Name: manufacture and drug delivery applications of silk nanoparticles
Uploaded: 2016-10-08T15:00:00
Description: Watch this Scientific Journal Video about Manufacture and Drug Delivery Applications of Silk Nanoparticles at JoVE.com

This research was supported by a Marie Curie FP7 Career Integration Grant 334134 within the seventh European Union Framework Program.

봄 빅스 모리 고치에서 리버스 엔지니어링 실크 해결 방법 1. 준비 참고 :이 방법은 다른 곳에서 12,27 설명 프로토콜을 기반으로합니다. <ol><li> 5mm X 5mm 조각으로 가위 말린 고치의 5g을 잘라. 어떤 오염 층을 제거합니다. </li><li> 탄산나트륨 4.24 g을 달아 증류수 끓는 물 2 L에 신중을 추가합니다. 주 :이 0.02 M 탄산나트륨 용액을 산출한다. </li><li> 끓는 탄산나트륨 용액에 절?...

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다양한 방법은, 초 임계 이산화탄소를 21 건조 모세관 미소 도트 (23)을 인쇄 (22)를 염석 스프레이 (20)를 혼합 한 폴리 비닐 알코올을 포함한 실크 나노 입자를 생성 할 수있는이 침전 24 nanoprecipitation 16,25 (참고 26에서 검토). 그러나 nanoprecipitation 인해 전반적인 단순성 실크 나노 입자를 생성하는 가장 인기있는 방법이다. 따라서 본 ?...

예를 들어, 단백질, 펩티드 및 작은 분자량 의약품 - - 세포 및 조직을 대상으로 나노 크기의 약물 전달 시스템은 종종 약물의 방출을 제어하는 ​​치료 및 페이로드의 다양한 세트를 전달하는데 사용된다. 이러한 치료 페이로드가 종종 리포좀 (덴드리머 포함) 수용성 고분자, 및 마이크로 및 나노 입자 (1) 등 다양한 거대 분자 약물 캐리어에 통합됩니다. (일반적으로 1 nm 내지 1000의 크기 범위에서) 나노 입자 널리 특히 항암 약물 전달 2, 잠재적 약물 담체로서 탐구 중이다. 약물 전달을위한 더 많은 나노 입자는 현재 임상 시험 (4)를 입력 할 수 있도록 일상적인 임상 3에 아브 락산의 성공적인 도입 (120 나노 미터 크기의 알부민 기반 나노 입자 파클리탁셀로드), 필드 촉매했다. 고형 종양은 일반적으로 그 N을 의미하는 가난한 림프 배수를 표시하고 새는 혈관이200 nm의 최대의 anoparticles 수동적으로 정맥 투여 후 이러한 종양을 대상으로한다. 이 수동 타겟팅 현상은 향상된 침투성 및 보존 (EPR) 효과라고 먼저 1986 5에보고되었다. EPR 효과가 주어진 약물 투여 할 때의 종양 미세 환경 내 약물 농도의 50 ~ 100 배 증가로 이어질 수 약물 페이로드 캐리어가없는 고분자 약물 전달체 방식이 아닌 무료로 약을 사용하여 제공됩니다. 항암 약물 전달을 위해 설계된 약물 - 로딩 된 나노 입자는 종양 미세 환경에 도달해야하고, 약물이 목적하는 치료 효과를 달성하기 위해 3 종종 일반적으로 세포 내 이입 흡수하여 특정 세포 내 구획을 입력한다. 세포 내 약물 전달을 위해 디자인 된 나노 입자는 세포 내로 게이트웨이뿐만 아니라 약제 내성 기전을 극복하기위한 경로로 엔도 시토 시스를 이용한다. 나노 입자에서 약물 방출은 종종 특히 오에 설계나노 입자 캐리어의 pH 응답 (리소좀의 pH 약 4.5) 캐리어 (7)에서 페이로드를 해방 약물 방출 또는 리소좀 효소에 대한 트리거 역할을 할 수 리소좀에서 ccur (즉, lysosomotropic 약물 전달) 6. 물질의 많은 다른 클래스는 나노 입자 (예를 들면, 금속 및 다양한 유기 및 무기 재료)를 생성하는데 사용될 수있다. 그러나, 바이오 폴리머는 그들의 알려진 생체 적합성, 생분해 성 및 낮은 독성 (8)의 매력 물질을 부상하고있다. 많은 바이오 폴리머는 알부민, 알긴산, 키토산과 실크를 포함하여, 탐구되고있다. 이들 중, 실크 약물 전달 시스템 (9)에 발전을위한 유망한 경쟁자로 떠오르고있다. 다양한 유형의 실크 (예를 들어, 봄 빅스 모리) (예를 들어, Nephila의 clavipes) 거미와 누에를 포함하는 절지 동물의 숫자에 의해 생성된다. 누에 실크 훨씬 더 exten를 사용sively 거미 실크보다 누에가 완전히 길 들여진 때문에 그 실크 따라서 재생 가능한 원료를 나타냅니다. 누에 실크는 식품의 약국 (FDA)은 특히 봉합 재료로서 인간의 사용을 위해 자료를 승인이다 이는 인간에서 강력한 안전 기록을 보유하고 생체 내에서 열 분해하는 것으로 알려져있다. 실크의 분해 프로필 12 개월 이상 (높은 결정 성 실크) 시간 (저 결정 실크)의 범위를 미세 조정할 수 있습니다. 실크 분해물 무독성이며, 본체 (10)에서 대사된다. 실크 구조가 제어 된 약물 방출을위한 좋은 재료로 만들어 작은 분자량 화합물 및 고분자 단백질 약물 (11)에 결합하는 능력을 부여한다. 단백질 약물 (예, 항체) 면역 시스템에 의한 변성, 응집, 단백질 분해 절단 및 클리어런스에 민감하다. 그러나, 실크 인해 나노 다시의 완충 능력에 치료 용 단백질을 안정화gions 및 나노 스케일 (11)에서 수분을 조정하는 능력. 이러한 독특한 기능은 물리적 보호를 제공하고 페이로드의 이동성 (11)을 줄이고 일반적으로 다른 (바이오) 폴리머로 볼 수 없습니다. 예를 실크 기반 하이드로 겔 (12), 영화 13 ~ 15과 나노 입자 (16, 17)에 대한 많은 항암 약물 전달 시스템, 지금 (18, 19 참조 검토)이 기능을 이용하기 위해 개발되었습니다 여기서, 실크 나노 입자는 오랜 기간에 걸쳐 크기 및 전하를 결정하는 것을 특징으로 하였다. 독소루비신, 임상 적으로 관련된 항암 약물, 약물 - 로딩 된 나노 입자 실크 처리 트리플 제외 인간 유방암 세포에서의 약물 충전 및 세포 독성 연구를위한 모델 약물로서 사용 하였다.

<table border="0" cellpadding="0" cellspacing="0" width="859">
	<colgroup>
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		<col />
		<col />
		<col />
	</colgroup>
	<tbody>
		<tr height="40">
			<td height="40" style="height:40px;width:216px;">Acetone</td>
			<td style="width:235px;">VWR International, Radnor, PA, USA</td>
			<td style="width:119px;">20066.33</td>
			<td style="width:291px;"></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;width:216px;">Automated Critical Point Dryer</td>
			<td style="width:235px;">Leica Microsystems, Wetzlar, Germany</td>
			<td style="width:119px;">EM CPD300</td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;width:216px;">Balancing&#160;</td>
			<td style="width:235px;">Mettler Toledo, Greifensee, Switzerland</td>
			<td style="width:119px;">NewClassic MS</td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;width:216px;">Black polystyrene microplate , 96 well</td>
			<td style="width:235px;">Sigma-Aldrich, St. Louis, MO, USA</td>
			<td style="width:119px;">3991</td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;width:216px;">Capillary cell (DTS 1070)</td>
			<td style="width:235px;">Malvern Instrument, Worcestershire, UK</td>
			<td style="width:119px;">DTS107</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:216px;">Carbon adhesive disc</td>
			<td style="width:235px;">Agar Scientific, Essex, UK</td>
			<td style="width:119px;">G3347N</td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;width:216px;">Centrifuge&#160;</td>
			<td style="width:235px;">Hermle Labortechnik, Wehingen, Germany</td>
			<td style="width:119px;">Z323K</td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;width:216px;">Centrifuge&#160;</td>
			<td style="width:235px;">Beckman Coulter, Brea, CA, USA</td>
			<td style="width:119px;">Avanti J-E, Rotor: J20</td>
			<td></td>
		</tr>
		<tr height="80">
			<td height="80" style="height:80px;width:216px;">Centrifuge&#160;</td>
			<td style="width:235px;">Beckman Coulter, Brea, CA, USA</td>
			<td style="width:119px;">Optima L-70K, Rotor: 50.2 Ti, Adaptor 303392</td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;width:216px;">Coater, low vacuum</td>
			<td style="width:235px;">Leica Microsystems, Wetzlar, Germany</td>
			<td style="width:119px;">EM ACE200</td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;width:216px;">Cuvettes, polystyrene, disposable</td>
			<td style="width:235px;">Fisher Scientific, Waltham, MA, USA</td>
			<td style="width:119px;">FB55147</td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;width:216px;">Doxorubixin&#160;</td>
			<td style="width:235px;">LC Laboratories, Boston, MA, USA</td>
			<td style="width:119px;">D4000</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:216px;">Electronic pipetting, Easypet&#160;</td>
			<td style="width:235px;">Eppendorf, Hamburg, Germany</td>
			<td style="width:119px;">N/A</td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;width:216px;">FE-SEM</td>
			<td style="width:235px;">Hitachi High-Technologies, Krefeld, Germany</td>
			<td style="width:119px;">SU6600</td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;width:216px;">Fetal Bovine Serum</td>
			<td style="width:235px;">Thermo Scientific, Waltham, MA, USA</td>
			<td style="width:119px;">16000-044</td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;width:216px;">Freeze dryer</td>
			<td style="width:235px;">Martin Christ, Osterode, Germany</td>
			<td style="width:119px;">Epsilon 2-4</td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;width:216px;">Heat inactivated Bombyx mori silk cocoons</td>
			<td style="width:235px;">Tajima Shoji, Kanagawa, Japan</td>
			<td style="width:119px;">N/A</td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;width:216px;">Hotplate with Stirrer</td>
			<td style="width:235px;">Bibby Scientific, Stanffordshire, UK</td>
			<td style="width:119px;">US 152</td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;width:216px;">Incubator</td>
			<td style="width:235px;">Memmert, Schwabach, Germany</td>
			<td style="width:119px;">INB 200</td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;width:216px;">Insulin, human recombinant, zinc solution</td>
			<td style="width:235px;">Thermo Scientific, Waltham, MA, USA</td>
			<td style="width:119px;">12585-014</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:216px;">Lithium bromide</td>
			<td style="width:235px;">Acros Organics, Geel, Belgium</td>
			<td style="width:119px;">AC199870025</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:216px;">MDA-MB-231</td>
			<td style="width:235px;">ATCC, Manassas, VA, U.S.A</td>
			<td style="width:119px;">N/A</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:216px;">Micropipette and tips</td>
			<td style="width:235px;">Eppendorf, Hamburg, Germany</td>
			<td style="width:119px;">N/A</td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;width:216px;">Microplate Reader</td>
			<td style="width:235px;">Molecular devices, Sunnyvale, CA, USA</td>
			<td style="width:119px;">SpectraMax M5</td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;width:216px;">Oak Ridge High-Speed Centrifuge Tubes, 50 ml</td>
			<td style="width:235px;">Thermo Scientific, Waltham, MA, USA</td>
			<td style="width:119px;">N/A</td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;width:216px;">Open-Top Thickwall Polycarbonate tube, 4 ml</td>
			<td style="width:235px;">Beckman Coulter, Brea, CA, USA</td>
			<td style="width:119px;">355645</td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;width:216px;">Penicilin/streptomycin&#160;</td>
			<td style="width:235px;">Thermo Scientific, Waltham, MA, USA</td>
			<td style="width:119px;">15140-122</td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;width:216px;">RPMI medium</td>
			<td style="width:235px;">Thermo Scientific, Waltham, MA, USA</td>
			<td style="width:119px;">11875-093</td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;width:216px;">Serological pipettes, 5 ml</td>
			<td style="width:235px;">Sigma-Aldrich, St. Louis, MO, USA</td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:216px;">Silicon wafers</td>
			<td style="width:235px;">Agar Scientific, Essex, UK</td>
			<td style="width:119px;">G3391</td>
			<td></td>
		</tr>
		<tr height="60">
			<td height="60" style="height:60px;width:216px;">Slide-A-Lyzer Dialysis cassettes, 3.5K MWCO, 15 ml</td>
			<td style="width:235px;">Thermo Scientific, Waltham, MA, USA</td>
			<td style="width:119px;">87724</td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;width:216px;">Sodium carbonate anhydrous</td>
			<td style="width:235px;">Fisher Scientific, Waltham, MA, USA</td>
			<td style="width:119px;">S/2840/62</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:216px;">Specimen stubs for SEM</td>
			<td style="width:235px;">Agar Scientific, Essex, UK</td>
			<td style="width:119px;">G301</td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;width:216px;">Ultrasonic homogenizer</td>
			<td style="width:235px;">Bandelin, Berlin, Germany</td>
			<td style="width:119px;">Sonoplus HD 2070</td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;width:216px;">UV transparent microplate, 96 well</td>
			<td style="width:235px;">Sigma-Aldrich, St. Louis, MO, USA</td>
			<td style="width:119px;">3635</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:216px;">Vortex</td>
			<td style="width:235px;">IKA, Staufen, Germany</td>
			<td style="width:119px;">Genius 3</td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;width:216px;">Zetasizer</td>
			<td style="width:235px;">Malvern Instrument, Worcestershire, UK</td>
			<td style="width:119px;">Nano ZS</td>
			<td></td>
		</tr>
		<tr height="20">
			<td colspan="2" height="20" style="height:20px;">Zetasizer Software version 7.11</td>
			<td style="width:119px;"></td>
			<td>DLS software</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:216px;">Micro Modulyo</td>
			<td style="width:235px;">&#160;Thermo Fisher</td>
			<td style="width:119px;">230</td>
			<td>Freeze drying system&#160;</td>
		</tr>
	</tbody>
</table>

manufacture and drug delivery applications of silk nanoparticles

실크 때문에 뛰어난 기계적 특성, 생체 적합성 및 생분해 성뿐만 아니라, 트리거에 응답하여 보호하고이어서 페이로드를 분리하는 능력 의용 제약 어플리케이션을위한 유망한 생체 고분자이다. 실크 다양한 재료 포맷으로 제형 화 될 수 있지만, 실크 나노 입자로서 유망한 약물 전달 시스템이 등장하고있다. 따라서,이 문서 안정 실크 나노 입자를 생성하는 데 사용될 수있는 재생 실크 용액을 수득 리버스 엔지니어링 실크 고치 절차를 다룬다. 이러한 나노 입자는 후속 적 특징 약물 로딩 잠재적 항암제 전달체로서 탐구. 간단히, 실크 고치은 역, 고치를 검 제거하여 첫 번째 설계 실크 용해 다음과 수성 실크 솔루션을 산출하기 위해 정리된다. 이어서, 재생 된 실크 용액 실크 나노 입자를 수득 nanoprecipitation을 받는다 - 간단하지만 강력한 방법즉, 균일 한 나노 입자를 생성합니다. 실크 나노 입자의 크기, 제타 전위, 수성 매체의 형태 및 안정성뿐만 아니라, 화학 요법 페이로드를 포획 및 인간 유방암 세포를 죽일 수있는 능력에 따른 특징이다. 전반적으로 설명하는 방법은 잠재적 인 나노 의학 등의 사용을 포함하여 쉽게 응용 프로그램의 무수한에 대해 탐색 할 수 균일 한 실크 나노 입자를 얻을 수 있습니다.

이전 17 설명 된대로 데이터를 통계적으로 분석 하였다. 학생의 t 검정 여러 샘플 페로 니의 다중 비교 사후 시험 다음 샘플 쌍과 분산 (ANOVA)의 일방 분석을 위해 사용 하였다. 다음과 같이 별표는 통계적 유의성을 의미한다 : * P &lt;0.05 ** P &lt;0.001. 모든 데이터는 독립적 인 실험의 수를 나타내는 표준 편차 (SD) 및 괄호 안의 숫자 ± 평균 값으로 표시하고 있습니다. </...

Watch this Scientific Journal Video about Manufacture and Drug Delivery Applications of Silk Nanoparticles at JoVE.com

Manufacture and Drug Delivery Applications of Silk Nanoparticles

나노 입자는 표시의 광범위한 유망 약물 전달 시스템이 등장하고있다. 여기, 우리는 역 설계 누에 나방 실크를 사용하여 실크 나노 입자를 제조 할 수있는 간단하면서도 강력한 방법을 설명합니다. 이들은 실크 나노 용이 치료 페이로드이어서 약물 전달 애플리케이션을 위해 탐색 될 수있다.

실크 나노 입자의 제조 및 약물 전달 응용 프로그램

Silk is a promising biopolymer for biomedical and pharmaceutical applications due to its outstanding mechanical properties, biocompatibility and ...

The overall goal of this procedure is to manufacture silk nanoparticles from a reverse-engineered aqueous silk solution, and subsequently use these silk nanoparticles for drug-delivery applications. So this method utilized nano-precipitation to generate silk-based particle, which can be used for a broad range of applications, such as drug delivery. The main advantage of this technique is simplicity and reproducibility to deliver uniform silk nanoparticles that are stable, and of a narrow-side distribution.To begin this procedure, bring two liters of distilled water to a boil. Then, carefully add 4.24 grams of sodium carbonate. Next, use scissors to cut five grams of dried cocoons into five-millimeter-by-five-millimeter pieces.Add the cocoon pieces to the sodium carbonate solution, and boil for 60 minutes to de-gum the silk fibers, stirring occasionally. Once boiling is complete, removed the de-gummed silk. Wash with one liter of distilled water for 20 minutes, three times.Then, squeeze the washed silk to remove excess liquid, untie the silk by hand, and place it in a fume hood to air dry overnight. After drying is complete, tightly pack five grams of air-dried silk fibers into the bottom of a 50-milliliter beaker. Then, prepare a fresh 9.3-molar solution of lithium bromide.Dissolve the silk in lithium bromide, adding four milliliters of lithium bromide per gram of silk. Cover the beaker with aluminum foil to prevent evaporation. Allow the silk to dissolve for four hours at 60 degrees Celsius, stirring occasionally.After dissolution is complete, wet a dialysis cassette in water for five minutes. Then, transfer 15 milliliters of the silk-lithium bromide solution into the dialysis cassette. Use a needle and syringe to remove any air bubbles.Dialyze against one liter of distilled water. Change the water at one, three, and six hours. Change the beaker with new water the following morning and evening, and one final time on the morning of the third day.After dialysis is complete, collect the silk solution from the cassette and centrifuge at 9, 500 times g for 20 minutes at five degrees Celsius. Recover the supernatant and repeat the centrifugation twice more. Then, remove, dry, and determine the total dry weight and concentration of the silk solution as outlined in the text protocol.Next, add a 5%weight-per-volume silk solution drop-wise to acetone, maintaining the acetone volume-per-volume concentration over 75. Fill the tubes completely to ensure that they do not collapse during centrifugation. Centrifuge the precipitate at 48, 000 times g for two hours at four degrees Celsius.Dislodge the pellet with a spatula and add 20 milliliters of distilled water. Use pipette tips to remove the pellet from the spatula and vortex it for 20 seconds. Then, sonicate the pellet using an ultrasonic probe at 30%amplitude for 30 seconds, two times.After sonication is complete, top up the centrifuge tube to capacity with distilled water. Repeat the centrifugation and resuspension at least two more times. Then, store at four degrees Celsius for future use.Begin by preparing a Doxorubicin solution as outlined in the text protocol. Then, mix two milliliters of a 0.2 micromole-per-milliliter Doxorubicin solution with 200 microliters of silk nanoparticles at 10, 30, or 50 milligrams per milliliter in a two-milliliter tube. Incubate the silk-Doxorubicin suspension at room temperature overnight on a rotating mixer.After drug-loading is complete, centrifuge the suspension at 194, 000 times g for 30 minutes. Wash the Doxorubicin-loaded silk nanoparticles with distilled water and repeat the centrifugation twice. Next, pull the supernatant and note the total volume.Pipette 200 microliters of the supernatant into a black microtiter plate. Using a fluorescence microplate reader, set the excitation wavelength to 485 nanometers and the emission wavelength to 590 nanometers, and record the fluorescence values. After culturing and seeding MDA-MB-231 cells, add freely diffusable Doxorubicin, silk nanoparticles, and silk nanoparticles loaded with Doxorubicin into the respective 96-well plate.Then, add MTT at 72 hours to determine cell viability and the half-maximal inhibitory concentration. Incubate for five hours. After incubation is complete, carefully drain the wells with a pipette.Add 100 microliters of dimethyl sulfoxide to dissolve the formazan. Using an absorbance microplate reader, measure the absorbance at 560 nanometers. Gather this data for three independent experiments.In this procedure, silk nanoparticles are manufactured from Bombyx mori cocoons. Doxorubicin is used as a clinically relevant chemotheraputic model drug for drug loading and in vitro cytotoxicity studies. The representative results for the Doxorubicin encapsulation efficiency for 2, 6, and 10 milligrams of silk nanoparticles can be seen here.The ability of drug-loaded silk nanoparticles to deliver Doxorubicin and subsequently kill cancer cells is assessed in vitro with human breast cancer MDA-MB-231 cells. At equivalent drug doses of 0.1 micrograms, freely diffusable Doxorubicin and Doxorubicin-loaded silk particles cause significant decreases in cell viability. Freely diffusable Doxorubicin shows a greater cytotoxicity than loaded silk nanoparticles due to differences in cell uptake.The quantitative measurements are corroborated by qualitative scanning-electron-microscope imaging. Control cultures show higher cellular density and a predominating mesenchymal phenotype. Similar results are seen in cultures exposed to silk nanoparticles.Cultures exposed to Doxorubicin show a markedly different phenotype, with a broad and spread-out morphology. Exposure to either freely diffusable Doxorubicin or Doxorubicin-loaded silk nanoparticles results in a substantial reduction of cell numbers. So one master delivers engineered silk solution, can be delivered within three days.And a silk nanoparticle can be manufactured within a day. Where attempting this procedure, is it important to remember to use an acetone-resistant centrifuge to insure nanoprecipitation and sample work-up. Furthermore, insure that the centrifuge tube are filled completely so that they don't collapse during the centrifugation.After watching this video, you should have a good understanding of how to manufacture a reverse-engineered silk solution and use this to generate the uniform silk nanoparticle with a narrow-side distribution.

Research

JoVE Journal

Bioengineering

Evaluation of Polymeric Gene Delivery Nanoparticles by Nanoparticle Tracking Analysis and High-throughput Flow Cytometry

고분자 유전자 배달에 대한 평가는 Nanoparticle 추적 분석 및 높은 처리량 유동 세포 계측법에 의해 나노 입자

Non-viral gene delivery using polymeric nanoparticles has emerged as an attractive approach for gene therapy to treat genetic diseases1 and as a ...

연구

생체공학

Self-reporting Scaffolds for 3-Dimensional Cell Culture

3 차원 세포 배양을위한 자기보고 발판

Culturing cells in 3D on appropriate scaffolds is thought to better mimic the in vivo microenvironment and increase cell-cell interactions. The resulting ...

Models and Methods to Evaluate Transport of Drug Delivery Systems Across Cellular Barriers

모델 및 세포 장벽에 걸쳐 약물 전달 시스템의 전송을 평가하는 방법

Sub-micrometer carriers (nanocarriers; NCs) enhance efficacy of drugs by improving solubility, stability, circulation time, targeting, and release. ...

Designing Silk-silk Protein Alloy Materials for Biomedical Applications

바이오 메디컬 응용 프로그램에 대한 실크 실크 단백질 합금 재료 설계

Fibrous proteins display different sequences and structures that have been used for various applications in biomedical fields such as biosensors, ...

Drug-induced Sensitization of Adenylyl Cyclase: Assay Streamlining and Miniaturization for Small Molecule and siRNA Screening Applications

아데 닐 레이트 사이 클라 제의 약물에 의한 과민성 : 작은 분자와 siRNA의 심사 응용 프로그램에 대한 분석 최적화 기술과 소형화

Sensitization of adenylyl cyclase (AC) signaling has been implicated in a variety of neuropsychiatric and neurologic disorders including substance abuse ...

Biofunctionalized Prussian Blue Nanoparticles for Multimodal Molecular Imaging Applications

멀티 모달 분자 이미징 애플리케이션을위한 Biofunctionalized 프 러시안 블루 나노 입자

Multimodal, molecular imaging allows the visualization of biological processes at cellular, subcellular, and molecular-level resolutions using multiple, ...

Engineered 3D Silk-collagen-based Model of Polarized Neural Tissue

편광 신경 조직의 엔지니어링 3D 실크 콜라겐 기반 모델

Despite huge efforts to decipher the anatomy, composition and function of the brain, it remains the least understood organ of the human body. To gain a ...

Solid Lipid Nanoparticles (SLNs) for Intracellular Targeting Applications

Solid Lipid Nanoparticles SLNs for Intracellular Targeting Applications

고체 지질 나노 입자 (SLNs) 세포 내 타겟팅을위한 응용 프로그램

Nanoparticle-based delivery vehicles have shown great promise for intracellular targeting applications, providing a mechanism to specifically alter ...

Mesenchymal Stromal Cell Culture and Delivery in Autologous Conditions: A Smart Approach for Orthopedic Applications

중간 엽 기질자가 조건에서 세포 배양 배송 : 정형 외과 응용 프로그램에 대한 스마트 접근

Human Mesenchymal Stromal Cells (hMSCs) are cultured in vitro with different media. Limits on their use in clinical settings, however, mainly depend on ...

Synthesis of Functionalized 10-nm Polymer-coated Gold Particles for Endothelium Targeting and Drug Delivery

기능성된 10 nm 폴리머-코팅 된 금 입자 피 대상에 대 한 및 약물 전달의 합성

Gold nanoparticles (AuNPs) have been used extensively in medical research due to their size, biocompatibility, and modifiable surface. Specific targeting ...