Decellularized bovine small intestinal submucosa-PCL/hydroxyapatite-based multilayer composite scaffold for hard tissue repair

Mater Sci Eng C Mater Biol Appl. 2019 Jan 1;94:788-797. doi: 10.1016/j.msec.2018.10.011. Epub 2018 Oct 2.

Abstract

This study involved the development of a multilayer osteogenic tissue scaffold by assembling decellularized bovine small intestinal submucosa (bSIS) layers, together with synthetic hydroxyapatite microparticles (HAp) and poly(ε-caprolactone) (PCL) as the binder. As a first step, the surface and mechanical properties of the developed scaffold was determined, after which the biocompatibility was evaluated through seeding with isolated rat bone marrow mesenchymal stem cells (BM-MSCs). Then, a 21-day culture study was performed to investigate the in vitro osteoinductive potential of the scaffold on BM-MSCs under standard and osteogenic culture conditions. The SEM findings indicated that a uniform multilayer and perforated structure was acquired; that the HAp microparticles were homogenously distributed within the structure; and that the PCL-bound laminar scaffold had structural integrity. Mechanical tests revealed that the scaffold maintained its mechanical stability for at least 21 days in culture, with no changes in the first-day maximum strength and maximum stress values of 625.123 ± 70.531 N and 6.57762 ± 0.742 MPa, respectively. MTT and SEM analyses together revealed that BM-MSCs preserved their viability and proliferated during a 14-day culture period on the multilayer scaffold. Immunofluorescence analyses indicated that cells on the scaffold differentiated into the osteogenic lineage, by the culture-time-dependent increase in osteogenic markers' expression, i.e. Alkaline phosphatase, Osteopontin, and Osteocalcin. It was also clear that, the osteoinductive effect by the composite scaffold on BM-MSCs could be achieved even without the use of any external osteogenic inducers.

Keywords: Bone tissue engineering; Decellularized bovine SIS; Hydroxyapatite; Multilayer hybrid template; Osteogenic scaffold.

MeSH terms

  • Alkaline Phosphatase / metabolism
  • Animals
  • Cattle
  • Cell Proliferation
  • Durapatite / chemistry*
  • Immunophenotyping
  • Intestinal Mucosa / cytology*
  • Intestine, Small / cytology*
  • Male
  • Mesenchymal Stem Cells / cytology
  • Mesenchymal Stem Cells / enzymology
  • Mesenchymal Stem Cells / ultrastructure
  • Osteopontin / metabolism
  • Polyesters / chemistry*
  • Rats
  • Tissue Scaffolds / chemistry*
  • Wound Healing*

Substances

  • Polyesters
  • Osteopontin
  • polycaprolactone
  • Durapatite
  • Alkaline Phosphatase