Name: an anesthesia, surgery, and harvest method for the evaluation of transpedicular screws using an in vivo porcine lumbar spine model
Uploaded: 2017-05-31T12:30:00.000+00:00
Duration: 9 min 7 s
Description: Watch this Scientific Journal Video about An Anesthesia, Surgery, and Harvest Method for the Evaluation of Transpedicular Screws Using an In Vivo Porcine Lumbar Spine Model at JoVE.com

이 연구는 전북 대학교 의과 대학 생물 의학 연구소 (CNUH-BRI)가 후원하는 보조금 (CNUH-BRI-2012-02-005)의 지원을 받았다.

전북 대학교 동물성 동물 관리 및 이용위원회는이 연구를 승인했다. 동물의 치료, 사용 및 취급은 모든 지침과 정책을 따랐습니다. 24 ° C에서 수술실을 유지하십시오. 1. 마취 <ol><li> 최소 1 주일 동안 실험군에서 12 개월 된 소형 돼지를 순화하십시오. 호흡 수, 심장 박동수 및 체온을 측정하여 임상 검사를 수행하십시오. 마취 수술 12 시간 전에 각 소형 돼지를 먹이지 마십시오. </li><li> 전립선 수술을 위해 귀 뒤쪽의 자궁 경부 근 부위에 atropine (0.05 mg / kg)과 ketamine (20 mg / kg) / xylaxine (2 mg / kg)을 주입하십시오. </li><li> 진정 후, 귀의 기저부 주변에 고무 밴드를 단단히 붙이고 국소 알코올로 귀를 닦으십시오. </li><li> 귀 정맥에 바늘 모양의 플라스틱 카테터를 놓고 고무 밴드를 제거합니다. 카테터가 올바르게 놓 였는지 확인하십시오. 독감heparinized 식염수와 카테 테르 sh와 테이프로 해결. </li><li> 기관 내 삽관을 위해서, 소형 돼지를 흉골 경련에 배치하십시오. 보조자의 도움을 받아 적절한 밧줄로 돼지 턱을 잡고 입을 벌리십시오. <ol><li> 후두 경의 팁을 인두 구멍으로 통과시켜 후궁을 연한 구개에서 옮깁니다. 후두경의 끝 부분을 사용하여 성대를 눈으로 확인하고 만기 중에 기관 내 튜브를 기관으로 전진시킵니다. </li><li> 적절한 삽관을위한 공기의 자유로운 통과를 느끼고 소형 돼지의 왼쪽과 오른쪽 양쪽에서 호흡 소리에 대한 가슴 청진을 확인하십시오. </li></ol></li><li> 10 mL 주사기를 사용하여 공기로 기관 내 튜브 공기의 적절한 부피를 채우고 접착 테이프를 사용하여 튜브를 주둥이에 고정하십시오. </li><li> 엔도르타를 통해 2.0 % 농도의 장기간 마취를위한 흡입 마취제 인 isoflurane을 제공하십시오.심한 삽관 튜브. 각막 및 안구 반사를 검사하여 마취를 확인하고 눈에 연고를 사용하여 건조를 방지하십시오. </li><li> 소형 돼지가 회복 될 때까지 적어도 5 분마다 마취 중 심장 혈관계, 호흡계 및 체온을 모니터링하십시오. </li><li> 척추 수술 30 분 전에 cephalosporin 항생제의 1 세대 인 cefazolin IV 30mg / kg을 천천히 바르십시오. </li><li> 항상성을 유지하기 위해 IV 라인을 사용하여 5 - 10 mL / kg / h의 따뜻한 (37 ° C) 식염수를 관리하고 통증을 조절하기 위해 펜타닐을 50 μg / kg / min으로 공급하십시오. </li><li> 척추 수술 후에 강한 삼키는 반사가 명백 해지면 발열 치료를하십시오. </li><li> 소형 돼지를 방으로 가져 가서 마취에서 회복 될 때까지 모니터하십시오. 소형 돼지가 충분히 의식이있을 때 음식과 물을 제공하십시오. </li><li> 첫 3 일 동안 통증 조절을 위해 carprofen 4.4 mg / kg과 함께 3 mg / kg enrofloxacin 항생제를 매일 사용하십시오. </li><li> 모니봉합사가 제거 될 때까지 매일 돼지에게 투여하십시오. </li></ol> 2. 척추 수술 <ol><li> 제조사의 지침에 따라, Transexicular screw와 posterior fixator system을 오토 클레이브하여 살균하십시오. </li><li> 돼지가 앙와위에있는 동안 면도기를 사용하여 중앙에서 왼쪽 또는 오른쪽으로 약 10cm 정도의 소형 돼지 뒷면을 면도하십시오. 포비돈 요오드 용액과 70 % 알코올로 피부를 닦으십시오. </li><li> 메스를 사용하여 허리 척추의 두 번째 가시 돌기에서 첫 번째 중앙 성 천장까지 경도의 정중선 절개를하십시오. 상처 부위의 피하 조직과 근막을 통해 해부합니다. </li><li> 코브 (Cobb) 엘리베이터를 사용하여 근막으로부터 subperiosteally paraspinal 근육을 올리십시오. 가시 돌기와 박판을 따라 해부학을하십시오. </li><li> 엔트리 포인트의 피상 피질을 엽니 다 (L3에서 유방 부속기 프로세스보다 열등합니다).burr 또는 rongeur로 양쪽에서 L5까지). </li><li> 가이드 핀을 상부 끝판과 평행하고 가시 돌기와 20 °의 각도로 개방 된 위치에 삽입하십시오. C-arm 또는 휴대용 postero-anterior / lateral X-ray를 사용하여 이상적인 출발점을 정의하십시오. </li><li> X- 레이에 따라 최대 25 mm까지 pedicle probe를 삽입하십시오. pedicle 발음 장치를 사용하여 pedicle과 body palpation으로 완전한 intraosseous 궤도를 확인하십시오. </li><li> 나사 헤드가 잘 장착 될 때까지 L3 ~ L5의 준비된 척추에 6 개의 척추 나사를 삽입하십시오. 임플란트 헤드의 측면 개구부를 원하는 방향으로 향하게하고 수평 위치를로드 궤도와 맞 춥니 다. </li><li> 척추 경 나사못 머리의 양쪽에 각각 두 개의 막대를 삽입하십시오. 유니버설 핸들을 사용하여 척추 경 나사못 헤드에 슬리브와 너트를 넣습니다. </li><li> 똑 바른 소켓 렌치를 사용하여 너트를 느슨하게 조이고 카운터 토크 렌치를 사용하여 너트를 단단히 조입니다. </li><li> 위치 확인 o이동식 postero-anterior / lateral X-ray를 사용하는 척추 경 나사. 참고 : 미니어처 돼지가 일어나기를 기다렸다가 뒷다리의 보행 패턴과 운동 기능을 확인하여 나사가 심하게 이식되었는지 여부를 확인합니다. </li><li> 흡입과 함께 관개 주사기를 사용하여 정상적인 염분 3 L로 수술 부위를 관개하십시오. </li><li> 수술 부위에 실리콘 배출구를 놓고 실리콘 팁을 꺼냅니다. 1.0 미터의 흡수성 봉합을 사용하여 paraspinal 근육과 피하를 닫습니다. 비 흡수성 나일론 2.0 미터법으로 피부를 닫으십시오. </li><li> 포비돈 요오드로 봉합 부위를 소독하고 멸균 된 거즈와 테이프를 사용하여 드레싱을하십시오. </li></ol> 3. 수확 절차 <ol><li> 수술 후 12 주에 xylazine (2 mg / kg)과 케타민 (ketamine) (10 mg / kg)을 귀 뒤쪽의 자궁 경부 근 지방에 주입하여 사전 투여한다. </li><li> 진정 작용 후 15 mg / kg KCl을 귀 정맥 내로 직접 투여하십시오안락사를위한 도뇨관. </li><li> 이전 수술 흉터 부위에 종 방향 정중선 절개를하십시오. 연조직과 paraspinal 근육을 해부하십시오. </li><li> 요추, 박편, 막대, 너트 및 횡단 과정의 가시 돌기를 L3에서 L5로 노출시킵니다. </li><li> 카운터 토크 렌치와로드를 사용하여 너트를 제거합니다. 진동 톱 (oscillating saw)을 사용하여 L2-3 디스크 공간과 L5-S1 디스크 공간을 자릅니다. </li><li> Cobb 엘리베이터와 타워 포 셉터로 L3-5 등뼈의 가운데와 앞부분의 양쪽을 해부합니다. 수확 후 척추를 즉시 검사 할 수 없다면 식염수로 적신 거즈에 싸서 -20 ° C에서 보관하십시오. </li></ol>

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H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+use+of+a+quadrupted+as+an+in+vivo+model+for+the+study+of+the+spine-biomechanical+considrations.">The use of a quadrupted as an in vivo model for the study of the spine-biomechanical considrations.</a> Eur Spine J. 11 (2), 137-144 (2002).</li><li>Aldini, N. 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돼지 척추의 경정맥 나사의 평가에는 많은 시간과 노력이 요구됩니다. 첫째, 소형 돼지는 큰 동물입니다. 동물 관리 및 마취를 위해 연구원은 전문적인 프로토콜이 필요합니다. 둘째, 수술은 인간의 수술과 비슷한 환경을 유지해야합니다. 돼지 척추에서 척추 경 나사를 평가하는 목적은 인간에게 적용 할 수있는 효율적인 나사를 개발하는 것입니다. 셋째, 척추 경 나사의 장기 안정성을 평가하는 ...

Transpedicular screw fixation is a gold-standard treatment for degenerative lumbar spine and bursting fracture because it involves three columns of the spine and achieves stabilization1,2. However, most patients who undergo such surgery also have osteoporosis3,4. Many studies have evaluated the fixation strength and the osseointegration status of transpedicular screws, because the loosening of pedicle screws currently in use has been reported in patients with osteoporosis5,6.
The porcine spine is similar to the human spine in terms of size. It is less expensive compared to a primate model7. Furthermore, an in vivo mechanical study has demonstrated that the quadruped porcine spine is essentially loaded in the same way as that of the human spine8, which is why many researchers use porcine spines for studies on the prevention of pedicle screw loosening. However, it takes several months to study pedicle screws in the porcine spine because identifying the long-term stability of pedicle screws takes times. In order to compare different types of screws in the vertebral body, it is necessary to insert the screws in similar positions. Therefore, researchers should be well-acquainted with proper anesthesia techniques, standardized surgical protocols, and harvest procedures before performing any experiments. Here, we describe a detailed method for anesthesia, surgery, and harvest for the evaluation of pedicle screw fixation using a porcine spine model, including ex vivo imaging, histology, and strength testing.

<table><tbody><tr><td>Miniature pig</td><td>OrientBio</td><td></td><td></td></tr><tr><td>Atropine</td><td>Jeil pharmaceutical</td><td>A04900241</td><td>Anesthesia</td></tr><tr><td>Over-the needle plastic catheter</td><td>BD</td><td>REF382412</td><td>Maintenance of IV line</td></tr><tr><td>Ketamine</td><td>Yuhan</td><td>A04502441</td><td>Anesthesia</td></tr><tr><td>Xylazine</td><td>Bayer Korea</td><td>A00800071</td><td>Anesthesia</td></tr><tr><td>Laryngoscope</td><td>Karl storz</td><td></td><td>Intubation</td></tr><tr><td>Endotracheal tube</td><td>Covidien</td><td></td><td>Intubation</td></tr><tr><td>Isoflurane</td><td>JW pharmaceutical Co</td><td>A02104781</td><td>Anesthesia</td></tr><tr><td>Eye ointment</td><td>Hanlim pharma</td><td>A37851721</td><td>Protection of pig&#039;s eye</td></tr><tr><td>Cefazolin</td><td>Donga pharma</td><td>A01503951</td><td>Antibiotics</td></tr><tr><td>Saline</td><td>JW pharmaceutical Co</td><td>A02151392</td><td>Maintenance of fluid homeostasis</td></tr><tr><td>Fentanyl</td><td>Hana pharm</td><td>C03200032</td><td>Pain control</td></tr><tr><td>Enrofloxacin</td><td>Bayer</td><td>93106-60-6&amp;nbsp;</td><td>Antibiotics</td></tr><tr><td>Morphine</td><td>Myungmoon pharma</td><td>C03700091</td><td>Pain control</td></tr><tr><td>Meloxicam</td><td>Boehringer Ingelheim</td><td>A07600711</td><td>Antibiotics</td></tr><tr><td>Povidone-iodine</td><td>Hyundai pharma</td><td></td><td>Wound dressing</td></tr><tr><td>Scalpel blade size 15</td><td>Braun</td><td>&amp;nbsp;I1 BB515</td><td>Skin incision</td></tr><tr><td>Cobb elevator</td><td>Codman</td><td>65-2546</td><td>Dissection of muscle</td></tr><tr><td>Burr</td><td>Medtronic</td><td></td><td>Making of starting point of screw</td></tr><tr><td>Rongeur</td><td>Aesculap</td><td>FO515R</td><td>Making of starting point of screw</td></tr><tr><td>Guide pin (K-wire)</td><td>CE</td><td>01067803</td><td>Guidance of screw trajectory</td></tr><tr><td>C-arm</td><td>GE</td><td>OEC 9800 plus</td><td>Guidance of screw trajectory</td></tr><tr><td>Portable X-ray</td><td>Siemens</td><td>Mobile XP hybrid</td><td>Guidance of screw trajectory</td></tr><tr><td>Pedicle probe</td><td>OtisBiotech</td><td>SPI-02-01</td><td>Guidance of screw trajectory</td></tr><tr><td>Pedicle sounding device</td><td>OtisBiotech</td><td>SPI-03-01</td><td>Guidance of screw trajectory</td></tr><tr><td>Pedicle screw</td><td>OtisBiotech</td><td>MS-40025</td><td></td></tr><tr><td>Posterior fixator systems</td><td>OtisBiotech</td><td></td><td></td></tr><tr><td>Rod&amp;nbsp;</td><td>OtisBiotech</td><td>ROD-60140</td><td>Rigid fixation between screws</td></tr><tr><td>Universal handle</td><td>OtisBiotech</td><td>SPI-08-01</td><td>To fix the screws to the rod</td></tr><tr><td>Straight socket wrench</td><td>OtisBiotech</td><td>SPI-06-01</td><td>To fix the screws to the rod</td></tr><tr><td>counter torque wrench</td><td>OtisBiotech</td><td>SPI-07-01</td><td>To fix the screws to the rod</td></tr><tr><td>Bulb irrigation syringe</td><td>Hyupsug medical</td><td>HS-IR-140</td><td>Irrigation</td></tr><tr><td>Silicone drain</td><td>Sewon medical</td><td>2205-006</td><td>To drain the fluid at the surgical site</td></tr><tr><td>3.0 metric absorbable suture</td><td>Ethicon</td><td>BA1673H</td><td>Muscle suture</td></tr><tr><td>2.0 metric nonabsorbable nylon suture</td><td>Ethicon</td><td>W1626T</td><td>Skin suture</td></tr><tr><td>Gauze</td><td>Kingphar Korea</td><td>KP120-06</td><td></td></tr><tr><td>Pentobarbital</td><td>Hanlim pharma</td><td>645301221</td><td>Euthanasia</td></tr><tr><td>Oscillating saw</td><td>Zimmer</td><td></td><td>Harvest spine</td></tr><tr><td>Tower forceps</td><td>Aesculap</td><td>BF461R</td><td>Harvest spine</td></tr></tbody></table>

an anesthesia, surgery, and harvest method for the evaluation of transpedicular screws using an in vivo porcine lumbar spine model

척추 경 나사 고정은 척추 질환의 치료에있어 가장 중요한 표준입니다. 그러나 척추 수술 후 척추 경 나사못을 느슨하게하는 문제에 대한 많은 연구가있어 심각한 문제입니다. 이 문제를 해결하기 위해 다양한 유형의 척추 경 나사못을 검사하여 척추골에서 좋은 고정 강도와 골 유착을 가진 환자를 확인했습니다. 돼지 척추는 해부학 적 크기, 기계적 특성 및 비용으로 인해 척추 경 나사의 평가에서 인간의 척추에 대한 좋은 대안입니다. 여러 연구에 따르면 척추 모형이 돼지 모형에서 효율적이라는보고가 있지만, 돼지 모형을 사용한 척추 경 나사 평가에 대한 자세한 프로토콜은 아직 없습니다. 여기, 우리 는 생체 내 돼지 허리 받침 모델을 사용하여 Transedicular 나사를 평가하기위한 상세한 방법을 설명합니다. 여기에 제공된 마취, 척추 수술 및 수확에 대한 기술 세부 정보는 t경 나사못 고정 모델.

생체 내 돼지 허리 받침 모델을 사용하여 Transedicular screw를 평가하기위한 마취, 수술 및 수확을위한 자세한 프로토콜이 여기에 설명되어 있습니다. 이 프로토콜은 기계적 테스트 ( 그림 1 ), 정량적 마이크로 CT 평가 ( 그림 2 ) 및 조직학 ( 그림 3 )을 포함한 여러 가지 다운 스트림 분석에 적합합니다. 대표적인 기계적 시험 ( <strong...

Watch this Scientific Journal Video about An Anesthesia, Surgery, and Harvest Method for the Evaluation of Transpedicular Screws Using an In Vivo Porcine Lumbar Spine Model at JoVE.com

An Anesthesia, Surgery, and Harvest Method for the Evaluation of Transpedicular Screws Using an In Vivo Porcine Lumbar Spine Model

여기에서는 생체 내 돼지 허리 받침 모델을 사용하여 경정맥 나사를 평가하는 방법을 소개합니다.

수술 용 나사를 이용한 마취, 수술 및 수확 방법<em&gt; In Vivo</em&gt; 돼지의 허리 척추 모델

Pedicle screw fixation is the gold standard for the treatment of spinal diseases. However, many studies have reported the issue of loosening pedicle ...

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An Anesthesia, Surgery, and Harvest Method for the Evaluation of Transpedicular Screws Using an In Vivo Porcine Lumbar Spine Model

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7.6K 조회수.  Chonbuk National University School of Medicine, Chonbuk National University Hospital.  여기에서는 생체 내 돼지 허리 받침 모델을 사용하여 경정맥 나사를 평가하는 방법을 소개합니다. 

비디오: An Anesthesia, Surgery, and Harvest Method for the Evaluation of Transpedicular Screws Using an In Vivo Porcine Lumbar Spine Model

In vivo Imaging of the Mouse Spinal Cord Using Two-photon Microscopy

In vivo imaging using two-photon microscopy 1 in mice that have been genetically engineered to express fluorescent proteins in specific cell types 2-3 has significantly broadened our knowledge of physiological and pathological processes in numerous tissues in vivo 4-7. In studies of the central nervous system (CNS), there has been a broad application of in vivo imaging in the brain, which has produced a plethora of novel and often unexpected findings about the behavior of cells such as neurons, astrocytes, microglia, under physiological or pathological conditions 8-17. However, mostly technical complications have limited the implementation of in vivo imaging in studies of the living mouse spinal cord. In particular, the anatomical proximity of the spinal cord to the lungs and heart generates significant movement artifact that makes imaging the living spinal cord a challenging task.
We developed a novel method that overcomes the inherent limitations of spinal cord imaging by stabilizing the spinal column, reducing respiratory-induced movements and thereby facilitating the use of two-photon microscopy to image the mouse spinal cord in vivo. This is achieved by combining a customized spinal stabilization device with a method of deep anesthesia, resulting in a significant reduction of respiratory-induced movements. This video protocol shows how to expose a small area of the living spinal cord that can be maintained under stable physiological conditions over extended periods of time by keeping tissue injury and bleeding to a minimum. Representative raw images acquired in vivo detail in high resolution the close relationship between microglia and the vasculature. A timelapse sequence shows the dynamic behavior of microglial processes in the living mouse spinal cord. Moreover, a continuous scan of the same z-frame demonstrates the outstanding stability that this method can achieve to generate stacks of images and/or timelapse movies that do not require image alignment post-acquisition. Finally, we show how this method can be used to revisit and reimage the same area of the spinal cord at later timepoints, allowing for longitudinal studies of ongoing physiological or pathological processes in vivo.

In vivo Imaging of the Mouse Spinal Cord Using Two-photon Microscopy

A minimally invasive protocol to stabilize the mouse spinal column and perform repetitive in vivo spinal cord imaging using two-photon microscopy is described. This method combines a spinal stabilization device and an anesthetic regimen to minimize respiratory-induced movements and produce raw imaging data that require no alignment or other post-processing.

두 광자 현미경을 사용하여 마우스 척수의 생체내 이미징에

In vivo imaging using two-photon microscopy 1 in mice that have been genetically engineered to express fluorescent proteins in specific cell types 2-3 has ...

연구

신경과학

A Procedure for Implanting a Spinal Chamber for Longitudinal In Vivo Imaging of the Mouse Spinal Cord

Studies in the mammalian neocortex have enabled unprecedented resolution of cortical structure, activity, and response to neurodegenerative insults by repeated, time-lapse in vivo imaging in live rodents. These studies were made possible by straightforward surgical procedures, which enabled optical access for a prolonged period of time without repeat surgical procedures. In contrast, analogous studies of the spinal cord have been previously limited to only a few imaging sessions, each of which required an invasive surgery. As previously described, we have developed a spinal chamber that enables continuous optical access for upwards of 8 weeks, preserves mechanical stability of the spinal column, is easily stabilized externally during imaging, and requires only a single surgery. Here, the design of the spinal chamber with its associated surgical implements is reviewed and the surgical procedure is demonstrated in detail. Briefly, this video will demonstrate the preparation of the surgical area and mouse for surgery, exposure of the spinal vertebra and appropriate tissue debridement, the delivery of the implant and vertebral clamping, the completion of the chamber, the removal of the delivery system, sealing of the skin, and finally, post-operative care. The procedure for chronic in vivo imaging using nonlinear microscopy will also be demonstrated. Finally, outcomes, limitations, typical variability, and a guide for troubleshooting are discussed.

A Procedure for Implanting a Spinal Chamber for Longitudinal In Vivo Imaging of the Mouse Spinal Cord

In this video, we describe a procedure for implanting a chronic optical imaging chamber over the dorsal spinal cord of a live mouse. The chamber, surgical procedure, and chronic imaging are reviewed and demonstrated.

종의 척추 상공 회의소를 주입하는 절차<em&gt; 생체</em&gt; 마우스 척수의 영상

Studies in the mammalian neocortex have enabled unprecedented resolution of cortical structure, activity, and response to neurodegenerative insults by ...

Laminotomy for Lumbar Dorsal Root Ganglion Access and Injection in Swine

Dorsal root ganglia (DRG) are anatomically well defined structures that contain all primary sensory neurons below the head. This fact makes DRG attractive targets for injection of novel therapeutics aimed at treating chronic pain. In small animal models, laminectomy has been used to facilitate DRG injection because it involves surgical removal of the vertebral bone surrounding each DRG. We demonstrate a technique for intraganglionic injection of lumbar DRG in a large animal species, namely, swine. Laminotomy is performed to allow direct access to DRG using standard neurosurgical techniques, instruments, and materials. Compared with more extensive bone removal via laminectomy, we implement laminotomy to conserve spinal anatomy while achieving sufficient DRG access. Intraoperative progress of DRG injection is monitored using a non-toxic dye. Following euthanasia on post-operative day 21, the success of injection is determined by histology for intraganglionic distribution of 4&#39;,6-diamidino-2-phenylindole (DAPI). We inject a biologically inactive solution to demonstrate the protocol. This method could be applied in future preclinical studies to target therapeutic solutions to DRG. Our methodology should facilitate testing the translatability of intraganglionic small animal paradigms in a large animal species. Additionally, this protocol may serve as a key resource for those planning preclinical studies of DRG injection in swine.

We describe a method for laminotomy in swine that provides access to lumbar dorsal root ganglia (DRG) for intraganglionic injection. Injection progress is monitored intraoperatively and histologically confirmed up to 21 days after surgery. This protocol could be used for future preclinical studies involving DRG injection.

Laminotomy 허리 지 루트 신경 절의 액세스 및 돼지에 주입

Dorsal root ganglia (DRG) are anatomically well defined structures that contain all primary sensory neurons below the head. This fact makes DRG attractive ...

Laminectomy and Spinal Cord Window Implantation in the Mouse

This protocol describes a method for spinal cord laminectomy and glass window implantation for in vivo imaging of the mouse spinal cord. An integrated digital vaporizer is utilized to achieve a stable plane of anesthesia at a low-flow rate of isoflurane. A single vertebral spine is removed, and a commercially available cover-glass is overlaid on a thin agarose bed. A 3D-printed plastic backplate is then affixed to the adjacent vertebral spines using tissue adhesive and dental cement. A stabilization platform is used to reduce motion artifact from respiration and heartbeat. This rapid and clamp-free method is well-suited for acute multi-photon fluorescence microscopy. Representative data are included for an application of this technique to two-photon microscopy of the spinal cord vasculature in transgenic mice expressing eGFP:Claudin-5 &#8212; a tight junction protein.

This protocol describes implantation of a glass window onto the spinal cord of a mouse to facilitate visualization by intravital microscopy.

마우스에 있는 Laminectomy 및 척수 창 이식

This protocol describes a method for spinal cord laminectomy and glass window implantation for in vivo imaging of the mouse spinal cord. An integrated ...

In Vivo Mouse Model of Spinal Implant Infection

Spine implant infections portend poor outcomes as diagnosis is challenging and surgical eradication is at odds with mechanical spinal stability. The purpose of this method is to describe a novel mouse model of spinal implant infection (SII) that was created to provide an inexpensive, rapid, and accurate in vivo tool to test potential therapeutics and treatment strategies for spinal implant infections.
In this method, we present a model of posterior-approach spinal surgery in which a stainless-steel k-wire is transfixed into the L4 spinous process of 12-week old C57BL/6J wild-type mice and inoculated with 1 x 103 CFU of a bioluminescent strain of Staphylococcus aureus Xen36 bacteria. Mice are then longitudinally imaged for bioluminescence in vivo on post-operative days 0, 1, 3, 5, 7, 10, 14, 18, 21, 25, 28, and 35. Bioluminescence imaging (BLI) signals from a standardized field of view are quantified to measure in vivo bacterial burden.
To quantify bacteria adhering to implants and peri-implant tissue, mice are euthanized and the implant and surrounding soft tissue are harvested. Bacteria are detached from the implant by sonication, cultured overnight and then colony forming units (CFUs) are counted. The results acquired from this method include longitudinal bacterial counts as measured by in vivo S. aureus bioluminescence (mean maximum flux) and CFU counts following euthanasia.
While prior animal models of instrumented spine infection have involved invasive, ex vivo tissue analysis, the mouse model of SII presented in this paper leverages noninvasive, real time in vivo optical imaging of bioluminescent bacteria to replace static tissue study. Applications of the model are broad and may include utilizing alternative bioluminescent bacterial strains, incorporating other types of genetically engineered mice to contemporaneously study host immune response, and evaluating current or investigating new diagnostic and therapeutic modalities such as antibiotics or implant coatings.

The protocol describes a novel in vivo mouse model of spinal implant infection where a stainless-steel k-wire implant is infected with bioluminescent Staphylococcus aureus Xen36. Bacterial burden is monitored longitudinally with bioluminescent imaging and confirmed with colony forming unit counts after euthanasia.

척추 임플란트 감염의 In Vivo 마우스 모델

Spine implant infections portend poor outcomes as diagnosis is challenging and surgical eradication is at odds with mechanical spinal stability. The ...

의학

Real-Time Assessment of Spinal Cord Microperfusion in a Porcine Model of Ischemia/Reperfusion

Spinal cord injury is a devastating complication of aortic repair. Despite developments for the prevention and treatment of spinal cord injury, its incidence is still considerably high and therefore, influences patient outcome. Microcirculation plays a key role in tissue perfusion and oxygen supply and is often dissociated from macrohemodynamics. Thus, direct evaluation of spinal cord microcirculation is essential for the development of microcirculation-targeted therapies and the evaluation of existing approaches in regard to spinal cord microcirculation. However, most of the methods do not provide real-time assessment of spinal cord microcirculation. The aim of this study is to describe a standardized protocol for real-time spinal cord microcirculatory evaluation using laser-Doppler needle probes directly inserted in the spinal cord. We used a porcine model of ischemia/reperfusion to induce deterioration of the spinal cord microcirculation. In addition, a fluorescent microsphere injection technique was used. Initially, animals were anesthetized and mechanically ventilated. Thereafter, laser-Doppler needle probe insertion was performed, followed by the placement of cerebrospinal fluid drainage. A median sternotomy was performed for exposure of the descending aorta to perform aortic cross-clamping. Ischemia/reperfusion was induced by supra-celiac aortic cross-clamping for a total of 48 min, followed by reperfusion and hemodynamic stabilization. Laser-Doppler Flux was performed in parallel with macrohemodynamic evaluation. In addition, automated cerebrospinal fluid drainage was used to maintain a stable cerebrospinal pressure. After completion of the protocol, animals were sacrificed, and the spinal cord was harvested for histopathological and microsphere analysis. The protocol reveals the feasibility of spinal cord microperfusion measurements using laser-Doppler probes and shows a marked decrease during ischemia as well as recovery after reperfusion. Results showed comparable behavior to fluorescent microsphere evaluation. In conclusion, this new protocol might provide a useful large animal model for future studies using real-time spinal cord microperfusion assessment in ischemia/reperfusion conditions.

Spinal cord microcirculation plays a pivotal role in spinal cord injury. Most methods do not allow real-time assessment of spinal cord microcirculation, which is essential for the development of microcirculation-targeted therapies. Here, we propose a protocol using Laser-Doppler-Flow Needle probes in a large animal model of ischemia/reperfusion.

허혈/재관류의 돼지 모델에서 척수 미세 퍼퍼퓨전의 실시간 평가

Spinal cord injury is a devastating complication of aortic repair. Despite developments for the prevention and treatment of spinal cord injury, its ...

Surgical Technique for Lumbar Spinal Catheter Insertion in Pigs Enabling Continuous Access to the Thecal Sac in a Terminal Setup

Pigs are increasingly used as a large animal model for pharmacologic CNS research due to the anatomical and physiological similarities between the porcine and human central nervous systems (CNS). However, accessing the cerebrospinal fluid (CSF) in larger pig breeds by conventional lumbar puncture techniques can be challenging due to an oblique orientation of the spinal spinous processes and a limited interlaminar space. Accordingly, an open surgical procedure for inserting a lumbar spinal catheter for continuous CSF sampling at the L4/L5 level in pigs is thoroughly described in this work. After positioning the pig and identifying the anatomical landmarks, a dorsal midline surgical incision is made to expose the spinous processes. By advancing the introducer needle, the spinal catheter is inserted inside the thecal sac of the spinal canal while leaving the bone structures of the spine intact. This method allows continuous infusion into or sampling from the porcine thecal sac with minimal bleeding or CSF leakage. The procedure is simple, time-efficient, and reproducible across different experimental setups, offering significant potential for various pre-clinical studies, including pharmacokinetic research, surgical training, and spinal cord injury models.

We present a technique for inserting a lumbar spinal catheter at the L4-L5 level in a 3-month-old Danish Landrace pig as part of a terminal research protocol, enabling continuous infusion or CSF sampling from the thecal sac.

돼지의 요추 척추 카테터 삽입을 위한 수술 기법으로 터미널 설정에서 종골낭에 지속적으로 접근할 수 있습니다.

Pigs are increasingly used as a large animal model for pharmacologic CNS research due to the anatomical and physiological similarities between the porcine ...

A Mouse Model of Lumbar Spine Instability

Intervertebral disc degeneration (IDD) is a common pathological change leading to low back pain. Appropriate animal models are desired for understanding the pathological processes and evaluating new drugs. Here, we introduced a surgically induced lumbar spine instability (LSI) mouse model that develops IDD starting from 1 week post operation. In detail, the mouse under anesthesia was operated by low back skin incision, L3&#8211;L5 spinous processes exposure, detachment of paraspinous muscles, resection of processes and ligaments, and skin closure. L4&#8211;L5 IVDs were chosen for the observation. The LSI model develops lumbar IDD by porosity and hypertrophy in endplates at an early stage, decrease in intervertebral disc volume, shrinkage in nucleus pulposus at an intermediate stage, and bone loss in lumbar vertebrae (L5) at a later stage. The LSI mouse model has the advantages of strong operability, no requirement of special equipment, reproducibility, inexpensive, and relatively short period of IDD development. However, LSI operation is still a trauma that causes inflammation within the first week post operation. Thus, this animal model is suitable for study of lumbar IDD.

We developed a lumbar intervertebral disc degeneration mouse model by resection of L3&#8211;L5 spinous processes along with supra- and inter-spinous ligaments and detachment of paraspinous muscles.

요추 척추 불안정의 마우스 모델

Intervertebral disc degeneration (IDD) is a common pathological change leading to low back pain. Appropriate animal models are desired for understanding ...

생물학

Pedicle Screw Placement Using an Augmented Reality Head-Mounted Display in a Porcine Model

This protocol helps assess the accuracy and workflow of an augmented reality (AR) hybrid navigation system using the Magic Leap head-mounted display (HMD) for minimally invasive pedicle screw placement. The cadaveric porcine specimens were placed on a surgical table and draped with sterile covers. The levels of interest were identified using fluoroscopy, and a dynamic reference frame was attached to the spinous process of a vertebra in the region of interest. Cone beam computerized tomography (CBCT) was performed, and a 3D rendering was automatically generated, which was used for the subsequent planning of the pedicle screw placements. Each surgeon was fitted with an HMD that was individually eye-calibrated and connected to the spinal navigation system.
Navigated instruments, tracked by the navigation system and displayed in 2D and 3D in the HMD, were used for 33 pedicle cannulations, each with a diameter of 4.5 mm. Postprocedural CBCT scans were assessed by an independent reviewer to measure the technical (deviation from the planned path) and clinical (Gertzbein grade) accuracy of each cannulation. The navigation time for each cannulation was measured. The technical accuracy was 1.0 mm &#177; 0.5 mm at the entry point and 0.8 mm &#177; 0.1 mm at the target. The angular deviation was 1.5&#176; &#177; 0.6&#176;, and the mean insertion time per cannulation was 141 s &#177; 71 s. The clinical accuracy was 100% according to the Gertzbein grading scale (32 grade 0; 1 grade 1). When used for minimally invasive pedicle cannulations in a porcine model, submillimeter technical accuracy and 100% clinical accuracy could be achieved with this protocol.

The augmented reality head-mounted display, Magic Leap, was used in combination with a conventional navigation system to place pedicle screws in a porcine model by adhering to a novel workflow. With a median insertion time of &lt;2.5 min, submillimeter technical accuracy and 100% clinical accuracy were achieved according to Gertzbein.

돼지 모델에서 증강 현실 헤드 마운트 디스플레이를 사용한 척추경 나사 배치

This protocol helps assess the accuracy and workflow of an augmented reality (AR) hybrid navigation system using the Magic Leap head-mounted display (HMD) ...

Laser Doppler Needle Probe Insertion: A Technique for Real-time Quantification of Blood Flow within the Spinal Cord in a Porcine Model

Source: Behem, C. R. et al., <a href="https://www.jove.com/v/62047">Real-Time Assessment of Spinal Cord Microperfusion in a Porcine Model of Ischemia/Reperfusion.</a> J. Vis. Exp. (2020).
This video demonstrates the placement of a laser Doppler needle probe in the spinal cord of a porcine model. This helps in the continuous quantification of blood supply to the spinal cord, an important assessment in spinal cord injury.

레이저 도플러 바늘 프로브 삽입: 돼지 모델에서 척수 내 혈류의 실시간 정량화를 위한 기술

Source: Behem, C. R. et al., Real-Time Assessment of Spinal Cord Microperfusion in a Porcine Model of Ischemia/Reperfusion. J. Vis. Exp. (2020).
This ...

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