Kolşisin Makalesi
Kolşisin Makalesi
Kolşisin Makalesi
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Zhang, J. Miao, Z. Zhao, X. Jin, L. Liu, L. Yu, C. Shen and J. Ding, J. Mater. Chem. B, 2020, DOI:
10.1039/C9TB02523E.
Volume 6
Number 3
21 January 2018
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PAPER
Wei Wei, Guanghui Ma et al.
Macrophage responses to the physical burden of cell-sized
shall the Royal Society of Chemistry be held responsible for any errors
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rsc.li/materials-b
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ARTICLE
Received 00th January 20xx, Liang Liua, Lin Yub,c,*, Chengxing Shena,*, Jiandong Dingb,c
Accepted 00th January 20xx
DOI: 10.1039/x0xx00000x Localized administration of anti-inflammatory agents benefits patients after myocardial infarction (MI) by
repressing/modulating inflammatory response of the MI region and thus accelerating repair of the impaired
www.rsc.org/
tissues. Colchicine (Col), an ancient natural drug, has excellent anti-inflammatory effects; however, its
utilization is strictly limited due to its severe systemic toxicity and narrow therapeutic window. In this
study, we developed an intramyocardial delivery system of Col using an injectable, thermosensitive
poly(lactide-co-glycolide)-poly(ethylene glycol)-poly(lactide-co-glycolide) (PLGA-PEG-PLGA)
polymer hydrogel as the vehicle for the treatment of MI while minimizing its systemic toxicity. The
aqueous PLGA-PEG-PLGA solution loaded with Col (Col@Gel) underwent a sol-gel transition at 35 °C
and maintained a gel state at body temperature. Col was released from the Col@Gel in an initial burst
followed by a sustained release manner for over 8 days. The in vitro cell tests showed that the Col@Gel
system significantly inhibited macrophage proliferation and migration. In a mouse model of MI, a single
intramyocardial administration of the Col@Gel effectively alleviated cardiac inflammation, inhibited
myocardial apoptosis and fibrosis, improved cardiac function and structure, and increased mouse survival
without inducing severe systemic toxicity, which was observed following intraperitoneal administration
of Col solution. These results suggested that the Col@Gel system is a reliable drug delivery system for the
sustained local release of Col and has great potential as an anti-inflammatory therapy for the treat of MI.
Acute myocardial infarction (MI) and follow-up induced heart matrix synthesis, leading to myocardial fibrosis, adverse ventricular
failure are one of the leading causes of death in many countries remodeling, and cardiac dysfunction.2-4 In view of the important roles
and remain a therapeutic challenge worldwide.1, 2 In addition to of inflammation in myocardial injury, increasing attention has been
ischemia and hypoxia-induced myocardial damage, post-MI paid to the potential benefits of anti-inflammatory therapies to
injury is exacerbated by excessive local inflammation, which repress/modulate local inflammation post-MI, thereby accelerating
myocardial repair, inhibiting left ventricular remodeling, and
improving patient prognosis.2, 5, 6
aDepartment of Cardiology, Shanghai Jiao Tong University Affiliated Sixth
People’s Hospital, Shanghai 200233, China. Colchicine (Col), an ancient natural drug extracted from Colchicum
E-mail: [email protected]
bState Key Laboratory of Molecular Engineering of Polymers, Department of autumnale, was first described and utilized in Egypt more than 3000
Macromolecular Science, Fudan University, Shanghai 200438, China.
E-mail: [email protected]
years ago.7 Col prevents mitotic spindle formation, arrests
cZhuhai Fudan Innovation Institute, Zhuhai, Guangdong 51900, China
microtubule polymerization and inhibits crucial inflammatory
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signaling pathways, such as inflammasomes, proinflammatory convenient synthesis, good reproducibility and wide recognition of
cytokines, as well as expression of adhesion molecules.7, 8 Therefore, safety.38, 39 It has been developed into a platform for the delivery of
Col has potent anti-inflammatory effects. Nowadays, Col is various kinds of drugs to treat systemic or regional diseases, including
recognized as a wonder drug for the treatment of refractory gout and anti-cancer treatments,40-42 anti-diabetic treatments,43-45 anti-
familial Mediterranean fever.7, 9 Col has also been tried to prevent osteopenia treatments,39 and others.46-50 For example, the local
Several studies found that the gout patients treated with Col exhibited Chen developed a tumor microenvironment-sensitive PLGA-PEG-
a reduced prevalence of MI.12, 13 Two recent clinic trials revealed that PLGA-doxorubicin conjugate hydrogel and a synergistic anticancer
the patients with acute MI receiving the systemic administration of effect was achieved after intratumoral injection of the conjugate
Col showed a decrease in left ventricular remodeling or infarct size.14, hydrogel containing docetaxel.51 Recently, the Hotta’s group revealed
15 More recently, in a mouse model of MI, oral ingestion or that the introduction of laponite clay nanoparticles to an aqueous
intraperitoneal injection of Col had beneficial effects on myocardial solution of PLGA-PEG-PLGA could facilely modulate the sol-gel
repair.8, 16 These findings opened up a new avenue for the old drug in transition temperature from 25 °C to 37 °C, while the degradation rate
new use and provided a promising strategy for the treatment of acute of composite hydrogel was mainly dependent on the LA/GA ratio.52
MI. Of note, the risk of systemic toxicity remains a considerable 53 It is worth pointing out that the application of PLGA-PEG-PLGA
hurdle for the application of Col. One valid alternative to avoid the hydrogel for MI treatment is rarely reported.
systemic toxicity of Col is to realize the regional sustained delivery of In this study, we developed a sustained delivery of Col based on an
the drug. Meanwhile, advances in echocardiography and injectable thermosensitive PLGA-PEG-PLGA hydrogel for
thoracoscopy technologies have made intramyocardial injection of myocardial repair after MI. First, a PLGA-PEG-PLGA triblock
drugs feasible. copolymer with rational molecular composition was designed and
Injectable hydrogels formed by in situ gelation have received synthesized to ensure its temperature-responsive sol-gel transition in
increasing attention as vehicles for drug delivery and as scaffolds for water below body temperature. The delivery system of Col (labeled
tissue repair due to the unique superiority of minimally invasive Col@Gel) was then fabricated using the PLGA-PEG-PLGA hydrogel
application of these systems.17-26 In particular, temperature-induced as the vehicle, as shown in Fig. 1A. Next, in vitro release behavior and
gelling materials composed of poly(ethylene glycol) effects of the Col@Gel on macrophage proliferation and migration
(PEG)/polyester,27-30 PEG/polypeptide,31-33 poly(N- were evaluated. Finally, a mouse model of MI was constructed and
isopropylacrylamide-co-2- hydroxyethyl methacrylate-co- the safety and efficacy of intramyocardial injection of Col@Gel for
methacrylate-polylactide) (poly(NIPAAm-co-HEMA- co- the post-MI treatment was confirmed by analysis of mouse weight,
MAPLA),34, 35 poly(phosphazene)s copolymers36, 37 are low-viscous survival, liver function, cardiac structure and function, and
liquids at ambient temperature, but can rapidly convert into standing myocardial inflammation, apoptosis, and fibrosis. Fig. 1B
in situ gels in response to physiological temperature. Therefore, schematically presents the creation of mouse model of MI and the use
various therapeutic agents can readily be loaded into these materials of Col@Gel after MI.
by blending them in the sol state without drug loss. Upon in vivo
injection, the in situ forming hydrogels can offer sustained release of Materials and Methods
the encapsulated drugs.
Materials
So far, the thermosensitive and biodegradable poly(lactide-co-
Poly(ethylene glycol) (PEG) with molecular weight (MW) 1500 and
glycolide)-poly(ethylene glycol)-poly(lactide-co-glycolide) (PLGA-
stannous octoate (Sn(Oct)2) were purchased from Sigma-Aldrich (St.
PEG-PLGA) hydrogel enjoys great popularity because of its
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Fig. 1 A) The fabrication of Col@Gel system. The Col@Gel system exhibited a free-flowing sol at ambient temperature (25 °C) and
spontaneously converted into a semi-solid hydrogel at body temperature. This sol-gel transition was reversible. B) The establishment of
mouse MI model and the subsequent administration of Col@Gel system into three sites around the infarct region after MI. MI was
induced by ligating the left anterior descending coronary artery (LAD).
Louis, MO, USA). DL-Lactide (LA) and glycolide (GA) were supplied 0.01 mol) was placed in a three-necked flask and dried under vacuum
by Hangzhou Mingzhong Biotechnology Co. Ltd. (China) and used as with stirring at 125 °C for 3 h to remove moisture. LA and GA at a
received. given amount were added to the flask and the mixture was incubated
All animal experimental procedures were approved by the at 80 °C for 30 min. After LA and GA had melted, 0.08 g of Sn(Oct)2
Institutional Authority for Laboratory Animal Care of Shanghai Jiao was added, and the reaction mixture was heated with stirring at 150
Tong University Affiliated Sixth People’s Hospital and conformed to °C for 12 h under an argon atmosphere. The temperature was then
the Guide for the Care and Use of Laboratory Animals (National cooled to 120 °C with stirring for 3 h under vacuum to eliminate the
Research Council, Eighth Edition, 2010) and the AVMA Guidelines unreacted monomers. Subsequently, the crude polymers were poured
for the Euthanasia of Animals (2013 Edition). Male C57BL/6 mice (6 into 80 °C water to remove water-soluble oligomers and unreacted
weeks of age) were supplied by SLAC Laboratory Animal Co. Ltd. monomers. This washing procedure was carried out for at least three
(Shanghai, China). Mice were housed under appropriate humidity and times. Afterwards, residual water was removed by freeze drying. The
temperature conditions and received fed standard chow. Tap water purified products were harvested and stored at −20 °C until use and
was available ad libitum. the yield was approximately 80%.
The structure and composition of PLGA-PEG-PLGA was analyzed
Synthesis and characterization of PLGA-PEG-PLGA via 1H-nuclear magnetic resonance (NMR) on a 500 MHz proton
A ring-opening copolymerization approach using LA and GA spectrometer (DMX500 spectrometer, Bruker). Deuterated
monomers was used to prepare PLGA-PEG-PLGA triblock chloroform with the internal standard tetramethylsilane was selected
copolymers and PEG 1500 and Sn(Oct)2 were selected as macro- as the solvent. The value of molar mass dispersity (ĐM) of the polymer
initiator and catalyst, respectively.39, 47 Briefly, PEG 1500 (15.0 g, was analyzed by gel permeation chromatography (GPC; Agilent
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1260). Tetrahydrofuran was selected as the eluent, the flow rate was monitored for up to 20 days. Male C57BL/6 mice were anesthetized
set at 1 mL/min and the measurement temperature was fixed at 35 °C. with chloral hydrate and then injected subcutaneously with 0.1 mL of
Monodisperse polystyrene microspheres were used as standards. aqueous polymer solution (25 wt %) into the dorsal region. 3 mice
were euthanized at each specified time point and the remaining
Measurement of sol-gel transition temperature hydrogels were dissected. The appearance of the injected gels was
min and then inverted by 180°. If the sample showed no evidence of Cell culture. The mouse macrophage cell line Raw264.7 was
flow within 30 s, it was considered to be a gel. supplied by the Cell Bank of the Chinese Academy of Sciences
The temperature-induced gelation behaviors of the aqueous polymer (Shanghai, China). The cells were incubated in DMEM medium
solutions in the presence or absence of Col were further measured (Gibco) containing fetal bovine serum (FBS, 10%) and penicillin-
using a dynamic rotational rheometer (Kinexus PRO, Malvern) with streptomycin (100 U/mL) under a 5% CO2 atmosphere at body
a cone plate (1° steel cone, 60 mm diameter). The time sweep program temperature.
was set at an oscillatory frequency of 10 rad/s and the temperature was In vitro cytotoxicity assay. The cytotoxicity of blank hydrogel and
increased from 15 °C to 50 °C at a heating rate of 1 °C/min. Col@Gel system against Raw264.7 cells was evaluated by measuring
cell viability with a Cell Counting Kit-8 (CCK-8, Dojondo) assay.
In vitro Col release Transwell chambers and a cell culture plate are shown in Fig. S1A.
The aqueous polymer solutions containing Col (0.25, 0.5 or 1 mg/mL) As presented in Fig. S1B, aqueous polymer solutions (25 wt%) with
were readily prepared by blending the aqueous polymer solution (25 or without Col (1.0 or 0 mg/mL) were added to Transwell chambers
wt%) with Col at 4 °C to form a homogenous solution. The Col-loaded at 100 μL/chamber and incubated at 37 C for 10 min to form
aqueous polymer solution maintained a free-flowing sol state at room hydrogels in situ. Raw264.7 cells were seeded into 24-well culture
temperature while a gel state at body temperature, which was labeled plates at a density of 1×104 cells/well in DMEM growth medium and
as Col@Gel. The in vitro release of Col from Col@Gel was incubated for 24 h. The cells were then washed for three times and
investigated and the method was implemented following the previous incubated in fresh DMEM. Next, the Transwell chambers containing
protocols.18, 45 Briefly, the samples (0.5 mL) were injected into 10 mL the gel samples were placed into the culture wells and the DMEM
vials, equilibrated at 4 °C for at least 12 h, and then incubated in a could get touch with the gel through the permeable membrane of
shaking water bath (50 rpm) at 37 °C for 10 min to form gels. Transwell chamber. After co-culture for 48 h, the cells were washed
Phosphate-buffered solution (PBS, pH 7.4) adding 0.025 wt % NaN3 with PBS, and CCK-8 reagent was then added to the wells in
was introduced at 5 mL/vial. At various time points, 3 mL of PBS was accordance with the instructions provided by the manufacturer.
withdrawn from the vials and replaced with 3 mL of prewarmed (37 Control cells were incubated in the same manner in the absence of gel-
°C) fresh PBS. The Col contents in the PBS samples were detected containing Transwell inserts, and their viability was set at 100% for
using a double beam UV-vis spectrophotometer (TU-1950). The UV the purpose of calculating relative cell viability. Samples were
absorbance differences at 354 nm between the Col-containing analyzed in triplicate.
hydrogel samples and the Col-free blank hydrogel samples were In vitro cell migration assay. In vitro migration of Raw264.7
converted into the amounts of Col released at different time points by macrophages was measured using a Transwell assay. As presented in
comparison with the standard curve. Fig. S1C, aqueous polymer solutions (25 wt%) with or without Col
(1.0 or 0 mg/mL) were added to cell culture plates and then kept at 37
In vivo hydrogel degradation
℃ to form gels in situ. Each well with 1 mL of hydrogel was incubated
Changes in visual appearance, mass, and MW of the PLGA-PEG-
in DMEM (1 mL) at 37 C for 24 h and subsequently the DMEM was
PLGA hydrogel after subcutaneous injection into mice were
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collected. Raw264.7 cells (1×104 cells in the 100 μL of DMEM retro-orbitally and centrifuged to obtain serums. Serum
collected) were added to the Transwell chambers, the chambers were concentrations of BNP, ALT, and AST were measured using a BNP
placed in new cell culture plates, and an additional 600 μL of fresh ELISA Assay Kit (FineTest, Wuhan, China) or a Catalyst One®
DMEM was added per well. After incubation of 48 h, the chambers Chemistry Analyzer (IDEXX, Westbrook, USA) according to the
were taken out, washed using PBS, and the cells on the membrane of manufacturers’ instructions.
Animal experiments and terminal deoxynucleotidyl transferase dUTP nick end labeling
In vivo application of the Col@Gel in a mouse model of MI. (TUNEL) staining, respectively.
Male C57BL/6 mice were randomized into four groups (15 per Analysis of tissue fibrosis. Myocardial tissue sections were stained
group): Sham, MI control, MI + Col@Gel, and MI + Col (i.p.). MI with Masson’s trichrome. Sections were observed using an
was induced by ligating the LAD of mouse. The Sham group OLYMPUS microscope (Japan). The fibrotic area within each tissue
underwent surgery but the LAD was not ligated. The MI control group section was measured and analyzed via ImageJ software 1.48
underwent MI surgery but did not receive any treatment. The (National Institutes of Health, Bethesda, MD, USA).
Col@Gel group received intramyocardial injections of 1 mg/mL Col- TUNEL assay. A TUNEL assay was employed to detect apoptotic
loaded polymer solution (10 μL) into three sites around the infarct cells in myocardial tissues. The sections were probed using an
region immediately after LAD ligation. The Col (i.p.) group received TUNEL detection kit (Beyotime Biotechnology, Shanghai, China)
a single intraperitoneal injection of Col solution at the same dose. according to the manufacturer’s recommendations. Three images of
Electrocardiography was performed using a Biological Signal the infarct zone in each section were acquired using an OLYMPUS
Acquisition System (Techman, Chengdu, China) during LAD microscope (Japan). The number of TUNEL-positive and total
ligation. cardiomyocyte nuclei in each zone were counted using ImageJ
The ST-segment elevation of electrocardiogram suggested that MI software, and the percentage of apoptotic cardiomyocytes was
was successfully modeled, as illustrated in Fig. S2. The mice were calculated.
monitored three times per day during the 4-week follow-up period. Immunofluorescence staining of macrophages. Myocardial
The weights per week and the death of mice post-MI were recorded. inflammation was assessed by counting the number of F4/80-positive
Echocardiography measurement. At day 28 after MI, the survival macrophages in tissue sections. After deparaffinization, antigen
mice were shaved in the anterior and left lateral thoracic regions and unmasking, and permeabilization, the myocardial sections were
then placed in the supine position. Ultrasound transonic gel was incubated overnight at 4 °C with anti-F4/80 antibody (1:700 dilution,
smeared on the thoracic regions to optimize visibility. Two- CST, Danvers, USA), washed, and incubated with a goat anti-mouse
dimensional and m-mode echocardiographic images were acquired Alexa Fluor 488-labeled antibody (1:400 dilution, Jackson, West
using echocardiography. Left ventricular internal dimensions were Grove, USA) for 1 h at 37 C in the dark. Cell nuclei were
recorded during systole (LVIDs) and diastole (LVIDd). The ejection counterstained with 4',6-diamidino-2-phenylindole (DAPI) for 10 min
fraction (EF) and fractional shortening (FS) were calculated from the at room temperature. Three images of the infarct zone in each section
measured parameters. were acquired using an OLYMPUS microscope and the number of
Brain natriuretic peptide (BNP) and liver function tests. To F4/80-positive cells per mm2 were counted using ImageJ software.
assess serum levels of BNP, a biomarker of heart failure,54, 55 and Tumor necrosis factor-α (TNF-α) assay. The myocardial content
alanine transaminase (ALT) and aspartate aminotransferase (AST), of TNF-α was evaluated using a Mouse TNF-α ELISA kit (Sigma-
two biomarkers of liver function,56 the mice were anesthetized using Aldrich). Another gang of male C57BL/6 mice that underwent surgery
isoflurane at day 28 post-treatment and blood samples were sampled and then received different treatments were likewise divided into four
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groups: Sham, MI control, MI + Col@Gel, and MI+Col (i.p.). 3 mice undergoes deformation, while G″ denotes the viscous behavior. At
per group were euthanized each week, the hearts were removed, and low temperatures, G″ was higher than G′, suggesting a free-flowing
the myocardial tissues in the infarct zone were excised and stored at - liquid state and good injectability. As the temperature increased, both
80°C until analysis. Then, the stored myocardial tissues were of them increased, but G′ increased more rapidly. The sol-gel
homogenized five times in ice-cold lysis buffer for 1 min each time transition point was observed at 35 °C, when G′ was equal to G″.
Statistical analysis
Data are presented as the mean ± standard deviation (SD). Group
means were compared via one-way analysis of variance or unpaired t-
test. A p value of <0.05 was considered statistically significant.
Results
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Fig. 4 In vivo degradation and biocompatibility of the thermosensitive PLGA-PEG-PLGA hydrogel. A) In vivo hydrogel maintenance.
The pictures were shoot at various time points after dorsal subcutaneous injection. B) Optical micrographs of HE-stained slices of
surrounding tissues at designated time points after subcutaneous administration of hydrogel. M: muscle; G: hydrogel; S: skin; F: fibrous
capsule. C) The mass percentage of remaining gel excised from C57BL/6 mice measured by the gravimetric method. D) GPC traces of
residual PLGA-PEG-PLGA polymers collected from the excised gels during the in vivo degradation.
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Fig. 5 A) The effect of Col@Gel on the proliferation of Raw264.7 macrophages in vitro after 48 h of incubation (n=3). The viability of
Raw264.7 cells that did not receive any treatment was defined as 100%. B) The effect of Col@Gel on the migration of Raw264.7 cells after 48
h of incubation (n=3). ***p<0.001. C) Representative micrographs of migrated Raw264.7 cells. Raw264.7 cells were stained purple using
crystal violet. The cells that received different treatments migrated from the upper surface of membrane of chamber to the lower surface of
membrane and then the lower surface of membrane was photographed after 48 h of incubation.
points. Apparently, the in vivo biodegradability and biocompatibility
of the PLGA-PEG-PLGA hydrogel supports the feasibility and safety
of clinical application as a long-acting drug release system.
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In vivo toxicity
The systemic toxicity of Col in vivo was assessed by recording the
changes in body weights and serum ALT and AST levels of mice. As
shown in Fig. 7A, the mice in the Col@Gel group gained a mean
weight increase of 3.42±0.5 g after MI for 4 weeks, which was not
significantly different from the gain in either the Sham group or the
MI group. The Col (i.p.) group gained significantly less weight than
the other three groups (2.95±0.4, p<0.05), illustrating the
intraperitoneal administration of Col solution caused the remarkably
systemic toxicity compared with the intramyocardially injected
Col@Gel. In a similar way, serum AST and ALT levels in the Col
(i.p.) group were significantly higher than those in the other three
groups (p<0.001), suggesting that the systemic exposure to Col
induced severe liver injury (Fig. 7B). It is worth noting that the slight
increases in AST and ALT levels in the MI and Col@Gel groups
compared with the Sham group may be attributed to the MI surgery
itself.
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Fig. 9 Changes of myocardial TNF-α levels of mice that contributed to the timely suppression of the overactive acute
Published on 31 December 2019. Downloaded on 1/3/2020 1:26:29 AM.
received different treatments after MI (n = 3 per time point). inflammatory response as well as the subsequent chronic
inflammation in the infarct region. In contrast, the anti-inflammation
effect was much weaker after a single intraperitoneal injection of Col
solution.
Myocardial fibrosis
peaked at 1 week post-MI and gradually declined thereafter. Both the
Col@Gel and Col (i.p.) treatments significantly reduced the
myocardial TNF-α content at each time point examined, and the use
of Col@Gel was far more effective in suppressing TNF-α production
than that of Col (i.p.) (p<0.05). Combined with the results presented
in Fig. 3, we believed that the sustained release of Col for up to 1 week
To detect MI-induced myocardial fibrosis, myocardial tissues were
stained with Masson’s trichrome to reveal collagen fibers. As
displayed in Fig. 10A, the fibrosis degree of myocardial tissue in the
infarction zone of mice in the MI group was significantly higher than
that in the Sham group, while that was significantly reduced in the
mice treated with Col@Gel. Specifically, the proportion of total tissue
area with evidence of fibrosis was 1.03±0.07% in the Sham group,
24.83±3.55% in the MI group (p<0.001 vs Sham), 11.53±2.50% in the
Col@Gel group (p<0.001 vs MI), and 19.47±2.93% in the Col (i.p.)
group (p<0.001 vs MI group) (Fig. 10B). Also, the fibrosis area of
myocardial tissue in the Col@Gel group was significantly lower than
that in the Col (i.p.) group, indicating that the intramyocardial
injection of Col@Gel and the subsequent sustained release of Col was
more effective in reducing myocardial fibrosis post-MI than the
Fig. 10 The myocardial fibrosis degree of mice that received intraperitoneal injection of Col solution.
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Finally, serum concentrations of BNP, a biomarker of congestive confirmed that oral or intraperitoneal administration of Col had anti-
heart failure, were measured at 4 weeks post-MI (Fig. 13). Compared inflammatory and anti-fibrotic effects on experimentally induced MI
with the Sham group, the MI group showed a significantly higher BNP in mice, but their findings on improvement of cardiac function were
level (p<0.001), as expected. However, the serum BNP levels were inconsistent.8, 16 Moreover, they were unable to draw a clear
significantly lower in both the Col@Gel- and Col (i.p.)-treated groups conclusion about the systemic toxicity of Col in their studies.
post-MI treatment.
In the current work, we constructed an injectable drug delivery
system, enabling the localized, sustained release of Col in the
myocardium. First, we prepared a thermosensitive hydrogel
composed of PLGA-PEG-PLGA polymers that was an injectable
liquid at room temperature and converted into a non-flowing gel
within 30 s when exposed to body temperature. Col was readily,
homogenously dispersed in the aqueous polymer solution by blending
them at low or ambient temperature. The loading of Col did not
influence the injectability and temperature-induced gelation of
Fig. 13 The serum BNP concentrations of mice that received
polymer/water system (Fig. 2). The acute stage in the first 7 days after
different treatments at 4 weeks post-operation. n=the survival mice
an MI is the crucial period for myocardial repair as well as the peak
number per group at 4 weeks post-MI, specially, nSham=15, nMI=8,
period of myocardial necrosis, apoptosis, and inflammation
nCol@Gel=13, nCol (i.p.)=11. ***p <0.001.
response.64, 65 The in vitro release of Col from the Col@Gel system
exhibited an initial burst followed by a sustained release pattern for
Discussion
over 8 days (Fig. 3), suggesting that the liberated Col could rapidly
Overactive, persistent tissue inflammatory response post-MI, which
exert the anti-inflammatory effect upon injection as well as
not only disturbs tissue repair but also induces myocardial apoptosis,
throughout the follow-up crucial acute stage post-MI. This feature was
necrosis, fibrosis and leads to ventricular remodeling and cardiac
well confirmed by sustained, effective inhibition of the expression of
insufficiency, is a major cause of myocardial injury.2-4 Therefore, the
the inflammatory cytokine TNF-α following the intramyocardial
timely suppression of excess early myocardial inflammatory response
administration of the Col@Gel system post-MI (Fig. 9).
is in favor of promoting and accelerating myocardial repair of patients
The in vitro cell tests showed that the Col released from the Col@Gel
after MI.2, 3, 61, 62 For example, some clinic studies have demonstrated
effectively inhibited the proliferation and migration of macrophages
that administration of anti-inflammatory drugs, such as canakinumab,
(Fig. 5), which make vital negative contributions to cardiac
an anti-IL-1β monoclonal antibody, has beneficial effects on the
remodeling post-MI.57, 58 On the basis of demonstrating the in vivo
prognosis of MI patients.2, 63 However, the studies on the anti-
biodegradability and biocompatibility of PLGA-PEG-PLGA hydrogel
inflammatory remedy for patients after MI are still limited compared
itself (Fig. 4), the effect of Col@Gel system on myocardial repair of
with the other therapies, such as antiplatelet and lipid-lowering
mice after infarction was evaluated. We found that a single
treatments.
intramyocardial administration of the Col@Gel post-MI and the
Col is a mitotic spindle poison utilized for more than 3000 years and
follow-up sustained release of Col significantly reduced the
has pleiotropic anti-inflammatory properties, but the widespread
accumulation of macrophages and inhibited myocardial apoptosis and
clinical use is precluded because of its severe toxicity profile and
fibrosis in the infarct region, lowered the expression of the heart
narrow therapeutic window.7, 10, 11 Recently, Fujisue and Akodad
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DOI: 10.1039/C9TB02523E
Journal Name ARTICLE
failure biomarker BNP, improved the cardiac structure and function, tests confirmed the safety and efficacy of Col@Gel for the treatment
and prolonged the survival of mice (Figs. 6, 8, 10-13). Apparently, of MI after intramyocardial injection were significantly superior to
intramyocardial injection of the Col@Gel imparted multiple benefits those of intraperitoneal injection of Col solution, including lower
at the most critical stage of treatment after MI. myocardial inflammation, apoptosis, and fibrosis, better heart
Furthermore, we compared the safety and efficacy of Col function and structure, higher animal survival and negligible systemic
This journal is © The Royal Society of Chemistry 2019 J. Name., 2019, 00, 1-3 | 13
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DOI: 10.1039/C9TB02523E
Paper Journal of Materials Chemistry B
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after infarction
Yu Chen‡, Jiayue Shi‡, Yaping Zhang, Jiajun Miao, Zhe Zhao, Xian Jin, Liang Liu, Lin
Published on 31 December 2019. Downloaded on 1/3/2020 1:26:29 AM.
promoted myocardial repair after infarction while minimizing the systemic toxicity of
colchicine.