Abstract
We cryopreserved mouse tooth germs with widely open cervical margins of the enamel organ to overcome difficulties in cryoprotectant permeation and tested their efficacy by transplanting them into recipient mice. The upper right first molar germs of 8-day-old donor mice were extracted and categorized into the following four groups according to cryopreservation time: no cryopreservation, 1 week, 1 month, and 3 months. The donor tooth germs were transplanted into the upper right first molar germ sockets of the 8-day-old recipient mice. The upper left first molars of the recipient mice were used as controls. The outcome of the transplantation was assessed at 1, 2, and 3 weeks after transplantation. Stereomicroscopic evaluation revealed that most of the transplanted teeth erupted by 3 weeks after transplantation. Micro-computed tomography analysis revealed root elongation in the transplanted groups as well as in the controls. There was no significant difference between the cryopreserved and non-cryopreserved transplanted teeth, but the roots of the cryopreserved teeth were significantly shorter than those of the control teeth. Histological examination revealed root and periodontal ligament formations in all the transplanted groups. These results suggest that the transplantation of cryopreserved tooth germs facilitates subsequent root elongation and tooth eruption.
Highlights
We cryopreserved mouse tooth germs with widely open cervical margins of the enamel organ to overcome difficulties in cryoprotectant permeation and tested their efficacy by transplanting them into recipient mice
The findings of the present study demonstrated that transplanted cryopreserved tooth germs develop roots and erupt
On the basis of these reports, cryopreservation appears to have little negative influence on Periodontal ligament (PDL) cells, whose viability and function were maintained after cryopreservation and contrasted with the poor viability and lower survival rates of pulp cells
Summary
We cryopreserved mouse tooth germs with widely open cervical margins of the enamel organ to overcome difficulties in cryoprotectant permeation and tested their efficacy by transplanting them into recipient mice. In the cryopreservation of mature teeth, difficulty has been reportedly in maintaining the biological viability of the pulp tissue owing to the cryoprotective agents barely penetrating the pulp cavity through the narrow apical foramen, which is surrounded by hard t issue[3,4,5] This finding is consistent with that of studies that showed that autotransplantation of cryopreserved immature teeth achieved higher success rates than that of mature teeth. No differences in pulpal regeneration and revascularization of the pulp cavity have been reported between cryopreserved mature apicoectomized and immature teeth after transplantation in d ogs[10] These observations led us to investigate the possibility of transplanting tooth germs cryopreserved prior to root formation. Cryopreserved tooth germ transplantation outcomes were quantitatively monitored using micro-computed tomography (CT) and qualitatively assessed using histological analysis in mice
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