Human periodontal ligament stem cells cultured onto cortico-cancellous scaffold drive bone regenerative process

Abstract

The purpose of this work was to test, in vitro and in vivo, a new tissue-engineered construct constituted by porcine cortico-cancellous scaffold (Osteobiol Dual Block) (DB) and xeno-free ex vivo culture of human Periodontal Ligament Stem Cells (hPDLSCs). hPDLSCs cultured in xeno-free media formulation preserved the stem cells' morphological features, the expression of stemness and pluripotency markers, and their ability to differentiate into mesenchymal lineage. Transmission electron microscopy analysis suggested that after one week of culture, both noninduced and osteogenic differentiation induced cells joined and grew on DB secreting extracellular matrix (ECM) that in osteogenic induced samples was hierarchically assembled in fibrils. Quantitative RT-PCR (qRT-PCR) showed the upregulation of key genes involved in the bone differentiation pathway in both differentiated and undifferentiated hPDLSCs cultured with DB (hPDLSCs/DB). Functional studies revealed a significant increased response of calcium transients in the presence of DB, both in undifferentiated and differentiated cells stimulated with calcitonin and parathormone, suggesting that the biomaterial could drive the osteogenic differentiation process of hPDLSCs. These data were confirmed by the increase of gene expression of L-type voltage-dependent Ca 2+ (VDCC L ), subunits α1C and α2D1 in undifferentiated cells in the presence of DB. In vivo implantation of the hPDLSCs/DB living construct in the mouse calvaria evidenced a precocious osteointegration and vascularisation process. Our results suggest consideration of DB as a biocompatible, osteoinductive and osteoconductive biomaterial, making it a promising tool to regulate cell activities in biological environments and for a potential use in the development of new custom-made tissue engineering.

Treatment of Periodontal Ligament Stem Cells with MOR and CBD Promotes Cell Survival and Neuronal Differentiation via the PI3K/Akt/mTOR Pathway

“…hPDLSCs retain all the features of MSCs, such as self-renewal, immunomodulatory, clonogenicity, and multi-tissue differentiation potential [21,22]. Improve and extend survival time of stem cells in transplantation, is one of the main objectives of research in regenerative medicine.…”

Stemness Maintenance Properties in Human Oral Stem Cells after Long-Term Passage

“…Ingenuity Pathway Analysis (IPA) software (Ingenuity Systems, Redwood City, CA, USA) was employed to infer biological functions of the selected gene datasets. IPA predicts functional characterization based on known gene functional interactions and ranks them by a significance score [27]. …”

MyD88/ERK/NFkB pathways and pro-inflammatory cytokines release in periodontal ligament stem cells stimulated by Porphyromonas gingivalis

“…7 Moreover, data are being collected on their capability of taking part to tissue regeneration when used with appropriate biomaterials. 8,9 Few studies have reported the effects of LPS on hPDLSCs, the aim of the present work was to characterize the molecular pathway triggered by LPS-G in hPDLSCs and to evaluate, in the same cells, the production of a wide array of both inflammatory and antiinflammatory molecules induced by LPS-G.…”