Biomedical Application of Low Molecular Weight Heparin/Protamine Nano/Micro Particles as Cell- and Growth Factor-Carriers and Coating Matrix

Abstract

Low molecular weight heparin (LMWH)/protamine (P) nano/micro particles (N/MPs) (LMWH/P N/MPs) were applied as carriers for heparin-binding growth factors (GFs) and for adhesive cells including adipose-derived stromal cells (ADSCs) and bone marrow-derived mesenchymal stem cells (BMSCs). A mixture of LMWH and P yields a dispersion of N/MPs (100 nm–3 μm in diameter). LMWH/P N/MPs can be immobilized onto cell surfaces or extracellular matrix, control the release, activate GFs and protect various GFs. Furthermore, LMWH/P N/MPs can also bind to adhesive cell surfaces, inducing cells and LMWH/P N/MPs-aggregate formation. Those aggregates substantially promoted cellular viability, and induced vascularization and fibrous tissue formation in vivo. The LMWH/P N/MPs, in combination with ADSCs or BMSCs, are effective cell-carriers and are potential promising novel therapeutic agents for inducing vascularization and fibrous tissue formation in ischemic disease by transplantation of the ADSCs and LMWH/P N/MPs-aggregates. LMWH/P N/MPs can also bind to tissue culture plates and adsorb exogenous GFs or GFs from those cells. The LMWH/P N/MPs-coated matrix in the presence of GFs may provide novel biomaterials that can control cellular activity such as growth and differentiation. Furthermore, three-dimensional (3D) cultures of cells including ADSCs and BMSCs using plasma-medium gel with LMWH/P N/MPs exhibited efficient cell proliferation. Thus, LMWH/P N/MPs are an adequate carrier both for GFs and for stromal cells such as ADSCs and BMSCs, and are a functional coating matrix for their cultures.

Low-molecular weight heparin protamine complex augmented the potential of adipose-derived stromal cells to ameliorate limb ischemia

“…ADSC/LH/P-MP aggregates could stimulate cell proliferation and migration, resulting in neovascularization in vivo [21]. ADSCs can potentially release multiple angiogenic growth factors that can be efficiently immobilized onto LH/P-MPs [8] or LH/P-MP-coated culture plates [22].…”

Heparinoid Complex-Based Heparin-Binding Cytokines and Cell Delivery Carriers

“…The present review focuses on PEC hydrogels that formed by the chemical interaction of chitosan and crosslinkers [137,138]. Heparinoids, which are conjugated to nano-materials, have been recently investigated for their chemical and biological properties and applications [35,37]. Heparinoid has been conjugated to the surface of nanoparticles (NPs), such as magnetic [139] and metallic NPs [140], heparin-coated nanoparticles coupled to hemoglobin [141], and biopolymers [142].…”

“…Furthermore, Syndecan-4 proteoliposomes enhance FGF-2-induced proliferation, migration, and neovascularization of ischemic muscles [143]. When biological molecules, such as functional proteins and DNA, are incorporated on or within NPs, they provide novel and enhanced activities with potential applications in therapeutics, biosensors, imaging, and drug delivery [35,37,144]. Those biomolecules can be passivated on NPs, thus improving their biocompatibility.…”

“…Electrostatic interactions between oppositely charged polyelectrolytes, such as low-molecular-weight heparin (LMWH) (MW: approximately 5000 Da) and protamine, generate PECs ( Figure 6A). When this interaction occurs in non-equivalent ratios, nonstoichiometric PECs are produced, which causes each PEC particle to carry an excess charge [37,145,146]. Proteins interact with both synthetic and natural PECs [26].…”

“…The above-mentioned activities of heparin-binding cytokines occur in the ECM through specific non-covalent interactions with, for example, ECM receptor molecules and PGs in which multiple GAGs are covalently attached [34]. Those localized interactions have inspired the development of biomaterials that enhance and regulate the heparin-binding cytokine activities for practical applications [35][36][37]. For example, biomaterials that are modified with heparinoids may exhibit increased stability and controlled release and activation.…”