Name: preparation and characterization of sdf-1α-chitosan-dextran sulfate nanoparticles
Uploaded: 2015-01-22T14:00:00.000+00:00
Duration: 12 min
Description: Watch this Scientific Journal Video about Preparation and Characterization of SDF-1ÃŽÂ±-Chitosan-Dextran Sulfate Nanoparticles at JoVE.com

HL671795, HL048743 및 HL108630 :이 작품은 NIH 보조금에 의해 지원되었다.

SDF-1α 글리 칸 나노 입자의 1. 준비 생체 내 전달의 목적으로 인해, 제조에 사용 된 모든 용기, 피펫 팁을 소독. <ol><li> 초순수에서 다음 원액을 준비 : 1 % 덱스 트란 황산; 1 M의 NaOH (PES 막으로 멸균 여과); 0.1 % (0.8 연속적 0.22 μm의 필터를 통해 필터 및 그 후에 NaOH로 pH가 5.5로 조정), 0.2 % 빙초산 키토산; 0.1 M ZnSO 4; 15 % 만니톨; 0.92 ㎎ / ㎖ SDF-1α (80 ° C에서 분취 액에 저장되고, 4 ℃에서 보관 일단 해동 C). </li><li> 0.22 μm의 필터 멤브레인을 통해 재고 솔루션을 소독. 리 물러의 amebocyte 용 해물 (LAL) 젤 응고 분석에 준비된 용액에 독소 수준을 평가합니다. 수준이 &lt;0.06 EU / ㎖ 있는지 확인합니다. </li><li> 자기 교반 막대를 함유하는 1.5 mL 유리 바이알에 0.18 ml의 초순수를 추가한다. 800 rpm으로 교반 속도를 설정합니다. 0.1 ml의 1 %를 추가덱스 트란 설페이트 및 2 분 동안 교반한다. 0.08 mg을 SDF-1α (0.92 Mg를 0.087 ㎖ / ml의 SDF-1α 용액)를 첨가하고, 20 분 동안 교반한다. </li><li> 0.33 ml의 0.1 % 키토산 적가하고 추가 5 분 동안 교반한다. 변경 볶음 최대 속도 (1 분 이상) 0.1 ML의 주사기로 천천히 0.1 ml의 0.1 M ZnSO 4를 추가합니다. </li><li> 800 rpm으로 교반 속도를 반환하고, 30 분 동안 교반한다. 0.4 ml의 15 % 만니톨을 추가하고 5 분 동안 교반한다. 1.5ml의 미세 원심 분리 튜브에 반응 혼합물을 전송. 10 분 동안 4 ° C에서 16,000 XG에 원심 분리기. 주 : 만니톨의 존재는 원심 분리 후 입자의 부유를 용이하게한다. 펠릿의 소형화에 따라 만니톨의 농도는 0 %에서 5 %로 변화 될 수있다. 원심 분리 후, 각 입자의 완전 재현 탁하여 최종 현탁액 응집체를 피하기 위해 중요하다. </li><li> 기음 뜨는 천천히 액체의 마지막 한 방울을 제거하기 위해 피펫을 사용합니다. 0.2 ml의 5 % 만니톨을 추가합니다. 0.5 ml의 펠렛을 일시 중단, 29 G의 옛 성은DLE 주사기. 1 ml의 5 % 만니톨을 추가합니다. 15 분 동안 16,000 XG에 원심 분리기. </li><li> 단계를 반복 1.6. </li><li> 뜨는을 기음. 0.2 ml의 5 % 만니톨 펠렛을 재현 탁. 4 ° C에서 입자 현탁액을 저장합니다. 주 : 입자 현탁액도 저장 -80 ° C에서 동결 또는 동결 건조 될 수있다. 서스펜션 포함 5 % 만니톨 동결 및 해동 후 또는 동결 건조 및 수분 보충 후 입자의 응집을 방지하기 위해 필수적이다. 만니톨 보존 후의 현탁액을 원심 분리에 의해 제거 될 수있다. </li></ol> 입자 크기와 제타 전위 2. 측정 입경 및 제타 전위를 소재리스트에 표시된 입자 분석기로 각각 동적 광산란 및 전기 영동 광 산란에 의해 분석된다. <ol><li> 축적 시간 : 다음 입자 크기 측정을위한 파라미터를 설정 (70); 반복 시간 : 4; 온도 : 25 ° C;희석제 : 물; 강도 조절 : 자동. </li><li> 물 (10 배 희석)와 SDF-1α-DS-CS 샘플을 희석. 이러한 에펜 도르프 베트로 일회용 UV 큐벳에 시료 100 ㎕를 넣습니다. 셀 홀더에 큐벳을 삽입합니다. &quot;최적&quot;(~ 10,000 CPS)에 도달하기 위해 강도 조정 기다립니다. 데이터 수집을 시작합니다. </li><li> 측정이 완료되면, 직경 (㎚) 및 다 분산도 지수 cumulants 결과를 기록한다. 4 반복 판독 각각으로부터 얻어진 결과를 평균치와 표준 편차를 계산한다. </li><li> 로드 제타 전위 측정 플로우 셀에 10 배 희석 한 시료의 입자 500 μL. 누적 시간 : 다음 측정을위한 매개 변수를 설정합니다 (10); 반복 시간 : 5; 온도 : 25 ° C; 희석제 : 물; 강도 조절 : 자동. 제타 전위 (MV)의 결과를 기록한다. (5) 판독 반복의 각각으로부터 얻어진 결과를 평균 및 표준을 계산 deviatioN. </li></ol> 입자의 SDF-1α의 3 정량화 <ol><li> 1.3 램 믈리 샘플 버퍼에 0.01, 0.02, 0.03, 0.04, 0.05 ㎎ / ㎖의 농도 자유 양식 SDF-1α를 희석. 40 μL의 1.3 배 램 믈리 샘플 버퍼 6 μL SDF-1α-DS-CS 샘플을 희석. (나눈다. 작은 분취 량으로 원액을 0.25 M 트리스 -HCl, pH가 7.5, 8 % SDS, 40 % 글리세롤, 0.05 ㎎ / 브로 모 페놀 블루,는 ml 8 % 2- 머 캅토 에탄올을 함유 4X 램 믈리 스톡 완충액을 준비하고 -20 ℃로 유지 단일 사용하기 위해 C를 °.) </li><li> 10 분 동안 100 ℃에서 샘플을 가열한다. 소용돌이 입자를 완전히 해리시키기 위해 10 분 동안 가열 시간 배 샘플 (최대 속도로 10 초 동안 각각). 2 분 동안 RT에 샘플을 냉각. 0.5 분 10,000 XG에 원심 분리기는 응축수를 가져올 수 있습니다. 소용돌이는 다시 잘 섞는다. 참고 :이 시점에서, 시료 용액이 눈에 보이는 침전물 존재와 명확해야한다. </li><li> 로드 10# 181; L 샘플 / 웰에 4-20 %의 SDS 젤. 20 분 동안 200 V의 전기를 실행합니다. 코마시 블루 단백질 얼룩 젤을 얼룩. </li><li> 분자 이미 저와 농도계 분석 소프트웨어를 사용하여 SDF-1α의 단백질 밴드 밀도를 검사합니다. 무료 SDF-1α 표준 구성된 표준 곡선에 대한 입자 SDF-1α의 양을 계산합니다. </li></ol> 생체 외 자료 분석 4. <ol><li> 1 칼슘과 마그네슘 (D-PBS)없이 둘 베코 인산염 완충 생리 식염수와 SDF-1α 글리 칸 입자를 혼합 : 1의 비율 (v / v)로. </li><li> 1.5 ml의 튜브에 0.05 ㎖의 분취 량으로 입자 현탁액을 나눕니다. 튜브의 바닥에 샘플을로드합니다. 기포를 도입 또는 액체의 표면을 방해하지 않도록. 파라 필름으로 튜브의 상단을 밀봉합니다. 주 : 이때, 액체는 튜브 믹서 후속 상하 회전시 하단에 남아있을 것이다. </li><li> 37에서 튜브를 회전# 176; C 회전 마이크로 튜브 믹서에. 지정된 시간에 튜브에서 분취 액을 제거하고 즉시 4 ° C에서 10 분 동안 16,000 XG에서 원심 분리. </li><li> 상층 액과 펠렛을 분리, 0.05 ml의 D-PBS로 펠렛을 재구성. -20 ° C에서 샘플을 저장합니다. 모든 샘플이 수집 된 후 전술 한 바와 같이, SDS 겔상에서 상등액과 펠릿을 조사. </li></ol> 5. 마이그레이션 분석 이 분석은 SDF-1α의 화성 활동을 측정합니다. 세포 표면의 수용체 (CXCR4)와 SDF-1α의 상호 작용은 SDF-1α의 그라데이션으로 세포의 이동을 야기한다. 이 분석에서, 세포는 낮은 잘으로하고 SDF-1α 솔루션 (낮은 우물에서 반투과성 막에 의해 분리) 상부 우물에로드됩니다. <ol><li> SDF-1α 희석 (유리 또는 입자 - 결합 된 형태) -3- FO에서 이주 완충액 (0.5 % 소 혈청 알부민을 함유하는 RPMI-1640 배지)100, 33, 11, 3.7, 1.2, 0.41, 0.14, 및 0.05 ng를 / ㎖의 최종 농도로 용액을 희석 LD. </li><li> 24 웰 플레이트에 웰에 희석 된 SDF-1α 솔루션 또는 마이그레이션 버퍼 만 (대조군)의 0.6 ML을 추가합니다. 잘 (입력 셀 제어)에 0.57 ml의 마이그레이션 버퍼를 추가합니다. 30 분 동안 37 ℃에서 배양한다. 이러한 낮은 잘 위에없이 Transwell (기공 크기 5 μm의 직경 6.5 mm)로 투과 세포 배양 삽입을 놓습니다. 트랜스 웰에 0.1 ml의 된 Jurkat 세포 (5 × 10 5)를 삽입로드. 잘 입력 세포 제어에 직접 0.03 ml의 세포를로드합니다. 5 % CO 2 인큐베이터에서 2 시간 동안 37 ℃에서 플레이트를 인큐베이션. </li><li> 트랜스 웰 인서트를 제거합니다. 4 ml의 폴리스티렌 튜브에 낮은 우물로 마이그레이션 한 세포를 전송합니다. 유량 계측기와 마이그레이션 된 세포를 계산합니다. </li><li> negat에 숫자의 감산 입력 후 세포 수 (X 웰 100/30 입력 셀 컨트롤 세포 수)의 백분율로 계산 이주컨트롤 (에만 이동 완충액을 함유하는 웰에 마이그레이션 셀) 필자. </li></ol>

<ol>
	<li>Delair, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Colloidal+polyelectrolyte+complexes+of+chitosan+and+dextran+sulfate+towards+versatile+nanocarriers+of+bioactive+molecules.">Colloidal polyelectrolyte complexes of chitosan and dextran sulfate towards versatile nanocarriers of bioactive molecules.</a> Eur J Pharm Biopharm. 78 (1), 10-18 (2011).</li><li>Morachis, J. M., Mahmoud, E. A., Almutairi, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Physical+and+chemical+strategies+for+therapeutic+delivery+by+using+polymeric+nanoparticles.">Physical and chemical strategies for therapeutic delivery by using polymeric nanoparticles.</a> Pharmacol Rev. 64 (3), 505-519 (2012).</li><li>Yue, Z. 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=Surface+charge+affects+cellular+uptake+and+intracellular+trafficking+of+chitosan-based+nanoparticles.">Surface charge affects cellular uptake and intracellular trafficking of chitosan-based nanoparticles.</a> Biomacromolecules. 12 (7), 2440-2446 (2011).</li><li>Ghadge, S. K., Muhlstedt, S., Ozcelik, C., Bader, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=SDF-1alpha+as+a+therapeutic+stem+cell+homing+factor+in+myocardial+infarction.">SDF-1alpha as a therapeutic stem cell homing factor in myocardial infarction.</a> Pharmacol Ther. 129 (1), 97-108 (2011).</li><li>Sharma, M., Afrin, F., Satija, N., Tripathi, R. P., Gangenahalli, G. U. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Stromal-derived+factor-1/CXCR4+signaling:+indispensable+role+in+homing+and+engraftment+of+hematopoietic+stem+cells+in+bone+marrow.">Stromal-derived factor-1/CXCR4 signaling: indispensable role in homing and engraftment of hematopoietic stem cells in bone marrow.</a> Stem Cells Dev. 20 (6), 933-946 (2011).</li><li>Sadir, R., Baleux, F., Grosdidier, A., Imberty, A., Lortat-Jacob, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Characterization+of+the+stromal+cell-derived+factor-1alpha-heparin+complex.">Characterization of the stromal cell-derived factor-1alpha-heparin complex.</a> J Biol Chem. 276 (11), 8288-8296 (2001).</li><li>Amara, A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Stromal+cell-derived+factor-1alpha+associates+with+heparan+sulfates+through+the+first+beta-strand+of+the+chemokine.">Stromal cell-derived factor-1alpha associates with heparan sulfates through the first beta-strand of the chemokine.</a> J Biol Chem. 274 (34), 23916-23925 (1999).</li><li>Sadir, R., Imberty, A., Baleux, F., Lortat-Jacob, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Heparan+sulfate/heparin+oligosaccharides+protect+stromal+cell-derived+factor-1+(SDF-1)/CXCL12+against+proteolysis+induced+by+CD26/dipeptidyl+peptidase+IV.">Heparan sulfate/heparin oligosaccharides protect stromal cell-derived factor-1 (SDF-1)/CXCL12 against proteolysis induced by CD26/dipeptidyl peptidase IV.</a> J Biol Chem. 279 (42), 43854-43860 (1074).</li><li>Yin, T., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=SDF-1alpha+in+glycan+nanoparticles+exhibits+full+activity+and+reduces+pulmonary+hypertension+in+rats.">SDF-1alpha in glycan nanoparticles exhibits full activity and reduces pulmonary hypertension in rats.</a> Biomacromolecules. 14 (11), 4009-4020 (2013).</li><li>Huang, M., Vitharana, S. N., Peek, L. J., Coop, T., Berkland, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Polyelectrolyte+complexes+stabilize+and+controllably+release+vascular+endothelial+growth+factor.">Polyelectrolyte complexes stabilize and controllably release vascular endothelial growth factor.</a> Biomacromolecules. 8 (5), 1607-1614 (2007).</li><li>McCall, R. L., Sirianni, R. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=PLGA+nanoparticles+formed+by+single-+or+double-emulsion+with+vitamin+E-TPGS.">PLGA nanoparticles formed by single- or double-emulsion with vitamin E-TPGS.</a> J Vis Exp. (82), 51015 (2013).</li><li>Carrillo-Conde, B. R., Roychoudhury, R., Chavez-Santoscoy, A. V., Narasimhan, B., Pohl, N. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=High-throughput+synthesis+of+carbohydrates+and+functionalization+of+polyanhydride+nanoparticles.">High-throughput synthesis of carbohydrates and functionalization of polyanhydride nanoparticles.</a> J Vis Exp. (65), 3967 (2012).</li><li>Xu, J., Amiji, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Therapeutic+gene+delivery+and+transfection+in+human+pancreatic+cancer+cells+using+epidermal+growth+factor+receptor-targeted+gelatin+nanoparticles.">Therapeutic gene delivery and transfection in human pancreatic cancer cells using epidermal growth factor receptor-targeted gelatin nanoparticles.</a> J Vis Exp. (59), e3612 (2012).</li><li>Lauten, E. H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Nanoglycan+complex+formulation+extends+VEGF+retention+time+in+the+lung.">Nanoglycan complex formulation extends VEGF retention time in the lung.</a> Biomacromolecules. 11 (7), 1863-1872 (2010).</li><li>Schatz, C., Domard, A., Viton, C., Pichot, C., Delair, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Versatile+and+efficient+formation+of+colloids+of+biopolymer-based+polyelectrolyte+complexes.">Versatile and efficient formation of colloids of biopolymer-based polyelectrolyte complexes.</a> Biomacromolecules. 5 (5), 1882-1892 (2004).</li></ol>

The authors declare that they have no competing financial interests.

전술 한 바와 같이, DS-CS 나노 입자 폴리 음이온 (DS)과 폴리 양이온 (CS) 분자간 전하 중성화를 통해 형성된다. 전하 상호 작용은 분자 충돌시 쉽게 발생하기 때문에, 혼합 중에 중합체 용액 및 교반 속도의 농도는 생성 된 입자의 크기가 중요하다. 일반적인 경향은 더 DS 및 CS 솔루션 (15) 및 작은 입자의 높은 교반 속도 결과를 희석하고 있다는 것이다. SDF-1α 글리 칸의 ?...

덱스 트란 황산 (DS)과 키토산 (CS)는 (DS)에 여러 대체 음으로 하전 된 황산 그룹과 폴리 사카 라이드, 또는 양전하 아민 그룹입니다 (CS를 탈 아세틸). 수용액에 혼합 할 때, 두 가지 다당류는 정전 기적 상호 작용을 통해 고분자 전해질 복합체를 형성한다. 얻어진 착물 수용액 (침전물)로부터 상분리한다 큰 응집체 또는 수분 산성 (콜로이드)의 소형 입자를 형성 할 수있다. 이러한 결과에 기여하는 구체적인 조건은 광범위하게 연구되어 왔으며, 최근의 리뷰에서 요약 한 상세히 도시되어있다. 이러한 조건 중에서, 수분 산성 입자를 제조하기위한 두 가지 기본적인 요구 반대로 대전 된 중합체는 1)이 크게 상이한 몰 질량을 가져야이고; 2) 비 화학량 론적 비율로 혼합 될 수있다. 이러한 조건에 의해 생성 된 전하 전하 중성 복합체 중합체 세그먼트를 허용중화는 분리 및 입자의 코어를 형성하고, 여분의 중합체는 외부 쉘을 형성하기 위해 하나한다. 이 프로토콜에 기재된 당쇄 입자는 폐 전달을 위해 의도되고, 음으로 하전 된 순으로 설계되고, 나노 사이즈의. 음극 표면 전하 입자 2,3의 세포 흡수의 가능성을 감소시킨다. 나노 사이즈의 입자는 원위기도 통로를 통해 용이하게한다. 이 목표를 달성하기 위해, 본 제조에 사용되는 DS의 양은 CS 초과 (중량비 3 : 1); 고 분자량은 DS (중량 평균 MW 500,000)와 저분자 CS (MW kDa의 범위 50-190, 75~85% 탈 아세틸)이 사용된다. SDF-1α는 화성 활동을 통해 원점 복귀 기능을 발휘 줄기 세포 호밍 ​​요인이다. SDF-1α는 원점과 골수 조혈 줄기 세포의 유지에 중요한 역할을하고 proge의 모집부상 수리 4,5의 말초 조직에 nitor 세포. SDF-1α는 단백질이 프로테아제 (CD26 / DPPIV) 불활로부터 보호, 헤파린 / 헤파 란 설페이트, 형태 다이머에 결합하고, 세포 표면 수용체를 통해 표적 세포와 상호 작용할 수 있도록 그 단백질 서열에 헤파린 결합 부위를 갖는다 6-8. DS는 헤파린 / 헤파 란 황산과 유사한 구조적 특성이있다; 따라서, DS에 대한 SDF-1α의 결합은 천연 고분자 리간드의 그것과 유사하다. 다음 프로토콜에서 우리는 SDF-1α-DS-CS 나노 입자의 제조 방법을 설명합니다. 절차는 이전에 9를 연구 된 제제 중 하나를 나타냅니다. 프로토콜은 원래 VEGF-DS-CS 나노 입자 (10)의 조사에서 적합하다. 소규모 제조 용이 동일한 스톡 용액 및 제조 조건으로 확장 할 수있는 기술된다. 제조 후, 입자 b를 특징Y는 크기, 제타 전위, SDF-1α 통합의 정도, 체외 방출 시간 및 통합 SDF-1α의 활동을 조사.

<table><tbody><tr><td>Dextran sulfate</td><td>Fisher</td><td>BP1585-100</td><td></td></tr><tr><td>Chitosan, low molecular weight&amp;nbsp;</td><td>Sigma</td><td>448869</td><td></td></tr><tr><td>Zinc sulfate heptahydrate</td><td>Sigma</td><td>204986</td><td></td></tr><tr><td>D-Mannitol</td><td>Sigma</td><td>M9546</td><td></td></tr><tr><td>UltraPure water&amp;nbsp;</td><td>Invitrogen&amp;nbsp;</td><td>10977-023</td><td></td></tr><tr><td>SDF-1&amp;alpha;</td><td>Prepared according to reference 8.</td><td></td><td></td></tr><tr><td>Syringe filter, PES membrane 0.22&amp;nbsp;&amp;#956;m</td><td>Millipore</td><td>SLGP033RS</td><td></td></tr><tr><td>Magnetic Micro Stirring Bars (2&amp;nbsp;x&amp;nbsp;7&amp;nbsp;mm)</td><td>Fisher&amp;nbsp;</td><td>14-513-63</td><td></td></tr><tr><td>Glass vial Kit; SUN-SRi</td><td>Fisher&amp;nbsp;</td><td>14-823-182</td><td></td></tr><tr><td>Delsa Nano C Particle Analyzer&amp;nbsp;</td><td>Backman Coulter</td><td></td><td></td></tr><tr><td>Eppendorf UVette Cuvets</td><td>Eppendorf</td><td>952010069</td><td></td></tr><tr><td>4&amp;ndash;20% Mini-PROTEAN TGX Gel</td><td>Bio-Rad</td><td>456-1096</td><td></td></tr><tr><td>GelCode Blue Safe Protein Stain</td><td>Fisher&amp;nbsp;</td><td>PI-24592</td><td></td></tr><tr><td>Molecular Imager VersaDoc MP 4000 System</td><td>BioRad</td><td>170-8640</td><td></td></tr><tr><td>Corning Transwell Permeable Supports</td><td>Corning</td><td>3421</td><td></td></tr><tr><td>Accuri C6 Flow Cytometer</td><td>BD Biosciences</td><td></td><td></td></tr><tr><td>Dulbecco&amp;rsquo;s phosphate buffered saline&amp;nbsp;</td><td>Sigma</td><td>D8537</td><td></td></tr><tr><td>Pyrogent plus kit</td><td>Fisher</td><td>NC9753738</td><td></td></tr></tbody></table>

preparation and characterization of sdf-1&#945;-chitosan-dextran sulfate nanoparticles

Chitosan (CS) and dextran sulfate (DS) are charged polysaccharides (glycans), which form polyelectrolyte complex-based nanoparticles when mixed under appropriate conditions. The glycan nanoparticles are useful carriers for protein factors, which facilitate the in vivo delivery of the proteins and sustain their retention in the targeted tissue. The glycan polyelectrolyte complexes are also ideal for protein delivery, as the incorporation is carried out in aqueous solution, which reduces the likelihood of inactivation of the proteins. Proteins with a heparin-binding site adhere to dextran sulfate readily, and are, in turn, stabilized by the binding. These particles are also less inflammatory and toxic when delivered in vivo. In the protocol described below, SDF-1&#945; (Stromal cell-derived factor-1&#945;), a stem cell homing factor, is first mixed and incubated with dextran sulfate. Chitosan is added to the mixture to form polyelectrolyte complexes, followed by zinc sulfate to stabilize the complexes with zinc bridges. The resultant SDF-1&#945;-DS-CS particles are measured for size (diameter) and surface charge (zeta potential). The amount of the incorporated SDF-1&#945; is determined, followed by measurements of its in vitro release rate and its chemotactic activity in a particle-bound form.

제조 된 SDF-1α-DS-CS 입자의 크기 및 제타 전위는 입자 분석기로 측정된다. 1 사이즈의 측정 분석을 보여준다. 네 개의 반복 된 측정에 의한 결과로부터 cumulants, SDF-1α-DS-CS 입자의 유체 역학적 평균 직경은 661 ± 8.2 (㎚)이고 분산도는 0.23 ± 0.02이다. 제타 전위 측정의 결과는도 2에 도시되어있다. 다섯 반복 측정에서, SDF-1α-DS-CS 입자의 제타 전위는 -24.8 ± 0.5 MV이?...

Watch this Scientific Journal Video about Preparation and Characterization of SDF-1ÃŽÂ±-Chitosan-Dextran Sulfate Nanoparticles at JoVE.com

Preparation and Characterization of SDF-1&#945;-Chitosan-Dextran Sulfate Nanoparticles

The objective of this protocol is to incorporate SDF-1&#945;, a stem cell homing factor, into dextran sulfate-chitosan nanoparticles. The resultant particles are measured for their size and zeta potential, as well as the content, activity, and in vitro release rate of SDF-1&#945; from the nanoparticles.

SDF-1α-키토산 덱스 트란 황산 나노 입자의 제조 및 특성

Chitosan (CS) and dextran sulfate (DS) are charged polysaccharides (glycans), which form polyelectrolyte complex-based nanoparticles when mixed under ...

preparation-characterization-sdf-1-chitosan-dextran-sulfate

Universidad San Sebastian

Research

JoVE Journal

Chemistry

12.3K 조회수.  Brigham and Women's Hospital and Harvard Medical School. The objective of this protocol is to incorporate SDF-1α, a stem cell homing factor, into dextran sulfate-chitosan nanoparticles. The resultant particles are measured for their size and zeta potential, as well as the content, activity, and in vitro release rate of SDF-1α from the nanoparticles.

비디오: Preparation and Characterization of SDF-1α-Chitosan-Dextran Sulfate Nanoparticles

Constructing a Collagen Hydrogel for the Delivery of Stem Cell-loaded Chitosan Microspheres

Multipotent stem cells have been shown to be extremely useful in the field of regenerative medicine1-3. However, in order to use these cells effectively for tissue regeneration, a number of variables must be taken into account. These variables include: the total volume and surface area of the implantation site, the mechanical properties of the tissue and the tissue microenvironment, which includes the amount of vascularization and the components of the extracellular matrix. Therefore, the materials being used to deliver these cells must be biocompatible with a defined chemical composition while maintaining a mechanical strength that mimics the host tissue. These materials must also be permeable to oxygen and nutrients to provide a favorable microenvironment for cells to attach and proliferate. Chitosan, a cationic polysaccharide with excellent biocompatibility, can be easily chemically modified and has a high affinity to bind with in vivo macromolecules4-5. Chitosan mimics the glycosaminoglycan portion of the extracellular matrix, enabling it to function as a substrate for cell adhesion, migration and proliferation. In this study we utilize chitosan in the form of microspheres to deliver adipose-derived stem cells (ASC) into a collagen based three-dimensional scaffold6. An ideal cell-to-microsphere ratio was determined with respect to incubation time and cell density to achieve maximum number of cells that could be loaded. Once ASC are seeded onto the chitosan microspheres (CSM), they are embedded in a collagen scaffold and can be maintained in culture for extended periods. In summary, this study provides a method to precisely deliver stem cells within a three dimensional biomaterial scaffold.

A major hurdle in current stem cell therapies is determining the most effective method to deliver these cells to host tissues. Here, we describe a chitosan-based delivery method that is efficient and simple in approach, while allowing adipose-derived stem cells to maintain their multipotency.

줄기 세포 - 로드된 키토산 Microspheres의 배달을 위해 콜라겐 히드로겔을 구축

Multipotent stem cells have been shown to be extremely useful in the field of regenerative medicine1-3. However, in order to use these cells effectively ...

연구

생체공학

Preparation and Characterization of Individual and Multi-drug Loaded Physically Entrapped Polymeric Micelles

Amphiphilic block copolymers like polyethyleneglycol-block-polylactic acid (PEG-b-PLA) can self-assemble into micelles above their critical micellar concentration forming hydrophobic cores surrounded by hydrophilic shells in aqueous environments. The core of these micelles can be utilized to load hydrophobic, poorly water soluble drugs like docetaxel (DTX) and everolimus (EVR). Systematic characterization of the micelle structure and drug loading capabilities are important before in vitro and in vivo studies can be conducted. The goal of the protocol described herein is to provide the necessary characterization steps to achieve standardized micellar products. DTX and EVR have intrinsic solubilities of 1.9 and 9.6 &#181;g/ml respectively Preparation of these micelles can be achieved through solvent casting which increases the aqueous solubility of DTX and EVR to 1.86 and 1.85 mg/ml, respectively. Drug stability in micelles evaluated at room temperature over 48 hr indicates that 97% or more of the drugs are retained in solution. Micelle size was assessed using dynamic light scattering and indicated that the size of these micelles was below 50 nm and depended on the molecular weight of the polymer. Drug release from the micelles was assessed using dialysis under sink conditions at pH 7.4 at 37 oC over 48 hr. Curve fitting results indicate that drug release is driven by a first order process indicating that it is diffusion driven.

The goal of this protocol is to describe the preparation and characterization of physically entrapped, poorly water soluble drugs in micellar drug delivery systems composed of amphiphilic block copolymers.

준비 및 개인의 특성과 다중 약물은 물리적으로 고분자 미셀을 포획로드

Amphiphilic block copolymers like polyethyleneglycol-block-polylactic acid (PEG-b-PLA) can self-assemble into micelles above their critical micellar ...

화학

Engineering a Bilayered Hydrogel to Control ASC Differentiation

Natural polymers over the years have gained more importance because of their host biocompatibility and ability to interact with cells in vitro and in vivo. An area of research that holds promise in regenerative medicine is the combinatorial use of novel biomaterials and stem cells. A fundamental strategy in the field of tissue engineering is the use of three-dimensional scaffold (e.g., decellularized extracellular matrix, hydrogels, micro/nano particles) for directing cell function. This technology has evolved from the discovery that cells need a substrate upon which they can adhere, proliferate, and express their differentiated cellular phenotype and function 2-3. More recently, it has also been determined that cells not only use these substrates for adherence, but also interact and take cues from the matrix substrate (e.g., extracellular matrix, ECM)4. Therefore, the cells and scaffolds have a reciprocal connection that serves to control tissue development, organization, and ultimate function. Adipose-derived stem cells (ASCs) are mesenchymal, non-hematopoetic stem cells present in adipose tissue that can exhibit multi-lineage differentiation and serve as a readily available source of cells (i.e. pre-vascular endothelia and pericytes). Our hypothesis is that adipose-derived stem cells can be directed toward differing phenotypes simultaneously by simply co-culturing them in bilayered matrices1. Our laboratory is focused on dermal wound healing. To this end, we created a single composite matrix from the natural biomaterials, fibrin, collagen, and chitosan that can mimic the characteristics and functions of a dermal-specific wound healing ECM environment.

This protocol focuses on utilizing the inherent ability of stem cells to take cue from their surrounding extracellular matrix and be induced to differentiate into multiple phenotypes. This methods manuscript extends our description and characterization of a model utilizing a bilayered hydrogel, composed of PEG-fibrin and collagen, to simultaneously co-differentiate adipose-derived stem cells1.

ASC의 분화를 제어하는​​ Bilayered 히드로겔 꾸민

Natural polymers over the years have gained more importance because of their host biocompatibility and ability to interact with cells in vitro and in ...

Synthesis and Characterization of Placental Chondroitin Sulfate A (plCSA)-Targeting Lipid-Polymer Nanoparticles

An effective cancer therapeutic method reduces and eliminates tumors with minimal systemic toxicity. Actively targeting nanoparticles offer a promising approach to cancer therapy. The glycosaminoglycan placental chondroitin sulfate A (plCSA) is expressed on a wide range of cancer cells and placental trophoblasts, and malarial protein VAR2CSA can specifically bind to plCSA. A reported placental chondroitin sulfate A binding peptide (plCSA-BP), derived from malarial protein VAR2CSA, can also specifically bind to plCSA on cancer cells and placental trophoblasts. Hence, plCSA-BP-conjugated nanoparticles could be used as a tool for targeted drug delivery to human cancers and placental trophoblasts. In this protocol, we describe a method to synthesize plCSA-BP-conjugated lipid-polymer nanoparticles loaded with doxorubicin (plCSA-DNPs); the method consists of a single sonication step and bioconjugate techniques. In addition, several methods for characterizing plCSA-DNPs, including determining their physicochemical properties and cellular uptake by placental choriocarcinoma (JEG3) cells, are described.

Synthesis and Characterization of Placental Chondroitin Sulfate A plCSA-Targeting Lipid-Polymer Nanoparticles

Here, we present a protocol for the synthesis of placental chondroitin sulfate A binding peptide (plCSA-BP)-conjugated lipid-polymer nanoparticles via single-step sonication and bioconjugate techniques. These particles constitute a novel tool for the targeted delivery of therapeutics to most human tumors and placental trophoblasts to treat cancers and placental disorders.

합성 및 Placental 콘의 황산 (plCSA)-지질 타겟팅-고분자 나노 입자

An effective cancer therapeutic method reduces and eliminates tumors with minimal systemic toxicity. Actively targeting nanoparticles offer a promising ...

암 연구학

Preparation and Photoacoustic Analysis of Cellular Vehicles Containing Gold Nanorods

Gold nanorods are attractive for a range of biomedical applications, such as the photothermal ablation and the photoacoustic imaging of cancer, thanks to their intense optical absorbance in the near-infrared window, low cytotoxicity and potential to home into tumors. However, their delivery to tumors still remains an issue. An innovative approach consists of the exploitation of the tropism of tumor-associated macrophages that may be loaded with gold nanorods in vitro. Here, we describe the preparation and the photoacoustic inspection of cellular vehicles containing gold nanorods. PEGylated gold nanorods are modified with quaternary ammonium compounds, in order to achieve a cationic profile. On contact with murine macrophages in ordinary Petri dishes, these particles are found to undergo massive uptake into endocytic vesicles. Then these cells are embedded in biopolymeric hydrogels, which are used to verify that the stability of photoacoustic conversion of the particles is retained in their inclusion into cellular vehicles. We are confident that these results may provide new inspiration for the development of novel strategies to deliver plasmonic particles to tumors.

We show the preparation and address the feasibility of cellular vehicles containing gold nanorods for the photoacoustic imaging of cancer.

금 나노 막대를 포함하는 세포 차량의 제조 및 광 음향 분석

Gold nanorods are attractive for a range of biomedical applications, such as the photothermal ablation and the photoacoustic imaging of cancer, thanks to ...

Preparation and Characterization of Novel HDL-mimicking Nanoparticles for Nerve Growth Factor Encapsulation

The objective of this article is to introduce preparation and characterization methods for nerve growth factor (NGF)-loaded, high-density, lipoprotein (HDL)-mimicking nanoparticles (NPs). HDLs are endogenous NPs and have been explored as vehicles for the delivery of therapeutic agents. Various methods have been developed to prepare HDL-mimicking NPs. However, they are generally complicated, time consuming, and difficult for industrial scale-up. In this study, one-step homogenization was used to mix the excipients and form the prototype NPs. NGF is a water-soluble protein of 26 kDa. To facilitate the encapsulation of NGF into the lipid environment of HDL-mimicking NPs, protamine USP was used to form an ion-pair complex with NGF to neutralize the charges on the NGF surface. The NGF/protamine complex was then introduced into the prototype NPs. Apolipoprotein A-I was finally coated on the surface of the NPs. NGF HDL-mimicking NPs showed preferable properties in terms of particle size, size distribution, entrapment efficiency, in vitro release, bioactivity, and biodistribution. With the careful design and exploration of homogenization in HDL-mimicking NPs, the procedure was greatly simplified, and the NPs were made scalable. Moreover, various challenges, such as separating unloaded NGF from the NPs, conducting reliable in vitro release studies, and measuring the bioactivity of the NPs, were overcome.

Simple homogenization was used to prepare novel, high-density, lipoprotein-mimicking nanoparticles to encapsulate nerve growth factor. Challenges, detailed protocols for nanoparticle preparation, in vitro characterization, and in vivo studies are described in this article.

신경 성장 인자 인 캡슐 레이션을위한 새로운 HDL 모방 나노 입자의 제조 및 특성 규명

The objective of this article is to introduce preparation and characterization methods for nerve growth factor (NGF)-loaded, high-density, lipoprotein ...

Uptake of New Lipid-coated Nanoparticles Containing Falcarindiol by Human Mesenchymal Stem Cells

Nanoparticles are the focus of an increased interest in drug delivery systems for cancer therapy. Lipid-coated nanoparticles are inspired in structure and size by low-density lipoproteins (LDLs) because cancer cells have an increased need for cholesterol to proliferate, and this has been exploited as a mechanism for delivering anticancer drugs to cancer cells. Moreover, depending on drug chemistry, encapsulating the drug can be advantageous to avoid degradation of the drug during circulation in vivo. Therefore, in this&#160;study, this design is used to fabricate lipid-coated nanoparticles of the anticancer drug falcarindiol, providing a potential new delivery system of falcarindiol in order to stabilize its chemical structure against degradation and improve its uptake by tumors. Falcarindiol nanoparticles, with a phospholipid and cholesterol monolayer encapsulating the purified drug core of the particle, were designed. The lipid monolayer coating consists of 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC), cholesterol (Chol), and 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[methoxy(polyethylene glycol)-2000] (DSPE PEG 2000) along with the fluorescent label DiI (molar ratios of 43:50:5:2). The nanoparticles are fabricated using the rapid injection method, which is a fast and simple technique to precipitate nanoparticles by good-solvent for anti-solvent exchange. It consists of a rapid injection of an ethanol solution containing the nanoparticle components into an aqueous phase. The size of the fluorescent nanoparticles is measured using dynamic light scattering (DLS) at 74.1 &#177; 6.7 nm. The uptake of the nanoparticles is tested in human mesenchymal stem cells (hMSCs) and imaged using fluorescence and confocal microscopy. The uptake of the nanoparticles is observed in hMSCs, suggesting the potential for such a stable drug delivery system for falcarindiol.

This article describes the encapsulation of falcarindiol in lipid-coated 74 nm nanoparticles. The cellular uptake of the nanoparticles&#160;by human stem cells into lipid droplets is monitored by fluorescent and confocal imaging. Nanoparticles are fabricated by the rapid injection method of solvent shifting, and their size is measured with the dynamic light scattering technique.

Falcarindiol 인간 중간 엽 줄기 세포에 의해 포함 된 새로운 지질 코팅 나노 입자의

Nanoparticles are the focus of an increased interest in drug delivery systems for cancer therapy. Lipid-coated nanoparticles are inspired in structure and ...

Preparation of Neutrally-charged, pH-responsive Polymeric Nanoparticles for Cytosolic siRNA Delivery

The success of siRNA as a targeted molecular medicine is dependent upon its efficient cytosolic delivery to cells within the tissue of pathology. Clinical success for treating previously &#8216;undruggable&#8217; hepatic disease targets with siRNA has been achieved. However, efficient tumor siRNA delivery necessitates additional pharmacokinetic design considerations, including long circulation time, evasion of clearance organs (e.g., liver and kidneys), and tumor penetration and retention. Here, we describe the preparation and in vitro physicochemical/biological characterization of polymeric nanoparticles designed for efficient siRNA delivery, particularly to non-hepatic tissues such as tumors. The siRNA nanoparticles are prepared by electrostatic complexation of siRNA and the diblock copolymer poly(ethylene glycol-b-[2-(dimethylamino)ethyl methacrylate-co-butyl methacrylate]) (PEG-DB) to form polyion complexes (polyplexes) where siRNA is sequestered within the polyplex core and PEG forms a hydrophilic, neutrally-charged corona. Moreover, the DB block becomes membrane-lytic as vesicles of the endolysosomal pathway acidify (&#60; pH 6.8), triggering endosomal escape and cytosolic delivery of siRNA. Methods to characterize the physicochemical characteristics of siRNA nanoparticles such as size, surface charge, particle morphology, and siRNA loading are described. Bioactivity of siRNA nanoparticles is measured using luciferase as a model gene in a rapid and high-throughput gene silencing assay. Designs which pass these initial tests (such as PEG-DB-based polyplexes) are considered appropriate for translation to preclinical animal studies assessing the delivery of siRNA to tumors or other sites of pathology.

Methods to prepare and characterize the physicochemical properties and bioactivity of neutrally-charged, pH-responsive siRNA nanoparticles are presented. Criteria for successful siRNA nanomedicines such as size, morphology, surface charge, siRNA loading, and gene silencing are discussed.

사이토 솔 릭 siRNA 전달을 위한 중성 충전, pH 반응 형 고분자 나노 입자의 제조

The success of siRNA as a targeted molecular medicine is dependent upon its efficient cytosolic delivery to cells within the tissue of pathology. Clinical ...

A Freeze-Thawing Method to Prepare Chitosan-Poly(vinyl alcohol) Hydrogels Without Crosslinking Agents and Diflunisal Release Studies

Chitosan-poly(vinyl alcohol) hydrogels can be produced by the freeze-thawing method without using toxic crosslinking agents. The applications of these systems are limited by their characteristics (e.g., porosity, flexibility, swelling capacity, drug loading and drug release capacity), which depend on the freezing conditions and the kind and ratio of polymers. This protocol describes how to prepare hydrogels from chitosan and poly(vinyl alcohol) at 50/50 w/w % of polymer composition and varying the freezing temperature (-4 &#176;C, -20 &#176;C, -80 &#176;C) and freeze-thawing cycles (4, 5, 6 freezing cycles). FT-IR spectra, SEM micrograph and porosimetry data of hydrogels were obtained. Also, the swelling capacity and drug loading and release of diflunisal were assessed. Results from SEM micrographs and porosimetry show that the pore size decreases, while the porosity increases at lower temperatures. The swelling percentage was higher at the minor freezing temperature. The release of diflunisal from the hydrogels has been studied. All the networks maintain the drug release for 30 h and it has been observed that a simple diffusion mechanism regulates the diflunisal release according to Korsmeyer-Peppas and Higuchi models.

A Freeze-Thawing Method to Prepare Chitosan-Polyvinyl alcohol Hydrogels Without Crosslinking Agents and Diflunisal Release Studies

The freezing-thawing method is used to produce chitosan-poly(vinyl alcohol) hydrogels without crosslinking agents. For this method, it is important to consider the freezing conditions (temperature, number of cycles) and polymer ratio, which can affect the properties and applications of the obtained hydrogels.

가교제와 디플루니살 방출 연구를 하지 않고 키토산-폴리(비닐 알코올) 하이드로겔을 준비하는 동결 해동 방법

Chitosan-poly(vinyl alcohol) hydrogels can be produced by the freeze-thawing method without using toxic crosslinking agents. The applications of these ...

Preparation and Characterization of Nanoliposomes for the Entrapment of Bioactive Hydrophilic Globular Proteins

Liposome nanocapsules have been applied for many purposes in the pharmaceutical, cosmetic, and food industries. Attributes of liposomes include their biocompatibility, biodegradability, non-immunogenicity, non-toxicity, and ability to entrap both hydrophilic and hydrophobic compounds. The classical hydration of thin lipid films in an organic solvent is applied herein as a technique to encapsulate tarin, a plant lectin, in nanoliposomes. Nanoliposome size, stability, entrapment efficiency, and morphological characterization are described in detail. The nanoliposomes are prepared using 1,2-dioleoyl-sn-glycerol-3-phosphoethanolamine (DOPE), 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[amino(polyethylene glycol)-2000] (ammonium salt; DSPE-MPEG 2000), and cholesterylhemisuccinate (CHEMS) as the main constituents. Lipids are first dissolved in chloroform to obtain a thin lipid film that is subsequently rehydrated in ammonium sulfate solution containing the protein to be entrapped and incubated overnight. Then, sonication and extrusion techniques are applied to generate nanosized unilamellar vesicles. The size and polydispersity index of the nanovesicles are determined by dynamic light scattering, while nanovesicle morphology is assessed by scanning electron microscopy. Entrapment efficiency is determined by the ratio of the amount of unencapsulated protein to original amount of initially loaded protein. Homogeneous liposomes are obtained with an average size of 155 nm and polydispersity index value of 0.168. A high entrapment efficiency of 83% is achieved.

This study describes classical hydration using the thin lipid film method for nanoliposome preparation followed by nanoparticle characterization. A 47 kDa-hydrophilic and globular protein, tarin, is successfully encapsulated as a strategy to improve stability, avoid fast clearance, and promote controlled release. The method can be adapted to hydrophobic molecules encapsulation.

생리 활성 친수성 구형 단백질의 함정에 대한 나노 리포좀의 준비 및 특성화

Liposome nanocapsules have been applied for many purposes in the pharmaceutical, cosmetic, and food industries. Attributes of liposomes include their ...

SDF-1α-키토산 덱스 트란 황산 나노 입자의 제조 및 특성

요약

더 많은 비디오 탐색

이 비디오의 챕터

줄기 세포 - 로드된 키토산 Microspheres의 배달을 위해 콜라겐 히드로겔을 구축

준비 및 개인의 특성과 다중 약물은 물리적으로 고분자 미셀을 포획로드

ASC의 분화를 제어하는 Bilayered 히드로겔 꾸민

합성 및 Placental 콘의 황산 (plCSA)-지질 타겟팅-고분자 나노 입자

금 나노 막대를 포함하는 세포 차량의 제조 및 광 음향 분석

신경 성장 인자 인 캡슐 레이션을위한 새로운 HDL 모방 나노 입자의 제조 및 특성 규명

Falcarindiol 인간 중간 엽 줄기 세포에 의해 포함 된 새로운 지질 코팅 나노 입자의

사이토 솔 릭 siRNA 전달을 위한 중성 충전, pH 반응 형 고분자 나노 입자의 제조

가교제와 디플루니살 방출 연구를 하지 않고 키토산-폴리(비닐 알코올) 하이드로겔을 준비하는 동결 해동 방법

생리 활성 친수성 구형 단백질의 함정에 대한 나노 리포좀의 준비 및 특성화

SDF-1α-키토산 덱스 트란 황산 나노 입자의 제조 및 특성

요약

더 많은 비디오 탐색

이 비디오의 챕터

줄기 세포 - 로드된 키토산 Microspheres의 배달을 위해 콜라겐 히드로겔을 구축

준비 및 개인의 특성과 다중 약물은 물리적으로 고분자 미셀을 포획로드

ASC의 분화를 제어하는​​ Bilayered 히드로겔 꾸민

합성 및 Placental 콘의 황산 (plCSA)-지질 타겟팅-고분자 나노 입자

금 나노 막대를 포함하는 세포 차량의 제조 및 광 음향 분석

신경 성장 인자 인 캡슐 레이션을위한 새로운 HDL 모방 나노 입자의 제조 및 특성 규명

Falcarindiol 인간 중간 엽 줄기 세포에 의해 포함 된 새로운 지질 코팅 나노 입자의

사이토 솔 릭 siRNA 전달을 위한 중성 충전, pH 반응 형 고분자 나노 입자의 제조

가교제와 디플루니살 방출 연구를 하지 않고 키토산-폴리(비닐 알코올) 하이드로겔을 준비하는 동결 해동 방법

생리 활성 친수성 구형 단백질의 함정에 대한 나노 리포좀의 준비 및 특성화

ASC의 분화를 제어하는 Bilayered 히드로겔 꾸민