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 절단 누에 고치 조각을 추가하고 60 분은 실크 섬유를 degum하는 끓인다. 균일 한 샘플 처리를 보장하기 위해 때때로 실크 저어. </li><li> 고무질 제거 실크를 제거하고 20 분 동안 증류수 1 L로 세척한다; 세정 공정을 3 회 이상 반복한다. </li><li> 세척 된 실크를 제거하고 물기를 제거한 다음 풀어 / 손으로 실크를 뽑아 잘 짠다. 건조 하룻밤을 방송하는 흄 후드에 묶이지 실크를 놓습니다. 이것은 일반적 고무질의 3.6 g을 수득ILK 섬유. </li><li> 다음 날, 5g 공기 건조 고무질 제거 실크 섬유를 무게와 50 ㎖ 비이커의 바닥에 단단히 실크 팩. </li><li> 신선한 9.3 M의 LiBr 용액을 준비합니다. 4 ml의의 LiBr 비율에 1g 실크를 사용하여의 LiBr의 실크 섬유를 녹여. 증발을 방지하고, 실크 완전히 60 ° C를 용해 할 수 있도록 알루미늄 호일로 실크의 LiBr 샘플을 커버한다. 이 단계는 4 시간까지 소요 때때로 교반하여 도움을 받는다. </li><li> 5 분 동안 물에서 투석 카세트 (3500 다 분자량 컷 - 오프)을 젖은. 15 주사 용액 15 ml의 투석 카세트에 실크의 LiBr 용액과 공기 방울을 제거하기 위해 바늘과 주사기를 사용한다. </li><li> 증류수 1 L에 대해 투석하고 (즉, 첫날 3 변경)하고 다시 다음 아침과 저녁에 (즉, 두 번째 날의 변경), 다시 1, 3, 6 시간에서 물을 변경 다음날 아침 (세 번째 날 즉, 1 변경). </li><li> 실크 soluti를 수집투석 카세트와 원심 분리기 9,500 X g에서 5 ° C에서 20 분 동안 용액으로부터합니다. 뜨는을 복구하고 두 번 더이 원심 분리 과정을 반복합니다. </li><li> 빈 무게 보트 (W1)의 무게를 결정하고 실크 액 1ml를 추가합니다. 다시 (W2) 무게를 기록하고 밤새 60 ºC에서 계량 보트를 떠나하여 샘플을 건조. 다음으로, 전체 건조 중량 (W3) (건조 실크와 무게 보트)를 결정합니다. 실크 용액의 농도 (/ w는 V)이다 %는 = (W3-W1은 / W2-W1은) × 100. </li></ol> 리버스 엔지니어링 실크 솔루션에서 실크 나노 입자의 2. 준비 <ol><li> A&gt; 75 % (v / v)의 아세톤 용액을 유지하면서 실크 솔루션 적하 (/ V w) 아세톤에 5 %를 추가합니다. 예를 들어, 5 % (9)를 가하여 (W / V) 실크 용액 34 ml를 적하 아세톤 (50 방울 / 분의 속도로 10 μL / 방울). </li><li> 4 ℃에서 2 시간 동안 48,000 XG에서 침전물을 원심 분리기. </li><li> 뜨는을 기음과 PELLE를 재현 탁제 주걱으로 펠렛을 빠지 후 증류수 20 ㎖를 첨가하여 증류수 t. 주걱에서 펠렛을 제거하기 위해 피펫 팁을 사용합니다. 30 초 동안 30 % 진폭으로 초음파 프로브를 사용하여 두 개의 초음파 사이클 다음 20 초간 증류수로 용량의 원심 분리 관 상단을 볼 텍싱 후. </li><li> 적어도 두 번 원심 분리 및 재 부유 단계를 반복합니다. </li><li> 사용까지 4 ° C에서 2.3 및 저장에 설명 된대로, 증류수 (6) 용액에 펠렛을 재현 탁. 세포 배양 연구에, 실크 나노 입자 주식은 감마 (17)를 조사 할 수 있습니다. </li></ol> 실크 나노 입자 농도 3. 결정 <ol><li> 4 ℃에서 2 시간 동안 48,000 XG에 원심 분리기 실크 나노 입자. </li><li> 30 초 동안 30 % 진폭으로 초음파 두 사이클이어서 증류수 3 mL를 모든 나노 입자를 수집한다. </li><li> 2 mL 및 1 ml의 많은으로 실크 나노 입자의 3 ㎖ 주식을 분할하고 홍보로 전송2 ml의 튜브 전자는-무게. 2 mL의 샘플의 총 중량을 기록한다. 사용까지 4 ° C에서 1 ml를 많이 저장; 이 1 ㎖의 시료 검량선을 생성하는 데 사용된다. </li><li> 다음 스냅 동결과 밤새 동결 건조기에서 2 ml의 실크 나노 입자를 많이 동결 건조. 동결 건조 후, 2 ㎖의 튜브를 닫고 다시 무게를 측정하고, 2 mL의 샘플에 원래 존재 실크 나노 입자 (mg)의 ​​양을 계산한다. </li><li> 5 점 검정 곡선 (0.04-7 ㎎ / ㎖)를 생성하기 위해 증류수 1ml를 실크 나노 스톡을 희석. 샘플 흡광도 최대를 초과하지 않았는지 확인하십시오. </li><li> 600 nm에서 각각의 표준 희석의 흡광도를 결정합니다. 이것은, 96 웰 플레이트 설치 프로그램을 사용하여 수행됩니다. 표준 곡선 농도 (㎎ / ㎖)에 비해 플롯 흡광도. 이어서 현탁액 실크 나노 입자의 농도를 결정하기 위해 통상적으로이 표준 곡선을 사용한다. </li></ol> 독소루비신로드 실크 나노 입자의 제조 4 <ol><li&gt; 독소루비신 용액의 조제 <ol><li> 증류수 8 ml의 독소루비신의 HCl 1.2 mg을 녹이고. </li><li> 116 μg의 / ㎖ (0.2 μmol / ㎖)의 작업 주식을 산출하기 위해 증류수로 10 ml의 최대합니다. </li></ol></li><li> 독소루비신로드 실크 나노 입자의 제조 <ol><li> 10, 30 또는 50 밀리그램 / 2 ML 튜브 실크 나노 입자 ml의 200 μL와 0.2 μmol / ㎖ 독소루비신 용액 2 ㎖를 섞는다. </li><li> 회전 믹서 (25 ° C) 실온에서 밤새 실크 독소루비신 서스펜션을 품어. </li><li> 다음, 30 분 동안 194,000 XG에서 실크 - 독소루비신 현탁액을 원심 분리기. 증류수로 독소루비신로드 실크 나노 입자를 세척하고 두 번 더이 절차를 반복합니다. </li><li> (이 샘플은 포위 효율을 결정하는 데 사용된다) 상층 액을 풀 전량 참고. </li><li> 증류수에 독소루비신로드 실크 나노 입자를 재현 탁 4 ºC 유엔에서 빛과 가게에서 보호사용 참깨. </li></ol></li><li> 캡슐화 효율 의약품로드의 결정 <ol><li> 피펫 검은 미세 적정 플레이트로 단계 4.2.4에서 뜨는 200 μL. </li><li> 고정 광전자 설정에서 독소루비신 관련 형광을 측정하기 위해 형광 마이크로 플레이트 리더를 사용. </li><li> 590 내지 485 nm의 발광 파장에 여기 파장을 설정 한 후 형광 값을 기록한다. </li><li> 독소루비신 교정 곡선을 생성합니다. 확인 측정 (고정 광전자 증 설정에 예) 동일한 기기 설정을 취득한다. 검량선을 사용하여, 결합 된 상청액 독소루비신 농도를 계산한다. 3 개의 독립적 인 실험에서이 측정을 반복합니다. </li><li> 캡슐화 효율을 결정하기 위해 식 (1)을 사용하여 : <img alt="식 (1)" src="/files/ftp_upload/54669/54669equation1.jpg" /></li></ol></li></ol> 실크 나노 5. 특성입자 <ol><li> 갓 준비 및 저장 실크 나노 입자의 크기와 제타 전위의 평가. <ol><li> 4 ° C, 25 ° C에서 증류수에 보관 실크 나노 입자. </li><li> 동적 광산란 (DLS)을 이용하여 0 일, 14 일 및 28 일에 실크 나노 입자의 크기 및 제타 전위를 측정한다. 단백질 17 증류수 1.33과 1.60로 굴절률을 설정합니다. 사용자 인터페이스 소프트웨어 입자 크기를 계산한다. </li></ol></li><li> 주사 전자 현미경에 의한 실크 나노 입자의 형태 학적 평가 (SEM). <ol><li> SEM의 스터브로 카본 접착제 디스크를 삽입하고,이어서, 실리콘 웨이퍼를 부착. </li><li> 1 ㎎ / ㎖의 농도로 실크 나노 입자를 희석. 피펫 실리콘 웨이퍼 상에 샘플 10㎕를가, -80 ° C에서 샘플을 고정하고, 밤새 제조자의 지시에 따라, 동결 건조 시스템을 사용하여 동결 건조. </li><li> 코트 골드와 샘플은 20 nm 두께까지 층저 진공 스퍼터 코터. 참고 : 악기 설정은 모델에 따라 다릅니다. 여기에 사용 된 모델은 완전히 자동화 된 사항이며 두께에 의해 작동합니다. </li><li> 5 kV로의 주사 전자 현미경 40,000 배의 배율 이미지 샘플. </li></ol></li></ol> 컨트롤의 생체 외 세포 독성 및 독소루비신로드 실크 나노 입자 6. <ol><li> 실크 나노 입자에 노출 후 세포 생존 능력. <ol><li> 10 % v / V를의 FBS와 RPMI 1640 문화 MDA-MB-231 세포. 조직 배양 접시에 세포는 폴리스티렌을 처리하고 37 ℃에서 가습 5 % CO 2 분위기에서 품어. 정기적으로 2-3 일마다 80 %의 합류에 하위 문화. </li><li> 96 웰 플레이트에 2 × 104 세포 / cm 2의 밀도로 MDA-MB-231 세포를 플레이트. 세포가 밤새 복구 할 수 있습니다. </li><li> μg의 자유 확산 독소루비신 0.001 (i)를 추가, (ⅱ) 0.001 mg의 실크 나노 입자를 0.1 mg의 실크 나노 입자96 웰 플레이트 (최종 부피 잘 당 100 μL)에 μg의 독소루비신 0.001-1로드. </li><li> 세포 생존 및 (3- (4,5- 디메틸 티아 졸 -2- 일) -2,5- 디 페닐 테트라 졸륨 브로마이드 PBS에서 (MTT 5 ㎎ / ㎖)에 72 시간을 추가하여 반 최대한 억제 농도 (IC 50)을 결정한다. 인큐베이션을 5 시간 동안주의 깊게 피펫으로 우물을 드레인 디메틸 설폭 100 ㎕로 포르 마잔을 용해. 560 nm에서 흡광도를 측정한다. 3 개의 독립적 인 실험에서이 측정을 반복합니다. 주 : 미처리 컨트롤의 흡광도 값은 100 %의 세포 생존율에 대한 기준 값으로 작용한다. </li></ol></li><li> 실크 나노 입자에 노출 된 세포의 SEM. <ol><li> 2 × 104 세포 / ㎠의 밀도로 멸균 된 유리 커버 슬립의 종자 MDA-MB-231 세포. 세포가 밤새 복구 할 수 있습니다. 72 시간 동안 원하는 치료 조건에 세포를 노출시킨다. </li><li> 디 씻어 30 분 동안 PBS의 2 % v / V를 글루 타르 알데히드와 세포를 수정잠잠 물 두 번, 에탄올 시리즈 탈수, 다른 곳에서 28 설명 된대로 중요한 점은, 샘플을 건조. </li><li> 금과 샘플 두꺼운 저 진공 스퍼터 코터를 사용하여 20 nm의 업층 코트 스퍼터. 참고 : 악기 설정은 모델에 따라 다릅니다. 여기에 사용 된 모델은 완전히 자동화 된 사항이며 두께에 의해 작동합니다. </li><li> 5 kV의 이미지 및 700 배 배율의 전자 가속을 이용하여 SEM 샘플. </li></ol></li></ol>

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The authors have nothing to disclose.

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.

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	<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 ...

manufacture-and-drug-delivery-applications-of-silk-nanoparticles

Research

JoVE Journal

Bioengineering

15.6K 조회수.  University of Strathclyde.  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. 

비디오: Manufacture and Drug Delivery Applications of Silk Nanoparticles

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

Non-viral gene delivery using polymeric nanoparticles has emerged as an attractive approach for gene therapy to treat genetic diseases1 and as a technology for regenerative medicine2. Unlike viruses, which have significant safety issues, polymeric nanoparticles can be designed to be non-toxic, non-immunogenic, non-mutagenic, easier to synthesize, chemically versatile, capable of carrying larger nucleic acid cargo and biodegradable and/or environmentally responsive. Cationic polymers self-assemble with negatively charged DNA via electrostatic interaction to form complexes on the order of 100 nm that are commonly termed polymeric nanoparticles. Examples of biomaterials used to form nanoscale polycationic gene delivery nanoparticles include polylysine, polyphosphoesters, poly(amidoamines)s and polyethylenimine (PEI), which is a non-degradable off-the-shelf cationic polymer commonly used for nucleic acid delivery1,3 . Poly(beta-amino ester)s (PBAEs) are a newer class of cationic polymers4 that are hydrolytically degradable5,6 and have been shown to be effective at gene delivery to hard-to-transfect cell types such as human retinal endothelial cells (HRECs)7, mouse mammary epithelial cells8, human brain cancer cells9 and macrovascular (human umbilical vein, HUVECs) endothelial cells10.
A new protocol to characterize polymeric nanoparticles utilizing nanoparticle tracking analysis (NTA) is described. In this approach, both the particle size distribution and the distribution of the number of plasmids per particle are obtained11. In addition, a high-throughput 96-well plate transfection assay for rapid screening of the transfection efficacy of polymeric nanoparticles is presented. In this protocol, poly(beta-amino ester)s (PBAEs) are used as model polymers and human retinal endothelial cells (HRECs) are used as model human cells. This protocol can be easily adapted to evaluate any polymeric nanoparticle and any cell type of interest in a multi-well plate format.

A protocol for nanoparticle tracking analysis (NTA) and high-throughput flow cytometry to evaluate polymeric gene delivery nanoparticles is described. NTA is utilized to characterize the nanoparticle particle size distribution and the plasmid per particle distribution. High-throughput flow cytometry enables quantitative transfection efficacy evaluation for a library of gene delivery biomaterials.

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

Culturing cells in 3D on appropriate scaffolds is thought to better mimic the in vivo microenvironment and increase cell-cell interactions. The resulting 3D cellular construct can often be more relevant to studying the molecular events and cell-cell interactions than similar experiments studied in 2D. To create effective 3D cultures with high cell viability throughout the scaffold the culture conditions such as oxygen and pH need to be carefully controlled as gradients in analyte concentration can exist throughout the 3D construct. Here we describe the methods of preparing biocompatible pH responsive sol-gel nanosensors and their incorporation into poly(lactic-co-glycolic acid) (PLGA) electrospun scaffolds along with their subsequent preparation for the culture of mammalian cells. The pH responsive scaffolds can be used as tools to determine microenvironmental pH within a 3D cellular construct. Furthermore, we detail the delivery of pH responsive nanosensors to the intracellular environment of mammalian cells whose growth was supported by electrospun PLGA scaffolds. The cytoplasmic location of the pH responsive nanosensors can be utilized to monitor intracellular pH (pHi) during ongoing experimentation.

Biocompatible pH responsive sol-gel nanosensors can be incorporated into poly(lactic-co-glycolic acid) (PLGA) electrospun scaffolds. The produced self-reporting scaffolds can be used for in&#160;situ monitoring of microenvironmental conditions whilst culturing cells upon the scaffold. This is beneficial as the 3D cellular construct can be monitored in real-time without disturbing the experiment.

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

Sub-micrometer carriers (nanocarriers; NCs) enhance efficacy of drugs by improving solubility, stability, circulation time, targeting, and release. Additionally, traversing cellular barriers in the body is crucial for both oral delivery of therapeutic NCs into the circulation and transport from the blood into tissues, where intervention is needed. NC transport across cellular barriers is achieved by: (i) the paracellular route, via transient disruption of the junctions that interlock adjacent cells, or (ii) the transcellular route, where materials are internalized by endocytosis, transported across the cell body, and secreted at the opposite cell surface (transyctosis). Delivery across cellular barriers can be facilitated by coupling therapeutics or their carriers with targeting agents that bind specifically to cell-surface markers involved in transport. Here, we provide methods to measure the extent and mechanism of NC transport across a model cell barrier, which consists of a monolayer of gastrointestinal (GI) epithelial cells grown on a porous membrane located in a transwell insert. Formation of a permeability barrier is confirmed by measuring transepithelial electrical resistance (TEER), transepithelial transport of a control substance, and immunostaining of tight junctions. As an example, ~200&#160;nm polymer NCs are used, which carry a therapeutic cargo and are coated with an antibody that targets a cell-surface determinant. The antibody or therapeutic cargo is labeled with 125I for radioisotope tracing and labeled NCs are added to the upper chamber over the cell monolayer for varying periods of time. NCs associated to the cells and/or transported to the underlying chamber can be detected. Measurement of free 125I allows subtraction of the degraded fraction. The paracellular route is assessed by determining potential changes caused by NC transport to the barrier parameters described above. Transcellular transport is determined by addressing the effect of modulating endocytosis and transcytosis pathways.

Many therapeutic applications require safe and efficient transport of drug carriers and their cargoes across cellular barriers in the body. This article describes an adaptation of established methods to evaluate the rate and mechanism of transport of drug nanocarriers (NCs) across cellular barriers, such as the gastrointestinal (GI) epithelium.

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

Fibrous proteins display different sequences and structures that have been used for various applications in biomedical fields such as biosensors, nanomedicine, tissue regeneration, and drug delivery. Designing materials based on the molecular-scale interactions between these proteins will help generate new multifunctional protein alloy biomaterials with tunable properties. Such alloy material systems also provide advantages in comparison to traditional synthetic polymers due to the materials biodegradability, biocompatibility, and tenability in the body. This article used the protein blends of wild tussah silk (Antheraea pernyi) and domestic mulberry silk (Bombyx mori) as an example to provide useful protocols regarding these topics, including how to predict protein-protein interactions by computational methods, how to produce protein alloy solutions, how to verify alloy systems by thermal analysis, and how to fabricate variable alloy materials including optical materials with diffraction gratings, electric materials with circuits coatings, and pharmaceutical materials for drug release and delivery. These methods can provide important information for designing the next generation multifunctional biomaterials based on different protein alloys.

Blending is an efficient approach to generate biomaterials with a broad range of properties and combined features. By predicting the molecular interactions between different natural silk proteins, new silk-silk protein alloy platforms with tunable mechanical resiliency, electrical response, optical transparency, chemical processability, biodegradability, or thermal stability can be designed.

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

Sensitization of adenylyl cyclase (AC) signaling has been implicated in a variety of neuropsychiatric and neurologic disorders including substance abuse and Parkinson&#39;s disease. Acute activation of G&#945;i/o-linked receptors inhibits AC activity, whereas persistent activation of these receptors results in heterologous sensitization of AC and increased levels of intracellular cAMP. Previous studies have demonstrated that this enhancement of AC responsiveness is observed both in vitro and in vivo following the chronic activation of several types of G&#945;i/o-linked receptors including D2 dopamine and &#956; opioid receptors. Although heterologous sensitization of AC was first reported four decades ago, the mechanism(s) that underlie this phenomenon remain largely unknown. The lack of mechanistic data presumably reflects the complexity involved with this adaptive response, suggesting that nonbiased approaches could aid in identifying the molecular pathways involved in heterologous sensitization of AC. Previous studies have implicated kinase and Gb&#947; signaling as overlapping components that regulate the heterologous sensitization of AC. To identify unique and additional overlapping targets associated with sensitization of AC, the development and validation of a scalable cAMP sensitization assay is required for greater throughput. Previous approaches to study sensitization are generally cumbersome involving continuous cell culture maintenance as well as a complex methodology for measuring cAMP accumulation that involves multiple wash steps. Thus, the development of a robust cell-based assay that can be used for high throughput screening (HTS) in a 384&#160;well format would facilitate future studies. Using two D2 dopamine receptor cellular models (i.e.&#160;CHO-D2L and HEK-AC6/D2L), we have converted our 48-well sensitization assay (&#62;20 steps 4-5 days) to a five-step, single day assay in 384-well format. This new format is amenable to small molecule screening, and we demonstrate that this assay design can also be readily used for reverse transfection of siRNA in anticipation of targeted siRNA library screening.

Persistent activation of inhibitory G protein-coupled receptors results in sensitization of adenylyl cyclase signaling. To identify the essential molecular pathways, nonbiased approaches are necessary; however, this strategy requires the development of a scalable cell-based cAMP sensitization assay. Herein, we describe a sensitization assay for small molecule and siRNA screening.

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

Multimodal, molecular imaging allows the visualization of biological processes at cellular, subcellular, and molecular-level resolutions using multiple, complementary imaging techniques. These imaging agents facilitate the real-time assessment of pathways and mechanisms in vivo, which enhance both diagnostic and therapeutic efficacy. This article presents the protocol for the synthesis of biofunctionalized Prussian blue nanoparticles (PB NPs) - a novel class of agents for use in multimodal, molecular imaging applications. The imaging modalities incorporated in the nanoparticles, fluorescence imaging and magnetic resonance imaging (MRI), have complementary features. The PB NPs possess a core-shell design where gadolinium and manganese ions incorporated within the interstitial spaces of the PB lattice generate MRI contrast, both in T1 and T2-weighted sequences. The PB NPs are coated with fluorescent avidin using electrostatic self-assembly, which enables fluorescence imaging. The avidin-coated nanoparticles are modified with biotinylated ligands that confer molecular targeting capabilities to the nanoparticles. The stability and toxicity of the nanoparticles are measured, as well as their MRI relaxivities. The multimodal, molecular imaging capabilities of these biofunctionalized PB NPs are then demonstrated by using them for fluorescence imaging and molecular MRI in vitro.

This protocol describes the synthesis of biofunctionalized Prussian blue nanoparticles and their use as multimodal, molecular imaging agents. The nanoparticles have a core-shell design where gadolinium or manganese ions within the nanoparticle core generate MRI contrast. The biofunctional shell contains fluorophores for fluorescence imaging and targeting ligands for molecular targeting.

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, ...

Solid Lipid Nanoparticles (SLNs) for Intracellular Targeting Applications

Nanoparticle-based delivery vehicles have shown great promise for intracellular targeting applications, providing a mechanism to specifically alter cellular signaling and gene expression. In a previous investigation, the synthesis of ultra-small solid lipid nanoparticles (SLNs) for topical drug delivery and biomarker detection applications was demonstrated. SLNs are a well-studied example of a nanoparticle delivery system that has emerged as a promising drug delivery vehicle. In this study, SLNs were loaded with a fluorescent dye and used as a model to investigate particle-cell interactions. The phase inversion temperature (PIT) method was used for the synthesis of ultra-small populations of biocompatible nanoparticles. A 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenylphenyltetrazolium bromide (MTT) assay was utilized in order to establish appropriate dosing levels prior to the nanoparticle-cell interaction studies. Furthermore, primary human dermal fibroblasts and mouse dendritic cells were exposed to dye-loaded SLN over time and the interactions with respect to toxicity and particle uptake were characterized using fluorescence microscopy and flow cytometry. This study demonstrated that ultra-small SLNs, as a nanoparticle delivery system, are suitable for intracellular targeting of different cell types.

Solid Lipid Nanoparticles SLNs for Intracellular Targeting Applications

In this study, a method for synthesizing ultra-small populations of biocompatible nanoparticles was described, as well as several in vitro methods by which to assess their cellular interactions.

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

Human Mesenchymal Stromal Cells (hMSCs) are cultured in vitro with different media. Limits on their use in clinical settings, however, mainly depend on potential biohazard and inflammation risks exerted by xenogeneic nutrients for their culture. Human derivatives or recombinant materials are the first choice candidates to reduce these reactions. Therefore, culture supplements and materials of autologous origin represent the best nutrients and the safest products.
Here, we describe a new protocol for the isolation and culture of bone marrow hMSCs in autologous conditions &#8212;&#160;namely,&#160;patient-derived serum as a supplement for the culture medium and fibrin as a scaffold for hMSC administration. Indeed, hMSC/fibrin clot constructs could be extremely useful for several clinical applications. In particular, we focus on their use in orthopedic surgery, where the fibrin clot derived from the donor&#39;s own blood allowed effective cell delivery and nutrient/waste exchanges. To ensure optimal safety conditions, it is of the utmost importance to avoid the risks of hMSC transformation and tissue overgrowth. For these reasons, the approach described in this paper also indicates a minimally ex vivo hMSC expansion, to reduce cell senescence and morphologic changes, and short-term osteo-differentiation before implantation, to induce osteogenic lineage specification, thus decreasing the risk of subsequent uncontrolled proliferation.

Culturing human Mesenchymal Stromal Cells (hMSCs) with autologous serum, reduces the risks of rejection by xenogeneic material and other negative effects. It also allows for the recovery of a subset of mesodermal progenitors, which can deliver fresh hMSCs. Embedding hMSCs in an autologous fibrin clot enables easy handling and effective surgical implantation.

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

Gold nanoparticles (AuNPs) have been used extensively in medical research due to their size, biocompatibility, and modifiable surface. Specific targeting and drug delivery are some of the applications of these AuNPs, but endothelial extracellular matrices&#39; defensive properties hamper particle uptake. To address this issue, we describe a synthesis method for ultrasmall gold nanoparticles to improve vascular delivery, with customizable functional groups and polymer lengths for further adjustments. The protocol yields 2.5 nm AuNPs that are capped with tetrakis(hydroxymethyl)phosphonium chloride (THPC). The replacement of THPC with hetero-functional polyethylene glycol (PEG) on the surface of the AuNP increases the hydrodynamic radius to 10.5 nm while providing various functional groups on the surface. The last part of the protocol includes an optional addition of a fluorophore to allow the AuNPs to be visualized under fluorescence to track nanoparticle uptake. Dialysis and lyophilization were used to purify and isolate the AuNPs. These fluorescent nanoparticles can be visualized in both in vitro and in vivo experiments due to the biocompatible PEG coating and fluorescent probes. Additionally, the size range of these nanoparticles render them an ideal candidate for probing the glycocalyx without disrupting normal vasculature function, which may lead to improved delivery and therapeutics.

We describe a method of synthesizing biocompatible 10-nm gold nanoparticles, functionalized by coating poly-ethylene glycol onto the surface. These particles can be used in vitro and in vivo for delivering therapeutics to nanoscale cellular and extracellular spaces that are difficult to access with conventional nanoparticle sizes.

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 ...

Comprehensive Evaluation of the Effectiveness and Safety of Placenta-Targeted Drug Delivery Using Three Complementary Methods

No effective treatments currently exist for placenta-associated pregnancy complications, and developing strategies for the targeted delivery of drugs to the placenta while minimizing fetal and maternal side effects remains challenging. Targeted nanoparticle carriers provide new opportunities to treat placental disorders. We recently demonstrated that a synthetic placental chondroitin sulfate A binding peptide (plCSA-BP) could be used to guide nanoparticles to deliver drugs to the placenta. In this protocol, we describe in detail a system for assessing the efficiency of drug delivery to the placenta by plCSA-BP that employs three separate methods used in combination: in vivo imaging, high-frequency ultrasound (HFUS), and high-performance liquid chromatography (HPLC). Using in vivo imaging, plCSA-BP-guided nanoparticles were visualized in the placentas of live animals, while HFUS and HPLC demonstrated that plCSA-BP-conjugated nanoparticles efficiently and specifically delivered methotrexate to the placenta. Thus, a combination of these methods can be used as an effective tool for the targeted delivery of drugs to the placenta and development of new treatment strategies for several pregnancy complications.

We describe a system that utilizes three methods to evaluate the safety and effectiveness of placenta-targeted drug delivery: in vivo imaging to monitor nanoparticle accumulation, high-frequency ultrasound to monitor placental and fetal development, and HPLC to quantify drug delivery to tissue.

3 보완 방법을 사용 하 여 태 반 대상 약물 전달의 안전과 효율성의 종합 평가

No effective treatments currently exist for placenta-associated pregnancy complications, and developing strategies for the targeted delivery of drugs to ...