In vitro and in vivo assessment of an innovative double layer scaffold for periodontal tissue engineering
Abstract
Periodontal disease is an inflammatory pathology highly prevalent in human and canine species1. It can result in tooth loss and in systemic implications, and the current therapies sometimes are ineffective2. Tissue Engineering (TE) strategies have paved the way for new therapies3,4.
Our goal is to develop a construct for periodontal regeneration based on culturing adipose stem cells (ASCs) onto a bi-layered scaffold comprising a starch+poly(e-caprolactone) (SPCL) membrane, which acts as a guided tissue regeneration membrane, and a SPCL fibre mesh functionalized with osteoconductive silanol groups5.
The SPCL membrane was obtained by solvent casting and then combined with a wet-spun fibre mesh (WSFM) with/without silanol groups. Canine ASCs (cASCs) were obtained from subcutaneous adipose tissue. The proliferation of cASCs seeded/cultured onto the scaffold was studied by dsDNA quantification and SEM. Osteogenic differentiation on the WSFM was assessed by ALP quantification, real time RT-PCR (osteoblastic markers) and histology (Alizarin Red and Lévai Laczkó stainings).
Further in this work, the performance of the SPCL-Si scaffolds was assessed in a mandibular rodent model and compared with collagen commercial membranes (positive control) and empty defects (negative control). After 8 weeks of implantation, the explants were processed by the Donath technique and the new bone formation quantified.
Cell culturing experiments showed that the developed materials provide a good support for cASCs adhesion and proliferation. ALP activity increasing until 21th day and also the calcium content revealing osteoconductivity and bioactivity of the SPCL scaffolds, particularly those functionalized with Si groups. Osteogenic markers, COLIA1, RUNX2 and Osteocalcin, were also expressed by cASCs cultured in the constructs.
Histomorphometrical analysis revealed that the SPCL-Si scaffolds induced higher bone formation compared to collagen materials and empty defects.
This bioactive double layer scaffold revealed the potential to be used in TE approach to reach periodontal regeneration.