פרויקט זה מומן על ידי אגודת המחקר הגרמני (מענק HE 4578/3-1) וחלקו על ידי האיחוד האירופי FP7-הפרויקט &quot;גמבה&quot; NMP3-SL-2010-245,993.

הכנת א ארנב תורם לבידוד של תאי גזע mesenchymal (חדר ניתוח) <ol style=";text-align:right;direction:rtl"><li style=";text-align:right;direction:rtl"> תאים מבודדים מזכר לבן (NZW) ארנבי ניו זילנד בגיל 4 חודשים וכ 3 ק&quot;ג משקל גוף. </li><li style=";text-align:right;direction:rtl"> לגרום הרדמה פרופופול על ידי (10 מ&quot;ג / ק&quot;ג משקל הגוף ד) ותקריב עם נתרן pentobarbital (משקל גוף מ&quot;ג / ק&quot;ג 100 ד). </li><li style=";text-align:right;direction:rtl"> לגלח את הפרווה מגפיים אחוריות, גב ובטן עם גוזז חשמלי ולשאוב את הפרווה. </li><li style=";text-align:right;direction:rtl"> לחטא את האזור המגולח ביסודיות עם אתנול 70%. </li><li style=";text-align:right;direction:rtl"> השתמש במלקחיים בוטים, מספריים חדים (או סכין מנתחים) וחותכים לרקמת עצם ורצועות. </li><li style=";text-align:right;direction:rtl"> עושה חתך לאורך משטח הגולגולת של הרגל והעגל. </li><li style=";text-align:right;direction:rtl"> משקף עור ורקמה התת עורית או על ידי נתיחה חדה או קהה. </li><li style=";text-align:right;direction:rtl"> שרירים ורצועות נפרדים מעצם השוק וירך. שמור על קיצוצים כקרובים לעצם ככל האפשר כדי להפוך את חיתוך נקי. אל תפריד בין עצם ירך מעצם השוק בשלב זה. </li><li style=";text-align:right;direction:rtl"> חותכים through מפרק הירך להפריד את ראש עצם הירך ממרחשת. </li><li style=";text-align:right;direction:rtl"> לרומם מורכב השוקה-הירך. </li><li style=";text-align:right;direction:rtl"> השתמש בלהב סכין המנתחים כדי לגרד את כל רקמות רכות שנותרו מהעצמות או לשפשף את העצמות עם רקמות בד סטריליים. בשלב זה, עצם הירך ועצם השוק עדיין מחוברות. </li><li style=";text-align:right;direction:rtl"> הסר את פיקת הברך על ידי חיתוך, ואז לחתוך רצועות מפרק הברך לעצמות נפרדות סוף סוף. </li><li style=";text-align:right;direction:rtl"> ספריי הופרדו עצמות עם 70% אתנול, לתת אוויר יבש ולמקם כל עצם לתוך צינור צנטריפוגה 50 מ&quot;ל עם תרבית תאים בינוניות (DMEM + 1% פניצילין סטרפטומיצין / (עט / סטרפטוקוקוס)) כדי לשמור עליהם לח. </li><li style=";text-align:right;direction:rtl"> לעבור עכשיו מתחת למכסת מנוע מינרית סטרילי תרבית תאי זרימה. </li></ol> ב &#39;פלאשינג של MSC ארנב מעצמות והרחבה (הוד תרבות תאים) <ol style=";text-align:right;direction:rtl"><li style=";text-align:right;direction:rtl"> לאסוף עצמות מצינורות ולמקם אותם לתוך 150 מנות מ&quot;מ באמצעות מלקחיים סטריליים. </li><li style=";text-align:right;direction:rtl"> להסיר גם עצם מסתיים עם מסור סטרילי וחתיכות לעבור למנות חדשות 150 מ&quot;מ. </li><li style=";text-align:right;direction:rtl"> ממלאי מזרק 10 מ&quot;ל עם Mediאום (DMEM), לצרף מחט מד 18 ולהכניס לתוך הפתח של מח העצם. </li><li style=";text-align:right;direction:rtl"> לאחר מכן, יש לשטוף את חלל מח עם מדיום כדי לשטוף את מח העצם לתוך הצלחת. לאחר מכן, יש לשטוף מהקצה השני, אם אפשר. במידת צורך, ראה את יותר מהקצוות. אם צריכה לשבור עצמות, פשוט לשטוף את החלק הפנימי של העצם. </li><li style=";text-align:right;direction:rtl"> לשאוב השעיה מדיום סלולרי לתוך המזרק ולשטוף את מח העצם שוב ושוב עד שההשעיה היא צף חופשי דרך חלל מח העצם ולא עצם נוסף מח קרישים יופיעו. </li><li style=";text-align:right;direction:rtl"> ברגע שמוח עצם שנאסף מכל העצמות, לשבש את גושי מח על ידי עובר דרך מחט מד 18: למלא מזרק עם המחט מחוברת ולאלץ החוצה לתוך מדיום. </li><li style=";text-align:right;direction:rtl"> לאחר מכן, לסנן את ההשעיה דרך פילטר תא לתוך צינור 50 מ&quot;ל. על מנת למנוע אובדן תא, לשטוף 2x צלחת התרבות עם מדיום 10 מ&quot;ל ולסנן גם כן. </li><li style=";text-align:right;direction:rtl"> צנטריפוגה השעיה על XG 500 במשך 5 דקות ב RT. </li><li style=";text-align:right;direction:rtl"> הסר supernatant ו resuspend התא פלet במדיום 10 מ&quot;ל (עט% + 1 / סטרפטוקוקוס DMEM). </li><li style=";text-align:right;direction:rtl"> תאי דם נפרדים מתאי דם היקפיים (mononuclear PBMC) ותאי גזע mesenchymal (MSC) באמצעות פתרון הפרדת Biocoll. </li><li style=";text-align:right;direction:rtl"> מלא 5 מ&quot;ל של Biocoll הפרדת פתרון לתוך שפופרת 15 מ&quot;ל ולהוסיף 5 מ&quot;ל של השעיה תא בזהירות על גבי וצנטריפוגות ב XG 800 עבור 20 דקות ב RT (ללא בלמים). </li><li style=";text-align:right;direction:rtl"> תוצאות אפשריות ראו איור 1: צפוף יותר מאשר להיות Biocoll, דם משקע תאים אדום בתחתית ואילו תאי גזע PBMC וmesenchymal להישאר בממשק. </li><li style=";text-align:right;direction:rtl"> בזהירות להסיר את הממשק לתוך צינור 15 מ&quot;ל ולשטוף עם 5 מ&quot;ל PBS. </li><li style=";text-align:right;direction:rtl"> צנטריפוגה כמתואר בשלב 22, resuspend ב 5 מ&quot;ל PBS ולחזור על 2-3x. </li><li style=";text-align:right;direction:rtl"> ואז, שוב בצנטריפוגה XG 350 עבור 10 דקות ב RT (עם בלמים). </li><li style=";text-align:right;direction:rtl"> Resuspend במדיום 10 מ&quot;ל ולספור תאים בhemocytometer. </li><li style=";text-align:right;direction:rtl"> פלייט תאים בצפיפות זריעה ראשונית של כ 5 x 10 6 מנות ב -150 מ&quot;מ. </li><li style=";text-align:right;direction:rtl"> לאחר 2-3 ימים, מחדשלהעביר תאים שאינם חסיד. ייתכן שתצטרך לשטוף עם PBS הראשון על מנת להסיר תא פסולת. הוסף בינונית מלא טרי (DMEM + עגל עוברי 10% בסרום (FCS) + 1% עט / סטרפטוקוקוס) לאחר מכן. </li><li style=";text-align:right;direction:rtl"> להאכיל את תאים כל 3-4 ימים (איור 2). </li><li style=";text-align:right;direction:rtl"> לאחר 5-10 ימים, מעבר התאים בפעם הראשונה. </li></ol> הכנת ג קרישי הפיברין במבחנה <ol style=";text-align:right;direction:rtl"><li style=";text-align:right;direction:rtl"> ביום של השתלה, לשחרר תאים חסיד מצלוחיות / מנות על ידי חשיפת 3 דקות-0.25% טריפסין-EDTA. תפסיק trypsinization ידי הוספה בינונית מלאה. </li><li style=";text-align:right;direction:rtl"> הפץ תאים בצינור פלקון 50 מ&quot;ל ולשטוף אותם פעמיים עם PBS. </li><li style=";text-align:right;direction:rtl"> לקבוע כדאיות תא ומספרים על ידי צביעת trypan הכחולה. </li><li style=";text-align:right;direction:rtl"> הוסף 50,000 תאים / צינור microcentrifuge ולאסוף כדורים על ידי צנטריפוגה בXG 500 במשך 5 דקות ב RT. הכן mastermix לקריש אחד לפחות יותר במידת צורך. </li><li style=";text-align:right;direction:rtl"> Resuspend התא גלולה ב 17 PBS μl ולערבב 25 μl של הרכיב של פיברינוגן TissuCOL-קיט עם ההשעיה MSC μl זה 17. </li><li style=";text-align:right;direction:rtl"> קח צלחת סטרילית עם חורים שנקדחו מראש (3x3.6 מ&quot;מ) בהתאם לחורי מקדח in vivo (איור 3). </li><li style=";text-align:right;direction:rtl"> ראשית, לחסן 4 פתרון תרומבין μl (500 IU / מ&quot;ל) לתוך חור אחד, ואחריו תוספת מיידית של 42 שוב 4 פתרון תרומבין μl על גבי תאי פיברינוגן השעיה וμl. אין לערבב את ההשעיה, כדי למנוע קרישה בקצה פיפטה. ראשית, נפח μl 50 של פיברינוגן-תא ההשעיה pipetted יבלוט שפה של החורים שנקדחו מראש ללא מסה בשל מתח הפנים. עם זאת, לאחר קרישה מלאה (לאחר 60 דקות) הקריש הוא מתכווץ ונכנס לתוך החור שנקדח מראש. </li><li style=";text-align:right;direction:rtl"> הסר את הקריש באמצעות מלקחיים בזהירות ומקום קהים לתוך צינור microcentrifuge עם PBS. להכין 2 קרישים / בעלי חיים. </li><li style=";text-align:right;direction:rtl"> קח את קרישים לחדר הניתוח. </li></ol> ההשרשה ד תאי גזע mesenchymal אלוגנאית בקרישי הפיברין<ol style=";text-align:right;direction:rtl"><li style=";text-align:right;direction:rtl"> לגרום הרדמה לארנב (NZW, זכר, 3.5-4.0 ק&quot;ג משקל גוף, ישן 5-6 חודשים) על ידי הזרקה של IV פרופופול (10 משקל גוף מ&quot;ג / ק&quot;ג). </li><li style=";text-align:right;direction:rtl"> לגלח את הברך להיות מופעלת על עם גוזז חשמלי וואקום הפרווה. כל ההליכים בשם לפני שמבוצעים בחדר ניתוח בהכנה, כדי למנוע זיהום של סביבת סטרילית של חדר הניתוח. </li><li style=";text-align:right;direction:rtl"> לאחר אינטובציה, לשמור על הרדמה עם 1.5 מ&quot;ג / ק&quot;ג / דקה propofol ו0.05 מ&quot;ג / ק&quot;ג / פנטניל דקות לוריד. לפקח הרדמה באמצעות capnography, oximetry דופק ודופק. </li><li style=";text-align:right;direction:rtl"> לחטא את הברך מגולח ביסודיות ולכסות את שאר ארנב עם רוטב סטרילי. </li><li style=";text-align:right;direction:rtl"> למשש את פיקת הברך ולבצע חתך בעור המדיאלי לפיקת הברך. </li><li style=";text-align:right;direction:rtl"> פתח את מפרק הברך על ידי arthrotomy parapatellar המדיאלי בתנאים סטריליים. נסו להימנע מחיתוך כל כלי דם שטחיים קטנים. </li><li style=";text-align:right;direction:rtl"> לעקור את פיקת הברך רוחבית (איור 4). </li><li style=";text-align:right;direction:rtl"> לאחר inspection של מפרק הברך לכל נגעי סחוס נלווים או סטיות משותפות, ליצור שני פגמי osteochondral (3 מ&quot;מ עמוק, דמות בצורת שמונה) בחריץ trochlear עם מקדחת הפעלה אוויר סטרילי (3.6 מ&quot;מ קוטר) עם עצירה-מכשיר (איור 5). </li><li style=";text-align:right;direction:rtl"> נקה את הפגמים ולשטוף אותם עם תמיסת מלח סטרילית. </li><li style=";text-align:right;direction:rtl"> לפני ההשתלה, למלא 20 μl של דבק הפיברין לפגמים ולהפיץ אותם באופן שווה על החלק התחתון של הפגם. </li><li style=";text-align:right;direction:rtl"> ואז להשתיל את קרישי עיתונות מצויד לתוך הפגם בצורת שמונה הדמות. </li><li style=";text-align:right;direction:rtl"> לאחר קרישה, מיקום מחדש של הפיקה בתוך חריץ trochlear ולכופף ולמתוח את הברך כמה פעמים. </li><li style=";text-align:right;direction:rtl"> לעקור את פיקת הברך רוחבית שוב ולבדוק אם פיברינוגן לתא, לקרישים עדיין נמצאים במקום. </li><li style=";text-align:right;direction:rtl"> החלף את פיקת הברך שוב ולסיים את המבצע עם סגירת פצע בשכבות עם תפרי כפתור אחת (Vicryl 4-0) ותפר עורית רציף (Monocryl 4-0) (שניהם עם תפר נספגחומר). </li><li style=";text-align:right;direction:rtl"> לבסוף, לאטום את הפצע עם תרסיס הלבשה חדירה לאדי מים. </li><li style=";text-align:right;direction:rtl"> לטיפול שלאחר ניתוח, הפצע הוא בדק ביום למשך 7 ימים. הארנבונים מקבלים לשיכוך כאבים שלאחר ניתוח Carprofen 4 מ&quot;ג / ק&quot;ג SC כל 24 שעות (במשך 4 ימים) וBuprenorphin 0.03 מ&quot;ג / ק&quot;ג SC כל 12 שעות (עבור 2.5 ימים). ייצוב של הברך (רוטב למשל) הוא לא הכרחי. </li></ol>

<ol>
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Dis. 63, 54-62 (2005).</li><li>Brittberg, M., 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=Treatment+of+deep+cartilage+defects+in+the+knee+with+autologous+chondrocyte+transplantation.">Treatment of deep cartilage defects in the knee with autologous chondrocyte transplantation.</a> N. Engl. J. Med. 331, 889-895 (1994).</li><li>Peterson, L., Vasiliadis, H. S., Brittberg, M., Lindahl, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Autologous+chondrocyte+implantation:+a+long-term+follow-up.">Autologous chondrocyte implantation: a long-term follow-up.</a> Am. J. Sports Med. 38, 1117-1124 (2010).</li><li>Gomoll, A. H., 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=The+subchondral+bone+in+articular+cartilage+repair:+current+problems+in+the+surgical+management.">The subchondral bone in articular cartilage repair: current problems in the surgical management.</a> Knee Surg. Sports Traumatol. Arthrosc. 18, 434-447 (2010).</li><li>Steinhagen, J., 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=Treatment+of+osteochondritis+dissecans+of+the+femoral+condyle+with+autologous+bone+grafts+and+matrix-supported+autologous+chondrocytes.">Treatment of osteochondritis dissecans of the femoral condyle with autologous bone grafts and matrix-supported autologous chondrocytes.</a> Int. Orthop. 34, 819-825 (2010).</li><li>Guo, X., 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=Repair+of+large+articular+cartilage+defects+with+implants+of+autologous+mesenchymal+stem+cells+seeded+into+beta-tricalcium+phosphate+in+a+sheep+model.">Repair of large articular cartilage defects with implants of autologous mesenchymal stem cells seeded into beta-tricalcium phosphate in a sheep model.</a> Tissue Eng. 10, 1818-1829 (2004).</li><li>Centeno, C. J., 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=Increased+knee+cartilage+volume+in+degenerative+joint+disease+using+percutaneously+implanted,+autologous+mesenchymal+stem+cells.">Increased knee cartilage volume in degenerative joint disease using percutaneously implanted, autologous mesenchymal stem cells.</a> Pain Physician. 11, 343-353 (2008).</li><li>Niederauer, G. G., 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=Evaluation+of+multiphase+implants+for+repair+of+focal+osteochondral+defects+in+goats.">Evaluation of multiphase implants for repair of focal osteochondral defects in goats.</a> Biomaterials. 21, 2561-2574 (2000).</li><li>Nagura, I., 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=Repair+of+osteochondral+defects+with+a+new+porous+synthetic+polymer+scaffold.">Repair of osteochondral defects with a new porous synthetic polymer scaffold.</a> J. Bone. Joint Surg. Br. 89, 258-264 (2007).</li><li>Schlichting, 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=Influence+of+scaffold+stiffness+on+subchondral+bone+and+subsequent+cartilage+regeneration+in+an+ovine+model+of+osteochondral+defect+healing.">Influence of scaffold stiffness on subchondral bone and subsequent cartilage regeneration in an ovine model of osteochondral defect healing.</a> The American Journal of Sports Medicine. 36, 2379-2391 (2008).</li><li>Schagemann, J. C., 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=Cell-laden+and+cell-free+biopolymer+hydrogel+for+the+treatment+of+osteochondral+defects+in+a+sheep+model.">Cell-laden and cell-free biopolymer hydrogel for the treatment of osteochondral defects in a sheep model.</a> Tissue Engineering. Part A. 15, 75-82 (2009).</li><li>Vogt, S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+influence+of+the+stable+expression+of+BMP2+in+fibrin+clots+on+the+remodelling+and+repair+of+osteochondral+defects.">The influence of the stable expression of BMP2 in fibrin clots on the remodelling and repair of osteochondral defects.</a> Biomaterials. 30, 2385-2392 (2009).</li><li>Schillinger, U., 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+fibrin+glue+composition+as+carrier+for+nucleic+acid+vectors.">A fibrin glue composition as carrier for nucleic acid vectors.</a> Pharm. Res. 25, 2946-2962 (2008).</li><li>Ahmed, T. A., Giulivi, A., Griffith, M., Hincke, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Fibrin+glues+in+combination+with+mesenchymal+stem+cells+to+develop+a+tissue-engineered+cartilage+substitute.">Fibrin glues in combination with mesenchymal stem cells to develop a tissue-engineered cartilage substitute.</a> Tissue Engineering. Part A. 17, 323-335 (2011).</li><li>Haleem, A. M., 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=The+Clinical+Use+of+Human+Culture-Expanded+Autologous+Bone+Marrow+Mesenchymal+Stem+Cells+Transplanted+on+Platelet-Rich+Fibrin+Glue+in+the+Treatment+of+Articular+Cartilage+Defects:+A+Pilot+Study+and+Preliminary+Results.">The Clinical Use of Human Culture-Expanded Autologous Bone Marrow Mesenchymal Stem Cells Transplanted on Platelet-Rich Fibrin Glue in the Treatment of Articular Cartilage Defects: A Pilot Study and Preliminary Results.</a> Cartilage. 1, 253-261 (2010).</li><li>Pape, D., Filardo, G., Kon, E., van Dijk, C. N., Madry, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Disease-specific+clinical+problems+associated+with+the+subchondral+bone.">Disease-specific clinical problems associated with the subchondral bone.</a> Knee Surg Sports Traumatol. Arthrosc. 18, 448-462 (2010).</li><li>Shirazi, R., Shirazi-Adl, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Computational+biomechanics+of+articular+cartilage+of+human+knee+joint:+effect+of+osteochondral+defects.">Computational biomechanics of articular cartilage of human knee joint: effect of osteochondral defects.</a> Journal of Biomechanics. 42, 2458-2465 (2009).</li><li>Jorgensen, C., Gordeladze, J., Noel, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Tissue+Engineering+through+autologous+mesenchymal+stem+cells.">Tissue Engineering through autologous mesenchymal stem cells.</a> Curr. Opin. Biotechnol. 15, 406-410 (2004).</li><li>Chen, F. H., Tuan, R. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mesenchymal+stem+cells+in+arthritic+diseases.">Mesenchymal stem cells in arthritic diseases.</a> Arthritis Res. Ther. 10, 223 (2008).</li><li>Le Blanc, K., Tammik, C., Rosendahl, K., Zetterberg, E., Ringden, O. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=HLA+expression+and+immunologic+properties+of+differentiated+and+undifferentiated+mesenchymal+stem+cells.">HLA expression and immunologic properties of differentiated and undifferentiated mesenchymal stem cells.</a> Exp. Hematol. 31, 890-896 (2003).</li></ol>

המחברים אין לחשוף.

בשנים האחרונות, את האפשרות של טיפול במומים מורכבים מפרק osteochondral - כגון אלה הנובעים מdissecans osteochondritis, נמק וטראומה - עם הנדסת רקמות גישות הפכה ליותר ויותר אטרקטיבי. בגופים פתולוגיים שהוזכרו קודם לכן, נזק לרקמות משתרעת על עצם subchondral וכוללת שתי רקמות המתאפיינות ביכולות ריפו...

<table border="1">
	<tbody>
		<tr>
			<td>Name of reagent/equipment</td>
			<td>Company</td>
			<td>Catalogue Number</td>
			<td>Comments</td>
		</tr>
		<tr>
			<td>DMEM</td>
			<td>Biochrom AG</td>
			<td>F 0415</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>FCS</td>
			<td>PAN Biotech GmbH</td>
			<td>0401</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>Propofol</td>
			<td>Fresenius Kabi</td>
			<td>&nbsp;</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>Penicillin/Streptomycin</td>
			<td>Biochrom AG</td>
			<td>A 2210</td>
			<td>1,000 units/10 &mu;g/&mu;l in 0.9% NaCl</td>
		</tr>
		<tr>
			<td>PBS Dulbecco (1X)</td>
			<td>Biochrom AG</td>
			<td>L1815</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>Ethanol (70%)</td>
			<td>Merck KGaA</td>
			<td>410230</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>Trypan Blue Solution (0.4%)</td>
			<td>Sigma-Aldrich</td>
			<td>T8154</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>Biocoll Separation Sol.</td>
			<td>Biochrom AG</td>
			<td>L6115</td>
			<td>Isotonic solution Density: 1,077 g/ml</td>
		</tr>
		<tr>
			<td>Trypsin-EDTA 0.05%</td>
			<td>Invitrogen GmbH</td>
			<td>25300-054</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>Fentanyl</td>
			<td>DeltaSelectGmBH</td>
			<td>1819340</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>NaCl solution (0.9%)</td>
			<td>BBraun</td>
			<td>8333A193</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>Syringes (Injekt)</td>
			<td>BBraun</td>
			<td>4606108V</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>Needles (Sterican)</td>
			<td>BBraun</td>
			<td>4657519</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>Forceps (blunt/sharp)</td>
			<td>Aesculap</td>
			<td>&nbsp;</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>Scissors</td>
			<td>Aesculap</td>
			<td>&nbsp;</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>Scalpels</td>
			<td>Feather Safety Razor Co</td>
			<td>02.001.30.022</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>Pipettes research</td>
			<td>Eppendorf</td>
			<td>&nbsp;</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>Bone Cutter</td>
			<td>Aesculap</td>
			<td>&nbsp;</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>Tissue culture dishes 100 mm/150 mm</td>
			<td>TPP AG</td>
			<td>93100/93150</td>
			<td>Growth area 60.1 mm2/147.8 mm2</td>
		</tr>
		<tr>
			<td>Tissue culture flasks 25/75 mm2</td>
			<td>TPP AG</td>
			<td>90025/90075</td>
			<td>25 mm2, 75 mm2</td>
		</tr>
		<tr>
			<td>Centrifuge Tubes (50 ml)</td>
			<td>TPP AG</td>
			<td>91050</td>
			<td>Gamma-sterilized</td>
		</tr>
		<tr>
			<td>CO2 Incubator</td>
			<td>Forma Scientific Inc.</td>
			<td>&nbsp;</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>Cell culture laminar flow hood Hera Safe</td>
			<td>Heraeus Instruments</td>
			<td>&nbsp;</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>Sterile saw</td>
			<td>Aesculap</td>
			<td>&nbsp;</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>Centrifuge Megafuge 2.0 R</td>
			<td>Heraeus Instruments</td>
			<td>&nbsp;</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>Hemocytometer</td>
			<td>Brand GmbH+Co KG</td>
			<td>717810</td>
			<td>Neubauer</td>
		</tr>
		<tr>
			<td>Air operated power drill</td>
			<td>Aesculap</td>
			<td>&nbsp;</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>TISSUCOL-Kit 1.0 ml Immuno</td>
			<td>Baxter</td>
			<td>2546648</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>Fibers (4-0 Monocryl, 4-0 Vicryl)</td>
			<td>Ethicon</td>
			<td>&nbsp;</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>Spray dressing (OpSite)</td>
			<td>Smith&amp;Nephew</td>
			<td>66004978</td>
			<td>Permeable for water vapor</td>
		</tr>
	</tbody>
</table>

treatment of osteochondral defects in the rabbit's knee joint by implantation of allogeneic mesenchymal stem cells in fibrin clots

The treatment of osteochondral articular defects has been challenging physicians for many years. The better understanding of interactions of articular cartilage and subchondral bone in recent years led to increased attention to restoration of the entire osteochondral unit. In comparison to chondral lesions the regeneration of osteochondral defects is much more complex and a far greater surgical and therapeutic challenge. The damaged tissue does not only include the superficial cartilage layer but also the subchondral bone. For deep, osteochondral damage, as it occurs for example with osteochondrosis dissecans, the full thickness of the defect needs to be replaced to restore the joint surface 1. Eligible therapeutic procedures have to consider these two different tissues with their different intrinsic healing potential 2. In the last decades, several surgical treatment options have emerged and have already been clinically established 3-6.
Autologous or allogeneic osteochondral transplants consist of articular cartilage and subchondral bone and allow the replacement of the entire osteochondral unit. The defects are filled with cylindrical osteochondral grafts that aim to provide a congruent hyaline cartilage covered surface 3,7,8. Disadvantages are the limited amount of available grafts, donor site morbidity (for autologous transplants) and the incongruence of the surface; thereby the application of this method is especially limited for large defects.
New approaches in the field of tissue engineering opened up promising possibilities for regenerative osteochondral therapy. The implantation of autologous chondrocytes marked the first cell based biological approach for the treatment of full-thickness cartilage lesions and is now worldwide established with good clinical results even 10 to 20 years after implantation 9,10. However, to date, this technique is not suitable for the treatment of all types of lesions such as deep defects involving the subchondral bone 11.
The sandwich-technique combines bone grafting with current approaches in Tissue Engineering 5,6. This combination seems to be able to overcome the limitations seen in osteochondral grafts alone. After autologous bone grafting to the subchondral defect area, a membrane seeded with autologous chondrocytes is sutured above and facilitates to match the topology of the graft with the injured site. Of course, the previous bone reconstruction needs additional surgical time and often even an additional surgery. Moreover, to date, long-term data is missing 12.
Tissue Engineering without additional bone grafting aims to restore the complex structure and properties of native articular cartilage by chondrogenic and osteogenic potential of the transplanted cells. However, again, it is usually only the cartilage tissue that is more or less regenerated. Additional osteochondral damage needs a specific further treatment. In order to achieve a regeneration of the multilayered structure of osteochondral defects, three-dimensional tissue engineered products seeded with autologous/allogeneic cells might provide a good regeneration capacity 11.
Beside autologous chondrocytes, mesenchymal stem cells (MSC) seem to be an attractive alternative for the development of a full-thickness cartilage tissue. In numerous preclinical in vitro and in vivo studies, mesenchymal stem cells have displayed excellent tissue regeneration potential 13,14. The important advantage of mesenchymal stem cells especially for the treatment of osteochondral defects is that they have the capacity to differentiate in osteocytes as well as chondrocytes. Therefore, they potentially allow a multilayered regeneration of the defect.
In recent years, several scaffolds with osteochondral regenerative potential have therefore been developed and evaluated with promising preliminary results 1,15-18. Furthermore, fibrin glue as a cell carrier became one of the preferred techniques in experimental cartilage repair and has already successfully been used in several animal studies 19-21 and even first human trials 22.
The following protocol will demonstrate an experimental technique for isolating mesenchymal stem cells from a rabbit's bone marrow, for subsequent proliferation in cell culture and for preparing a standardized in vitro-model for fibrin-cell-clots. Finally, a technique for the implantation of pre-established fibrin-cell-clots into artificial osteochondral defects of the rabbit's knee joint will be described.

הטכניקה כירורגית תאר מאפשרת בידוד מוצלח והשתלה של תאי גזע מזנכימיים אלוגניות לפגם osteochondral מלאכותי. ההתקנה הניסיונית הביאה לשילוב מוצלח של השתל לתוך הסחוס שמסביב. הפגם היה מלא על ידי רקמת תיקון עם מאפיינים דומים biomechanical ועמידות ד?...

Watch this Scientific Journal Video about Treatment of Osteochondral Defects in the Rabbit's Knee Joint by Implantation of Allogeneic Mesenchymal Stem Cells in Fibrin Clots at JoVE.com

Treatment of Osteochondral Defects in the Rabbit&#039;s Knee Joint by Implantation of Allogeneic Mesenchymal Stem Cells in Fibrin Clots

טכניקה ניסויית לטיפול בפגמי osteochondral במפרק הברך של הארנב מתוארת. ההשתלה של תאי גזע מזנכימיים אלוגניות לפגמי osteochondral מספקת פיתוח מבטיח בתחום הנדסת רקמות. הכנת הפיברין-תא קרישים<em&gt; במבחנה</em&gt; מציע שיטה סטנדרטית להשתלה.

טיפול במומי Osteochondral במפרק הברך של הארנב על ידי השתלה של תאי גזע mesenchymal אלוגנאית בקרישי הפיברין

The treatment of osteochondral articular defects has been challenging physicians for many years. The better understanding of interactions of articular ...

treatment-osteochondral-defects-rabbit-s-knee-joint-implantation

Research

JoVE Journal

Medicine

Treatment of Osteochondral Defects in the Rabbit's Knee Joint by Implantation of Allogeneic Mesenchymal Stem Cells in Fibrin Clots

17.3K Views.  Klinikum rechts der Isar der Technischen Universität München. טכניקה ניסויית לטיפול בפגמי osteochondral במפרק הברך של הארנב מתוארת. ההשתלה של תאי גזע מזנכימיים אלוגניות לפגמי osteochondral מספקת פיתוח מבטיח בתחום הנדסת רקמות. הכנת הפיברין-תא קרישים במבחנה מציע שיטה סטנדרטית להשתלה.

Video: Treatment of Osteochondral Defects in the Rabbit's Knee Joint by Implantation of Allogeneic Mesenchymal Stem Cells in Fibrin Clots

Isolation, Characterization and Comparative Differentiation of Human Dental Pulp Stem Cells Derived from Permanent Teeth by Using Two Different Methods

Developing wisdom teeth are easy-accessible source of stem cells during the adulthood which could be obtained by routine orthodontic treatments. Human pulp-derived stem cells (hDPSCs) possess high proliferation potential with multi-lineage differentiation capacity compare to the ordinary source of adult stem cells1-8; therefore, hDPSCs could be the good candidates for autologous transplantation in tissue engineering and regenerative medicine. Along with these benefits, possessing the mesenchymal stem cells (MSC) features, such as immunolodulatory effect, make hDPSCs more valuable, even in the case of allograft transplantation6,9,10. Therefore, the primary step for using this source of stem cells is to select the best protocol for isolating hDPSCs from pulp tissue. In order to achieve this goal, it is crucial to investigate the effect of various isolation conditions on different cellular behaviors, such as their common surface markers &amp; also their differentiation capacity.
Thus, here we separate human pulp tissue from impacted third molar teeth, and then used both existing protocols based on literature, for isolating hDPSCs,11-13 i.e. enzymatic dissociation of pulp tissue (DPSC-ED) or outgrowth from tissue explants (DPSC-OG). In this regards, we tried to facilitate the isolation methods by using dental diamond disk. Then, these cells characterized in terms of stromal-associated Markers (CD73, CD90, CD105 &amp; CD44), hematopoietic/endothelial Markers (CD34, CD45 &amp; CD11b), perivascular marker, like CD146 and also STRO-1. Afterwards, these two protocols were compared based on the differentiation potency into odontoblasts by both quantitative polymerase chain reaction (QPCR) &amp; Alizarin Red Staining. QPCR were used for the assessment of the expression of the mineralization-related genes (alkaline phosphatase; ALP, matrix extracellular phosphoglycoprotein; MEPE &amp; dentin sialophosphoprotein; DSPP).14

The method described isolation and characterization of human Dental Pulp Stem Cells (hDPSCs) by using either enzymatic dissociation of pulp (DPSC-ED) or direct outgrowth of stem cells from pulp tissue explants (DPSC-OG). Then followed by in vitro comparative differentiation of both types of hDPSCs into odontoblasts.

בידוד, אפיון והשוואתי התמיינות של תאי גזע אנושיים שיניים זולות נגזרה שיניים קבועות על ידי שימוש בשתי שיטות שונות

Developing wisdom teeth are easy-accessible source of stem cells during the adulthood which could be obtained by routine orthodontic treatments. Human ...

Implantation of Fibrin Gel on Mouse Lung to Study Lung-specific Angiogenesis

Recent significant advances in stem cell research and bioengineering techniques have made great progress in utilizing biomaterials to regenerate and repair damage in simple tissues in the orthopedic and periodontal fields. However, attempts to regenerate the structures and functions of more complex three-dimensional (3D) organs such as lungs have not been very successful because the biological processes of organ regeneration have not been well explored. It is becoming clear that angiogenesis, the formation of new blood vessels, plays key roles in organ regeneration. Newly formed vasculatures not only deliver oxygen, nutrients and various cell components that are required for organ regeneration but also provide instructive signals to the regenerating local tissues. Therefore, to successfully regenerate lungs in an adult, it is necessary to recapitulate the lung-specific microenvironments in which angiogenesis drives regeneration of local lung tissues. Although conventional in vivo angiogenesis assays, such as subcutaneous implantation of extracellular matrix (ECM)-rich hydrogels (e.g., fibrin or collagen gels or Matrigel - ECM protein mixture secreted by Engelbreth-Holm-Swarm mouse sarcoma cells), are extensively utilized to explore the general mechanisms of angiogenesis, lung-specific angiogenesis has not been well characterized because methods for orthotopic implantation of biomaterials in the lung have not been well established. The goal of this protocol is to introduce a unique method to implant fibrin gel on the lung surface of living adult mouse, allowing for the successful recapitulation of host lung-derived angiogenesis inside the gel. This approach enables researchers to explore the mechanisms by which the lung-specific microenvironment controls angiogenesis and alveolar regeneration in both normal and pathological conditions. Since implanted biomaterials release and supply physical and chemical signals to adjacent lung tissues, implantation of these biomaterials on diseased lung can potentially normalize the adjacent diseased tissues, enabling researchers to develop new therapeutic approaches for various types of lung diseases.

Recapitulation of the organ-specific microenvironment, which stimulates local angiogenesis, is indispensable for successful regeneration of damaged tissues. This report demonstrates a novel method to implant fibrin gels on the lung surface of living mouse in order to explore how the lung-specific microenvironment modulates angiogenesis and alveolar regeneration in adult mouse.

השרשה של הפיברין ג&#39;ל על עכבר ריאות לחקר אנגיוגנזה ריאות ספציפית

Recent significant advances in stem cell research and bioengineering techniques have made great progress in utilizing biomaterials to regenerate and ...

In Vivo Imaging and Tracking of Technetium-99m Labeled Bone Marrow Mesenchymal Stem Cells in Equine Tendinopathy

Recent advances in the application of bone marrow mesenchymal stem cells (BMMSC) for the treatment of tendon and ligament injuries in the horse suggest improved outcome measures in both experimental and clinical studies. Although the BMMSC are implanted into the tendon lesion in large numbers (usually 10 - 20 million cells), only a relatively small number survive (&#60;10%) although these can persist for up to 5 months after implantation. This appears to be a common observation in other species where BMMSC have been implanted into other tissues and it is important to understand when this loss occurs, how many survive the initial implantation process and whether the cells are cleared into other organs. Tracking the fate of the cells can be achieved by radiolabeling the BMMSC prior to implantation which allows non-invasive in vivo imaging of cell location and quantification of cell numbers.
This protocol describes a cell labeling procedure that uses Technetium-99m (Tc-99m), and tracking of these cells following implantation into injured flexor tendons in horses. Tc-99m is a short-lived (t1/2 of 6.01 hr) isotope that emits gamma rays and can be internalized by cells in the presence of the lipophilic compound hexamethylpropyleneamine oxime (HMPAO). These properties make it ideal for use in nuclear medicine clinics for the diagnosis of many different diseases. The fate of the labeled cells can be followed in the short term (up to 36 hr) by gamma scintigraphy to quantify both the number of cells retained in the lesion and distribution of the cells into lungs, thyroid and other organs. This technique is adapted from the labeling of blood leukocytes and could be utilized to image implanted BMMSC in other organs.

In Vivo Imaging and Tracking of Technetium-99m Labeled Bone Marrow Mesenchymal Stem Cells in Equine Tendinopathy

This protocol describes the radiolabeling of equine mesenchymal stem cells and their implantation into tendon injuries in the horse in order to determine cell survival and tissue distribution using gamma scintigraphy.

In vivo ההדמיה ומעקב של טכנציום-99m תווית תאי גזע מח העצם mesenchymal בסוסי Tendinopathy

Recent advances in the application of bone marrow mesenchymal stem cells (BMMSC) for the treatment of tendon and ligament injuries in the horse suggest ...

Studying Pancreatic Cancer Stem Cell Characteristics for Developing New Treatment Strategies

Pancreatic ductal adenocarcinoma (PDAC) contains a subset of exclusively tumorigenic cancer stem cells (CSCs) which have been shown to drive tumor initiation, metastasis and resistance to radio- and chemotherapy. Here we describe a specific methodology for culturing primary human pancreatic CSCs as tumor spheres in anchorage-independent conditions. Cells are grown in serum-free, non-adherent conditions in order to enrich for CSCs while their more differentiated progenies do not survive and proliferate during the initial phase following seeding of single cells. This assay can be used to estimate the percentage of CSCs present in a population of tumor cells. Both size (which can range from 35 to 250 micrometers) and number of tumor spheres formed represents CSC activity harbored in either bulk populations of cultured cancer cells or freshly harvested and digested tumors 1,2. Using this assay, we recently found that metformin selectively ablates pancreatic CSCs; a finding that was subsequently further corroborated by demonstrating diminished expression of pluripotency-associated genes/surface markers and reduced in vivo tumorigenicity of metformin-treated cells. As the final step for preclinical development we treated mice bearing established tumors with metformin and found significantly prolonged survival. Clinical studies testing the use of metformin in patients with PDAC are currently underway (e.g., NCT01210911, NCT01167738, and NCT01488552). Mechanistically, we found that metformin induces a fatal energy crisis in CSCs by enhancing reactive oxygen species (ROS) production and reducing mitochondrial transmembrane potential. In contrast, non-CSCs were not eliminated by metformin treatment, but rather underwent reversible cell cycle arrest. Therefore, our study serves as a successful example for the potential of in vitro sphere formation as a screening tool to identify compounds that potentially target CSCs, but this technique will require further in vitro and in vivo validation to eliminate false discoveries.

Pancreatic cancer stem cells (CSCs) can be expanded in vitro using the anchorage-independent sphere culture technique, which represents a powerful tool to study CSC biology and can serve as the first step to develop novel CSC-targeting therapies. Here the methodology for expanding, analyzing and targeting of pancreatic CSCs is provided.

לומד סרטן לבלב גזע מאפייני תא לפיתוח טיפול חדש אסטרטגיות

Pancreatic ductal adenocarcinoma (PDAC) contains a subset of exclusively tumorigenic cancer stem cells (CSCs) which have been shown to drive tumor ...

Preparation, Procedures and Evaluation of Platelet-Rich Plasma Injection in the Treatment of Knee Osteoarthritis

Knee osteoarthritis (KOA) is one of the most frequently encountered diseases in the orthopedic department. Existing non-surgical treatments have a limited effect on the repair of cartilage and on bone regeneration. Platelet-rich plasma (PRP) is an autologous bioactive substance that can repair cartilage injury and accelerate bone regeneration effectively. However, reporting of PRP preparation protocols in clinical studies is highly inconsistent, with the majority of studies providing insufficient information to allow the protocol to be reproduced. We describe a repeatable method of preparing PRP visually, the treatment of KOA using PRP intra-articular injection, and methods of evaluating the outcome. PRP was prepared using manual double centrifugation. The PRP layer was extracted from peripheral blood and used for knee joint cavity injection. Evaluations included assessments of blood platelet concentrations and clinical outcomes. Preparation of PRP by manual centrifugation requires less apparatus and is less costly than plasma filtration or centrifugation using equipment. The centrifugation time of our double centrifugation method was 6 and 5 minutes for the respective centrifugations at forces of 800 and 1400 x g, respectively, to allow for the consistent preparation of standardized PRP. However, a manual method is susceptible to operator error, and PRP batch preparation is not available. Intra-articular injection of PRP proved to be an effective treatment for knee osteoarthritis. The entire treatment procedure took less than 30 minutes, the blood platelet concentration of PRP could be standardized, and treatment was proven to be effective when evaluated by follow-up.

Knee osteoarthritis is frequently seen in the orthopedic department. We introduce in detail the entire knee osteoarthritis treatment process with platelet-rich plasma injection, including preparation, procedures, and evaluation.

הכנה, נהלים והערכת הזרקת הפלסמה טסיות דם עשיר בטיפול של דלקת מפרקים ניוונית בברך

Knee osteoarthritis (KOA) is one of the most frequently encountered diseases in the orthopedic department. Existing non-surgical treatments have a limited ...

Using a Knee Arthrometer to Evaluate Tissue-specific Contributions to Knee Flexion Contracture in the Rat

Normal knee range of motion (ROM) is critical to well-being and allows one to perform basic activities such as walking, climbing stairs and sitting. Lost ROM is called a joint contracture and results in increased morbidity. Due to the difficulty of reversing established knee contractures, early detection is important, and hence, knowing risk factors for their development is essential. The rat represents a good model with which the effect of an intervention can be studied due to the similarity of rat knee anatomy to that of humans, the rat's ability to tolerate long durations of knee immobilization in flexion, and because mechanical data can be correlated with histologic and biochemical analysis of knee tissue.
Using an automated arthrometer, we demonstrate a validated, precise, reproducible, user-independent method of measuring the extension ROM of the rat knee joint at specific torques. This arthrometer can be used to determine the effects of interventions on knee joint ROM in the rat.

The goal of the protocol is to measure the extension range of motion of the rat knee. The effects of various diseases that increase the stiffness of the knee joint and the effectiveness of treatments can be quantified.

באמצעות Arthrometer של הברך כדי להעריך את תרומתה רקמות ספציפיות הברך הקופסית כיפוף בחולדה

Normal knee range of motion (ROM) is critical to well-being and allows one to perform basic activities such as walking, climbing stairs and sitting. Lost ...

Isolation and Cultivation of Mandibular Bone Marrow Mesenchymal Stem Cells in Rats

Here we present an efficient method for isolating and culturing mandibular bone marrow mesenchymal stem cells (mBMSCs) in vitro to rapidly obtain numerous high-quality cells for experimental requirements. mBMSCs could be widely used in therapeutic applications as tissue engineering cells in case of craniofacial diseases and cranio-maxillofacial regeneration in the future due to the excellent self-renewal ability and multi-lineage differentiation potential. Therefore, it is important to obtain mBMSCs in large numbers.
In this study, bone marrow was flushed from the mandible and primary mBMSCs were isolated through whole bone marrow adherent cultivation. Furthermore, CD29+CD90+CD45&#8722; mBMSCs were purified through fluorescent cell sorting. The second generation of purified mBMSCs were used for further study and displayed potential in differentiating into osteoblasts, adipocytes, and chondrocytes. Utilizing this in vitro model, one can obtain a high number of proliferative mBMSCs, which may facilitate the study of the biological characteristics, the subsequent reaction to the microenvironment, and other applications of mBMSCs.

This article presents a method that combines whole bone marrow adherence and flow cytometry sorting for isolating, cultivating, sorting, and identifying bone marrow mesenchymal stem cells from rat mandibles.

בידוד וטיפוח של תאי גזע מסנכימליים במח עצם מנדיבול בחולדות

Here we present an efficient method for isolating and culturing mandibular bone marrow mesenchymal stem cells (mBMSCs) in vitro to rapidly obtain numerous ...

Orthopedic Robot-Assisted Femoral Neck System in the Treatment of Femoral Neck Fracture

Cannulated screw fixation is the main therapy for femoral neck fractures, especially in young patients. The traditional surgical procedure uses C-arm fluoroscopy to place the screw freehand and requires several guide wire adjustments, which increases the operation time and radiation exposure. Repeated drilling can also cause damage to the blood supply and bone quality of the femoral neck, which can be followed by complications such as screw loosening, nonunion, and femoral head necrosis. In order to make fixation more precise and reduce the incidence of complications, our team applied robot-assisted orthopedic surgery for screw placement using the femoral neck system to modify the traditional procedure. This protocol introduces how to import a patient's X-ray information into the system, how to perform screw path planning in software, and how the robotic arm assists in screw placement. Using this method, the surgeons can place the screw successfully the first time, improve the accuracy of the procedure, and avoid radiation exposure. The whole protocol includes the diagnosis of femoral neck fracture; the collection of intraoperative X-ray images; screw path planning in the software; precise placement of the screw under the assistance of the robotic arm by the surgeon; and verification of the implant placement.

This article introduces a method of robot-assisted orthopedic surgery for screw placement during the treatment of femoral neck fracture using the femoral neck system, which allows for more accurate screw placement, improved surgical efficiency, and fewer complications.

מערכת צוואר הירך אורתופדית בסיוע רובוט לטיפול בשבר בצוואר הירך

Cannulated screw fixation is the main therapy for femoral neck fractures, especially in young patients. The traditional surgical procedure uses C-arm ...

הקמת מודל KOA בארנבים בשיטת וידמן שונה

Application of Acupotomy in a Knee Osteoarthritis Model in Rabbit

Knee osteoarthritis (KOA) is one of the most frequently encountered diseases in the orthopedic department, which seriously reduces the quality of life of people with KOA. Among several pathogenic factors, the biomechanical imbalance of the knee joint is one of the main causes of KOA. Acupotomology believes that restoring the mechanical balance of the knee joint is the key to treating KOA. Clinical studies have shown that acupotomy can effectively reduce pain and improve knee mobility by reducing adhesion, contracture of soft tissues, and stress concentration points in muscles and tendons around the knee joint. 
In this protocol, we used the modified Videman method to establish a KOA model by immobilizing the left hindlimb in a straight position. We have outlined the method of operation and the precautions related to acupotomy in detail and evaluated the efficacy of acupotomy in conjunction with the theory of "Modulating Muscles and Tendons to Treat Bone Disorders" through the detection of the mechanical properties of quadriceps femoris and tendon, as well as cartilage mechanics and morphology. The results show that acupotomy has a protective effect on cartilage by adjusting the mechanical properties of the soft tissues around the knee joint, improving the cartilage stress environment, and delaying cartilage degeneration.

מחבר זרקור: חקירת מנגנון הפעולה של אקופוטומיה בטיפול בדלקת מפרקים ניוונית בברך

In this protocol, a knee osteoarthritis model was prepared using the modified Videman method, and the operation procedures and precautions of acupotomy are detailed. The effectiveness of acupotomy has been demonstrated by testing the mechanical properties of quadriceps femoris and tendon and the mechanical and morphological properties of cartilage.

יישום של אקופוטומיה במודל אוסטיאוארתריטיס ברך בארנב

Knee osteoarthritis (KOA) is one of the most frequently encountered diseases in the orthopedic department, which seriously reduces the quality of life of ...