Name: treatment of osteochondral defects in the rabbit's knee joint by implantation of allogeneic mesenchymal stem cells in fibrin clots
Uploaded: 2013-05-21T13:00:00
Description: 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

Dieses Projekt wurde von der Deutschen Forschungsgemeinschaft (Zuschuss HE 4578/3-1) und teilweise durch das FP7 EU-Projekt &quot;GAMBE&quot; NMP3-SL-2010-245993 finanziert.

A. Herstellung einer Donor Kaninchen zur Isolierung von mesenchymalen Stammzellen (Chirurgie Zimmer) <ol><li> Die Zellen werden von männlichen New Zealand White (NZW) Kaninchen im Alter von 4 Monaten und ca. 3 kg Körpergewicht isoliert. </li><li> Induce Narkose durch Propofol (10 mg / kg Körpergewicht iv) und opfern mit Natrium-Pentobarbital (100 mg / kg Körpergewicht iv). </li><li> Shave Fell von Hinterbeine, Rücken und Bauch mit einem elektrischen Haarschneider und absaugen Fell. </li><li> Desinfizieren Sie die...

<ol>
	<li>Kon, E., 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=Novel+nano-composite+multilayered+biomaterial+for+osteochondral+regeneration:+a+pilot+clinical+trial.">Novel nano-composite multilayered biomaterial for osteochondral regeneration: a pilot clinical trial.</a> The American Journal of Sports Medicine. 39, 1180-1190 (2011).</li><li>Kon, E., 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=Orderly+osteochondral+regeneration+in+a+sheep+model+using+a+novel+nano-composite+multilayered+biomaterial.">Orderly osteochondral regeneration in a sheep model using a novel nano-composite multilayered biomaterial.</a> Journal of Orthopaedic Research: Official Publication of the Orthopaedic Research Society. 28, 116-124 (2010).</li><li>Hangody, 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=Autologous+osteochondral+grafting--technique+and+long-term+results.">Autologous osteochondral grafting--technique and long-term results.</a> Injury. 39, 32-39 (2008).</li><li>Marcacci, 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=Arthroscopic+autologous+osteochondral+grafting+for+cartilage+defects+of+the+knee:+prospective+study+results+at+a+minimum+7-year+follow-up.">Arthroscopic autologous osteochondral grafting for cartilage defects of the knee: prospective study results at a minimum 7-year follow-up.</a> The American Journal of Sports Medicine. 35, 2014-2021 (2007).</li><li>Ochs, B. 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=Remodeling+of+articular+cartilage+and+subchondral+bone+after+bone+grafting+and+matrix-associated+autologous+chondrocyte+implantation+for+osteochondritis+dissecans+of+the+knee.">Remodeling of articular cartilage and subchondral bone after bone grafting and matrix-associated autologous chondrocyte implantation for osteochondritis dissecans of the knee.</a> The American Journal of Sports Medicine. 39, 764-773 (2011).</li><li>Aurich, 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=Autologous+chondrocyte+transplantation+by+the+sandwich+technique.+A+salvage+procedure+for+osteochondritis+dissecans+of+the+knee.">Autologous chondrocyte transplantation by the sandwich technique. A salvage procedure for osteochondritis dissecans of the knee.</a> Unfallchirurg. 110, 176-179 (2007).</li><li>Williams, R. J., Ranawat, A. S., Potter, H. G., Carter, T., Warren, R. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Fresh+stored+allografts+for+the+treatment+of+osteochondral+defects+of+the+knee.">Fresh stored allografts for the treatment of osteochondral defects of the knee.</a> The Journal of Bone and Joint Surgery. American Volume. 89, 718-726 (2007).</li><li>Szerb, I., Hangody, L., Duska, Z., Kaposi, N. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mosaicplasty:+long-term+follow-up.">Mosaicplasty: long-term follow-up.</a> Bull. Hosp. Jt. 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>

In den letzten Jahren, die Möglichkeit der Behandlung von komplexen Gelenk osteochondralen Defekten - wurde mit Tissue Engineering Ansätze mehr und mehr attraktiv - wie jene, die aus ÖD, Osteonekrose und Trauma. In den zuvor genannten pathologischen Entitäten erstreckt Gewebeschäden an der subchondralen Knochens und umfasst zwei Geweben von verschiedenen intrinsischen Kapazitäten Heilung 1 gekennzeichnet. Es gibt ein zunehmendes Interesse an der Rolle des subchondralen Knochen für den pathogenen Prozes...

<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.

Die beschriebene Operationstechnik ermöglicht eine erfolgreiche Isolierung und die Implantation von allogenen mesenchymalen Stammzellen in einer künstlichen osteochondralen Defekt. Der experimentelle Aufbau ergab eine erfolgreiche Integration des Implantates in den umgebenden Knorpel. Der Defekt wurde durch Reparatur Gewebe mit ähnlichen biomechanischen Eigenschaften und ähnliche Haltbarkeit im Vergleich zum umgebenden Knorpel gefüllt. Das Fibrin-Clot-Zellen wurde in vitro an e...

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's Knee Joint by Implantation of Allogeneic Mesenchymal Stem Cells in Fibrin Clots

Ein experimentelles Verfahren für die Behandlung von osteochondralen Defekten in das Kaninchen Kniegelenk beschrieben. Die Implantation von allogenen mesenchymalen Stammzellen in osteochondralen Defekten stellt eine viel versprechende Entwicklung auf dem Gebiet des Tissue Engineering. Die Herstellung von Fibrin-Gerinnsel-Zelle<em&gt; In vitro</em&gt; Bietet eine standardisierte Methode für die Implantation.

Behandlung von osteochondralen Defekten in des Kaninchens Kniegelenk durch Implantation von allogenen mesenchymalen Stammzellen in Fibringerinnseln

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

Research

JoVE Journal

Medicine

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

Isolation, Charakterisierung und vergleichende Differenzierung humaner Dental Pulp Stammzellen aus bleibenden Zähne mit zwei verschiedenen Methoden abgeleitet

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

Forschung

Medizin

Slow-release Drug Delivery through Elvax 40W to the Rat Retina: Implications for the Treatment of Chronic Conditions

Slow-Release-Drug Delivery durch Elvax 40W, um die Ratte Retina: Implikationen für die Behandlung von chronischen Erkrankungen

Diseases of the retina are difficult to treat as the retina lies deep within the eye. Invasive methods of drug delivery are often needed to treat these ...

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

Die Implantation von Fibrin Gel auf Maus Lung Lung spezifischen Angiogenese Studieren

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

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

In-vivo-Imaging und Verfolgung von Technetium-99m markierten Knochenmark mesenchymale Stammzellen in Equine Tendinopathie

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

Studieren Bauchspeicheldrüsenkrebs Stammzelleigenschaften für die Entwicklung neuer Behandlungsstrategien

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

Rapid and Efficient Generation of Recombinant Human Pluripotent Stem Cells by Recombinase-mediated Cassette Exchange in the AAVS1 Locus

Rapid and Efficient Generation of Recombinant Human Pluripotent Stem Cells by Recombinase-mediated Cassette Exchange in the AAVS1 Locus

Eine schnelle und effiziente Erzeugung von rekombinanten humanen pluripotenten Stammzellen durch Rekombinase-vermittelten Kassettenaustausch in der<em&gt; AAVS1</em&gt; Locus

Even with the revolution of gene-targeting technologies led by CRISPR-Cas9, genetic modification of human pluripotent stem cells&#160;(hPSCs)&#160;is ...

Network Analysis of Foramen Ovale Electrode Recordings in Drug-resistant Temporal Lobe Epilepsy Patients

Netzwerkanalyse von Foramen Ovale Elektrode Recordings in Resistente Temporallappen Epilepsie-Patienten

Approximately 30% of epilepsy patients are refractory to antiepileptic drugs. In these cases, surgery is the only alternative to eliminate/control ...

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

Vorbereitung, Verfahren und Bewertung von Platelet-Rich Plasma-Injektion bei der Behandlung von Knie-Arthrose

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

Verwenden ein Knie Arthrometer auszuwertende gewebespezifischen Beiträge zum Knie Beugung Kontraktur in der Ratte

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

Isolierung und Kultivierung von MandibularKnochenmark Mesenchymal Stammzellen bei Ratten

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