The Interaction of Cationic Polymers and Their Bisphosphonate Derivatives with Hydroxyapatite

Abstract

Conjugating proteins with bisphosphonates (BPs), a class of molecules with exceptional affinity to hydroxyapatite (HA), is a feasible means to impart bone affinity to protein-based therapeutic agents. To increase the targeting effectiveness while minimizing protein modification, a polymeric linker containing multiple copies of BPs could be constructed for protein conjugation and targeting to bone. Towards this goal, poly(L-lysine) (PLL) and poly(ethylenimine) (PEI) were utilized as the polymeric backbones to incorporate a BP, namely 2-(3-mercaptopropylsulfanyl)-ethyl-1,1-bisphosphonic acid (thiolBP), by using N-hydroxysuccinimidyl polyethylene glycol maleimide and succinimidyl-4-(N-maleimidomethyl)-cyclohexane-1-carboxylate, respectively. In vitro and in vivo mineral affinity of the polymer-BP conjugates were determined in comparison with the unmodified polymers. The in vitro results indicated strong binding of the cationic polymers to HA in their unmodified form. BP conjugation did not enhance the inherent mineral affinity of the polymers; in contrast, certain modifications negatively affected the polymers' binding to the HA. In vivo results from a subcutaneous implant model in rats also showed no significant difference in mineral affinity of the BP modified and unmodified PEI. We conclude that thiolBP conjugation to the cationic polymers PLL and PEI was not beneficial for increasing the mineral affinity of the polymeric molecules. The strong interaction between the cationic polymers and HA may make the polymers suitable for imparting mineral affinity to bone-acting therapeutics.

Preparation of BMP-2 Containing Bovine Serum Albumin (BSA) Nanoparticles Stabilized by Polymer Coating

“…Recombinant human bone morphogenetic protein-2 (BMP-2) was expressed in Escherichia coli and purified as described before (22). Fluorescein isothiocyanate (FITC) was obtained from Pierce (Rockford, IL, USA) and it was used to label the polymers PLL and PEI according to previously described methodology (23). Dulbecco's Modified Eagle Medium (DMEM), Hank's Balanced Salt Solution (HBSS), GlutaMax-1 (GM), penicillin (10,000 U/mL) and streptomycin (10,000 µg/mL) were from Invitrogen (Carlsbad, CA, USA).…”

BSA Nanoparticles for siRNA Delivery: Coating Effects on Nanoparticle Properties, Plasma Protein Adsorption, andIn VitrosiRNA Delivery

“…Previously reported methods were used to conjugate PLL to 1 kDa PEG [15]. Briefly, PLL was dissolved in 0.1 M phosphate buffer (PB, pH 7.4) to a final concentration of 2.4 mg/mL.…”

Bisphosphonate‐functionalized poly(β‐amino ester) network polymers

“…30 They have strong affinity for the bone mineral, calcium hydroxyapatite, enabling them to chelate calcium ions and prevent bone dissolution. 31,32 BP conjugated small molecule drugs, [33][34][35][36][37] imaging agents, 38,39 proteins, 40,41 and polymers [42][43][44][45][46][47][48] are used for bone targeting. The polymer-drug conjugates with bisphosphonic acid groups are expected to both improve efficiency of drugs and reduce side effects by increasing the solubility of drugs and its blood-circulation time.…”