a novel treatment for metastatic lymph nodes using lymphatic delivery and photothermal therapy

by:CAI YI JIE     2019-10-08
Delivery of anti-system
Cancer drugs usually result in only a fraction of the amount of doses accumulated at the target site.
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Cancer drugs through the lymph network can achieve more efficient drug delivery for the treatment of lymph node metastasis.
For the first time, we showed the polymer gold nanoparticles (PAuNRs)
It can be effectively transported from the axillary lymph nodes to the tumor-
It contains appropriate axillary lymph nodes and can effectively treat lymph node metastasis.
In a mouse model of metastasis, lymph-
PAuNRs delivered and nearby
Infrared laser irradiation, control the skin temperature by cooling.
Unlike intravenous injections, lymphatic injections deliver PAuNRs at high concentrations in a short period of time.
The results show that there is a possibility of anti-lymphatic administration
Cancer drugs that transfer lymph nodes can inhibit the growth of tumors and can develop into a new treatment.
Animal experiments were conducted in accordance with guidelines approved by the Northeastern University Institutional Animal Care and Use Committee.
Using seeds to synthesize aunts with a high aspect ratio
Introduction techniques using the Ye method.
, Slightly modified.
In the typical reaction, 0.
Add 3645g of sds to 10 ml of water and heat it to 40 °c to completely dissolve the powder. 0.
250 mL of HAuCl solution (0. 01u2009M)
Add to the sds solution and gently stir for 30 minutes.
For this solution, 0 freshly prepared.
6 mL first-class wax.
Add 01 u2009 M NaBH 2 u2009 min under intense stirring to produce a light brown solution used as a seed solution.
The solution was kept at 30 °c for 30 minutes before use.
Prepare the growth solution, 0.
Sds and 1 of 037 CUCM.
234g NaOL, dissolved in 250 mL of warm water (~50u2009°C)
In a 1 liter cone
The solution was allowed to cool down to 30 °c and 4mm AgNO solution of different volumes was added.
At 30 °c, the mixture remains in its original state for 15 minutes, followed by the addition of ml HAuCl solution.
After stirring for 60 min, the solution becomes colorless (700u2009rpm)
A minimum volume of HCl is then introduced to adjust the pH.
After slowly stirring again at 400 rpm for 15 minutes, 1. 25u2009mL of 0.
064 u2009 M ascorbic acid (AA)
Adding, the solution was stirred vigorously for 30 µs.
Finally, a small amount of seed solution is injected into the growth solution.
Stir the resulting mixture at 30 °c and keep it undisturbed at 12 °c at 30 °c to allow the nano-stick to grow.
The final product was separated by centrifuge for 30 min at 7,000 rpm and then removing the supernatant.
No size and/or shape
Selective separation was carried out.
Used mPEG before-SH (MW 5,000)
, Centrifuge the 500 µμ l nano-Rod for 10 µmin at 8,000 µrpm to remove any excess sds.
Then 500 μl 1 mPEG mm mPEG-was added-
SH and 100 μl 1X-PBS were stirred for 24 hours under magnetic stirring.
Finally, mPEG-
After centrifugal removal of any unbound mPEG-, SH functional gold is obtained-
SH and ultrasonic treatment for 10 minutes.
The Nano stick was re-
Dispersed in 1X-PBS solution for further analysis and representation.
The absorption spectrum is mainly based on jasco v-
770 NIR spectrometer.
TEM using jeol jem-
2100F 200 kV high resolution transmission electron microscope at acceleration voltage.
Measurements were made using the Photal ELS-aligned Tower potentialZ2MH instruments.
Centrifugal was performed using Eppendorf 5415 u2009 r and KUBOTA 7780II centrifuge.
Breast cancer cells in C3H/He mice (FM3A-Luc)
Genes that stably express the firefly fluorescent enzyme gene are used after three passwords.
Culture cells in RPMI1640 medium (Sigma-Aldrich)
Add 10% calories
Inactivated fetal bovine serum, 0.
5% G418 Geneticin and 1% L-
(Inclusive) ammonia-penicillin-agricultural penicillin (Sigma-Aldrich).
Cells were incubated in a mixture of 5% CO 2 and 95% air at 37 °c until 80% confluent.
On the day of inoculation, no contamination was confirmed using the fungal alarm detection kit (Lonza Rockland)
According to the agreement of the manufacturer.
Maintain cells in RPMI
1640 medium supplemented with 10% calories
Inactivated fetal bovine serum, 0.
5% G418 Geneticin and 1% L-
(Inclusive) ammonia-penicillin-agricultural penicillin (Sigma-Aldrich).
For incubation with PAuNRs, cells were in triplicate with a density of 5,000 vivo cells/Wells at 48-
Microplate.
After 24 hours of incubation (
37 °c, 5% carbon dioxide)
, Cells are exposed to PAuNRs at different concentrations (
Dilute in 10 μ l Medium)
In triplicate for an hour.
Remove the medium and wash the cells twice with PBS.
Fresh medium was added (500u2009μL/well)
Plate incubation for 24 hours.
Cell toxicity was evaluated using a standard MTT method. MXH10/Mo-/ (MXH10/Mo/lpr)mice (
Age 12-14 weeks)
It was bred under a specific pathogen.
Free conditions for animal research institute, Northeastern University School of Medicine, Sendai, Miyagi, Japan.
MXH10/Mo/lpr mice developed systemic lymph node swelling with a diameter of up to 10mm at the age of 2.
5-3 months of age by accumulating a lot of lpr-
T cells in lymph nodes.
These mice did not express Fas genes involved in apoptosis.
These mice retain the basic internal structure of the lymph nodes, including the structure of the cortex, subcortex, and marrow regions.
PALN metastasis was induced by injection 3.
3 × 10 cells are suspended in a mixture of 20 μ l PBS and 40 μ l 400 mg/ml matrix gel (
Collaborative biomedical products
Into the unilateral SiLN (=u200923).
Intranodal was vaccinated into SiLN for the next one using 27 u2009 G needles
Frequency ultrasound imaging system (VEVO770; VisualSonics)with a 25-MHz frequency converter (RMV-710B;
Axial resolution of 70 μm;
Focal length, 15mm; VisualSonics).
After removing the needle, keep the needle in the same position for 1 minute to solidify the matrix gel.
Vaccination Day is defined as 0 days.
The transfer to PALN was assessed using a biomarker imaging system (IVIS Lumina; PerkinElmer)
Every 3 days. inoculation.
Background luciferase activity is ~ 4X10 photon/second.
Mice with luciferase activity greater than background level in PALN (
~ 5 Stars x 10 stars photons/s)
It is considered a transfer mouse.
The probability of a transfer in PALN is about 87% (
Event/23 mice).
Fluorescent protein of Mg/kg (Promega)
Injected by abdominal cavity.
After 10 min, Firefly bio-glow of 1 min was measured using IVIS.
Transfer mice were divided into 4 groups: control group (=5), PAuNRs group (=5), Laser group (=6)
Laser Group (=7).
PTT with PAuNRs was carried out by laser irradiation. The anti-
Evaluation of tumor effect of PTT on malignant cells by continuous wave Nd: YVO air-
Cooled NIR laser (2. 5u2009±u20090. 5u2009W/cm;
Wavelength 1064nm;
TEM beam diameter, 0. 6u2009mm; CYD-010-TUBC, Neoarc). PAuNRs (
40 μg/mL, 120 μ l)
Injected into the AALNs of the tumor-
Mice carrying MXH10/Mo/lpr.
Before the temperature control system is irradiated, the skin temperature cools to
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