In Vitro and In Vivo Osteogenesis of Porcine Skin-Derived Mesenchymal Stem Cell-like Cells with a Demineralized Bone and Fibrin Glue Scaffold

Abstract

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.