Immunomodulatory fibrous hyaluronic acid-Fmoc-diphenylalanine-based hydrogel induces bone regeneration

J Clin Periodontol. 2023 Feb;50(2):200-219. doi: 10.1111/jcpe.13725. Epub 2022 Sep 27.

Abstract

Aim: To investigate the potential of an ultrashort aromatic peptide hydrogelator integrated with hyaluronic acid (HA) to serve as a scaffold for bone regeneration.

Materials and methods: Fluorenylmethyloxycarbonyl-diphenylalanine (FmocFF)/HA hydrogel was prepared and characterized using microscopy and rheology. Osteogenic differentiation of MC3T3-E1 preosteoblasts was investigated using Alizarin red, alkaline phosphatase and calcium deposition assays. In vivo, 5-mm-diameter calvarial critical-sized defects were prepared in 20 Sprague-Dawley rats and filled with either FmocFF/HA hydrogel, deproteinized bovine bone mineral, FmocFF/Alginate hydrogel or left unfilled. Eight weeks after implantation, histology and micro-computed tomography analyses were performed. Immunohistochemistry was performed in six rats to assess the hydrogel's immunomodulatory effect.

Results: A nanofibrous FmocFF/HA hydrogel with a high storage modulus of 46 KPa was prepared. It supported osteogenic differentiation of MC3T3-E1 preosteoblasts and facilitated calcium deposition. In vivo, the hydrogel implantation resulted in approximately 93% bone restoration. It induced bone deposition not only around the margins, but also generated bony islets along the defect. Elongated M2 macrophages lining at the periosteum-hydrogel interface were observed 1 week after implantation. After 3 weeks, these macrophages were dispersed through the regenerating tissue surrounding the newly formed bone.

Conclusions: FmocFF/HA hydrogel can serve as a cell-free, biomimetic, immunomodulatory scaffold for bone regeneration.

Keywords: biomaterials; bone regeneration; hyaluronic acid; immunomodulation; self-assembling peptides.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Bone Regeneration
  • Calcium / pharmacology
  • Cattle
  • Hyaluronic Acid* / pharmacology
  • Hyaluronic Acid* / therapeutic use
  • Hydrogels* / chemistry
  • Hydrogels* / pharmacology
  • Osteogenesis
  • Periosteum
  • Rats
  • Rats, Sprague-Dawley
  • Tissue Scaffolds / chemistry
  • X-Ray Microtomography

Substances

  • Hydrogels
  • Hyaluronic Acid
  • diphenylalanine
  • Calcium