1.An Important Role of Macrophages for Wound Margin Regeneration in a Murine Flap Model
Ho Ryun WON ; Chorong SEO ; Hye Young LEE ; Jin ROH ; Chul Ho KIM ; Jeon Yeob JANG ; Yoo Seob SHIN
Tissue Engineering and Regenerative Medicine 2019;16(6):667-674
BACKGROUND: Macrophages have been known to have diverse roles either after tissue damage or during the wound healing process; however, their roles in flap wound healing are poorly understood. In this study, we aimed to evaluate how macrophages contribute to the flap wound regeneration.METHODS: A murine model of a pedicled flap was generated, and the time-course of the wound healing process was determined. Especially, the interface between the flap and the residual tissue was histopathologically evaluated. Using clodronate liposome, a macrophage-depleting agent, the functional role of macrophages in flap wound healing was investigated. Coculture of human keratinocyte cell line HaCaT and monocytic cell line THP-1 was performed to unveil relationship between the two cell types.RESULTS: Macrophage depletion significantly impaired flap wound healing process showing increased necrotic area after clodronate liposome administration. Interestingly, microscopic evaluation revealed that epithelial remodeling between the flap tissue and residual normal tissue did not occurred under the lack of macrophage infiltration. Coculture and scratch wound healing assays indicated that macrophages significantly affected the migration of keratinocytes.CONCLUSION: Macrophages play a critical role in the flap wound regeneration. Especially, epithelial remodeling at the flap margin is dependent on proper macrophage infiltration. These results implicate to support the cellular mechanisms of impaired flap wound healing.
Cell Line
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Clodronic Acid
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Coculture Techniques
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Humans
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Keratinocytes
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Liposomes
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Macrophages
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Regeneration
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Surgical Flaps
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Wound Healing
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Wounds and Injuries
2.Adjuvant Effect of IV Clodronate on the Delay of Bone Metastasis in High-Risk Prostate Cancer Patients: A Prospective Study.
Paulo RODRIGUES ; Flavio O HERING ; Alex MELLER
Cancer Research and Treatment 2011;43(4):231-235
PURPOSE: High-risk prostate cancer patients undergoing treatment often experience biochemical recurrence. The use of bisphosphonates as an adjuvant treatment delays skeletal events, yet whether or not bisphosphonates also delay metastastic development remains to be determined. MATERIALS AND METHODS: A total of 140 high-risk prostate cancer patients who were undergoing definitive treatment and who had clinically organ-confined disease and who suffered from biochemical recurrence were administered intravenous (IV) clodronate. The patients were treated with a radical retropubic prostatectomy (RP) or curative radiotherapy (RTx). Upon androgen deprivation therapy initiation, tri-monthly IV clodronate was added to the treatment to prevent bone demineralization. Twenty-six out of 60 operated cases and 45 out of 80 irradiated cases received bisphosphonate. The length of time until the first bone metastasis was recorded and analyzed. RESULTS: No statistical difference was found for the type of primary treatment (RP or RTx) on the time to the first bone metastasis (95% confidence interval [CI], 0.40 to 2.43; p=0.98). However, there was a clear advantage favoring the group that received bisphosphonate (p<0.001). The addition of bisphosphonate delayed the appearance of the first bone metastasis by seven-fold (95% CI, 3.1 to 15.4; p<0.001). CONCLUSION: Treatment with tri-monthly IV clodronate delayed the time to the first bone metastasis in high-risk prostate cancer patients who were experiencing an increase in the prostate specific antigen level after definitive treatment.
Androgen Antagonists
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Clodronic Acid
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Diphosphonates
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Hormone Antagonists
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Humans
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Imidazoles
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Neoplasm Metastasis
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Nitro Compounds
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Osteoporosis
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Prospective Studies
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Prostate
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Prostate-Specific Antigen
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Prostatectomy
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Prostatic Neoplasms
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Recurrence
3.The investigation of macrophage infiltration in the early phase of ischemic acute renal failure in mice.
Soo Jeong YU ; Dong Jin OH ; Suk Hee YU
The Korean Journal of Internal Medicine 2008;23(2):64-71
BACKGROUND/AIMS: Inflammation plays a key role in ischemic acute renal failure (ARF). The present study investigated the infiltration of macrophages in the early phase of ischemic ARF in mice. METHODS: Ischemic ARF was induced by renal clamping for 22 min, while the control mice underwent sham surgery (no clamping). The serum creatinine and blood urea nitrogen (BUN) levels were measured in the control and post-ischemia mice. Immunofluorescence staining was used to measure the number of CD 11b-positive cells in the kidney tissue sections to determine the amount of post-ischemic macrophage infiltration. Lipo-Cl2MBP (clodronate) for macrophages depletion was injected via a tail vein 5 d before ischemia induction and again 2 d before ischemia induction. RESULTS: The study found that the post-ischemia mice had higher levels of serum creatinine and BUN at 16 and 24 h compared to the controls. Immunofluorescence staining showed there were more macrophages in the post-ischemic tissue at 2, 8, 16 and 24 h compared to the control tissue, and that most of these macrophages were located in the outer medulla. The mice treated with clodronate prior to ischemia induction were found to have lower levels of serum creatinine compared to those mice that weren't treated with clodronate. CONCLUSIONS: There was significant infiltration of macrophages from the early phase of ischemic ARF, and this peaked at 16-24 h. Macrophage depletion using clodronate was protective against ischemic ARF.
Animals
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Antigens, CD11b
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Blood Urea Nitrogen
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Clodronic Acid
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Creatinine/blood
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Fluorescent Antibody Technique
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Inflammation/*physiopathology
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Ischemia/*complications/pathology/physiopathology
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Kidney Failure, Acute/blood/etiology/*pathology/physiopathology
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Kidney Medulla/*pathology
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*Macrophages
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Male
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Mice
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Mice, Inbred C57BL
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Perfusion
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Time Factors
4.Macrophage Depletion by Clodronate Liposomes Suppresses Neointimal Formation After Carotid Artery Injury in Apolipoprotein E-Deficient Mice.
Soon Jun HONG ; Tae Hoon AHN ; Wan Joo SHIM ; Seong Mi PARK ; Jong Il CHOI ; Jae Suk PARK ; Sang Yup LIM ; Do Sun LIM ; Chang Gyu PARK ; Hong Seog SEO
Korean Circulation Journal 2008;38(5):244-249
BACKGROUND AND OBJECTIVES: Clodronate liposomes deplete phagocytic cells, thereby suppressing inflammation after vascular injury. We compared the effect of clodronate liposomes on macrophage depletion and neointimal formation in apolipoprotein E-deficient mice [ApoE (-) mice]. MATERIALS AND METHODS: ApoE (-) mice were randomly assigned to the clodronate liposomes group (Clodronate Group, n=7) and the vehicle liposomes group (Control Group, n=7). Clodronate (0.1 mL/10 g) was injected via the tail vein starting 2 days (d-2) before left common carotid artery injury. RESULTS: The percentage of blood monocytes was subsequently decreased after clodronate injection (14.0+/-7.4% at baseline, 6.8+/-4.9% at 24 hours and 0.7+/-0.3% at 1 week after the clodronate liposome injection). The percentage of macrophages in the plaque area was significantly lower in the clodronate group at week 2 (32.0+/-6.5 vs. 68.7+/-7.6%, respectively, p<0.05) and at week 4 (37.3+/-8.5 vs. 62.6+/-9.4%, respectively, p<0.05). The interleukin (IL)-6 and tumor necrosis factor (TNF)-alpha concentrations were significantly decreased in the clodronate group at week 4 (12.3+/-2.5 vs. 22.9+/-3.5 pg/mL, respectively, p<0.05 for IL-6 and 16.6+/-2.2 vs. 43.6+/-6.1 pg/mL, respectively, p<0.05 for TNF-alpha). The plaque volume was significantly greater in the control group at week 2 (0.345+/-0.063 vs. 0.153+/-0.053 mm2, respectively, p<0.05) and at week 4 (0.320+/-0.027 vs. 0.167+/-0.070 mm2, respectively, p<0.05). CONCLUSION: Intravenous administration of clodronate liposomes depleted monocytes and macrophages, and so this reduced the inflammatory markers and neointimal formation in ApoE (-) mice.
Administration, Intravenous
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Animals
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Apolipoproteins
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Apolipoproteins E
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Carotid Arteries
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Carotid Artery Injuries
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Carotid Artery, Common
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Clodronic Acid
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Inflammation
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Interleukin-6
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Interleukins
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Liposomes
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Macrophages
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Mice
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Monocytes
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Phagocytes
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Tumor Necrosis Factor-alpha
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Vascular System Injuries
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Veins
5.Experimental study for thrombocytopenic purpura therapy by targeting macrophages in liver and spleen.
Zhong-Hua TAN ; Pei-Yong LI ; Yi-Hua ZHU ; Xiao-Wen JI ; Zhen-Yu WU
Journal of Experimental Hematology 2007;15(1):103-107
The study purpose was to explore whether dichloromethylene diphosphonate (Cl(2)MDP)-loaded gelatin particles can induce the depletion of macrophage in reticuloendothelial system of liver and spleen or can depress the immunity of macrophage in SD rat models of immune thrombocytopenic purpura (ITP) to treat the ITP rats. New Zealand rabbits were immunized with platelets of SD rats to prepare rabbit anti-rat platelet serum, and the serum was intravenously injected into SD rats to produce the ITP model. In experimental ITP models, 150 microl of anti-platelet serum was intravenously injected into SD rats per 24 hours. The platelet counts maintained pathological level and were persistently less than 50 x 10(9)/L in the models during experiment process. The MTT test of macrophage RAW264.7 was carried out by means of Cl(2)MDP-loaded gelatin particles in vitro. After intravenous injection of a group dose of Cl(2)MDP-gelatin particles, the platelet counts of the rats were measured at the time of 4 hours, 24 hours, 48 hours, 72 hours and 96 hours, respectively, and bleeding times were detected in 24 hours. The results showed that Cl(2)MDP-loaded gelatin particles increased the platelet counts of ITP models to mean of 180 x 10(9)/L, a physiological level in 24 hours after injection, and kept this platelet level through whole process of 120 hours. Furthermore, rats pre-treated with Cl(2)MDP-loaded gelatin particles avoided the decrease of platelet counts significantly when they were injected anti-platelet serum. It is concluded that Cl(2)MDP-loaded gelatin particles restrain multiplication of macrophage RAW264.7, and promptly, effectively restore platelet counts of ITP models to physiological level in a dose dependent manner. So, the targeting therapy of drug-loaded gelatin particles offers a new idea and approach to treat ITP, and this strategy is worthy of further studies.
Animals
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Clodronic Acid
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administration & dosage
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Drug Carriers
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Drug Delivery Systems
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Gelatin
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administration & dosage
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Liver
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cytology
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drug effects
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Macrophages
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drug effects
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physiology
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Particle Size
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Purpura, Thrombocytopenic, Idiopathic
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therapy
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Rabbits
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Rats
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Rats, Sprague-Dawley
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Spleen
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cytology
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drug effects