A comparison of the in vitro mineralisation and dentinogenic potential of mesenchymal stem cells derived from adipose tissue, bone marrow and dental pulp

Abstract

Stem-cell-based therapies provide a biological basis for the regeneration of mineralised tissues. Stem cells isolated from adipose tissue (ADSCs), bone marrow (BMSCs) and dental pulp (DPSCs) have the capacity to form mineralised tissue. However, studies comparing the capacity of ADSCs with BMSCs and DPSCs for mineralised tissue engineering are lacking, and their ability to regenerate dental tissues has not been fully explored. Characterisation of the cells using fluorescence-activated cell sorting and semi-quantitative reverse transcription PCR for MSC markers indicated that they were immunophenotypically similar. Alizarin red (AR) staining and micro-computed tomography (µCT) analyses demonstrated that the osteogenic potential of DPSCs was significantly greater than that of BMSCs and ADSCs. Scanning electron microscopy and AR staining showed that the pattern of mineralisation in DPSC cultures differed from ADSCs and BMSCs, with DPSC cultures lacking defined mineralised nodules and instead forming a diffuse layer of low-density mineral. Dentine matrix components (DMCs) were used to promote dentinogenic differentiation. Their addition to cultures resulted in increased amounts of mineral deposited in all three cultures and significantly increased the density of mineral deposited in BMSC cultures, as determined by µCT analysis. Addition of DMCs also increased the relative gene expression levels of the dentinogenic markers dentine sialophosphoprotein and dentine matrix protein 1 in ADSC and BMSC cultures. In conclusion, DPSCs show the greatest potential to produce a comparatively high volume of mineralised matrix; however, both dentinogenesis and mineral volume was enhanced in ADSC and BMSC cultures by DMCs, suggesting that these cells show promise for regenerative dental therapies.

Production of Human Dental Pulp Cells with a Medicinal Manufacturing Approach

“…DT ¼ CT=doubling numberðnf ¼ final number of cells at 80% confluence; ni ¼ initial number of cells; and CT ¼ culture timeÞ (2) Karyotyping Post-thaw P4-HDPCs were exposed for 3 hours to 0.7% colcemid (Life Technologies) diluted in the culture medium, and then cells were detached and centrifuged. The pellet was then resuspended in 0.075 mol/L KCl for 2 minutes at room temperature.…”

“…In this context, stem/progenitor cells appear to be particularly appropriate because of their high expansion ability and differentiation potential both in vitro and in vivo (1). If bone marrow and adipose tissue are considered potential sources of stem/progenitor cells, painful collection protocols, the decline of the amount of stem/ progenitor cells with age, the necessity of general anesthesia, reduced proliferation capacity, and risk of morbidity at the collection site encourage the search for alternative candidates (1,2). Human impacted third molars are frequently removed for therapeutic reasons and the loose connective tissue they contain; the dental pulp appears to be a valuable source of stem/progenitor cells for pulp/dentin and bone engineering.…”

<i>In vivo</i> evaluation of chitosan-glycerol gel scaffolds seeded with stem cells for full-thickness mandibular bone regeneration

“…Various studies have widely demonstrated the ability of adipose-derived stem cells (ADSCs) to differentiate into mesenchymal-derived cell lines (adipocytes, osteoblasts, chondroblasts, and endothelial cells) and not mesenchymal cell lines (skeletal muscle cells, cardiac cells, and neuronal cells) (12)(13)(14)(15). Numerous papers have been published in recent years on bone regeneration for skeletal and mandibular repair using ADSCs (16)(17)(18) …”

Therapeutic potential of dental stem cells

“…DPSCs have shown the greatest potential to produce a high volume of mineralized matrix, suggesting that these cells also show promise for use in regenerative dental therapies. 28 Dental pulp progenitors have not been clearly identified but some data suggest that pericytes, which are able to differentiate into osteoblasts, could also differentiate into odontoblasts. 29 …”