Periosteum as a source of mesenchymal stem cells: the effects of TGF-β3 on chondrogenesis

Abstract

INTRODUCTION:Numerous experimental efforts have been undertaken to induce the healing of lesions within articular cartilage by re-establishing competent repair tissue. Adult mesenchymal stem cells have attracted attention as a source of cells for cartilage tissue engineering. The purpose of this study was to investigate chondrogenesis employing periosteal mesenchymal cells.METHODS:Periosteum was harvested from patients who underwent orthopedic surgeries. Mesenchymal stem cells were characterized through flow cytometry using specific antibodies. The stem cells were divided into four groups. Two groups were stimulated with transforming growth factor β3 (TGF-β3), of which one group was cultivated in a monolayer culture and the other was cultured in a micromass culture. The remaining two groups were cultivated in monolayer or micromass cultures in the absence of TGF-β3. Cell differentiation was verified through quantitative reverse transcription-polymerase chain reaction (RT-PCR) and using western blot analysis.RESULT:In the groups cultured without TGF-β3, only the cells maintained in the micromass culture expressed type II collagen. Both the monolayer and the micromass groups that were stimulated with TGF-β3 expressed type II collagen, which was observed in both quantitative RT-PCR and western blot analysis. The expression of type II collagen was significantly greater in the micromass system than in the monolayer system.CONCLUSION:The results of this study demonstrate that the interactions between the cells in the micromass culture system can regulate the proliferation and differentiation of periosteal mesenchymal cells during chondrogenesis and that this effect is enhanced by TGF-β3.

Elucidating Multiscale Periosteal Mechanobiology: A Key to Unlocking the Smart Properties and Regenerative Capacity of the Periosteum?

“…63 In the PDC density study, the level of type II collagen mRNA was significantly higher in PDCs cultured in a high-density, micromass culture system compared to cells cultured in a monolayer. 73 Furthermore, PDCs cultured in a micromass culture system in the absence of TGF-b3, a chondrogenic growth factor also exhibited type II collagen expression, indicating that mechanical signaling alone can cause PDCs to differentiate. 73 Culture of periosteal explants in spinner flask bioreactors resulted in a fourfold increase in cartilage yield compared to static culture conditions.…”

“…73 Furthermore, PDCs cultured in a micromass culture system in the absence of TGF-b3, a chondrogenic growth factor also exhibited type II collagen expression, indicating that mechanical signaling alone can cause PDCs to differentiate. 73 Culture of periosteal explants in spinner flask bioreactors resulted in a fourfold increase in cartilage yield compared to static culture conditions. 54 The majority of tissue growth occurred in the longitudinal direction, indicating PDCs are sensitive to the direction of applied shear stress.…”

“…49 Culture density of PDCs has been found to affect the level of type II collagen mRNA expression, a marker for chondrogenesis. 73 Studies using a model MSC cell line demonstrate that increasing cell seeding density per se exerts volume changing dilatational stress on the cell as a whole and shape changing deviatoric stress on the nucleus of stem cells. 63 In the PDC density study, the level of type II collagen mRNA was significantly higher in PDCs cultured in a high-density, micromass culture system compared to cells cultured in a monolayer.…”

Current perspectives in stem cell research for knee cartilage repair

“…Periosteum progenitor MSCs88 and umbilical cord blood MSCs89 can both be induced towards the chondrogenic lineage in the presence of TGF-β. Periosteum progenitor MSCs have been successfully applied to repair models of cartilage defects in vivo 52,57,90.…”

[ARTIGO RETRATADO] Revisão dos artigos sobre ortopedia recentemente publicados na imprensa científica brasileira

“…The heparanase-2 isoform may be involved in normal metabolic processes, as evidenced by its higher expression in the control intervertebral discs relative to the expression of heparanase-1. Mara et al 22 investigated chondrogenesis employing periosteal mesenchymal cells harvested from patients who underwent orthopedic surgeries. Mesenchymal stem cells were characterized through flow cytometry using specific antibodies and were divided into four groups, two stimulated with transforming growth factor β3 (TGF-β3), one cultivated in a monolayer culture one in a micromass culture.…”