Periodontal disease, highly prevalent in human and canine species, is an inflammatory pathology which can result in tooth loss and in systemic implications. The current therapies are ineffective in avoiding the epithelium growth, bone resorption and ankilosis, which inhibit the formation of a functional periodontal ligament. Tissue Engineering (TE) strategies, combining scaffolds and mesenchymal stem cells, has paved the way for new therapies.
Our goal is to develop an innovative 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 groups.
The SPCL membrane was obtained by solvent casting and then combined with a wet-spun fibre mesh (WSFM) with/without silanol groups. Bi-layered scaffolds were characterized by scanning electron microscopy (SEM), tensile tests, Fourrier Transmission Infra-red (FTIR), and enzymatic degradation assays.
Canine ASCs (cASCs) were obtained from subcutaneous adipose tissue harvested upon programmed surgeries. 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).
SEM revealed a good adherence between the layers, roughness and fibres interconnection. FTIR confirmed the presence of Si-O-Si and Si-OH bonds in functionalized WSFM. Also, the scaffold exhibited a suitable mechanical properties and degradability to be applied in an in vivo environment.
Culturing experiments showed that materials provide a good support for ASCs according to DNA increasing and SEM. ALP activity increasing until 21th day and also the calcium content revealing osteoconductivity and bioactivity.
This work showed that cASCs onto this SPCL bioactive scaffold are a promising TE approach to reach periodontal regeneration, namely, in its osseous component.