The effect of platelet rich concentrate on the regenerative capacity of mesenchymal stromal cells and its potential application in cartilage repair / Shani Samuel
The use of regenerative promoters such as mesenchymal stromal cells (MSC) constitutes one of the novel approaches to enhance tissue repair. However, these cells need to be greatly expanded and differentiated prior to their clinical use. Platelet rich concentrate (PRC) has recently emerged as a po...
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| Format: | Thesis |
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2017
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| Online Access: | http://studentsrepo.um.edu.my/10362/ http://studentsrepo.um.edu.my/10362/1/shani.pdf |
| Summary: | The use of regenerative promoters such as mesenchymal stromal cells (MSC) constitutes
one of the novel approaches to enhance tissue repair. However, these cells need to be
greatly expanded and differentiated prior to their clinical use. Platelet rich concentrate
(PRC) has recently emerged as a potentially valuable adjunct that could considerably
enhance the therapeutic potential of MSC. Although there are several lines of evidence
indicating that PRC enhanced proliferation and differentiation of MSC when combined
with lineage-specific culture media, the extent of contribution of the platelet itself has not
been previously demonstrated. This thesis aimed to investigate the effect of PRC in serum
free medium (without the influence of additional differentiation media) in enhancing the
proliferation and differentiation capacity of MSC in vitro and their regenerative potential
to repair focal cartilage defect in vivo. The first study involved optimisation of PRC
preparation and investigating the effect of a single application of PRC in inducing the
proliferation and differentiation of human bone marrow-derived MSC (hMSC) within a
very short time span. The results showed that hMSC proliferation was greatest when the
cells were cultured in medium containing 15% PRC. At this optimal dose, PRC was able
to induce hMSC differentiation to the common mesenchymal lineages i.e. osteogenic,
adipogenic and chondrogenic, within 8 days of culture duration, without the need to be
activated and the aid of any external lineage-specific growth factors in the culture
medium. The study was further extended to compare the expression of lineage-specific
markers both at the mRNA and protein levels of cells undergoing osteogenesis,
adipogenesis and chondrogenesis under the influence of PRC to those cultured in the
standard lineage-specific differentiation media during 24 days of culture (i.e. typical
duration to achieve complete cellular differentiation to the common mesenchymal
lineages). The results indicate that PRC induced osteogenesis and chondrogenesis at a
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greater extent compared to the respective standard differentiation media. Chondrogenic
hMSCs in the PRC group showed lower expression of hypertrophic genes. The final study
was conducted to determine the applicability and the potential advantageous role of PRC
in promoting chondrogenesis of MSC in vivo to enhance cartilage repair in a rabbit model
of focal cartilage defect. Rabbits (6-7 months old) were subjected to full thickness focal
cartilage defects limited to the chondral surface. At 3 and 6 month post-transplantation,
animals in the PRC+allogenic rabbit MSC (rbMSC) group had significantly higher
morphological score (ICRS) and histological score (O’Driscoll) compared to rbMSC and
PRC group alone. Strong safranin O and collagen type II staining and high
glycosaminoglycan content confirmed the formation of a hyaline-like cartilage in the
PRC+rbMSC group. Collectively, these results demonstrate that PRC alone in serum free
medium enhanced MSC regenerative capacity both in vitro and in vivo. PRC on its own
could be used as an adjunct to provide sufficient pool of pre-differentiated MSC for
potential clinical application in musculoskeletal tissue regeneration, particularly for the
repair of focal cartilage injury |
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