Human dental mesenchymal stem cells isolated from the dental follicle, pulp, and root apical papilla of extracted wisdom teeth have been known to exhibit successful and potent neurogenic differentiation capacity. In particular, human dental pulp-derived stem cells (hDPSCs) stand out as the most prominent source for in vitro neuronal differentiation. In this study, to evaluate the in vivo peripheral nerve regeneration potential of hDPSCs and differentiated neuronal cells from DPSCs (DF-DPSCs), a total of 1 × 10 hDPSCs or DF-hDPSCs labeled with PKH26 tracking dye and supplemented with fibrin glue scaffold and collagen tubulization were transplanted into the sciatic nerve resection (5-mm gap) of rat models. At 12 weeks after cell transplantation, both hDPSC and DF-hDPSC groups showed notably increased behavioral activities and higher muscle contraction forces compared with those in the non-cell transplanted control group. In immunohistochemical analysis of regenerated nerve specimens, specific markers for angiogenesis, axonal fiber, and myelin sheath increased in both the cell transplantation groups. Pretransplanted labeled PKH26 were also distinctly detected in the regenerated nerve tissues, indicating that transplanted cells were well-preserved and differentiated into nerve cells. Furthermore, no difference was observed in the nerve regeneration potential between the hDPSC and DF-hDPSC transplanted groups. These results demonstrate that dental pulp tissue is an excellent stem cell source for nerve regeneration, and in vivo transplantation of the undifferentiated hDPSCs could exhibit sufficient and excellent peripheral nerve regeneration potential.
Porcine skin-derived mesenchymal stem cells (pSMSCs) were evaluated on their biological MSC characterizations and differentiation into mesenchymal lineages, along with in vitro and in vivo neural inductions. Isolated pSMSCs showed plate-adherent growth, expression of various MSC-marker proteins and transcriptional factors, and differentiation potential into mesenchymal lineages. Neuron-like cell morphology and various neural markers were highly detected at 6 h and 24 h after in vitro neural induction of pSMSCs, but their neuron-like characteristics disappeared as induction time extended to 48 and 72 h. To evaluate the in vivo peripheral nerve regeneration potential of pSMSCs, a total of 5 × 10(6) autologous pSMSCs labelled with tracking dye, supplemented with fibrin glue scaffold and collagen tubulization, were transplanted into the peripheral nerve defected miniature pigs. At 2 and 4 weeks after cell transplantation, well-preserved transplanted cells and remarkable in vivo nerve regeneration, including histologically complete nerve bundles, were observed in the regenerated nerve tissues. Moreover, S-100 protein and p75 nerve growth factor receptor were more highly detected in regenerated nerve fibres compared to non-cell grafted control fibres. These results suggest that autologous pSMSCs transplanted with fibrin glue scaffold can induce prominent nerve regeneration in porcine peripheral nerve defect sites.
In vitro and in vivo osteogenesis of skin-derived mesenchymal stem cell-like cells (SDMSCs) with a demineralized bone (DMB) and fibrin glue scaffold were compared. SDMSCs isolated from the ears of adult miniature pigs were evaluated for the expression of transcriptional factors (Oct-4, Sox-2, and Nanog) and MSC marker proteins (CD29, CD44, CD90, and vimentin). The isolated SDMSCs were cocultured in vitro with a mixed DMB and fibrin glue scaffold in a nonosteogenic medium for 1, 2, and 4 weeks. Osteonectin, osteocalcin, and Runx2 were expressed during the culture period and reached maximum at 2 weeks after in vitro coculture. von Kossa-positive bone minerals were also noted in the cocultured medium at 4 weeks. Autogenous porcine SDMSCs (1 x 10(7)) labeled with a tracking dye, PKH26, were grafted into the maxillary sinus with a DMB and fibrin glue scaffold. In the contralateral side, only a scaffold was grafted without SDMSCs (control). In vivo osteogenesis was evaluated from two animals euthanized at 2 and 4 weeks after grafting. In vivo PKH26 staining was detected in all the specimens at both time points. Trabecular bone formation and osteocalcin expression were more pronounced around the grafted materials in the SDMSC-grafted group compared with the control group. New bone generation was initiated from the periphery to the center of the grafted material. The number of proliferating cells increased over time and reached a peak at 4 weeks in both in vivo and in vitro specimens. These findings suggest that autogenous SDMSC grafting with a DMB and fibrin glue scaffold can serve as a predictable alternative to bone grafting in the maxillary sinus floor.
SSEH is an uncommon clinical condition, and a manifestation of SSEH with anterior spinal artery syndrome is also rare. Furthermore, an emergency operation after IV thrombolytic treatment is an extraordinary situation.
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