PurposeMost studies on immune tolerance of mesenchymal stem cells (MSCs) have been performed using MSCs derived from bone marrow, cord blood, or adipose tissue. MSCs also exist in the craniofacial area, specifically in teeth. The purpose of this study was to evaluate the mechanisms of immune tolerance of dental pulp-derived MSC (DP-MSC) in vitro and in vivo.Materials and MethodsWe isolated DP-MSCs from human dental pulp and co-cultured them with CD4+ T-cells. To evaluate the role of cytokines, we blocked TGF-β and IL-10, separately and together, in co-cultured DP-MSCs and CD4+ T-cells. We analyzed CD25 and FoxP3 to identify regulatory T-cells (Tregs) by fluorescence-activated cell sorting (FACS) and real-time PCR. We performed alloskin grafts with and without DP-MSC injection in mice. We performed mixed lymphocyte reactions (MLRs) to check immune tolerance.ResultsCo-culture of CD4+ T-cells with DP-MSCs increased the number of CD4+CD25+FoxP3+ Tregs (p<0.01). TGF-β or/and IL-10 blocking suppressed Treg induction in co-cultured cells (p<0.05). TGF-β1 mRNA levels were higher in co-cultured DP-MSCs and in co-cultured CD4+ T-cells than in the respective monocultured cells. However, IL-10 mRNA levels were not different. There was no difference in alloskin graft survival rate and area between the DP-MSC injection group and the non-injection group. Nonetheless, MLR was reduced in the DP-MSC injected group (p<0.05).ConclusionDP-MSCs can modulate immune tolerance by increasing CD4+CD25+FoxP3+ Tregs. TGF-β1 and IL-10 are factors in the immune-tolerance mechanism. Pure DP-MSC therapy may not be an effective treatment for rejection, although it may module immune tolerance in vivo.

BackgroundSource material used to fill calvarial defects includes autologous bones and synthetic alternatives. While autologous bone is preferable to synthetic material, autologous reconstruction is not always feasible due to defect size, unacceptable donor-site morbidity, and other issues. Today, advanced three-dimensional (3D) printing techniques allow for fabrication of titanium implants customized to the exact need of individual patients with calvarial defects. In this report, we present three cases of calvarial reconstructions using 3D-printed porous titanium implants.MethodsFrom 2013 through 2014, three calvarial defects were repaired using custommade 3D porous titanium implants. The defects were due either to traumatic subdural hematoma or to meningioma and were located in parieto-occipital, fronto-temporo-parietal, and parieto-temporal areas. The implants were prepared using individual 3D computed tomography (CT) data, Mimics software, and an electron beam melting machine. For each patient, several designs of the implant were evaluated against 3D-printed skull models. All three cases had a custom-made 3D porous titanium implant laid on the defect and rigid fixation was done with 8 mm screws.ResultsThe custom-made 3D implants fit each patient's skull defect precisely without any dead space. The operative site healed without any specific complications. Postoperative CTs revealed the implants to be in correct position.ConclusionAn autologous graft is not a feasible option in the reconstruction of large calvarial defects. Ideally, synthetic materials for calvarial reconstruction should be easily applicable, durable, and strong. In these aspects, a 3D titanium implant can be an optimal source material in calvarial reconstruction.

It is a traditional teaching that the levator aponeurosis is the main transmitter of the levator palpebrae muscle. However, there are several points that raise doubts in this fundamental concept of the levator aponeurosis as being the primary interconnecting mechanism in upper lid elevation. Despite the structural integrity of the levator complex, drooping of the upper eyelids is seen to develop in situations such as Horner's syndrome and in times of excessive fatigue and sleepiness. Amid the controversy in the literature regarding the specific role of the levator aponeurosis in the lid-elevating mechanism, we have observed that the levator aponeurosis fails to make constant attachment to the tarsal plate. This has led us to speculate on the possible role of the posterior lamella - Müller's muscle - as the primary transmitter of the levator muscle action to the tarsal plate.

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PurposeZZMicrotia is congenital anomaly of external ear and the reconstruction method for the external ear of microtia patient was based on autogenous costal cartilage framework. The application of 3D printing technique in medical science has made more possibility of human tissue restoration, and we tried to apply this technique in auricular reconstruction field. Materials and MethodsZZAs for unilateral microtia patient, the contralateral side ear is normal and reconstructive surgeon tried to mimic it for reconstruction of affected ear. So, we obtained facial CT scan of microtia patient and made mirror image of normal side ear. Moreover, to make the 3D scaffold based on the mirror image of normal ear and to apply this scaffold for the auricular reconstruction surgery, we included auriculocephalic sulcus and anterior fixation part.ResultsZZWe could successfully obtain mirror image of normal ear, auriculocephalic sulcus and anterior fixation part for 3D scaffold printing. ConclusionsZZUsing this CT image processing and 3D printing technique, we will be able to make the scaffold for auricular reconstruction of unilateral microtia patient, and perform auricular reconstruction in near future. Key WordsZZ3D CT ㆍ3D printing ㆍAuricular reconstruction ㆍMicrotia.