 동물을 희생에서 흉골 절개술 및 장기 노출되면 식도를 제거합니다. 슬라이드 얼룩헤 마톡 실린 및 사프란 15. 컴퓨터 분석을 위해 슬라이드를 디지털화하고, 동물 군 (15)을 비교합니다. </li></ol></li></ol>

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L., Annese, V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=New+endoscopic+imaging+techniques+in+surveillance+of+inflammatory+bowel+disease.">New endoscopic imaging techniques in surveillance of inflammatory bowel disease.</a> World Journal of Gastrointestinal Endoscopy. 7 (3), 230-236 (2015).</li><li>Nagyova, M., Slovinska, L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+comparative+study+of+PKH67,+DiI,+and+BrdU+labeling+techniques+for+tracing+rat+mesenchymal+stem+cells.">A comparative study of PKH67, DiI, and BrdU labeling techniques for tracing rat mesenchymal stem cells.</a> In Vitro Cellular &amp; Developmental Biology. Animal. 50 (7), 656-663 (2014).</li><li>Jain, D., Singhal, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Esophageal+Stricture+Prevention+after+Endoscopic+Submucosal+Dissection.">Esophageal Stricture Prevention after Endoscopic Submucosal Dissection.</a> Clinical Endoscopy. 49 (3), 241-256 (2016).</li><li>Lua, G. W., Tang, J., Liu, F., Li, Z. S. <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+Esophageal+Strictures+After+Endoscopic+Submucosal+Dissection:+A+Promising+Therapy+Using+Carboxymethyl+Cellulose+Sheets.">Prevention of Esophageal Strictures After Endoscopic Submucosal Dissection: A Promising Therapy Using Carboxymethyl Cellulose Sheets.</a> Digestive Diseases and Sciences. 61 (6), 1763-1769 (2016).</li><li>Sakaguchi, Y., Tsuji, 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=Triamcinolone+Injection+and+Shielding+with+Polyglycolic+Acid+Sheets+and+Fibrin+Glue+for+Postoperative+Stricture+Prevention+after+Esophageal+Endoscopic+Resection:+A+Pilot+Study.">Triamcinolone Injection and Shielding with Polyglycolic Acid Sheets and Fibrin Glue for Postoperative Stricture Prevention after Esophageal Endoscopic Resection: A Pilot Study.</a> The American Journal of Gastroenterology. 111 (4), 581-583 (2016).</li><li>Sakurai, T., Miyazaki, S., Miyata, G., Satomi, S., Hori, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Autologous+buccal+keratinocyte+implantation+for+the+prevention+of+stenosis+after+EMR+of+the+esophagus.">Autologous buccal keratinocyte implantation for the prevention of stenosis after EMR of the esophagus.</a> Gastrointestinal endoscopy. 66 (1), 167-173 (2007).</li><li>Nieponice, A., McGrath, 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=An+extracellular+matrix+scaffold+for+esophageal+stricture+prevention+after+circumferential+EMR.">An extracellular matrix scaffold for esophageal stricture prevention after circumferential EMR.</a> Gastrointestinal Endoscopy. 69 (2), 289-296 (2009).</li><li>Honda, M., Hori, 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=Use+of+adipose+tissue-derived+stromal+cells+for+prevention+of+esophageal+stricture+after+circumferential+EMR+in+a+canine+model.">Use of adipose tissue-derived stromal cells for prevention of esophageal stricture after circumferential EMR in a canine model.</a> Gastrointestinal endoscopy. 73 (4), 777-784 (2011).</li><li>Barret, M., Pratico, C. 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=Amniotic+membrane+grafts+for+the+prevention+of+esophageal+stricture+after+circumferential+endoscopic+submucosal+dissection.">Amniotic membrane grafts for the prevention of esophageal stricture after circumferential endoscopic submucosal dissection.</a> PloS One. 9 (7), 100236 (2014).</li></ol>

저자는 공개 아무것도 없어.

이 돼지 모델에서, ADSC 시트 이식 기술적 성공 및 생체 pCLE 평가 셀 생착 모니터링시켰다. 임상 내시경, 방사선 학적, 조직 학적 평가는 확장 된 ESD 후 식도 협착 예방에 내시경 ADSC 시트의 효과를 보여 주었다. 인디고 카민 염료를 함유하는 시트 ADSC 글리세롤 용액의 내시경 이식 재생 의학에 혁신적인 방법이다. Ohki 등. 제 내시경 이식 절차를 설명했다. 이들 연구...

피상적 식도 종양 관리 새로운 내시경 기술의 발달로 변경되었다. 요즘, 내시경 절제술은 제 치료제이다. 사실상, 종양은 동일한 결과 1, 2, 3과 수술보다 낮은 이환율 및 사망률과 연관된다. 내시경 점막 절제술 (EMR) 및 내시경 적 점막하 절제술 (ESD)는 가장 널리 사용되는 기술이다. 확장 된 표면 종양의 경우, ESD가 바람직하다. EMR에 비해 ESD 관계없이 병변의 크기, 일괄 resectioning 욕실 수, 4, 5, 6을 형성. ESD의 주요 지연 합병증은 일반적으로 절제 후 1 2 주 사이에 발생하는 식도 협착 형성된다. 최근 발표 된 연구는 협착 형성이 상관된다 보여절제의 크기입니다. 일본 내시경 학회 ESD들이 사건의 90 % 이상에 협착 개발과 관련된 심각한 공급 문제와 삶의 질에 큰 저하에 대한 책임 때문에 식도 주위의 ¾보다 큰 크기를 피하는 것이 좋습니다. 식도 협착의 예방 도전 남아 있습니다. 협착 형성에 관여하는 메커니즘은 부분적으로 만 알려져있다. (1) 염증성 세포 모집 및 (2) 과도한 섬유화 개발 7 : 형성을 협착은 두 개의 서로 다른 메커니즘의 관계에서 발생할 것으로 보인다. 몇 가지 예방 치료가 제안되었다. 그러나, 결과는 약간의 이익과 심각한 부작용이 8, 9, 불만족했다. 최근, 일본 팀, Ohki 등. 상기 esoph로자가 구강 점막 세포의 단층 세포 시트를 이식하는 것이 제안ageal 흉터. 이식 ESD (10), (11) 직후에 수행 하였다. 그들은 제 개과 모델 후 환자에서이 혁신적인 접근법의 효과를 입증 하였다. 지방 조직 유래 기질 세포 (ADSCs)는 재생 의학에 약속되어있다. 여러 분야에서의 응용 프로그램은 특히 상처 치유 과정에서 흥미로운 결과를 보여 주었다. 세포가 쉽게 분리되어 및 간엽 아닌 중간 엽 계통 (12, 13), (14)에 항염 로컬 면역 조절 효과, 혈관 신생을 유도하고, 분화 능력과 연관되기 때문에 ADSC 요법은 여러 장점을 제공한다. 이전 연구에서 우리 팀은 식도 협착 이전에 대한 이중 ADSC 장 내시경 이식의 효과를 입증돼지 모델 (15)의 확장 된 ESD 후 ention. 이 글에서, ADSC 장 건설 및 내시경 이식 기술의 보고서가 제공됩니다.

<table><tbody><tr><td>Transparent endoscopic cap Q180 compatible</td><td>Olympus Optical Co</td><td></td><td></td></tr><tr><td>GIF-Q180 gastroscope&amp;nbsp;</td><td>Olympus Optical Co</td><td></td><td></td></tr><tr><td>Videoscope System Exera II&amp;nbsp;</td><td>Olympus Optical Co</td><td></td><td></td></tr><tr><td>Injection needle 18 G</td><td>Olympus Optical Co</td><td></td><td></td></tr><tr><td>Electrosurgery unit ERBE ICC 350&amp;nbsp;</td><td>ERBE Technology</td><td></td><td></td></tr><tr><td>Indigo carmin 1%</td><td>Life</td><td></td><td></td></tr><tr><td>Endoscopic hybrid knife</td><td>Life</td><td></td><td></td></tr><tr><td>Minisonde Z pCLE green probe</td><td>Mauna Kea Technology</td><td></td><td>You must learn how to use the probe. The manipulation could be difficult.</td></tr><tr><td>Fetal bovine serum</td><td>Sigma Aldrich</td><td>12105C</td><td></td></tr><tr><td>Trypsin</td><td>Sigma Aldrich</td><td>T146</td><td></td></tr><tr><td>Alpha minimum essential medium</td><td>Thermo Fisher</td><td>22561-021</td><td></td></tr><tr><td>Phosphate-Buffered Salines</td><td>Thermo Fisher</td><td>10010-023</td><td></td></tr><tr><td>PKH67 dye kit</td><td>Sigma Aldrich</td><td>Mini67-KT</td><td></td></tr><tr><td>12-well temperature responsive cell culture dish</td><td>Upcell Thermo Scientific</td><td>174900</td><td>Feel the weel with 4 mL standard medium culture 30 min before seeding cells</td></tr><tr><td>Esomeprazole 40 mg&amp;nbsp;</td><td>Biogaran</td><td></td><td></td></tr><tr><td>Moprhine sulfate 50 mg/mL</td><td>Lavoisier</td><td></td><td></td></tr><tr><td>Amoxicilline 1 g</td><td>Biogaran</td><td></td><td></td></tr><tr><td>Ketamine 250 mg/5 mL</td><td>Panpharma</td><td></td><td></td></tr><tr><td>Propofol 10 mg/mL</td><td>Fresenius</td><td></td><td></td></tr><tr><td>Hydrophobic paper</td><td>Carrefour</td><td></td><td></td></tr></tbody></table>

adsc-sheet transplantation to prevent stricture after extended esophageal endoscopic submucosal dissection

지난 몇 년 동안, 세포 시트의 구조는 특히 재건 수술 절차, 재생 의학에서 폭 넓은 관심을 촉발하고있다. 다양한 생체 물질과 지방 조직 유래 기질 세포 (ADSCs)를 조합 한 다양한 기술의 개발은 뼈, 연골, 그리고 쥐, 돼지, 인간 ADSCs에서 지방 조직과 같은 조직 공학적 대체, 수많은 종류의 건설을 주도하고있다. 확장 식도 내시경 점막하 절제술 (ESD)은 식도 협착 형성 할 책임이있다. 협착 방지를 사용할 수없는 효율적인 치료와 함께 도전 남아있다. 이전의 연구는 송곳니 모델과 인간 점막 세포 시트를 이식 효과를보고 하였다. ADSCs는 중간 엽과 비 중간 엽 계통에 항 염증, 지방의 면역 변조 효과, 혈관 신생 유도 및 분화 능력을 기인한다. 이 원래의 연구는 내시경 TRA 설명ADSC의 조직 공학적 구조의 nsplantation은 돼지 모델에서 식도 협착을 방지합니다. ADSC 구조는 종이지지 막 서로에 계층이 동종 ADSC 시트로 구성되었다. ADSCs는 프로브 기반 공 초점 레이저 endomicroscopy (pCLE) 모니터링을 허용하는 PKH67의 형광으로 표지 하였다. 이식 당일 5 cm 식도 협착을 유발하는 것으로 알려져 헤미 둘레 ESD 수행 하였다. 동물은 즉시 내시경 4 ADSC 구조로 이식되었다. ADSC 구조의 완전한 접착 부드러운 적용 10 분 후에 얻어졌다. 동물은 모든 동물이 성공적으로 이식 날 (28)에 희생되었다. 이식은 양의 pCLE 평가와 함께 3 일에 확인되었다. 이식 된 동물에 비해 대조군의 주요 섬유 성 조직 개발 빈번한 소화 문제, 및 감소 된 체중 증가와 심한 협착을 개발 하였다. allogeni 우리의 모델에서, 이식C 이중 세포 시트로 구성 ADSCs는 확장 된 ESD 후 성공적으로 강하게 하부 식도 협착 속도와 관련된했다.

ADSCs 및 ADSC 시트를 획득하는 절차의 문화는도 1에 도시된다. 도 2는 용지지지 막상에서 서로 적층 된 두 ADSC 시트로 이루어지는 그래프트의 구조를 나타낸다. ADSCs는 이전에 pCLE와 생체 이식 모니터링 수 있도록 PKH67의 형광으로 표지 하였다. 그림 3은 5 cm 및 헤미 - 원주 식도 흉터의 결과로, 확장 된 식도 내시경 점막하 박?...

Watch this Scientific Journal Video about ADSC-sheet Transplantation to Prevent Stricture after Extended Esophageal Endoscopic Submucosal Dissection at JoVE.com

ADSC-sheet Transplantation to Prevent Stricture after Extended Esophageal Endoscopic Submucosal Dissection

이 연구는 내시경 지방 조직 유래 기질 세포 (ADSC)의 성공적인 방법을보고하는 돼지 모델의 확장 내시경 점막하 절제술 (ESD) 후 식도 협착 방지를위한 - 시트 이식.

ADSC 매 이식은 확장 식도 내시경 점막하 절개 후 협착을 방지하는

In past years, the cell-sheet construct has spurred wide interest in regenerative medicine, especially for reconstructive surgery procedures. The ...

adsc-sheet-transplantation-to-prevent-stricture-after-extended

Research

JoVE Journal

Medicine

8.4K 조회수.  Université Paris Descartes Sorbonne Paris Cité.  이 연구는 내시경 지방 조직 유래 기질 세포 (ADSC)의 성공적인 방법을보고하는 돼지 모델의 확장 내시경 점막하 절제술 (ESD) 후 식도 협착 방지를위한 - 시트 이식. 

비디오: ADSC-sheet Transplantation to Prevent Stricture after Extended Esophageal Endoscopic Submucosal Dissection

An Ivor Lewis Esophagectomy Designed to Minimize Anastomotic Complications and Optimize Conduit Function

We describe a novel esophagogastric anastomotic technique (&ldquo;side-to-side: staple line-on-staple line&rdquo;, STS) for intrathoracic anastomoses designed to create a large diameter anastomosis while simultaneously maintaining conduit blood supply. This technique aims to minimize the incidence of anastomotic leaks and strictures, which is a frequent source of morbidity and occasional mortality after esophagectomy. We analyze the results of this STS technique on 368 patients and compared outcomes to 112 patients who underwent esophagogastric anastomoses using an end-to-end stapler (EEA) over an 8-year time interval at our institution.
The STS technique involves aligning the remaining intrathoracic esophagus over the tip of the lesser curve staple line of a stomach tube, created as a replacement conduit for the esophagus. A linear stapling device cuts through and restaples the conduit staple line to the lateral wall of the esophagus in a side-to-side fashion. The open common lumen is then closed in two layers of sutures.
There was a total of 12 (3.8%) anastomotic leaks in patients who underwent STS esophagogastric anastomosis. Two of eight patients (25%) had anastomotic leaks after esophagectomy for end-stage achalasia as compared to a 2.8% leak rate (10/336) after esophagectomy for other conditions. Eighteen (5.2%) patients required a median of 2 dilatations for anastomotic stricture after STS anastomosis. Supplemental jejunostomy feedings were required in only 11.1% of patients undergoing STS anastomoses following hospital discharge. In contrast, patients undergoing EEA anastomoses demonstrated anastomotic leak and stricture rates of 16.1% and 14.3% respectively (p&lt;0.01). Time analysis of postoperative contrast studies following the STS technique typically demonstrated a straight/uniform diameter conduit with essentially complete contrast emptying into the small bowel within 3 minutes in 88.4% of patients.
The incidence of esophagogastric anastomotic leaks and strictures were extremely low using this novel anastomotic technique. Additionally we believe that based on time and qualitative analyses of postoperative contrast studies, this technique appears to optimize postoperative upper gastrointestinal tract function; however, further comparative studies are needed.

We describe a novel intrathoracic esophagogastric anastomotic technique designed to create a large diameter anastomosis while simultaneously maintaining conduit blood supply to minimize the incidence of anastomotic leaks and strictures. Construction and orientation of the stomach conduit designed to optimize upper gastrointestinal tract function is also described.

We describe a novel esophagogastric anastomotic technique (&ldquo;side-to-side: staple line-on-staple line&rdquo;, STS) for intrathoracic anastomoses ...

연구

암 연구학

Conditional Reprogramming of Pediatric Human Esophageal Epithelial Cells for Use in Tissue Engineering and Disease Investigation

Identifying and expanding patient-specific cells in culture for use in tissue engineering and disease investigation can be very challenging. Utilizing various types of stem cells to derive cell types of interest is often costly, time consuming and highly inefficient. Furthermore, undesired cell types must be removed prior to using this cell source, which requires another step in the process. In order to obtain enough esophageal epithelial cells to engineer the lumen of an esophageal construct or to screen therapeutic approaches for treating esophageal disease, native esophageal epithelial cells must be expanded without altering their gene expression or phenotype. Conditional reprogramming of esophageal epithelial tissue offers a promising approach to expanding patient-specific esophageal epithelial cells. Furthermore, these cells do not need to be sorted or purified and will return to a mature epithelial state after removing them from conditional reprogramming culture. This technique has been described in many cancer screening studies and allows for indefinite expansion of these cells over multiple passages. The ability to perform esophageal screening assays would help revolutionize the treatment of pediatric esophageal diseases like eosinophilic esophagitis by identifying the trigger mechanism causing the patient's symptoms. For those patients who suffer from congenital defect, disease or injury of the esophagus, this cell source could be used as a means to seed a synthetic construct for implantation to repair or replace the affected region.

Expansion of human pediatric esophageal epithelial cells utilizing conditional reprogramming provides investigators with a patient-specific population of cells that can be utilized for engineering esophageal constructs for autologous implantation to treat defects or injury and serve as a reservoir for therapeutic screening assays.

조직 공학 및 질병 조사에있는 사용을위한 소아 인간의 식도 상피 세포의 조건부 재 프로그래밍

Identifying and expanding patient-specific cells in culture for use in tissue engineering and disease investigation can be very challenging. Utilizing ...

발생학

Creation and Transplantation of an Adipose-derived Stem Cell (ASC) Sheet in a Diabetic Wound-healing Model

Artificial skin has achieved considerable therapeutic results in clinical practice. However, artificial skin treatments for wounds in diabetic patients with impeded blood flow or with large wounds might be prolonged. Cell-based therapies have appeared as a new technique for the treatment of diabetic ulcers, and cell-sheet engineering has improved the efficacy of cell transplantation. A number of reports have suggested that adipose-derived stem cells (ASCs), a type of mesenchymal stromal cell (MSC), exhibit therapeutic potential due to their relative abundance in adipose tissue and their accessibility for collection when compared to MSCs from other tissues. Therefore, ASCs appear to be a good source of stem cells for therapeutic use. In this study, ASC sheets from the epididymal adipose fat of normal Lewis rats were successfully created using temperature-responsive culture dishes and normal culture medium containing ascorbic acid. The ASC sheets were transplanted into Zucker diabetic fatty (ZDF) rats, a rat model of type 2 diabetes and obesity, that exhibit diminished wound healing. A wound was created on the posterior cranial surface, ASC sheets were transplanted into the wound, and a bilayer artificial skin was used to cover the sheets. ZDF rats that received ASC sheets had better wound healing than ZDF rats without the transplantation of ASC sheets. This approach was limited because ASC sheets are sensitive to dry conditions, requiring the maintenance of a moist wound environment. Therefore, artificial skin was used to cover the ASC sheet to prevent drying. The allogenic transplantation of ASC sheets in combination with artificial skin might also be applicable to other intractable ulcers or burns, such as those observed with peripheral arterial disease and collagen disease, and might be administered to patients who are undernourished or are using steroids. Thus, this treatment might be the first step towards improving the therapeutic options for diabetic wound healing.

Creation and Transplantation of an Adipose-derived Stem Cell ASC Sheet in a Diabetic Wound-healing Model

Adipose-derived stem cells (ASCs) are easily isolated and harvested from the fat of normal rats. ASC sheets can be created using cell-sheet engineering and can be transplanted into Zucker diabetic fatty rats exhibiting full-thickness skin defects with exposed bone and then covered with a bilayer of artificial skin.

생성 및 당뇨병 치유 하는 부상을 모델에서 줄기 세포 지방이 많은 파생 (ASC) 시트의 이식

Artificial skin has achieved considerable therapeutic results in clinical practice. However, artificial skin treatments for wounds in diabetic patients ...

의학

Tissue-Engineered Graft for Circumferential Esophageal Reconstruction in Rats

The use of biocompatible materials for circumferential esophageal reconstruction is a technically challenging task in rats and requires an optimal implant technique with nutritional support. Recently, there have been many attempts at esophageal tissue engineering, but the success rate has been limited due to difficulty in early epithelization in the special environment of peristalsis. Here, we developed an artificial esophagus that can improve the regeneration of the esophageal mucosa and muscle layers through a two-layered tubular scaffold, a mesenchymal stem cell-based bioreactor system, and a bypass feeding technique with modified gastrostomy. The scaffold is made of polyurethane (PU) nanofibers in a cylindrical shape with a three-dimensional (3D) printed polycaprolactone strand wrapped around the outer wall. Prior to transplantation, human-derived mesenchymal stem cells were seeded into the lumen of the scaffold, and bioreactor cultivation was performed to enhance cellular reactivity. We improved the graft survival rate by applying surgical anastomosis and covering the implanted prosthesis with a thyroid gland flap, followed by temporary nonoral gastrostomy feeding. These grafts were able to recapitulate the findings of initial epithelialization and muscle regeneration around the implanted sites, as demonstrated by histological analysis. In addition, increased elastin fibers and neovascularization were observed in the periphery of the graft. Therefore, this model presents a potential new technique for circumferential esophageal reconstruction.

Esophageal reconstruction is a challenging procedure, and development of a tissue-engineered esophagus that enables regeneration of esophageal mucosa and muscle and that can be implanted as an artificial graft is necessary. Here, we present our protocol to generate an artificial esophagus, including scaffold manufacturing, bioreactor cultivation, and various surgical techniques.

쥐의 원주 식도 재건을 위한 조직 공학 이식편

The use of biocompatible materials for circumferential esophageal reconstruction is a technically challenging task in rats and requires an optimal implant ...

생체공학

Transplantation of Adipose Tissue-Derived Stem Cell Sheet to Reduce Leakage After Partial Colectomy in A Rat Model

Anastomotic leakage is a disastrous complication after colorectal surgery. Although current methods for leakage prevention have different levels of clinical efficacy, they are until now imperfect solutions. Stem cell therapy using ASC sheets could provide a solution to this problem. ASCs are considered as promising candidates for promoting tissue healing because of their trophic and immunomodulatory properties. Here, we provide methods to produce high-density ASC sheets, that are transplanted onto a colorectal anastomosis in a rat model to reduce the leakage. ASCs formed cell sheets in thermo-responsive culture dishes that could be easily detached. On the day of the transplantation, a partial colectomy with a 5-suture colorectal anastomosis was performed. Animals were immediately transplanted with 1 ASC sheet per rat. ASC sheets adhered spontaneously to the anastomosis without any glue, suture, or any biomaterial. Animal groups were sacrificed 3 and 7 days postoperatively. Compared to transplanted animals, the incidence of anastomotic abscesses and leakage was higher in control animals. In our model, the transplantation of ASC sheets after colorectal anastomosis was successful and associated with a lower leakage rate.

The preparation and transplantation of adipose tissue derived stem cell (ASC) sheets onto insufficiently sutured colorectal anastomoses in a rat model is presented. This novel application shows that ASC sheets can reduce colorectal anastomosis leakage.

쥐 모델에 부분 Colectomy 후 누설을 줄이기 위해 지방 조직 유래 줄기 세포의 이식

Anastomotic leakage is a disastrous complication after colorectal surgery. Although current methods for leakage prevention have different levels of ...

Vessel-sparing Excision and Primary Anastomosis

Urethroplasty is considered to be the standard treatment for urethral strictures since it provides excellent long-term success rates. For isolated short bulbar or posterior urethral strictures, urethroplasty by excision and primary anastomosis (EPA) is recommended. As EPA only requires the excision of the narrowed segment and the surrounding spongiofibrosis, a full-thickness transection of the corpus spongiosum, as performed in the traditional transecting EPA (tEPA), is usually unnecessary. Jordan et al. introduced the idea of a vessel-sparing approach in 2007, aiming to reduce surgical trauma, especially to the dual arterial blood supply of the urethra, and, thus, potentially reducing the risk of postoperative erectile dysfunction or glans ischemia. This approach could also be beneficial for subsequent urethral interventions such as redo urethroplasty using a free graft, in which a well-vascularized graft bed is imperative. Nevertheless, these potential benefits are only assumptions as prospective studies comparing the functional outcome of both techniques with validated questionnaires are currently lacking. Moreover, vessel-sparing EPA (vsEPA) should at least be able to provide similar surgical outcomes as tEPA. The aim of this paper is to give an elaborate, step-by-step overview of how to manage patients with isolated short bulbar or posterior urethral strictures with vsEPA. The main objective of this manuscript is to outline the surgical technique and to report the representative surgical outcome. A total of 117 patients were managed according to the described protocol. The analysis was performed on the entire patient cohort and on the bulbar (n = 91) and posterior (n = 26) vsEPA group separately. Success rates were 93.4% and 88.5% for the bulbar and posterior vsEPA, respectively. To conclude, vsEPA, as outlined in the protocol, provides excellent success rates with low complication rates for isolated short bulbar and posterior urethral strictures.

Here, we present an elaborate and efficient protocol to treat isolated short bulbar or posterior urethral strictures with vessel-sparing excision and primary anastomosis.

선박을 살려주는 절단 및 기본 문 합

Urethroplasty is considered to be the standard treatment for urethral strictures since it provides excellent long-term success rates. For isolated short ...

Transpupillary-Guided Trans-Scleral Transplantation of Subretinal Grafts in a Retinal Degeneration Mouse Model

Transplantation of photoreceptor cells and retinal pigment&#160;epithelial (RPE) cells provide a potential therapy for retinal degeneration diseases. Subretinal transplantation of therapeutic donor cells into mouse recipients is challenging due to the limited surgical space allowed by the small volume of the mouse eye. We developed a trans-scleral surgical transplantation platform with direct transpupillary vision guidance to facilitate the subretinal delivery of exogenous cells in mouse recipients. The platform was tested using retinal cell suspensions and three-dimensional retinal sheets collected from rod-rich Rho::EGFP mice and cone-rich OPN1LW-EGFP;NRL-/- mice, respectively. Live/dead assay showed low cell mortality for both forms of donor cells. Retinal grafts were successfully delivered into the subretinal space of a mouse model of retinal degeneration, Rd1/NS, with minimum surgical complications as detected by multimodal confocal scanning laser ophthalmoscope (cSLO) imaging. Two months post-transplantation, histological staining demonstrated evidence of advanced maturation of the retinal grafts into &#39;adult&#39; rods and cones (by robust Rho::EGFP, S-opsin, and OPN1LW:EGFP expression, respectively) in the subretinal space. Here, we provide a surgical platform that can enable highly accurate subretinal delivery with a low rate of complications in mouse recipients. This technique offers precision and relative ease of skill acquisition. Furthermore, the technique could be used not only for studies of subretinal cell transplantation but also for other intraocular therapeutic studies including gene therapies.

This protocol presents a transpupillary vision-guided trans-scleral approach to safely and precisely deliver subretinal cellular grafts, with a low rate of surgical complications, in mouse recipients with or without retinal degeneration.

망막 변성 마우스 모델에서 망막하 이식편의 Transpupillary-Guided Trans-Scleral Transplantation

Transplantation of photoreceptor cells and retinal pigment&#160;epithelial (RPE) cells provide a potential therapy for retinal degeneration diseases. ...

신경과학

Use of the Scissor-Type Knife During the Peroral Endoscopy Myotomy Procedure for the Treatment of Achalasia

Peroral endoscopic myotomy (POEM) is one of the first-line treatment modalities along with pneumatic dilation and Heller myotomy for patients with achalasia. Endoscopists, especially trainees during the learning phase, commonly face difficulty in tissue plane dissection and selective myotomy while working near the esophagogastric junction, with increased risks of inadvertent injury, unexpected bleeding, and inadequate myotomy. To minimize the technical difficulty and improve the safety of POEM, we describe a protocol for using a scissor-type knife for the main steps of POEM, including mucosal incision, submucosal tunneling, myotomy, and hemostasis. The standard techniques used with the scissor-type knife involve grasping the target tissue, and then dissection or coagulation. The confirmation of the cutting line after grasping improves the accuracy and reliability of dissection, which is particularly useful for the selective myotomy of the internal circular muscle. Meanwhile, the scissor-type knife provides enhanced hemostatic capability and enables hemostasis and pre-coagulation without the device exchange for hemostatic forceps. Evaluation of the clinical outcomes in three patients who successfully received POEM using the scissor-type knife revealed no perioperative adverse events. At the 3-month follow-up, all patients achieved clinical success with postoperative Eckardt scores ranging from 0 to 1. In conclusion, the use of a scissor-type knife could minimize the technical difficulty and improve the safety of the POEM procedures, which may be suitable for trainees during the learning phase.

To minimize the technical difficulty and improve the safety of peroral endoscopic myotomy (POEM), we describe a protocol for using a scissor-type knife for the main steps of POEM, including mucosal incision, submucosal tunneling, myotomy, and hemostasis.

이완불능증 치료를위한 구강 내시경 근절개술 절차 중 가위 형 칼 사용

Peroral endoscopic myotomy (POEM) is one of the first-line treatment modalities along with pneumatic dilation and Heller myotomy for patients with ...

Robotic Myotomy and Partial Fundoplication for Achalasia

Laparoscopic Heller myotomy is currently considered the standard definitive treatment of achalasia. With the advancements in technology, robotic Heller myotomy has emerged as an alternative approach to traditional laparoscopy due to three-dimensional (3D) visualization, fine motor control, and improved ergonomics provided by the robot. 
Although there is a lack of randomized controlled trials, robotic-assisted Heller myotomy seems to be associated with lower rates of intraoperative perforations compared to the laparoscopic approach. A robotic approach may also improve surgical outcomes by providing a more complete myotomy. 
Here, we describe the detailed steps of robotic myotomy and partial fundoplication for achalasia.

Surgical myotomy with a partial fundoplication may be used in selected patients as a definitive treatment for achalasia. This article provides a step-by-step description of a robotic myotomy and partial fundoplication in a 32-year-old patient with megaesophagus.

이완불능증에 대한 로봇 근절개술 및 부분 위저부주름술

Laparoscopic Heller myotomy is currently considered the standard definitive treatment of achalasia. With the advancements in technology, robotic Heller ...

Robotic D3 Partial Duodenal Resection with Primary Side-to-Side Anastomosis

Duodenal stenosis is a condition that can be related to several diseases, being either intrinsic, such as neoplasm and inflammatory stenosis, or extrinsic, such as pancreatic pseudocyst, superior mesenteric artery syndrome, and foreign bodies. Current treatments range from endoscopic approaches, such as endoscopic resection and stent placement, to surgical approaches, including duodenal resection, pancreaticoduodenectomy, and gastrointestinal bypass. Minimally invasive robot-assisted surgery is gaining importance due to its potential to decrease surgical stress, intraoperative blood loss, and postoperative pain, while its instruments and 3D-vision facilitate fine dissection and intra-abdominal suturing, all leading to a reduced time to functional recovery and shorter hospital stay. We present a case of a 75-year-old female who underwent robotic D3 partial duodenal resection with primary side-to-side duodeno-jejunal anastomosis for a 5 cm adenoma with focal high-grade dysplasia.

This protocol presents a case of a robotic partial duodenal resection with primary side-to-side duodeno-jejunal reconstruction in a patient with a 5 cm duodenal stenosis. This is done at the third duodenal segment (D3) after an endoscopic mucosal resection (EMR) for a duodenal polyp.