Novel approach with intratracheal administration of microgelatin hydrogel microspheres incorporating basic fibroblast growth factor for rescue of rats.

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Novel approach with intratracheal administration of microgelatin hydrogel microspheres incorporating basic fibroblast growth factor for rescue of rats with monocrotaline-induced pulmonary hypertension Keiichi Hirose, MD, PhD, Akira Marui, MD, PhD, Yoshio Arai, MD, PhD, Toshihiro Kushibiki, PhD, Yu Kimura, MEng, Hisashi Sakaguchi, MD, Huang Yuang, MD, B.I.R. Shyamal Chandra, MD, Tadashi Ikeda, MD, PhD, Shigeru Amano, MD, PhD, Yasuhiko Tabata, PhD, DMSc, Dpharm, Masashi Komeda, MD, PhD The Journal of Thoracic and Cardiovascular Surgery Volume 136, Issue 5, Pages 1250-1256 (November 2008) DOI: 10.1016/j.jtcvs.2008.05.038 Copyright © 2008 The American Association for Thoracic Surgery Terms and Conditions

Figure 1 Release profile of the radioactivity remaining of basic fibroblast growth factor (bFGF) after intratracheal administration. The x-axis shows the time course and the y-axis shows the percentage of the radioactivity remaining in the lung. mGHMs/bFGF group (filles circles, solid line), Gelatin hydrogel microspheres incorporating iodine 125–labeled bFGF were injected into the trachea by using a pulmonary drug delivery device; free-FGF group (open circles, dotted line): free iodine 125–labeled bFGF solution was injected into the trachea with the same device. The Journal of Thoracic and Cardiovascular Surgery 2008 136, 1250-1256DOI: (10.1016/j.jtcvs.2008.05.038) Copyright © 2008 The American Association for Thoracic Surgery Terms and Conditions

Figure 2 Survival curves after a single intratracheal administration into rats with progressive pulmonary hypertension. Survival curves are generated by means of Kaplan–Meier analysis. These results suggest that a single intratracheal administration of microgelatin hydrogels microspheres incorporating basic fibroblast growth factor (mGHMs/bFGF) improved survival of rats with progressive pulmonary hypertension. mGHMs group (open circles, small dotted line), Monocrotaline (MCT)–injected rats (a single subcutaneous injection of monocrotaline) treated with intratracheal administration of plain mGHMs; free-FGF group (open circles, large dotted line), MCT-injected rats treated with intratracheal administration of free-bFGF; mGHMs/bFGF group (closed circles, solid line), MCT-injected rats treated with intratracheal administration of mGHMs/bFGF (sustained-release bFGF); control group, healthy rats treated with intratracheal administration of 0.1 mL of saline (no circles, thin dotted line). The Journal of Thoracic and Cardiovascular Surgery 2008 136, 1250-1256DOI: (10.1016/j.jtcvs.2008.05.038) Copyright © 2008 The American Association for Thoracic Surgery Terms and Conditions

Figure 3 Right ventricular (RV)/left ventricular (LV) pressure ratio and RV/LV plus septum (LV+S) weight ratio. Panel A shows the RV/LV pressure ratio for the evaluation of pulmonary hypertension 3 weeks after the treatment. Panel B shows the RV/LV+S weight ratio for the evaluation of RV hypertrophy. Control group, Healthy rats treated with intratracheal administration of 0.1 mL of saline; vehicle group, monocrotaline (MCT)– injected rats (receiving a single subcutaneous injection of monocrotaline) treated with intratracheal administration of plain microgelatin hydrogel microspheres (mGHMs); free-FGF group, MCT–injected rats treated with intratracheal administration of free basic fibroblast growth factor (bFGF); mGHMs/bFGF group, MCT-injected rats treated with intratracheal administration of microgelatin hydrogel microspheres incorporating basic fibroblast growth factor (mGHMs/bFGF; sustained-release bFGF). The Journal of Thoracic and Cardiovascular Surgery 2008 136, 1250-1256DOI: (10.1016/j.jtcvs.2008.05.038) Copyright © 2008 The American Association for Thoracic Surgery Terms and Conditions

Figure 4 Micrographs of the lung tissue. Micrographs of the lung tissue (stained with von Willebrand factor, 100× magnification) 3 weeks after treatment in rates with progressive pulmonary hypertension. Histologic sections of the lung tissue stained with von Willebrand factor confirmed the increase in the number of vessels and capillaries. Micrograph I (left upper, control group), Healthy rats treated with intratracheal administration of 0.1 mL saline; Micrograph II (left lower, vehicle group), monocrotaline (MCT)–injected rats (receiving a single subcutaneous injection of monocrotaline) treated with intratracheal administration of plain microgelatin hydrogel microspheres (mGHMs); Micrograph III (right upper, free-FGF group), MCT-injected rats treated with intratracheal administration of free basic fibroblast growth factor (bFGF); Micrograph IV (right lower, mGHMs/bFGF group), MCT-injected rats treated with intratracheal administration of microgelatin hydrogel microspheres incorporating basic fibroblast growth factor (mGHMs/bFGF; sustained-release bFGF). The Journal of Thoracic and Cardiovascular Surgery 2008 136, 1250-1256DOI: (10.1016/j.jtcvs.2008.05.038) Copyright © 2008 The American Association for Thoracic Surgery Terms and Conditions

Figure 5 The number of vessels and capillaries. The left panel (A) shows the number of vessels (diameter >50 μm) per square millimeter, and the right panel (B) shows the number of small vessels, capillaries, or both (diameter <50 μm) per square millimeter 3 weeks after treatment in rats with progressive pulmonary hypertension. Control group, Healthy rats treated with intratracheal administration of 0.1 mL saline; vehicle group, monocrotaline (MCT)–injected rats (receiving a single subcutaneous injection of monocrotaline) treated with intratracheal administration of plain microgelatin hydrogel microspheres (mGHMs); free-FGF group, MCT-injected rats treated with intratracheal administration of free basic fibroblast growth factor (bFGF); mGHMs/bFGF group, MCT-injected rats treated with intratracheal administration of microgelatin hydrogel microspheres incorporating basic fibroblast growth factor (mGHMs/bFGF; sustained-release bFGF). The Journal of Thoracic and Cardiovascular Surgery 2008 136, 1250-1256DOI: (10.1016/j.jtcvs.2008.05.038) Copyright © 2008 The American Association for Thoracic Surgery Terms and Conditions