Top

Published in:

01-12-2012 | Original Article

Delivery system for autologous growth factors fabricated with low-molecular-weight heparin and protamine to attenuate ischemic hind-limb loss in a mouse model

Authors: Shingo Nakamura, Megumi Takikawa, Masayuki Ishihara, Takefumi Nakayama, Satoko Kishimoto, Susumu Isoda, Yuichi Ozeki, Masahiro Sato, Tadaaki Maehara

Published in: Journal of Artificial Organs | Issue 4/2012

Abstract

Frozen and thawed platelet-rich plasma (PRP) contains high concentrations of various growth factors, such as fibroblast growth factor (FGF)-2, vascular endothelial growth factor, and hepatocyte growth factor. We previously reported that low-molecular-weight heparin/protamine microparticles (LH/P MPs) are useful as biodegradable carriers for the controlled release of FGF-2. In this study, we examined the ability of PRP/LH/P MPs to prevent limb loss in an induced ischemic hind-limb model that used adult BALB/c-nu/nu male mice. One day after inducing ischemia, intramuscular injections of a PRP/LH/P MPs solution were administered into several sites of the ischemic hind limb. Seven days and onward after the injections, the PRP/LH/P MPs-treated and PRP-treated groups recovered from ischemia, as reflected by the improved oxygen saturation. In the PRP-treated group, however, the level of recovery of oxygen saturation after ischemia decreased after 14 days. From the 21st day onward, there was a significant difference between those two groups. In the LH/P MPs-treated group, a partial recovery occurred only in the early period. The saline-treated group (i.e., the control) and the noninjection group (i.e., ischemia only) exhibited no recovery. The limb survival rate at 1 year in the ischemia-induced mice injected with PRP/LH/P MPs was approximately 25 % (two of eight mice) but was absent in the other groups.

Abraham JA, Mergia J, Whang L, Tumolo A, Friedman J, Hjerrild KA, Gospodarowicz D, Fiddes JC. Nucleotide sequence of a bovine clone encoding the angiogenic protein, basic fibroblast growth factor. Science. 1986;234:545–8.CrossRef

Takeshita S, Tsurumi Y, Couffinahal T, Asahara T, Bauters C, Symes J, Ferrara N, Isner JM. Gene transfer of naked DNA encoding for three isoforms of vascular endothelial growth factor stimulates collateral development in vivo. Lab Invest. 1996;75:487–501.PubMed

VanBelle E, Witzenbichler B, Chen D, Silver M, Chang L, Schwall R, Isner JM. Potentiated angiogenic effect of scatter factor/hepatocyte growth factor via induction of vascular endothelial growth factor. The case for paracrine amplification of angiogenesis. Circulation. 1998;97:381–90.CrossRef

Cote MF, Laroche G, Gagnon E, Chevallier P, Doillon CJ. Denatured collagen as support for a FGF-2 delivery system: physicochemical characterizations and in vitro release kinetics and bioactivity. Biomaterials. 2004;25:3761–72.PubMedCrossRef

Young S, Wong M, Tabata Y, Mikos AG. Gelatin as a delivery vehicle for the controlled release of bioactive molecules. J Control Release. 2005;109:256–74.PubMedCrossRef

DeBlois C, Cote MF, Doillon CJ. Heparin–fibroblast growth factor–fibrin complex: in vitro and in vivo applications to collagen-based materials. Biomaterials. 1994;15:665–72.PubMedCrossRef

Nakamura S, Nambu M, Ishizuka T, Hattori H, Kanatani Y, Takase B, Kishimoto S, Amano Y, Aoki H, Kiyosawa T, Ishihara M, Maehara T. Effect of controlled release of fibroblast growth factor-2 from chitosan/fucoidan micro complex hydrogel on in vitro and in vivo vascularization. J Biomed Mater Res A. 2008;85:619–27.PubMed

Kocher AA, Schuster MD, Szabolcs MJ, Takuma S, Burkhoff D, Wang J, Homma S, Edwards NM, Itescu S. Neovascularization of ischemic myocardium by human bone-marrow-derived angioblasts prevents cardiomyocyte apoptosis, reduces remodeling and improves cardiac function. Nat Med. 2001;7:430–6.PubMedCrossRef

Fuchs S, Baffour R, Zhou YF, Shou M, Pierre A, Tio FO, Weissman NJ, Leon MB, Epstein SE, Kornowski R. Transendocardial delivery of autologous bone marrow enhances collateral perfusion and regional function in pigs with chronic experimental myocardial ischemia. J Am Coll Cardiol. 2001;37:1726–32.PubMedCrossRef

10.

Nakamura S, Kishimoto S, Nakamura S, Nambu M, Fujita M, Tanaka Y, Mori Y, Tagawa M, Maehara T, Ishihara M. Fragmin/protamine microparticles as cell carriers to enhance viability of adipose-derived stromal cells and their subsequent effect on in vivo neovascularization. J Biomed Mater Res A. 2010;92:1614–22.PubMed

11.

Pittenger MF, Martin BJ. Mesenchymal stem cells and their potential as cardiac therapeutics. Circ Res. 2004;95:9–20.PubMedCrossRef

12.

Kalka C, Masuda H, Takahashi T, Kalka-Moll WM, Silver M, Kearney M, Li T, Isner JM, Asahara T. Transplantation of ex vivo expanded endothelial progenitor cells for therapeutic neovascularization. Proc Natl Acad Sci USA. 2000;97:3422–7.PubMedCrossRef

13.

Shyu KG, Manor O, Magner M, Yancopoulos GD, Isner JM. Direct intramuscular injection of plasmid DNA encoding angiopoietin-1 but not angiopoietin-2 augments revascularization in the rabbit ischemic hindlimb. Circulation. 1998;98:2081–7.PubMedCrossRef

14.

Taniyama Y, Morishita R, Aoki M, Nakagami H, Yamamoto K, Yamazaki K, Matsumoto K, Nakamura T, Kaneda Y, Ogihara T. Therapeutic angiogenesis induced by human hepatocyte growth factor gene in rat and rabbit hind limb ischemia models: preclinical study for treatment of peripheral arterial disease. Gene Ther. 2001;8:181–9.PubMedCrossRef

15.

Morishita R, Makino H, Aoki M, Hashiya N, Yamasaki K, Azuma J, Taniyama Y, Sawa Y, Kaneda Y, Ogihara T. Phase I/IIa clinical trial of therapeutic angiogenesis using hepatocyte growth factor gene transfer to treat critical limb ischemia. Arterioscler Thromb Vasc Biol. 2011;31:713–20.PubMedCrossRef

16.

Marx RE. Platelet-rich plasma (PRP): what is PRP and what is not PRP? Implant Dent. 2001;10:225–8.PubMedCrossRef

17.

Man D, Plosker H, Winland-Brown JE. The use of autologous platelet-rich plasma (platelet gel) and autologous platelet-poor plasma (fibrin glue) in cosmetic surgery. Plast Reconstr Surg. 2001;107:229–37.PubMedCrossRef

18.

Bhanot S, Alex JC. Current applications of platelet gels in facial plastic surgery. Fac Plast Surg. 2002;18:27–33.CrossRef

19.

Pietramaggiori G, Kaipainen A, Czeczuga JM, Wagner CT, Orgill DP. Freeze-dried platelet-rich plasma shows beneficial healing properties in chronic wounds. Wound Repair Regen. 2006;14:573–80.PubMedCrossRef

20.

Sugimori E, Shintani S, Ishikawa K, Hamakawa H. Effects of apatite foam combined with platelet-rich plasma on regeneration of bone defects. Dent Mater J. 2006;25:591–6.PubMedCrossRef

21.

Abuzeni PZ, Alexander RW. Enhancement of autologous fat transplantation with platelet rich plasma. Int J Radiat Oncol Biol Phys. 2010;78:888–96.CrossRef

22.

Sadati KS, Alexander RW, Corrado AC. Platelet-rich plasma (PRP) utilized to promote greater graft volume retention in autologous fat grafting. Am J Cosm Surg. 2006;23:203–11.

23.

Powell DM, Chang E, Farrior EH. Recovery from deep-plane rhytidectomy following unilateral wound treatment with autologous platelet gel: a pilot study. Arch Fac Plast Surg. 2001;3:245–50.CrossRef

24.

Kurita J, Miyamoto M, Ishii Y, Aoyama J, Takagi G, Naito Z, Tabata Y, Ochi M, Shimizu K. Enhanced vascularization by controlled release of platelet-rich plasma impregnated in biodegradable gelatin hydrogel. Ann Thorac Surg. 2011;92:837–44.PubMedCrossRef

25.

Ishihara M, Obara K, Ishizuka T, Fujita M, Sato M, Masuoka K, Saito Y, Yura H, Matsui T, Hattori H, Kikuchi M, Kurita A. Controlled release of fibroblast growth factors and heparin from photocrosslinked chitosan hydrogels and subsequent effect on in vivo vascularization. J Biomed Mater Res A. 2003;64:551–9.PubMedCrossRef

26.

Salmivirta M, Lidholt K, Lindahl U. Heparan sulfate: a piece of information. FASEB J. 1996;10:1270–9.PubMed

27.

Lindahl U, Lidholt K, Spillmann D, Kjellen L. More to “heparin” than anticoagulation. Thromb Res. 1994;75:1–32.PubMedCrossRef

28.

Hirsh J, Warkentin TE, Shaughnessy SG, Anand SS, Halperin JL, Raschke R, Granger C, Ohman EM, Dalen JE. Heparin and low-molecular-weight heparin, mechanisms of action, pharmacokinetics, dosing, monitoring, efficacy, and safety. Chest. 2001;119:64S–94S.PubMedCrossRef

29.

Wolzt M, Weltermann A, Nieszpaur-Los M, Schneider B, Fassolt A, Lechner K, Eichler HG, Kyrle PA. Studies on the neutralizing effects of protamine on unfractionated and low molecular weight heparin (Fragmin) at the site of activation of the coagulation system in man. Thromb Haemost. 1995;73:439–43.PubMed

30.

Pan M, Suarez de Lezo J, Medina A, Romero M, Hernández E, Segura J, Melian F, Wangüemert F, Landin M, Benítez F, Amat M, Velasco F, Torres A. In-laboratory removal of femoral sheath following protamine administration in patients having intracoronary stent implantation. Am J Cardiol. 1997;80:1336–8.PubMedCrossRef

31.

Nakamura S, Kanatani Y, Kishimoto S, Nakamura S, Ohno C, Horio T, Masanori F, Hattori H, Tanaka Y, Kiyosawa T, Maehara T, Ishihara M. Controlled release of FGF-2 using fragmin/protamine microparticles and effect on neovascularization. J Biomed Mater Res A. 2009;91:814–23.PubMed

32.

Couffinhal T, Silver M, Zheng LP, Kearney M, Witzenbichler B, Isner JM. Mouse model of angiogenesis. Am J Pathol. 1998;152:1667–79.PubMed

33.

Cho SW, Moon SH, Lee SH, Kang SW, Kim J, Lim JM, Kim HS, Kim BS, Chung HM. Improvement of postnatal neovascularization by human embryonic stem cell derived endothelial-like cell transplantation in a mouse model of hindlimb ischemia. Circulation. 2007;116:2409–19.PubMedCrossRef

34.

Fujita M, Ishihara M, Shimizu M, Obara K, Nakamura S, Kanatani Y, Morimoto Y, Takase B, Matsui T, Kikuchi M, Maehara T. Therapeutic angiogenesis induced by controlled release of fibroblast growth factor-2 from injectable chitosan/non-anticoagulant heparin hydrogel in a rat hindlimb ischemia model. Wound Repair Regen. 2007;15:58–65.PubMedCrossRef

35.

Ring FJ. Thermal imaging today and its relevance to diabetes. J Diabetes Sci Technol. 2010;4:857–62.PubMed

36.

Eppley BL, Pietrzak WS, Blanton M. Platelet-rich plasma: a review of biology and applications in plastic surgery. Plast Reconstr Surg. 2006;118:147e–59e.PubMedCrossRef

37.

Zimmermann R, Arnold D, Strasser E, Ringwald J, Schlegel A, Wiltfang J, Eckstein R. Sample preparation technique and white cell content influence the detectable levels of growth factors in platelet concentrates. Vox Sang. 2003;85:283–9.PubMedCrossRef

38.

Takikawa M, Nakamura S, Nakamura S, Nambu M, Ishihara M, Fujita M, Kishimoto S, Doumoto T, Yanagibayashi S, Azuma R, Yamamoto N, Kiyosawa T. Enhancement of vascularization and granulation tissue formation by growth factors in human platelet-rich plasma-containing fragmin/protamine microparticles. J Biomed Mater Res B Appl Biomater. 2011;97:373–80.PubMed

39.

Eppley BL, Woodell JE, Higgins J. Platelet quantification and growth factor analysis from platelet-rich plasma: implications for wound healing. Plast Reconstr Surg. 2004;114:1502–8.PubMed

40.

Weibrich G, Kleis WK, Hafner G, Hitzler WE. Growth factor levels in platelet-rich plasma and correlations with donor age, sex, and platelet count. J Craniomaxillofac Surg. 2002;30:97–102.PubMedCrossRef

41.

Ishihara M, Ono K. Structure and function of heparin and heparan sulfate; heparinoid library and modification of FGF-activities. Trends Glycosci Glycotechnol. 1998;10:223–33.CrossRef

42.

Mori Y, Nakamura S, Kishimoto S, Kawakami M, Suzuki S, Matsui T, Ishihara M. Preparation and characterization of low-molecular-weight heparin/protamine nanoparticles (LMW-H/P NPs) as FGF-2 carrier. Int J Nanomed. 2010;7:147–55.CrossRef

43.

Kishimoto S, Hattori H, Nakamura S, Amano Y, Kanatani Y, Tanaka Y, Mori Y, Harada Y, Tagawa M, Ishihara M. Expansion and characterization of human bone marrow-derived mesenchymal stem cells cultured on fragmin/protamine microparticle-coated matrix with fibroblast growth factor-2 in low serum medium. Tissue Eng Part C Methods. 2009;15:523–7.PubMedCrossRef

44.

Chen FM, Zhang M, Wu ZF. Toward delivery of multiple growth factors in tissue engineering. Biomaterials. 2001;31:6279–86.CrossRef

Title: Delivery system for autologous growth factors fabricated with low-molecular-weight heparin and protamine to attenuate ischemic hind-limb loss in a mouse model
Authors: Shingo Nakamura
Megumi Takikawa
Masayuki Ishihara
Takefumi Nakayama
Satoko Kishimoto
Susumu Isoda
Yuichi Ozeki
Masahiro Sato
Tadaaki Maehara
Publication date: 01-12-2012
Publisher: Springer Japan
Published in: Journal of Artificial Organs / Issue 4/2012
Print ISSN: 1434-7229
Electronic ISSN: 1619-0904
DOI: https://doi.org/10.1007/s10047-012-0658-0

At a glance: The ONWARDS insulin icodec trials

Springer Medicine

Delivery system for autologous growth factors fabricated with low-molecular-weight heparin and protamine to attenuate ischemic hind-limb loss in a mouse model

Abstract

At a glance: The ONWARDS insulin icodec trials

Springer Medicine

Abstract

Please log in to get access to this content

Other articles of this Issue 4/2012

Long-term results of tissue-engineered small-caliber vascular grafts in a rat carotid arterial replacement model

Effect of metal surface characteristics on the adhesion performance of the integrated low-level energies method of adhesion

Atrial septum defect closure device in a beating heart, from the perspective of a researcher in artificial organs

Survey of blood pump diaphragm damage in the NIPRO-ventricular assist device

Regulatory frameworks for cell therapy products in Japan

Comparison of two spike areas in bileaflet mechanical valve closing sound