Name: computed tomography and optical imaging of osteogenesis-angiogenesis coupling to assess integration of cranial bone autografts and allografts
Uploaded: 2015-12-22T13:00:00
Description: Watch this Scientific Journal Video about Computed Tomography and Optical Imaging of Osteogenesis-angiogenesis Coupling to Assess Integration of Cranial Bone Autografts and Allografts at JoVE.com

The authors acknowledge funding from the NIDCR (Grant No. DE019902) and from the Israeli Science Foundation (Grant No. 382/13).

Protokollen følger retningslinjerne i den institutionelle dyr pleje og brug udvalg (IACUC) i Det hebraiske Universitet i Jerusalem, Israel (Request nr MD-12-13524-4), en AAALAC godkendt anlæg, og af Cedars-Sinai Medical Center IACUC (Request nr 3770). Dyrene blev behandlet i streng overholdelse NIH retningslinjer. 1. Fremstilling af Bone Allografter <ol><li> Euthanize 7- til 8 uger gamle Balb / C-mus, eller hvilken som helst stamme forskellige fra modtageren under anvendelse af standardm...

<ol>
	<li>Finkemeier, C. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Bone-grafting+and+bone-graft+substitutes.">Bone-grafting and bone-graft substitutes.</a> J Bone Joint Surg Am. 84-A (3), 454-464 (2002).</li><li>Kanczler, J. M., Oreffo, R. O. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Osteogenesis+and+angiogenesis:+the+potential+for+engineering+bone.">Osteogenesis and angiogenesis: the potential for engineering bone.</a> Eur Cell Mater. 15, 100-114 (2008).</li><li>Schipani, E., Maes, C., Carmeliet, G., Semenza, G. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Regulation+of+osteogenesis-angiogenesis+coupling+by+HIFs+and+VEGF.">Regulation of osteogenesis-angiogenesis coupling by HIFs and VEGF.</a> J Bone Miner Res. 24 (8), 1347-1353 (2009).</li><li>Huang, 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=Spatiotemporal+Analyses+of+Osteogenesis+and+Angiogenesis+via+Intravital+Imaging+in+Cranial+Bone+Defect.">Spatiotemporal Analyses of Osteogenesis and Angiogenesis via Intravital Imaging in Cranial Bone Defect.</a> J Bone Miner Res. , (2015).</li><li>Kimelman-Bleich, N., 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+use+of+a+synthetic+oxygen+carrier-enriched+hydrogel+to+enhance+mesenchymal+stem+cell-based+bone+formation+in+vivo.">The use of a synthetic oxygen carrier-enriched hydrogel to enhance mesenchymal stem cell-based bone formation in vivo.</a> Biomaterials. 30 (27), 4639-4648 (2009).</li><li>Iris, B., 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=Molecular+imaging+of+the+skeleton:+quantitative+real-time+bioluminescence+monitoring+gene+expression+in+bone+repair+and+development.">Molecular imaging of the skeleton: quantitative real-time bioluminescence monitoring gene expression in bone repair and development.</a> J Bone Miner Res. 18 (3), 570-578 (2003).</li><li>Bouxsein, M. 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=Guidelines+for+assessment+of+bone+microstructure+in+rodents+using+micro-computed+tomography.">Guidelines for assessment of bone microstructure in rodents using micro-computed tomography.</a> J Bone Miner Res. 25 (7), 1468-1486 (2010).</li><li>Lim, E., Modi, K. D., Kim, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=In+vivo+bioluminescent+imaging+of+mammary+tumors+using+IVIS+spectrum.">In vivo bioluminescent imaging of mammary tumors using IVIS spectrum.</a> J Vis Exp. (26), (2009).</li><li>Kallai, 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=Microcomputed+tomography-based+structural+analysis+of+various+bone+tissue+regeneration+models.">Microcomputed tomography-based structural analysis of various bone tissue regeneration models.</a> Nat Protoc. 6 (1), 105-110 (2011).</li><li>Fleming, J. T., 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=Bone+blood+flow+and+vascular+reactivity.">Bone blood flow and vascular reactivity.</a> Cells Tissues Organs. 169 (3), 279-284 (2001).</li><li>Dhillon, R. 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=PTH-enhanced+structural+allograft+healing+is+associated+with+decreased+angiopoietin-2-mediated+arteriogenesis,+mast+cell+accumulation,+and+fibrosis.">PTH-enhanced structural allograft healing is associated with decreased angiopoietin-2-mediated arteriogenesis, mast cell accumulation, and fibrosis.</a> J Bone Miner Res. 28 (3), 586-597 (2013).</li><li>Nebuloni, L., Kuhn, G. A., Vogel, J., Muller, R. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+novel+in+vivo+vascular+imaging+approach+for+hierarchical+quantification+of+vasculature+using+contrast+enhanced+micro-computed+tomography.">A novel in vivo vascular imaging approach for hierarchical quantification of vasculature using contrast enhanced micro-computed tomography.</a> PLoS One. 9 (1), e86562 (2014).</li><li>Zhang, 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=Periosteal+progenitor+cell+fate+in+segmental+cortical+bone+graft+transplantations:+implications+for+functional+tissue+engineering.">Periosteal progenitor cell fate in segmental cortical bone graft transplantations: implications for functional tissue engineering.</a> J Bone Miner Res. 20 (12), 2124-2137 (2005).</li><li>Movahed, R., Pinto, L. P., Morales-Ryan, C., Allen, W. R., Wolford, L. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Application+of+cranial+bone+grafts+for+reconstruction+of+maxillofacial+deformities.">Application of cranial bone grafts for reconstruction of maxillofacial deformities.</a> Proc (Bayl Univ Med Cent). 26 (3), 252-255 (2013).</li><li>Putters, T. F., Schortinghuis, J., Vissink, A., Raghoebar, G. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+prospective+study+on+the+morbidity+resulting+from+calvarial+bone+harvesting+for+intraoral+reconstruction.">A prospective study on the morbidity resulting from calvarial bone harvesting for intraoral reconstruction.</a> Int J Oral Maxillofac Surg. , (2015).</li><li>Kline, R. M., Wolfe, S. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Complications+associated+with+the+harvesting+of+cranial+bone+grafts.">Complications associated with the harvesting of cranial bone grafts.</a> Plast Reconstr Surg. 95 (1), 5-13 (1995).</li><li>Hassanein, 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=Effect+of+calvarial+burring+on+resorption+of+onlay+cranial+bone+graft.">Effect of calvarial burring on resorption of onlay cranial bone graft.</a> J Craniofac Surg. 23 (5), 1495-1498 (2012).</li><li>Yin, J., Jiang, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Completely+resorption+of+autologous+skull+flap+after+orthotopic+transplantation:+a+case+report.">Completely resorption of autologous skull flap after orthotopic transplantation: a case report.</a> Int J Clin Exp Med. 7 (4), 1169-1171 (2014).</li><li>Schuss, P., 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=Bone+flap+resorption:+risk+factors+for+the+development+of+a+long-term+complication+following+cranioplasty+after+decompressive+craniectomy.">Bone flap resorption: risk factors for the development of a long-term complication following cranioplasty after decompressive craniectomy.</a> J Neurotrauma. 30 (2), 91-95 (2013).</li><li>Ben Arav, A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Adeno-associated+virus-coated+allografts:+a+novel+approach+for+cranioplasty.">Adeno-associated virus-coated allografts: a novel approach for cranioplasty.</a> J Tissue Eng Regen Med. 6 (10), e43-e50 (2012).</li><li>Ito, 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=Remodeling+of+cortical+bone+allografts+mediated+by+adherent+rAAV-RANKL+and+VEGF+gene+therapy.">Remodeling of cortical bone allografts mediated by adherent rAAV-RANKL and VEGF gene therapy.</a> Nat Med. 11 (3), 291-297 (2005).</li><li>Sheyn, D., 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=PTH+promotes+allograft+integration+in+a+calvarial+bone+defect.">PTH promotes allograft integration in a calvarial bone defect.</a> Mol Pharm. 10 (12), 4462-4471 (2013).</li><li>Jain, R. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Molecular+regulation+of+vessel+maturation.">Molecular regulation of vessel maturation.</a> Nat Med. 9 (6), 685-693 (2003).</li><li>Reginato, S., Gianni-Barrera, R., Banfi, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Taming+of+the+wild+vessel:+promoting+vessel+stabilization+for+safe+therapeutic+angiogenesis.">Taming of the wild vessel: promoting vessel stabilization for safe therapeutic angiogenesis.</a> Biochem Soc Trans. 39 (6), 1654-1658 (2011).</li><li>Moutsatsos, I. 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=Exogenously+regulated+stem+cell-mediated+gene+therapy+for+bone+regeneration.">Exogenously regulated stem cell-mediated gene therapy for bone regeneration.</a> Mol Ther. 3 (4), 449-461 (2001).</li><li>Deckers, M. 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=Bone+morphogenetic+proteins+stimulate+angiogenesis+through+osteoblast-derived+vascular+endothelial+growth+factor.">Bone morphogenetic proteins stimulate angiogenesis through osteoblast-derived vascular endothelial growth factor.</a> A. Endocrinology. 143 (4), 1545-1553 (2002).</li><li>Cornejo, A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effect+of+adipose+tissue-derived+osteogenic+and+endothelial+cells+on+bone+allograft+osteogenesis+and+vascularization+in+critical-sized+calvarial+defects.">Effect of adipose tissue-derived osteogenic and endothelial cells on bone allograft osteogenesis and vascularization in critical-sized calvarial defects.</a> Tissue Eng Part A. 18 (15-16), 1552-1561 (2012).</li></ol>

Formålet med multimodale billedteknik er beskrevet her, er at gøre det muligt omhyggelig undersøgelse af angiogenese-osteogenese aksen i forbindelse med kraniel knogletransplantation. Neovaskularisering blev filmede med en μCT protokol, som tillod en nøjagtig høj opløsning 3D demonstration af det vaskulære træ fodring hele kranie defekt. μCT data kan let analyseres under anvendelse af avancerede værktøjer såsom IPL software. For eksempel er tykkelsen analyse vist i figur 3C viste, at de væ...

Craniofacial knogletab på grund af traumer, tumorresektion, dekompressiv kraniotomi og medfødt defekt sjældent helbreder sig selv og præsenterer et klart udækket behandlingsbehov. Autologe knogletransplantater og allogene knogletransplantater er flittigt brugt til at behandle disse tilstande 1. Det er almindeligt accepteret, at osteogenese tæt er kombineret med angiogenese 2,3. Således bør fuldstændig undersøgelse af en planlagt terapi for knogleregenerering indeholde en omfattende undersøgelse af vaskulære træ danner hele defektstedet. Der er flere tilgængelige metoder til at karakterisere vaskularisering i forsknings-modeller. Den vaskulære træ kan undersøges ved histologisk analyse. Da histologi afhængig sektionering væv, er der stor sandsynlighed for, at det resulterende billede vil blive forvrænget. For at løse dette problem, kan intravital mikroskopi udføres for at billedet intakt blodkar 4; Men denne metode erbegrænset til en plan afbildning. μCT scanning af prøver taget fra et dyr perfunderet med kontrastmiddel giver 3D-afbildning af det vaskulære netværk, der giver næring til regenerering site 5. Denne fremgangsmåde giver mulighed for en meget detaljeret demonstration af et organ s kar som helhed, samt en omhyggelig analyse af blodkar distribution. Endvidere μCT muliggør differentiering mellem forskellige diametre af blodkar, der kendetegner de forskellige undertyper af blodkar. Vi antager, at implantation af en calvarial autograft vil inducere større neovaskularisering end implantation af en allograft, og dette tal steg neovaskularisering vil føre til gengæld til øget knogle formation.To forfølge denne hypotese anvendte vi en række teknikker. Vi undersøgte mønstre af nydannede vaskulære træ ved at udføre en μCT-analyse. Vi målte blodperfusion anvendelse af et blod-pool fluorescerende probe. Næste, vi æslersed knoglevæv mineralisering af FLI en hydroxyapatit-rettet probe og μCT analyse. Endelig har vi overvåget stamcelle rekruttering og differentiering, der udfører BLI i transgen mus, hvor luciferase udtrykkes i osteocalcin-positive celler.

<table border="0" cellpadding="0" cellspacing="0" width="614">
	<tbody>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">C57BL/C&#160; Mice</td>
			<td style="width:112px;">Harlan laboratories</td>
			<td align="right" style="width:119px;">57</td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">FVB/n Mice</td>
			<td style="width:112px;">Harlan laboratories</td>
			<td align="right" style="width:119px;">862</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Phenobarbital</td>
			<td style="width:112px;">West waro</td>
			<td style="width:119px;">NDC 0641-0477-25</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Rodent hair clipper</td>
			<td style="width:112px;">Wahl animal</td>
			<td style="width:119px;">8786-451A</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Scalpel 11</td>
			<td style="width:112px;">Miltex</td>
			<td align="right" style="width:119px;">27111504</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Dental micro motor</td>
			<td style="width:112px;">marathon III</td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">5mm trephine</td>
			<td style="width:112px;">Fine Science tools</td>
			<td style="width:119px;">18004-50</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Hair removing cream</td>
			<td style="width:112px;">Veet</td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">KetaVed (Ketamine)</td>
			<td style="width:112px;">Vedco</td>
			<td style="width:119px;">NDC 50989-996-06</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Domitor</td>
			<td style="width:112px;">Zoetis</td>
			<td style="width:119px;">NADA 141-267</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">carprofen</td>
			<td style="width:112px;">Norbrook</td>
			<td style="width:119px;">02000/4229</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Eye ointment</td>
			<td style="width:112px;">Puralube</td>
			<td style="width:119px;">NDC 17033-211-38</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Operating binocular</td>
			<td style="width:112px;">Kent scientific</td>
			<td style="width:119px;">KSCXTS-1121</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Fine scissors&#160;</td>
			<td style="width:112px;">Fine Science tools</td>
			<td style="width:119px;">14060-11</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Curve tweezers</td>
			<td style="width:112px;">Fine Science tools</td>
			<td style="width:119px;">11274-20</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Spoon shaped spatula</td>
			<td style="width:112px;">Fine Science tools</td>
			<td style="width:119px;">10090-13</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Tisseel Fibin gel kit&#160;</td>
			<td style="width:112px;">Baxter</td>
			<td align="right" style="width:119px;">718971</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">needle holder</td>
			<td style="width:112px;">Fine Science tools</td>
			<td style="width:119px;">12060-01</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">vicryl suture 4-0</td>
			<td style="width:112px;">Ethicon</td>
			<td style="width:119px;">J392H</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Antisedan</td>
			<td style="width:112px;">Zoetis</td>
			<td style="width:119px;">NADA#141033</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Heparin</td>
			<td style="width:112px;">Sigma</td>
			<td style="width:119px;">H3393</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">20ml luerlock&#160;</td>
			<td style="width:112px;">BD</td>
			<td align="right" style="width:119px;">302830</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">23G scalp vein set (butterfly needle)</td>
			<td style="width:112px;">BD</td>
			<td align="right" style="width:119px;">367342</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Hemostat</td>
			<td style="width:112px;">Fine Science tools</td>
			<td style="width:119px;">13008-12</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Syringe pump</td>
			<td style="width:112px;">Harvard apparatus</td>
			<td style="width:119px;">PHD 2000</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">3sec gel glue&#160;</td>
			<td style="width:112px;">Scotch</td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">rodent dissection board</td>
			<td style="width:112px;">Leica</td>
			<td style="width:119px;">38DI02313</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Microfil MV-122</td>
			<td style="width:112px;">flow-tech</td>
			<td style="width:119px;">MV-122</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">uCT40 scanner</td>
			<td style="width:112px;">Scanco</td>
			<td style="width:119px;">uCT40</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">TCA6%</td>
			<td style="width:112px;">Sigma</td>
			<td style="width:119px;">T6399</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Osteosense 680</td>
			<td style="width:112px;">PerkinElmar</td>
			<td style="width:119px;">NEV10020EX</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Angiosense750</td>
			<td style="width:112px;">PerkinElmar</td>
			<td style="width:119px;">NEV10011</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Oxigen 100% medical grade</td>
			<td style="width:112px;"></td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">isoflurane (furane)</td>
			<td style="width:112px;">Baxter</td>
			<td align="right" style="width:119px;">1001936040</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">IVIS kinetics</td>
			<td style="width:112px;">Xenogen</td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Beetle luciferin</td>
			<td style="width:112px;">Promega</td>
			<td style="width:119px;">E160A</td>
			<td></td>
		</tr>
	</tbody>
</table>

computed tomography and optical imaging of osteogenesis-angiogenesis coupling to assess integration of cranial bone autografts and allografts

En stor parameter bestemme succes af en knogle-podning procedure er vaskularisering af området omkring transplantatet. Vi antager, at implantation af en knogle autotransplantat ville fremkalde en større knogle regeneration af rigelige blodkar formation. For at undersøge virkningen af ​​implantatet på neovaskularisering ved defektstedet, udviklede vi en mikro-computertomografi (μCT) tilgang til karakterisering nydannede blodkar, hvilket indebærer systemisk perfusion af dyret med en polymeriserende kontrastmiddel. Denne metode gør det muligt detaljeret vaskulær analyse af et organ i sin helhed. Derudover blev blod perfusion vurderet ved anvendelse fluorescensimagografi (FLI) af en blodbåren fluorescerende middel. Knogledannelse blev kvantificeret ved anvendelse af en FLI hydroxyapatit-målrettet probe og μCT analyse. Stamceller rekruttering blev overvåget ved bioluminescens imaging (BLI) af transgene mus, som udtrykker luciferase under styring af osteocalcin promotoren.Her beskriver og demonstrerer forberedelse af allotransplantatet, calvariale defekt kirurgi vi, μCT scanning protokoller for neovaskularisering studiet og knogledannelse analyse (herunder in vivo perfusion af kontrastmiddel), og protokollen til dataanalyse. 3D høj opløsning analyse af vaskulatur signifikant større angiogenese i dyr med implanterede autotransplantater, navnlig med hensyn til arteriole formation. Følgelig blodperfusion var signifikant højere i autograft gruppen af den 7. dag efter operationen. Vi observerede overlegen knoglemineralisering og måles større knogledannelse hos dyr, der modtog autotransplantater. Autograft implantation induceret resident stamcelle rekruttering til graft-værtsknoglen sutur, hvor cellerne differentieret til knogledannende celler mellem 7. og 10. postoperative dag. Dette fund betyder, at øget knogledannelse kan tilskrivesaugmented vaskulære fodring, der kendetegner autotransplantat implantation. Fremgangsmåderne afbildet kan tjene som et optimalt værktøj til at studere knogleregenerering i form af tæt afgrænset knogledannelse og neovaskularisering.

Neovaskularisering blev vurderet ved μCT volumetrisk analyse og ved hjælp af en FLI fluorescerende blodbårne agent at kvantificere blodperfusion. Syv dage efter operationen, μCT scanning viste en signifikant højere volumen af små og mellemstore diameter blodkar hos mus, der havde modtaget autotransplantater end i mus, der havde modtaget allotransplantater høstet fra C57BL / 6 (figur 3A). Interessant nok i autograft-gruppen den nydannede vaskulære træ syntes at nå hele defekte område, mens blo...

Watch this Scientific Journal Video about Computed Tomography and Optical Imaging of Osteogenesis-angiogenesis Coupling to Assess Integration of Cranial Bone Autografts and Allografts at JoVE.com

Computed Tomography and Optical Imaging of Osteogenesis-angiogenesis Coupling to Assess Integration of Cranial Bone Autografts and Allografts

Implantation af autologe og allogene knogletransplantater udgør accepterede tilgange til behandling af svære kraniofacial knogletab. Endnu virkningen af ​​graft sammensætning på samspillet mellem neovaskularisering, celledifferentiering og knogledannelse er uklar. Vi præsenterer en multimodal billedbehandling protokol til formål at belyse angiogenese-osteogenese indbyrdes afhængighed på graft nærhed.

Computed Tomography og optisk billeddannelse af Osteogenesis-angiogenese Kobling at Vurdere Integration af kranieknogler autografter og Allografter

A major parameter determining the success of a bone-grafting procedure is vascularization of the area surrounding the graft. We hypothesized that ...

Research

JoVE Journal

Bioengineering

Three-dimensional Optical-resolution Photoacoustic Microscopy

Optical microscopy, providing valuable insights at the cellular and organelle levels, has been widely recognized as an enabling biomedical technology. As ...

Attaching Biological Probes to Silica Optical Biosensors Using Silane Coupling Agents

Fastgørelse Biologiske Prober til silica Optiske biosensorer Anvendelse silankoblingsmidler

In order to interface with biological environments, biosensor platforms, such as the popular Biacore system (based on the Surface Plasmon Resonance (SPR) ...

Integrated Photoacoustic Ophthalmoscopy and Spectral-domain Optical Coherence Tomography

Integreret Photoacoustic oftalmoskopi og Spectral-domænet Optical Coherence Tomography

Both the clinical diagnosis and fundamental investigation of major ocular diseases greatly benefit from various non-invasive ophthalmic imaging ...

Wideband Optical Detector of Ultrasound for Medical Imaging Applications

Wideband optisk detektor af ultralyd til medicinsk billedbehandling

Optical sensors of ultrasound are a promising alternative to piezoelectric techniques, as has been recently demonstrated in the field of optoacoustic ...

An Improved Mechanical Testing Method to Assess Bone-implant Anchorage

En forbedret Mekanisk testning metode til at vurdere Bone-implantat Anchorage

Recent advances in material science have led to a substantial increase in the topographical complexity of implant surfaces, both on a micro- and a ...

In Vivo Quantitative Assessment of Myocardial Structure, Function, Perfusion and Viability Using Cardiac Micro-computed Tomography

In Vivo Quantitative Assessment of Myocardial Structure, Function, Perfusion and Viability Using Cardiac Micro-computed Tomography

In vivo kvantitativ vurdering af Myokardie Struktur, funktion, Perfusion og levedygtighed Brug Cardiac Micro-computertomografi

The use of Micro-Computed Tomography (MicroCT) for in vivo studies of small animals as models of human disease has risen tremendously due to the fact that ...

Proper Positioning and Restraint of a Rat Hind Limb for Focused High Resolution Imaging of Bone Micro-architecture Using In Vivo Micro-computed Tomography

Proper Positioning and Restraint of a Rat Hind Limb for Focused High Resolution Imaging of Bone Micro-architecture Using In Vivo Micro-computed Tomography

Korrekt positionering og fiksering af en rotte Hind lemmer for fokuseret høj opløsning billeddannelse af knogle-mikroarkitektur ved hjælp af In Vivo mikro-beregnet tomografi

The use of in vivo micro-computed tomography (&#181;CT) is a powerful tool which involves the non-destructive imaging of internal structures at high ...

Multimodal Volumetric Retinal Imaging by Oblique Scanning Laser Ophthalmoscopy (oSLO) and Optical Coherence Tomography (OCT)

Multimodal Volumetric Retinal Imaging by Oblique Scanning Laser Ophthalmoscopy oSLO and Optical Coherence Tomography OCT

Multimodal volumetriske Retinal Imaging af skrå Scanning Laser oftalmoskopi (oSLO) og optisk kohærens tomografi (OCT)

While fluorescence imaging is widely used in ophthalmology, a large field of view (FOV) three-dimensional (3D) fluorescence retinal image is still a big ...

Use of Micro X-ray Computed Tomography with Phosphotungstic Acid Preparation to Visualize Human Fibromuscular Tissue

Brug af mikro røntgen-computertomografi med Phosphotungstic syre præparat til at visualisere humant Fibromuskulær væv

Manual dissection and histological observation are common methods used to investigate human tissues. However, manual dissection can damage delicate ...

Optical Imaging of Isolated Murine Ventricular Myocytes

Optisk billeddannelse af isolerede murine ventrikulære Myocytter

The ability to isolate adult cardiac myocytes has permitted researchers to study a variety of cardiac pathologies at the single cell level. While advances ...