Nanostructure controlled sustained delivery of human growth hormone using injectable, biodegradable, pH/temperature responsive nanobiohybrid hydrogel

Abstract

The clinical efficacy of a therapeutic protein, the human growth hormone (hGH), is limited by its short plasma half-life and premature degradation. To overcome this limitation, we proposed a new protein delivery system by the self-assembly and intercalation of a negatively charged hGH onto a positively charged 2D-layered double hydroxide nanoparticle (LDH). The LDH–hGH ionic complex, with an average particle size of approximately 100 nm, retards hGH diffusion. Nanobiohybrid hydrogels (PAEU/LDH–hGH) were prepared by dispersing the LDH–hGH complex into a cationic pH- and temperature-sensitive injectable PAEU copolymer hydrogel to enhance sustained hGH release by dual ionic interactions. Biodegradable copolymer hydrogels comprising poly(β-amino ester urethane) and triblock poly(ε-caprolactone-lactide)-poly(ethylene glycol)-poly-(ε-caprolactone-lactide) (PCLA-PEG-PCLA) were synthesized and characterized. hGH was self-assembled and intercalated onto layered LDH nanoparticles through an anion exchange technique. X-ray diffraction and zeta potential results showed that the LDH–hGH complex was prepared successfully and that the PAEU/LDH–hGH nanobiohybrid hydrogel had a disordered intercalated nanostructure. The biocompatibility of the nanobiohybrid hydrogel was confirmed by an in vitro cytotoxicity test. The in vivo degradation of pure PAEU and its nanobiohybrid hydrogels was investigated and it showed a controlled degradation of the PAEU/LDH nanobiohybrids compared with the pristine PAEU copolymer hydrogel. The LDH–hGH loaded injectable hydrogels suppressed the initial burst release of hGH and extended the release period for 13 days in vitro and 5 days in vivo. The developed nanohybrid hydrogel has the potential for application as a protein carrier to improve patient compliance.