Bu çalışma Prostat Kanseri Araştırma Prenses Margaret Hastanesi Vakfı, Ulusal Bilim ve Kanada&#39;da, Sağlık Araştırma, Yenilik Kanada Vakfı için Kanada Enstitüsü Mühendislik Araştırma Konseyi ve Joey ve Toby Tanenbaum / Brezilya Topu Başkanı, tarafından desteklenen bir SUNY Araştırma Vakfı ve at Buffalo Üniversitesi&#39;nden başlangıç ​​desteği vermek.

1.. Pyropheophorbide-lipid sentezi <ol><li> 4, 98.7 mg 16:00 lysophophatidylcholine (1-palmitoil-2-hidroksi-sn-glisero, (... Zheng ve arkadaşları, Bioconj Chem, 2002, 13 -392, daha önce tarif edildiği gibi Spirulina Pacifica alglerden elde edilmiş) 200 mmol pyropheophorbide birleştirin Lipid: EDC --3-fosfokolin, Avanti Polar Lipids # 855675), EDC 76.3 mg, 48.7 mg DMAP (4 (dimetilamino)-5 ml amilen piridin) 1:1:2:2 Pyro bir oranı için kloroform stabilize : DMPA. </li><li> 24 saat argon altında oda sıcaklığında, reaksiyon karışımı karıştırılır. 40 ° C&#39;ye ısıtılarak bir döner buharlaştırıcı kullanılarak düşük basınç altında kloroform buharlaştınn (Piro sn-1 ya da sn-2 konumunda bağlı) piro-lipid izomerler karışımı elde edilir. </li><li> 1 mM CaCI2, 50 mM Tris (pH 8),% 0.5 Triton X-100 ve 5 mg / ml &#39;lik bir konsantrasyonda% 10 metanol içinde süspanse piro-lipid izomer karışımı. Bal arısı zehiri de elde edilen fosfolipaz A2 (PLA2) Ekle0.1 mg / ml konsantrasyon ve sn-1-piro lipid sindirimi ve saf sn-2-piro lipid izomer üretmek için 24 saat boyunca 37 ° C&#39;de çözelti inkübe edin. </li><li> Ek kloroform 2 hacim ve metanol özü 1.25 sayılarını ve daha sonra 40 ° C&#39;de düşük basınç altında döner buharlaştırma ile çözücünün çıkması DCM içinde% 1 MeOH yarılma ürünü yeniden süspanse edin ve izomerik olarak saf piro-lipid için küçük bir diol silika sütunu üzerinde saflaştırılması. </li></ol> 2. Porphysomes hazırlanması <ol><li> Pyropheophorbide-lipid, distearoil-sn-glisero-3-fosfoetanolamin-N-metoksi (polietilen glikol) (PEG-2000-PE) ve kloroform içerisinde kolesterol ayrı ayrı çözülür, ve sırasıyla konsantrasyonları kaydedin. </li><li> 65 mol% porfirin-yağ,% 5 mol PEG-2000-PE ve% 30 mol kolesterol birleştirilmesi ile 5 mg / tüpün toplam lipid ağırlığa sahip bir 12X75 mm borosilikat test tüpünde porphysome lipid filmi hazırlayın. </li><li> Nitro akışı altında lipid filmler Kurugen gaz ve bunları 1 saat daha vakumda kurutuldu sağlar. Gelecek hidrasyon ve ekstrüzyon kadar argon altında -20 ° C de lipid filmler saklayın. </li><li> Her bir lipid filmi tüpüne 1 ml fosfat tamponlu serum fizyolojik (PBS, 150 mM NaCl, 10 mM fosfat, pH 7.4) ilave edin. Tamamen donmuş kadar sıvı azot içinde test tüpü dondurma ve numune her eriyene kadar sonra 65 ° C su banyosunda tüpü eritin. Vortex 30 sn &#39;lik bir toplam (her vorteks arasında 65 ° C su banyosu içinde 5 saniye yeniden ısıtma ile 10 saniye her biri 3x) için boru. </li><li> Hiçbir büyük agregalar, hidratlı çözelti içerisinde gözlenebilir kadar ihtiyaç duyulan üzerine 5 ya da daha fazla döngü için donma-çözülme tekrarlayın. Bittiğinde buz üzerinde porphysome süspansiyon bırakın. </li><li> Dolaşan su banyosu 65 ° C&#39;de termostatlı bir sirkülasyon pompasına thermobarrel ekstrüzyon bağlan içeri yerleştirilmiş iki yığın 100 nm polikarbonat filtre ile yüksek basınçlı ekstrüzyon cihazı (10 mi) bir araya getirin ve yüksek pres için bir azot tankına ekstruder sistemi bağlamakEmin kaynağı. </li><li> 9 inç cam transfer pipet kullanarak ekstruder odasına porphysome süspansiyonu aktarın. 200 ve 300 psig arasında bir okuma başlangıç ​​ekstrüzyon basıncı olarak görülmektedir kadar nitrojen tankı regülatör üzerindeki kadranı çevirerek basıncını ayarlayın. Yeterli porphysome akış hızı elde edilir ve basınç 800 psig (5440 kPa), maksimum ulaşana kadar kademeli olarak basınç düzenleyici vanasını açın. </li><li> İlk çekme geçiş tamamlandıktan sonra, basınç kontrol valfı kapatmak ve ekstruder üzerinde basınç tahliye vanası açılarak ekstrüderin basınç serbest bırakın. Yeni presten geçirilmiş porphysome solüsyonu ile ekstrüder doldurun ve nihai çözelti, homojen bir ekstrüzyon sağlamak için en az 10 kat için ekstrüzyon tekrarlayın. </li><li> UV-Vis spektrometre ile, metanol içinde seyreltilmiş bir numunenin emilimin ölçülmesiyle porphysome nihai çözeltinin konsantrasyonunu belirleyin. Aşağıdaki denklem zekâ kullanarak konsantrasyonunu hesaplayınh 97,000 M yok olma katsayısı -1 cm -1 pyropheophorbide-lipid için 410 nm&#39;de. </li></ol><img alt="Denklem 1" src="/files/ftp_upload/50536/50536eq1.jpg" /><ol start="10"><li> Dinamik ışık saçılması ile bir Malvern Zetasizer kullanarak ZS90 porphysomes nihai boyut dağılımı ölçün. PBS içinde seyreltin porphysome çözeltisi ve ortalama sonuçlar için 15 çalışan ile üç ölçümler. </li><li> Spektroflorometre kullanılarak (% 1 Triton X-100, 1 mM) söndürüldü porphysome floresan emisyonu (PBS içinde 1 mM) ve unquenched porphysome karşılaştırarak porphysome floresans su verme belirler. (Tirton X-100) ilave edilmesinden önce ve sonra, deterjan spektrumlarını almak ve maksimum floresans okuma sinyalinin normalize. 410 nm ve 600-800 nm olarak emisyon dalgaboyu aralığı olarak uyarma ayarlayın. </li><li> Ileride kullanılana kadar 4 ° C de, argon altında, porphysomes çözüm tutun ve alüminyum f ışıktan korumakyağ. </li></ol> 3. Hayvan Modeli Ksenograft hazırlanması <ol><li> Kültür KB RPMI-1640 hücreleri (% 10 FBS ile) ve 37 ° C de% 5 CO2 ve nemli bir atmosferde hücreleri sürdürmek </li><li> Standart hücre kültürü tekniği kullanılarak hasat KB hücreleri. Enjeksiyon zamanına kadar kısa süreli depolama için buz üzerinde fosfat tamponlu salin (PBS) içinde hücre süspansiyonu (20,000 hücre / ml) tutun. </li><li> Çıplak fareler üzerinde KB ksenograftlarını hazırlayın. Standart diyet ve yatak (Harlan Laboratuvarları verilir; Diyet: bunlar (NU (NCr)-Foxn1 nu 6-7 haftalık eski Kategori # Crl): 20 g olduğunda çıplak farenin Charles River Laboratories&#39;den temin edildi Işınlanmış LM-485, # 7912; Yatak: Mısır koçanı yatak, ¼ &quot;, # 7097). Tüm hayvan taşıma araçları (şırınga, forseps, ve hayvan ped) deneylerden önce sterilize edin. % 100 oksijen taşıyıcı gazı içinde% 2 izofluran v / v ile fareler anestezi. Tüm hayvan protokolleri kurum (Üniversite Sağlık Ne tarafından onaylananğ bu durumda) hayvan bakımı ve kullanımı komitesi. </li><li> Farelerin sağ böğrüne deri altından PBS içinde süspansiyon haline getirilmiş, tümör hücreleri enjekte edilir. Fareler, tümör, 4-5 mm ve 2-3 mm kalınlığında bir çapa eriştiğinde, 10 gün sonra kullanılmak üzere hazırdır. </li></ol> 4.. In vivo Fototermal Tedavi <ol><li> Intravenöz kuyruk damarından (200 ul hacimde 750 nmol pyro içeren) porphysomes enjekte edilir. Porphysomes lazer ışını 24 saat sonrası enjeksiyon Davranış. </li><li> Bu bölümde, bir kapalı lazer güvenlik odadaki tüm işlemleri yürütmek. Lazer ışınlama sırasında lazer güvenlik gözlükleri takın. </li><li> Lazer ışınlama ekipmanı kurmak. Bir stand üzerinde bir kelepçe ile lazer fiber ve bir difüzör tutun. Düşük seviyede lazer gücünü ayarlamak ve lazer açın. (DPSS Lazer, LaserGlow Technologies, Toronto, Kanada), AP kullanarak 750 mW (1.18 W / cm 2) için, lazer gücü kalibre çapı 9 mm lik bir ışınlama alanı elde etmek için lazer fiber yüksekliğini ayarlamakower sonra metre ve hayvanları hazırlamak için lazeri kapatın. </li><li> Burun konisi ile% 100 oksijen taşıyıcı gaz içinde% 2 izofluran v / v ile hayvanların anestezisi yok) izofluran bir insan sağlığı açısından risk tehlike belirgin atma. Hayvan hayvanın arka pençeleri forseps veya parmak ile ayak tutam yaparak derin yeterli anestezi olduğunu onaylayın. Onlar bu prosedüre tepkisiz zaman hayvanlar lazer ışınlama için hazırız. </li><li> Bir kızılötesi termal kamera (Mikroshot, LumaSense Technologies) tümör sıcaklığını izlemek. Iyice odak ve bir ön PTT Görüntünün yanı sıra PTT ışınlama başlamadan önce düzenli bir beyaz ışık görüntü almak, yakın bir tümöre kamerayı yerleştirin. </li><li> Yine lazer açın ve 750 mW ışık gücünü artırmak. Tümör tamamen kapalı almak için lazer ışınının merkezinde tümör getirin ve ışınlama zamanlama başlar. Bir dakikalık bir lazer tedavisi için tümör sıcaklık görüntü her 5 sn atın. 1 dakika ile lazer kapatın. Stop% 2 izofluran gaz kaynağı, anestezi hayvan kurtarır kadar bekleyiniz ve tekrar kafese koydu. </li><li> Üniversite Sağlık Ağı tarafından onaylanmış hayvan protokollere göre 12:12 saat aydınlık ve karanlık döngüsü altında normotermi hayvan odasında fareler tutun. Buprenorfin (0.05 mg / kg, yara nedeniyle ağrıyı azaltmak için lazer ışınlaması bir hafta boyunca deri altına günde iki kez enjekte edilir. Her iki gün, bir kumpas kullanarak tümör çapını ölçmek ve bir fotoğraf çekin. Kullanarak tümör hacmini hesaplayın denklem aşağıdaki gibidir: V = π / 6 · a · bir uzun çapı ve b kısa biri, b, 2. </li><li> Bu laboratuvarda tarafından belirlenen uç nokta olarak tümörün uzun çapı, 10 mm ulaştığında CO2 odası içinde koyarak hayvanlar kurban. CO2 tedavi sonrası servikal dislokasyon gerçekleştirin. Sağkalım eğrisi çizilir. </li></ol>

<ol>
	<li>Lovell, J. F., 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=Porphysome+nanovesicles+generated+by+porphyrin+bilayers+for+use+as+multimodal+biophotonic+contrast+agents.">Porphysome nanovesicles generated by porphyrin bilayers for use as multimodal biophotonic contrast agents.</a> Nat. Mater. 10, 324-332 (2011).</li><li>Lovell, J. F., 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=Enzymatic+regioselection+for+the+synthesis+and+biodegradation+of+porphysome+nanovesicles.">Enzymatic regioselection for the synthesis and biodegradation of porphysome nanovesicles.</a> Angew Chem Int Ed Engl. 51, 2429-2433 (2012).</li><li>Liu, T. W., MacDonald, T. D., Shi, J., Wilson, B. C., Zheng, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Intrinsically+Copper-64-Labeled+Organic+Nanoparticles+as+Radiotracers.">Intrinsically Copper-64-Labeled Organic Nanoparticles as Radiotracers.</a> Angewandte Chemie. , (2012).</li><li>Zheng, 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=Low-density+lipoprotein+reconstituted+by+pyropheophorbide+cholesteryl+oleate+as+target-specific+photosensitizer.">Low-density lipoprotein reconstituted by pyropheophorbide cholesteryl oleate as target-specific photosensitizer.</a> Bioconjug Chem. 13, 392-396 (2002).</li><li>Chen, B., Pogue, B. W., Hasan, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Liposomal+delivery+of+photosensitising+agents.">Liposomal delivery of photosensitising agents.</a> Expert Opin Drug Deliv. 2, 477-487 (2005).</li><li>Jin, C. S., Zheng, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Liposomal+nanostructures+for+photosensitizer+delivery.">Liposomal nanostructures for photosensitizer delivery.</a> Lasers Surg Med. 43, 734-748 (2011).</li><li>Olson, F., Hunt, C. A., Szoka, F. C., Vail, W. J., Papahadjopoulos, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Preparation+of+liposomes+of+defined+size+distribution+by+extrusion+through+polycarbonate+membranes.">Preparation of liposomes of defined size distribution by extrusion through polycarbonate membranes.</a> Biochim Biophys Acta. 557, 9-23 (1979).</li><li>Berger, N., Sachse, A., Bender, J., Schubert, R., Brandl, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Filter+extrusion+of+liposomes+using+different+devices:+comparison+of+liposome+size,+encapsulation+efficiency,+and+process+characteristics.">Filter extrusion of liposomes using different devices: comparison of liposome size, encapsulation efficiency, and process characteristics.</a> Int J Pharm. 223, 55-68 (2001).</li><li>Jesorka, A., Orwar, O. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Liposomes:+technologies+and+analytical+applications.">Liposomes: technologies and analytical applications.</a> Annu Rev Anal Chem (Palo Alto Calif). 1, 801-832 (2008).</li><li>Lapinski, M. M., Castro-Forero, A., Greiner, A. J., Ofoli, R. Y., Blanchard, G. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Comparison+of+liposomes+formed+by+sonication+and+extrusion:+rotational+and+translational+diffusion+of+an+embedded+chromophore.">Comparison of liposomes formed by sonication and extrusion: rotational and translational diffusion of an embedded chromophore.</a> Langmuir. 23, 11677-11683 (2007).</li><li>Liu, D., Huang, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Size+Homogeneity+of+a+Liposome+Preparation+is+Crucial+for+Liposome+Biodistribution+In+Vivo.">Size Homogeneity of a Liposome Preparation is Crucial for Liposome Biodistribution In Vivo.</a> Journal of Liposome Research. 2, 57-66 (1992).</li><li>Zhang, Y., Lovell, J. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Porphyrins+as+Theranostic+Agents+from+Prehistoric+to+Modern+Times.">Porphyrins as Theranostic Agents from Prehistoric to Modern Times.</a> Theranostics. 2, 905-915 (2012).</li></ol>

Biz ifşa hiçbir şey yok.

Yüksek bir ilaç yükünü muhafaza ederken, ilaç verme teknolojilerin geliştirilmesi olarak, çok fonksiyonlu nanopartiküller ilaç verme aracın doğru izleme için geniş araştırmalar halen devam etmektedir. Porfirin yüklü lipozomlar, daha iyi farmakokinetik özellikler ve porfirin doğrudan uygulanması daha verimli teslimat için geliştirilmiştir, ancak engeller porfirin sınırlı yükleme miktarı ve plazma proteinlerine 5,6 lipozomlardan porfirin hızlı yeniden dağılımı dahil olmak üze...

Porphysomes multimodal görüntüleme ve tedavi 1 yeteneğine sahip son zamanlarda geliştirilen yeni çok fonksiyonlu nanovesicles vardır. Bunlar, kendi kendine toplanan porfirin çift tabakaları oluşturulur ve büyük sönüm katsayısı ve benzersiz yapısı bağımlı floresans kendini söndürme sonuçlar üretir (83 den fazla, 000/porphysome parçacık) porfirin son derece yüksek bir yoğunluğa, içerirler. Porphysomes iyi in vivo farmakokinetik ve biyolojik dağılım özellikleri vardır: sistemli uygulanmasından sonra 12 saat kadar bir kan yarı ömrü sergilemektedir ve pasif olarak 24 saat sonra enjeksiyon sonrası 2 7.5% ID / g ksenograft tümörlerinin birikir. Onların eşsiz yapısı ve fizikokimyasal özellikleri multimodal görüntüleme ve görüntü kılavuzluğunda tedavi için porphysome iyi bir aday. Her şeyden önce, porfirin içeren porphysomes tümör birikimi 1 üzerine tümörlerin floresan görüntüleme için içsel olarak uygundurlar.Buna ek olarak, her bir porfirin, bu nedenle, kolay porphysomes PET görüntüleme 3 için bu tür 64Cu gibi radyoizotop ile etiketlenebilir kenetleme radyoizotoplar için sabit bir sitesi vardır. Porphysomes de eşsiz foto akustik görüntüleme ve PTT yeteneklerini sergilemek böylece porphysome yapısı, sağlam olduğunda Ayrıca, emilen ışık enerjisi lazer ışınlama maruz kalma altında termal dağıldı. Bu porphysomes intravenöz enjeksiyonundan sonra 24 saat, porphysome-birikmiş tümör Lazer ışını, hızlı bir sıcaklık artışı ve güçlü fototermal tümör ablasyon neden olduğu gösterilmiştir. Bu porphysomes altın nano partiküller (AuNPs) gibi yüksek ekstinksiyon katsayısı ile etkin fototermal arttırıcılar olduğu gösterilmiştir 1. Öte yandan, AuNPs dahil olmak üzere diğer inorganik fototermal maddeler ile karşılaştırıldığında, porphysomes nedeniyle organik doğası Biyogüvenlikle olağanüstü bir avantaj göstermektedir. Porphysomes enzimatik biyolojik olarak parçalanabilir ve minimal akut toksiklik neden1000 mg / kg 1 kadar yüksek dozda intravenöz farelerde y. Dahası, lipozomların benzer porphysomes büyük sulu çekirdek pasif ya da aktif olabilir terapötik ya da görüntüleme maddeleri ile yüklenir. Optik özellikleri ve porphysomes arasında biyouyumluluk biyofotonik görüntüleme ve tedavi için organik nanopartiküllerin multimodal potansiyel göstermektedir. Bu yazıda, pyropheophorbide-lipid konjügatlarının sentezi yöntemi, üretim ve yüksek basınçlı ekstrüzyon kullanılarak porphysomes karakterizasyonu bir yöntem sunar. Fareler üzerindeki in vivo PTT tümörde porphysome etkin PTT etkinliğini göstermek için de yapılır bir subkütan ksenograft tümör modeli ile tedavisi.

<table border="1">
	<tbody>
		<tr>
			<td>Name of Equipment</td>
			<td>Company</td>
			<td>Catalog Number</td>
			<td>Comments (optional)</td>
		</tr>
		<tr>
			<td>Rotary evaporator</td>
			<td>Thermo-Savant</td>
			<td>SPD131DDA</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>Votex</td>
			<td>Scientific Industries</td>
			<td>SI-0236</td>
			<td>Model number: G560</td>
		</tr>
		<tr>
			<td>High pressure extruder</td>
			<td>LIPEX, Northern Lipids Inc.</td>
			<td>T.001</td>
			<td>10 ml Thermobarrel Extruder</td>
		</tr>
		<tr>
			<td>Heated bath circulators (Thermostatted circulator)</td>
			<td>Thermo SCIENTIFIC</td>
			<td>SC100-S5P</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>Polycarbonate filters</td>
			<td>Avanti Polar Lipids</td>
			<td>610005</td>
			<td>Pore size 100 nm, and membrane diameter 19 mm</td>
		</tr>
		<tr>
			<td>Zetasizer</td>
			<td>Malvern Instruments</td>
			<td>ZS90</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>UV/Visible Spectrophototmeter</td>
			<td>Varian Australia</td>
			<td>Cary 50 Bio UV/ Visible Spectrophotometer</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>Spectrofluorometer</td>
			<td>HORIBA Scientific</td>
			<td>FluoroMax-4</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>Transmission Electron Microscopy (TEM)</td>
			<td>Hitachi</td>
			<td>H-7000</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>Cell culture incubator</td>
			<td>SANYO</td>
			<td>MCO-18AIC</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>Powermeter</td>
			<td>Thorlabs</td>
			<td>PM100D</td>
			<td>with sensor S142C</td>
		</tr>
		<tr>
			<td>671nm Laser</td>
			<td>LaserGlow Technologies</td>
			<td>LRS-0671_PFN-02000-05</td>
			<td>S/N:10097270</td>
		</tr>
		<tr>
			<td>Infrared Thermometer</td>
			<td>Mikroshot, LUMASENSE Technologies</td>
			<td>9102409</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>Laser protective eye goggles</td>
			<td>LaserGlow Technologies</td>
			<td>AGF6602XX</td>
			<td>Optical Density: 1.5+ at 630-700 nm</td>
		</tr>
	</tbody>
</table>

one minute, sub-one-watt photothermal tumor ablation using porphysomes, intrinsic multifunctional nanovesicles

Yakın zamanda doğal olarak, çok fonksiyonlu olarak nanovesicles porphysomes geliştirdi. Bir foto-, α pyropheophorbide, bir fosfolipide konjuge edilmiş ve daha sonra lipozoma benzer kesecikler küresel kendi kendine toplandı. Nedeniyle porfirin-lipid çift tabakası içinde porfirin son derece yüksek yoğunluğa, büyük porphysomes sönme katsayıları, yapı bağımlı floresans kendini söndürme ve mükemmel fototermal etkinliğini oluşturulur. Bizim formülasyonda, porphysomes yüksek basınçlı ekstrüzyon kullanılarak sentez edildi ve 120 nm civarında bir ortalama parçacık boyutuna gösterilir. Porphysomes yirmi dört saat sonrası, damar içine enjeksiyon, tümörün lokal sıcaklık hızlı bir şekilde 750 mW (1.18 W / cm 2), 671 nm lazer ışını bir dakika maruz kaldıktan sonra 62 ° C&#39;ye 30 ° C artmıştır. , Tümörün komple bir ısı ablasyon sonra, oluşan ve iyileşmiş 2 hafta içinde, kontrol gruplarındaki tümörler büyümeye devam etti ve tüm tanımlanan en ulaştı eschars3 hafta içinde d noktasıdır. Bu veriler porphysomes kuvvetli fototermal tedavisi (PTT) ajanları olarak da kullanılabilir şeklini göstermektedir.

Pyropheophorbide istikrarlı bir lipid monomer (Şekil 1a) halinde fosfolipide konjuge ve konjügatlar, bir yüksek basınç membran bir çekici kullanılarak ekstrüzyon yöntemi ile porphysomes oluşturmak için kendini monte edilir. Bu, nispeten düşük akış hızı ile ekstrüzyon ilk 3 döngüleri sırasında porphysome-lipid süspansiyonu ekstrüde etmek genellikle zordur. Daha ekstrüzyon tekrarlanır olarak, ekstrüde çözeltinin akış hızı giderek artar, ve gerekirse hafif basınç azalt?...

Watch this Scientific Journal Video about One Minute, Sub-One-Watt Photothermal Tumor Ablation Using Porphysomes, Intrinsic Multifunctional Nanovesicles at JoVE.com

One Minute, Sub-One-Watt Photothermal Tumor Ablation Using Porphysomes, Intrinsic Multifunctional Nanovesicles

Biz böylece etkili fototermal tedavi ajanları işleyen, yapı-bağımlı floresan kendini söndürme ve benzersiz fototermal özelliklere sahip porphysomes denilen yeni içsel fonksiyonlu nanovesicles geliştirdi. Bu yüksek basınçlı ekstrüzyon porphysomes kullanılarak formüle edilebilir ve bir ksenograft tümör modelinde bunların fototermal tedavi etkinliğini araştırmıştır.

One Minute, Porphysomes Sub-Bir-Watt Fototermal Tümör Ablasyon, İçsel fonksiyonlu Nanovesicles

We recently developed porphysomes as intrinsically multifunctional nanovesicles. A photosensitizer, pyropheophorbide &alpha;, was conjugated to a ...

one-minute-sub-one-watt-photothermal-tumor-ablation-using-porphysomes

Research

JoVE Journal

Bioengineering

10.7K Görüntüleme.  University of Toronto.  Biz böylece etkili fototermal tedavi ajanları işleyen, yapı-bağımlı floresan kendini söndürme ve benzersiz fototermal özelliklere sahip porphysomes denilen yeni içsel fonksiyonlu nanovesicles geliştirdi. Bu yüksek basınçlı ekstrüzyon porphysomes kullanılarak formüle edilebilir ve bir ksenograft tümör modelinde bunların fototermal tedavi etkinliğini araştırmıştır. 

Video: One Minute, Sub-One-Watt Photothermal Tumor Ablation Using Porphysomes, Intrinsic Multifunctional Nanovesicles

Contrast Ultrasound Targeted Treatment of Gliomas in Mice via Drug-Bearing Nanoparticle Delivery and Microvascular Ablation

We are developing minimally-invasive contrast agent microbubble based therapeutic approaches in which the permeabilization and/or ablation of the microvasculature are controlled by varying ultrasound pulsing parameters.  Specifically, we are testing whether such approaches may be used to treat malignant brain tumors through drug delivery and microvascular ablation.  Preliminary studies have been performed to determine whether targeted drug-bearing nanoparticle delivery can be facilitated by the ultrasound mediated destruction of "composite" delivery agents comprised of 100nm poly(lactide-co-glycolide) (PLAGA) nanoparticles that are adhered to albumin shelled microbubbles. We denote these agents as microbubble-nanoparticle composite agents (MNCAs). When targeted to subcutaneous C6 gliomas with ultrasound, we observed an immediate 4.6-fold increase in nanoparticle delivery in MNCA treated tumors over tumors treated with microbubbles co-administered with nanoparticles and a 8.5 fold increase over non-treated tumors. Furthermore, in many cancer applications, we believe it may be desirable to perform targeted drug delivery in conjunction with ablation of the tumor microcirculation, which will lead to tumor hypoxia and apoptosis. To this end, we have tested the efficacy of non-theramal cavitation-induced microvascular ablation, showing that this approach elicits tumor perfusion reduction, apoptosis, significant growth inhibition, and necrosis. Taken together, these results indicate that our ultrasound-targeted approach has the potential to increase therapeutic efficiency by creating tumor necrosis through microvascular ablation and/or simultaneously enhancing the drug payload in gliomas.

Insonation of microbubbles is a promising strategy for tumor ablation at reduced time-averaged acoustic powers, as well as for the targeted delivery of therapeutics. The purpose of the present study is to develop low duty cycle ultrasound pulsing strategies and nanocarriers to maximize non-thermal microvascular ablation and payload delivery to subcutaneous C6 gliomas.

Kontrast Ultrason Gliomlar, Tedavi Hedeflenen İlaç-Rulman Nanopartikül Teslimat ve Mikrovasküler Ablasyon ile Fare

We are developing minimally-invasive contrast agent microbubble based therapeutic approaches in which the permeabilization and/or ablation of the ...

Biyomühendislik

Optical Frequency Domain Imaging of Ex vivo Pulmonary Resection Specimens: Obtaining One to One Image to Histopathology Correlation

Lung cancer is the leading cause of cancer-related deaths1. Squamous cell and small cell cancers typically arise in association with the conducting airways, whereas adenocarcinomas are typically more peripheral in location. Lung malignancy detection early in the disease process may be difficult due to several limitations: radiological resolution, bronchoscopic limitations in evaluating tissue underlying the airway mucosa and identifying early pathologic changes, and small sample size and/or incomplete sampling in histology biopsies. High resolution imaging modalities, such as optical frequency domain imaging (OFDI), provide non-destructive, large area 3-dimensional views of tissue microstructure to depths approaching 2 mm in real time (Figure 1)2-6. OFDI has been utilized in a variety of applications, including evaluation of coronary artery atherosclerosis6,7 and esophageal intestinal metaplasia and dysplasia6,8-10.
Bronchoscopic OCT/OFDI has been demonstrated as a safe in vivo imaging tool for evaluating the pulmonary airways11-23 (Animation). OCT has been assessed in pulmonary airways16,23 and parenchyma17,22 of animal models and in vivo human airway14,15. OCT imaging of normal airway has demonstrated visualization of airway layering and alveolar attachments, and evaluation of dysplastic lesions has been found useful in distinguishing grades of dysplasia in the bronchial mucosa11,12,20,21. OFDI imaging of bronchial mucosa has been demonstrated in a short bronchial segment (0.8 cm)18. Additionally, volumetric OFDI spanning multiple airway generations in swine and human pulmonary airways in vivo has been described19. Endobronchial OCT/OFDI is typically performed using thin, flexible catheters, which are compatible with standard bronchoscopic access ports. Additionally, OCT and OFDI needle-based probes have recently been developed, which may be used to image regions of the lung beyond the airway wall or pleural surface17.
While OCT/OFDI has been utilized and demonstrated as feasible for in vivo pulmonary imaging, no studies with precisely matched one-to-one OFDI:histology have been performed. Therefore, specific imaging criteria for various pulmonary pathologies have yet to be developed. Histopathological counterparts obtained in vivo consist of only small biopsy fragments, which are difficult to correlate with large OFDI datasets. Additionally, they do not provide the comprehensive histology needed for registration with large volume OFDI. As a result, specific imaging features of pulmonary pathology cannot be developed in the in vivo setting. Precisely matched, one-to-one OFDI and histology correlation is vital to accurately evaluate features seen in OFDI against histology as a gold standard in order to derive specific image interpretation criteria for pulmonary neoplasms and other pulmonary pathologies. Once specific imaging criteria have been developed and validated ex vivo with matched one-to-one histology, the criteria may then be applied to in vivo imaging studies. Here, we present a method for precise, one to one correlation between high resolution optical imaging and histology in ex vivo lung resection specimens. Throughout this manuscript, we describe the techniques used to match OFDI images to histology. However, this method is not specific to OFDI and can be used to obtain histology-registered images for any optical imaging technique. We performed airway centered OFDI with a specialized custom built bronchoscopic 2.4 French (0.8 mm diameter) catheter. Tissue samples were marked with tissue dye, visible in both OFDI and histology. Careful orientation procedures were used to precisely correlate imaging and histological sampling locations. The techniques outlined in this manuscript were used to conduct the first demonstration of volumetric OFDI with precise correlation to tissue-based diagnosis for evaluating pulmonary pathology24. This straightforward, effective technique may be extended to other tissue types to provide precise imaging to histology correlation needed to determine fine imaging features of both normal and diseased tissues.

Optical Frequency Domain Imaging of Ex vivo Pulmonary Resection Specimens: Obtaining One to One Image to Histopathology Correlation

A method to image ex vivo pulmonary resection specimens with optical frequency domain imaging (OFDI) and obtain precise correlation to histology is described, which is essential to developing specific OFDI interpretation criteria for pulmonary pathology. This method is applicable to other tissue types and imaging techniques to obtain precise imaging to histology correlation for accurate image interpretation and assessment. Imaging criteria established with this technique would then be applicable to image assessment in future in vivo studies.

Optik Frekans Domain Görüntüleme<em&gt; Ex vivo</em&gt; Pulmoner Rezeksiyon Örnekler: Histopatoloji Korelasyon için Birebir Görüntü Alınması

Lung cancer is the leading cause of cancer-related deaths1. Squamous cell and small cell cancers typically arise in association with the conducting ...

Viral Nanoparticles for In vivo Tumor Imaging

The use of nanomaterials has the potential to revolutionize materials science and medicine. Currently, a number of different nanoparticles are being investigated for applications in imaging and therapy. Viral nanoparticles (VNPs) derived from plants can be regarded as self-assembled bionanomaterials with defined sizes and shapes. Plant viruses under investigation in the Steinmetz lab include icosahedral particles formed by Cowpea mosaic virus (CPMV) and Brome mosaic virus (BMV), both of which are 30 nm in diameter. We are also developing rod-shaped and filamentous structures derived from the following plant viruses: Tobacco mosaic virus (TMV), which forms rigid rods with dimensions of 300 nm by 18 nm, and Potato virus X (PVX), which form filamentous particles 515 nm in length and 13 nm in width (the reader is referred to refs. 1 and 2 for further information on VNPs).
From a materials scientist's point of view, VNPs are attractive building blocks for several reasons: the particles are monodisperse, can be produced with ease on large scale in planta, are exceptionally stable, and biocompatible. Also, VNPs are "programmable" units, which can be specifically engineered using genetic modification or chemical bioconjugation methods 3. The structure of VNPs is known to atomic resolution, and modifications can be carried out with spatial precision at the atomic level4, a level of control that cannot be achieved using synthetic nanomaterials with current state-of-the-art technologies.
In this paper, we describe the propagation of CPMV, PVX, TMV, and BMV in Vigna ungiuculata and Nicotiana benthamiana plants. Extraction and purification protocols for each VNP are given. Methods for characterization of purified and chemically-labeled VNPs are described. In this study, we focus on chemical labeling of VNPs with fluorophores (e.g. Alexa Fluor 647) and polyethylene glycol (PEG). The dyes facilitate tracking and detection of the VNPs 5-10, and PEG reduces immunogenicity of the proteinaceous nanoparticles while enhancing their pharmacokinetics 8,11. We demonstrate tumor homing of PEGylated VNPs using a mouse xenograft tumor model. A combination of fluorescence imaging of tissues ex vivo using Maestro Imaging System, fluorescence quantification in homogenized tissues, and confocal microscopy is used to study biodistribution. VNPs are cleared via the reticuloendothelial system (RES); tumor homing is achieved passively via the enhanced permeability and retention (EPR) effect12. The VNP nanotechnology is a powerful plug-and-play technology to image and treat sites of disease in vivo. We are further developing VNPs to carry drug cargos and clinically-relevant imaging moieties, as well as tissue-specific ligands to target molecular receptors overexpressed in cancer and cardiovascular disease.

Viral Nanoparticles for In vivo Tumor Imaging

Plant viral nanoparticles (VNPs) are promising platforms for applications in biomedicine. Here, we describe the procedures for plant VNP propagation, purification, characterization, and bioconjugation. Finally, we show the application of VNPs for tumor homing and imaging using a mouse xenograft model and fluorescence imaging.

Viral Nanopartiküller<em&gt; In vivo</em&gt; Tümör Görüntüleme

The use of nanomaterials has the potential to revolutionize materials science and medicine. Currently, a number of different nanoparticles are being ...

Construction of a Preclinical Multimodality Phantom Using Tissue-mimicking Materials for Quality Assurance in Tumor Size Measurement

World Health Organization (WHO) and the Response Evaluation Criteria in Solid Tumors (RECIST) working groups advocated standardized criteria for radiologic assessment of solid tumors in response to anti-tumor drug therapy in the 1980s and 1990s, respectively. WHO criteria measure solid tumors in two-dimensions, whereas RECIST measurements use only one-dimension which is considered to be more reproducible 1, 2, 3,4,5. These criteria have been widely used as the only imaging biomarker approved by the United States Food and Drug Administration (FDA) 6. In order to measure tumor response to anti-tumor drugs on images with accuracy, therefore, a robust quality assurance (QA) procedures and corresponding QA phantom are needed.
To address this need, the authors constructed a preclinical multimodality (for ultrasound (US), computed tomography (CT) and magnetic resonance imaging (MRI)) phantom using tissue-mimicking (TM) materials based on the limited number of target lesions required by RECIST by revising a Gammex US commercial phantom 7. The Appendix in Lee et al. demonstrates the procedures of phantom fabrication 7. In this article, all protocols are introduced in a step-by-step fashion beginning with procedures for preparing the silicone molds for casting tumor-simulating test objects in the phantom, followed by preparation of TM materials for multimodality imaging, and finally construction of the preclinical multimodality QA phantom. The primary purpose of this paper is to provide the protocols to allow anyone interested in independently constructing a phantom for their own projects. QA procedures for tumor size measurement, and RECIST, WHO and volume measurement results of test objects made at multiple institutions using this QA phantom are shown in detail in Lee et al. 8.

This paper describes in-house procedures of constructing a preclinical multimodality phantom made of tissue-mimicking (TM) materials for quality assurance (QA) of tumor size measurement in animal imaging modalities such as ultrasound (US), computed tomography (CT) and magnetic resonance imaging (MRI).

Tümör Boyutu Ölçüm Kalite Güvencesi için Doku-taklit eden malzemeler kullanarak Klinik öncesi modelli Phantom İnşaatı

World Health Organization (WHO) and the Response Evaluation Criteria in Solid Tumors (RECIST) working groups advocated standardized criteria for ...

Production of Large Numbers of Size-controlled Tumor Spheroids Using Microwell Plates

Tumor spheroids are increasingly recognized as an important in vitro model for the behavior of tumor cells in three dimensions. More physiologically relevant than conventional adherent-sheet cultures, they more accurately recapitulate the complexity and interactions present in real tumors. In order to harness this model to better assess tumor biology, or the efficacy of novel therapeutic agents, it is necessary to be able to generate spheroids reproducibly, in a controlled manner and in significant numbers.
The AggreWell system consists of a high-density array of pyramid-shaped microwells, into which a suspension of single cells is centrifuged. The numbers of cells clustering at the bottom of each microwell, and the number and ratio of distinct cell types involved depend only on the properties of the suspension introduced by the experimenter. Thus, we are able to generate tumor spheroids of arbitrary size and composition without needing to modify the underlying platform technology. The hundreds of microwells per square centimeter of culture surface area in turn ensure that extremely high production levels may be attained via a straightforward, nonlabor-intensive process. We therefore expect that this protocol will be broadly useful to researchers in the tumor spheroid field.

We introduce a protocol for the generation of large numbers (thousands to hundreds of thousands) of uniform size- and composition-controlled tumor spheroids, using commercially available microwell plates.

Microwell Tabaklar kullanma büyüklüğü kontrollü Tümör küremsi Büyük Sayılar üretimi

Tumor spheroids are increasingly recognized as an important in vitro model for the behavior of tumor cells in three dimensions. More physiologically ...

PLGA Nanoparticles Formed by Single- or Double-emulsion with Vitamin E-TPGS

Poly(lactic-co-glycolic acid) (PLGA) is a biocompatible member of the aliphatic polyester family of biodegradable polymers. PLGA has long been a popular choice for drug delivery applications, particularly since it is already FDA-approved for use in humans in the form of resorbable sutures. Hydrophobic and hydrophilic drugs are encapsulated in PLGA particles via single- or double-emulsion. Briefly, the drug is dissolved with polymer or emulsified with polymer in an organic phase that is then emulsified with the aqueous phase. After the solvent has evaporated, particles are washed and collected via centrifugation for lyophilization and long term storage. PLGA degrades slowly via hydrolysis in aqueous environments, and encapsulated agents are released over a period of weeks to months. Although PLGA is a material that possesses many advantages for drug delivery, reproducible formation of nanoparticles can be challenging; considerable variability is introduced by the use of different equipment, reagents batch, and precise method of emulsification. Here, we describe in great detail the formation and characterization of microparticles and nanoparticles formed by single- or double-emulsion using the emulsifying agent vitamin E-TPGS. Particle morphology and size are determined with scanning electron microscopy (SEM). We provide representative SEM images for nanoparticles produced with varying emulsifier concentration, as well as examples of imaging artifacts and failed emulsifications. This protocol can be readily adapted to use alternative emulsifiers (e.g. poly(vinyl alcohol), PVA) or solvents (e.g.&#160;dichloromethane, DCM).

We describe the production and characterization of nanoparticles and microparticles composed of poly(lactic-co-glycolic acid) using vitamin E-TPGS as an emulsifier. By varying formulation parameters such as the concentration of emulsifier, it is possible to produce nanoparticles with mean diameters ranging from 220 nm to&#160;1.98 &#181;m.

Poly(lactic-co-glycolic acid) (PLGA) is a biocompatible member of the aliphatic polyester family of biodegradable polymers. PLGA has long been a popular ...

Longitudinal Measurement of Extracellular Matrix Rigidity in 3D Tumor Models Using Particle-tracking Microrheology

The mechanical microenvironment has been shown to act as a crucial regulator of tumor growth behavior and signaling, which is itself remodeled and modified as part of a set of complex, two-way mechanosensitive interactions. While the development of biologically-relevant 3D tumor models have facilitated mechanistic studies on the impact of matrix rheology on tumor growth, the inverse problem of mapping changes in the mechanical environment induced by tumors remains challenging. Here, we describe the implementation of particle-tracking microrheology (PTM) in conjunction with 3D models of pancreatic cancer as part of a robust and viable approach for longitudinally monitoring physical changes in the tumor microenvironment, in situ. The methodology described here integrates a system of preparing in vitro 3D models embedded in a model extracellular matrix (ECM) scaffold of Type I collagen with fluorescently labeled probes uniformly distributed for position- and time-dependent microrheology measurements throughout the specimen. In vitro tumors are plated and probed in parallel conditions using multiwell imaging plates. Drawing on established methods, videos of tracer probe movements are transformed via the Generalized Stokes Einstein Relation (GSER) to report the complex frequency-dependent viscoelastic shear modulus, G*(&#969;). Because this approach is imaging-based, mechanical characterization is also mapped onto large transmitted-light spatial fields to simultaneously report qualitative changes in 3D tumor size and phenotype. Representative results showing contrasting mechanical response in sub-regions associated with localized invasion-induced matrix degradation as well as system calibration, validation data are presented. Undesirable outcomes from common experimental errors and troubleshooting of these issues are also presented. The 96-well 3D culture plating format implemented in this protocol is conducive to correlation of microrheology measurements with therapeutic screening assays or molecular imaging to gain new insights into impact of treatments or biochemical stimuli on the mechanical microenvironment.

Particle-tracking microrheology can be used to non-destructively quantify and spatially map changes in extracellular matrix mechanical properties in 3D tumor models.

Partikül izleme Microrheology kullanarak 3D Tümör Modelleri Ekstrasellüler Matriks Rijitliğinin Boyuna Ölçümü

The mechanical microenvironment has been shown to act as a crucial regulator of tumor growth behavior and signaling, which is itself remodeled and ...

Real-time Monitoring of High Intensity Focused Ultrasound (HIFU) Ablation of In Vitro Canine Livers Using Harmonic Motion Imaging for Focused Ultrasound (HMIFU)

Harmonic Motion Imaging for Focused Ultrasound (HMIFU) is a technique that can perform and monitor high-intensity focused ultrasound (HIFU) ablation. An oscillatory motion is generated at the focus of a 93-element and 4.5 MHz center frequency HIFU transducer by applying a 25 Hz amplitude-modulated signal using a function generator. A 64-element and 2.5 MHz imaging transducer with 68kPa peak pressure is confocally placed at the center of the HIFU transducer to acquire the radio-frequency (RF) channel data. In this protocol, real-time monitoring of thermal ablation using HIFU with an acoustic power of 7 W on canine livers in vitro is described. HIFU treatment is applied on the tissue during 2 min and the ablated region is imaged in real-time using diverging or plane wave imaging up to 1,000 frames/second. The matrix of RF channel data is multiplied by a sparse matrix for image reconstruction. The reconstructed field of view is of 90&#176; for diverging wave and 20 mm for plane wave imaging and the data are sampled at 80 MHz. The reconstruction is performed on a Graphical Processing Unit (GPU) in order to image in real-time at a 4.5 display frame rate. 1-D normalized cross-correlation of the reconstructed RF data is used to estimate axial displacements in the focal region. The magnitude of the peak-to-peak displacement at the focal depth decreases during the thermal ablation which denotes stiffening of the tissue due to the formation of a lesion. The displacement signal-to-noise ratio (SNRd) at the focal area for plane wave was 1.4 times higher than for diverging wave showing that plane wave imaging appears to produce better displacement maps quality for HMIFU than diverging wave imaging.

Real-time Monitoring of High Intensity Focused Ultrasound HIFU Ablation of In Vitro Canine Livers Using Harmonic Motion Imaging for Focused Ultrasound HMIFU

This article describes real-time monitoring of HIFU ablation in canine liver with high frame rate ultrasound imaging using diverging and plane wave imaging. Harmonic Motion Imaging for Focused Ultrasound is used to image the decrease of acoustic radiation force induced displacement in the ablated region.

Yüksek Yoğunluk Gerçek zamanlı izleme Ultrason (HIFU) Ablasyonu Odaklı<em&gt; İn Vitro</em&gt; Odaklı Ultrason Harmonik Hareket Görüntüleme kullanma Köpek karaciğerler (HMIFU)

Harmonic Motion Imaging for Focused Ultrasound (HMIFU) is a technique that can perform and monitor high-intensity focused ultrasound (HIFU) ablation. An ...

Tissue Engineering by Intrinsic Vascularization in an In Vivo Tissue Engineering Chamber

In reconstructive surgery, there is a clinical need for an alternative to the current methods of autologous reconstruction which are complex, costly and trade one defect for another. Tissue engineering holds the promise to address this increasing demand. However, most tissue engineering strategies fail to generate stable and functional tissue substitutes because of poor vascularization. This paper focuses on an in vivo tissue engineering chamber model of intrinsic vascularization where a perfused artery and a vein either as an arteriovenous loop or a flow-through pedicle configuration is directed inside a protected hollow chamber. In this chamber-based system angiogenic sprouting occurs from the arteriovenous vessels and this system attracts ischemic and inflammatory driven endogenous cell migration which gradually fills the chamber space with fibro-vascular tissue. Exogenous cell/matrix implantation at the time of chamber construction enhances cell survival and determines specificity of the engineered tissues which develop. Our studies have shown that this chamber model can successfully generate different tissues such as fat, cardiac muscle, liver and others. However, modifications and refinements are required to ensure target tissue formation is consistent and reproducible. This article describes a standardized protocol for the fabrication of two different vascularized tissue engineering chamber models in vivo.

Tissue Engineering by Intrinsic Vascularization in an In Vivo Tissue Engineering Chamber

This is a guideline for constructing in vivo vascularized tissue using a microsurgical arteriovenous loop or a flow-through pedicle configuration inside a tissue engineering chamber. The vascularized tissues generated can be employed for organ regeneration and replacement of tissue defects, as well as for drug testing and disease modeling.

Bir ıntrinsic Vaskülarizasyon tarafından Doku Mühendisliği<em&gt; İn Vivo</em&gt; Doku Mühendisliği Odası

In reconstructive surgery, there is a clinical need for an alternative to the current methods of autologous reconstruction which are complex, costly and ...

A Rapid and Chemical-free Hemoglobin Assay with Photothermal Angular Light Scattering

Photo-thermal angular light scattering (PT-AS) is a novel optical method for measuring the hemoglobin concentration ([Hb]) of blood samples. On the basis of the intrinsic photothermal response of hemoglobin molecules, the sensor enables high-sensitivity, chemical-free measurement of [Hb]. [Hb] detection capability with a limit of 0.12 g/dl over the range of 0.35 - 17.9 g/dl has been demonstrated previously. The method can be readily implemented using inexpensive consumer electronic devices such as a laser pointer and a webcam. The use of a micro-capillary tube as a blood container also enables the hemoglobin assay with a nanoliter-scale blood volume and a low operating cost. Here, detailed instructions for the PT-AS optical setup and signal processing procedures are presented. Experimental protocols and representative results for blood samples in anemic conditions ([Hb] = 5.3, 7.5, and 9.9 g/dl) are also provided, and the measurements are compared with those from a hematology analyzer. Its simplicity in implementation and operation should enable its wide adoption in clinical laboratories and resource-limited settings.

A photo-thermal angular light scattering (PT-AS) sensor enables the rapid and chemical-free hemoglobin assay of nanoliter-scale blood samples. Here, details of the PT-AS setup and a measurement protocol for the hemoglobin concentration in blood are provided. Representative results for anemic blood samples are also presented.

Fototermal Eğik ışık saçılımı ile hızlı ve Kimyasal içermeyen Hemoglobin Deneyi

Photo-thermal angular light scattering (PT-AS) is a novel optical method for measuring the hemoglobin concentration ([Hb]) of blood samples. On the basis ...