Adipose-derived stem cells for dental tissue engineering

Davies, Owen (2014). Adipose-derived stem cells for dental tissue engineering. University of Birmingham. Ph.D.

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Abstract

Mesenchymal stem cells are valuable for regenerative dental research. However, the use of adipose-derived cells (ADCs) within regenerative dentistry remains relatively unexplored. This project aimed to determine an optimal method for the isolation of rat ADCs, to evaluate the influence of cell selection and cryo-storage on MSC phenotype, and compare the relative stemness and dentinogenic capacity of ADCs with bone marrow (BMDCs) and dental pulp-derived cells (DPDCs). Digestion with type-I collagenase for 30 minutes at 37ºC released the greatest number of viable and proliferative ADCs from inguinal adipose tissue. FACS and sqRT-PCR profiling indicated that ADCs shared similar levels of MSC markers (e.g. CD73, CD90, CD105) with BMDCS and DPDCs. The expression of MSC markers was also increased following cryo-storage for all cell types. Alizarin red staining, SEM and micro-CT analyses indicated that the osteogenic differentiation capacity of ADCs appeared lower than that of BMDCs and DPDCs. The FACS procedure reduced cell viability and CD29/CD90 cells had limited osteogenic differentiation capacity when compared to unsorted cell populations. Dentine matrix component (DMCs) supplementation (1 µg/mL) increased the volume of mineralised deposits in ADC, BMDC and DPDC cultures, as well as the expression of odontogenic markers (DMP1 and DSPP) in ADC and BMDC cultures. In conclusion ADCs have an odontogenic capacity, although this may be limited when compared with BMDCs and DPDCs. These findings indicate that when compared with BMDCs or DPDCs, ADCs may have a comparatively limited applicability for dental tissue engineering. However, ADCs can be isolated in comparatively large numbers with relatively little patient discomfort when compared with BMDCs and DPDCs, and these and previous studies have indicated that ADCs can be induced towards a dentinogenic phenotype.

Type of Work:

Thesis (Doctorates > Ph.D.)

Award Type:

Doctorates > Ph.D.

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