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An in vitro investigation into the role of bone marrow‑derived mesenchymal stem cells in the control of disc degeneration.

Hu J, Deng G, Tian Y, Pu Y, Cao P, Yuan W - Mol Med Rep (2015)

Bottom Line: The degenerative and apoptotic indexes were significantly reduced when NP cells were co‑cultured with BMSCs.In conclusion, the anti‑apoptosis and the migration, in addition to mitochondrial transfer associated with BMSC treatments in IVDD, were investigated in vitro in the present study.The interaction between stimulated NP cells and BMSCs is likely involved in to simulating the in vivo process of stem cell‑mediated repair.

View Article: PubMed Central - PubMed

Affiliation: Department of Orthopedic Surgery, Changzheng Hospital, Shanghai 200023, P.R. China.

ABSTRACT
Excessive apoptosis and high expression levels of interleukin‑1β (IL‑1β) in disc cells have been reported to serve important roles in intervertebral disc degeneration (IVDD). Previous studies investigating mesenchymal stem cells (MSCs) have indicated potential for their use in the treatment of IVDD. However, the therapeutic potential and anti‑apoptotic ability of MSCs remains to be fully elucidated. The present study aimed to establish an in vitro model for bone marrow‑derived MSC (BMSC) therapy by investigating the anti‑apoptotic effects, in addition to the migration of BMSCs to nucleus pulposus (NP) cells stimulated by IL‑1β. A co-culture system of BMSCs and NP cells was founded. Following inflammatory stimulation, the NP cells exhibited increased indexes for inflammation‑induced degeneration. The degenerative and apoptotic indexes were significantly reduced when NP cells were co‑cultured with BMSCs. Compared with the indirect co-culture group, the direct co-culture group exhibited an improved capacity for anti-apoptosis. In addition, IL‑1β‑stimulated NP cells attracted and mediated the migration of BMSCs. Mitochondrial transfer from BMSCs to NP cells by tunneling nanotubes was also observed. In conclusion, the anti‑apoptosis and the migration, in addition to mitochondrial transfer associated with BMSC treatments in IVDD, were investigated in vitro in the present study. The interaction between stimulated NP cells and BMSCs is likely involved in to simulating the in vivo process of stem cell‑mediated repair.

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Evaluation of apoptotic incidence. (A) Following double staining with annexin V-fluorescein isothiocyanate (phycoerythrin for direct co-culture) and propidium iodide, 0 ng/ml resulted in a normal apoptosis rate of the NP cells. The apoptotic rates of the NP cells were increased following different levels of IL-1β stimulation, then were reduced when indirectly co-cultured with BMSCs, with a more marked reduction observed in the direct co-culture. (B) The apoptotic incidences of the NP cells in different groups were compared (*P<0.05). (C) The directly co-cultured BMSCs were minimally impacted. NP, nucleus pulposus; IL-1β, interleukin-1β; BMSCs, bone marrow-derived mesenchymal stem cells.
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f4-mmr-12-04-5701: Evaluation of apoptotic incidence. (A) Following double staining with annexin V-fluorescein isothiocyanate (phycoerythrin for direct co-culture) and propidium iodide, 0 ng/ml resulted in a normal apoptosis rate of the NP cells. The apoptotic rates of the NP cells were increased following different levels of IL-1β stimulation, then were reduced when indirectly co-cultured with BMSCs, with a more marked reduction observed in the direct co-culture. (B) The apoptotic incidences of the NP cells in different groups were compared (*P<0.05). (C) The directly co-cultured BMSCs were minimally impacted. NP, nucleus pulposus; IL-1β, interleukin-1β; BMSCs, bone marrow-derived mesenchymal stem cells.

Mentions: Flow cytometric analysis of annexin V-FITC PE/PI staining demonstrated that the level of apoptosis was significantly increased in the NP cells stimulated with inflammatory factors. In addition, the apoptotic rate increased in line with the increase in inflammatory factor concentration. These results indicated that co-culturing NP cells with BMSCs significantly reduced the apoptotic rate of different levels of IL-1β. This suggested that direct co-culture may have improved the anti-apoptotic effects compared with the indirect co-culture (Fig. 4A and B). Notably, the apoptotic rate of the added BMSCs in the direct co-culture group was markedly low (Fig. 4C).


An in vitro investigation into the role of bone marrow‑derived mesenchymal stem cells in the control of disc degeneration.

Hu J, Deng G, Tian Y, Pu Y, Cao P, Yuan W - Mol Med Rep (2015)

Evaluation of apoptotic incidence. (A) Following double staining with annexin V-fluorescein isothiocyanate (phycoerythrin for direct co-culture) and propidium iodide, 0 ng/ml resulted in a normal apoptosis rate of the NP cells. The apoptotic rates of the NP cells were increased following different levels of IL-1β stimulation, then were reduced when indirectly co-cultured with BMSCs, with a more marked reduction observed in the direct co-culture. (B) The apoptotic incidences of the NP cells in different groups were compared (*P<0.05). (C) The directly co-cultured BMSCs were minimally impacted. NP, nucleus pulposus; IL-1β, interleukin-1β; BMSCs, bone marrow-derived mesenchymal stem cells.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4581747&req=5

f4-mmr-12-04-5701: Evaluation of apoptotic incidence. (A) Following double staining with annexin V-fluorescein isothiocyanate (phycoerythrin for direct co-culture) and propidium iodide, 0 ng/ml resulted in a normal apoptosis rate of the NP cells. The apoptotic rates of the NP cells were increased following different levels of IL-1β stimulation, then were reduced when indirectly co-cultured with BMSCs, with a more marked reduction observed in the direct co-culture. (B) The apoptotic incidences of the NP cells in different groups were compared (*P<0.05). (C) The directly co-cultured BMSCs were minimally impacted. NP, nucleus pulposus; IL-1β, interleukin-1β; BMSCs, bone marrow-derived mesenchymal stem cells.
Mentions: Flow cytometric analysis of annexin V-FITC PE/PI staining demonstrated that the level of apoptosis was significantly increased in the NP cells stimulated with inflammatory factors. In addition, the apoptotic rate increased in line with the increase in inflammatory factor concentration. These results indicated that co-culturing NP cells with BMSCs significantly reduced the apoptotic rate of different levels of IL-1β. This suggested that direct co-culture may have improved the anti-apoptotic effects compared with the indirect co-culture (Fig. 4A and B). Notably, the apoptotic rate of the added BMSCs in the direct co-culture group was markedly low (Fig. 4C).

Bottom Line: The degenerative and apoptotic indexes were significantly reduced when NP cells were co‑cultured with BMSCs.In conclusion, the anti‑apoptosis and the migration, in addition to mitochondrial transfer associated with BMSC treatments in IVDD, were investigated in vitro in the present study.The interaction between stimulated NP cells and BMSCs is likely involved in to simulating the in vivo process of stem cell‑mediated repair.

View Article: PubMed Central - PubMed

Affiliation: Department of Orthopedic Surgery, Changzheng Hospital, Shanghai 200023, P.R. China.

ABSTRACT
Excessive apoptosis and high expression levels of interleukin‑1β (IL‑1β) in disc cells have been reported to serve important roles in intervertebral disc degeneration (IVDD). Previous studies investigating mesenchymal stem cells (MSCs) have indicated potential for their use in the treatment of IVDD. However, the therapeutic potential and anti‑apoptotic ability of MSCs remains to be fully elucidated. The present study aimed to establish an in vitro model for bone marrow‑derived MSC (BMSC) therapy by investigating the anti‑apoptotic effects, in addition to the migration of BMSCs to nucleus pulposus (NP) cells stimulated by IL‑1β. A co-culture system of BMSCs and NP cells was founded. Following inflammatory stimulation, the NP cells exhibited increased indexes for inflammation‑induced degeneration. The degenerative and apoptotic indexes were significantly reduced when NP cells were co‑cultured with BMSCs. Compared with the indirect co-culture group, the direct co-culture group exhibited an improved capacity for anti-apoptosis. In addition, IL‑1β‑stimulated NP cells attracted and mediated the migration of BMSCs. Mitochondrial transfer from BMSCs to NP cells by tunneling nanotubes was also observed. In conclusion, the anti‑apoptosis and the migration, in addition to mitochondrial transfer associated with BMSC treatments in IVDD, were investigated in vitro in the present study. The interaction between stimulated NP cells and BMSCs is likely involved in to simulating the in vivo process of stem cell‑mediated repair.

Show MeSH
Related in: MedlinePlus