A Novel Dressing Method to Achieve Early Natural Tracheostoma Closure.

Tracheotomy is a routine surgical procedure in oral and maxillofacial surgery. After decannulation, spontaneous tracheostoma closure is usually expected. However, wound healing is often delayed, requiring 1 to 2 weeks for healing and resulting in the need for surgical closure. Although many reports have described the surgical closure of a tracheostoma, few reports have focused on the dressing methods for closure of tracheal openings after decannulation. Herein, the authors report a new tracheostoma closure method that does not rely on surgical closure or the adhesive strength of the tape. The authors' conventional dressing method was to place gauze over the tracheostoma after decannulation and apply pressure through elastic tape or with a film dressing to seal the tracheostoma and achieve natural closure by reducing the leakage of air and tracheal secretions. However, the conventional method cannot completely prevent the leakage of air and tracheal secretions. We developed a novel method to achieve early closure by markedly reducing the leakage by partially inserting the gauze into the tracheostoma.

Evaluation of Facial Soft Tissue in Patients With Cleft Lip and Palate Corrected Using Maxillary Protraction Appliances.

ABSTRACT: Noncontact optical surface scanners have been used to evaluate facial soft tissues. Appropriate evaluation of patients with cleft lip and palate requires comprehensible assessment of the changes in their pre- and post-orthodontic soft tissue and facial growth during chairside assistance. The authors developed a new scanning system that required a shorter measurement time than conventional modalities. The system was implemented on a mannequin and a 6-year-old patient. Seven midfacial landmarks were identified on their faces. The authors measured these landmarks 5 times daily. An experienced orthodontist evaluated and recorded the scores. The scores obtained from the mannequin had a variation of within 0.2 mm, while those obtained from the patient varied within 0.8 mm, except that of the inferior limit of the lips. The study findings suggest that the new laser scanning system can accurately measure facial soft tissue. Further studies should fix patients' head at a definite position for more accurate measurements. An appropriately angled laser sensor would eliminate distortions, thereby increasing the measurement validity.

Bone Regeneration Potential of Human Dental Pulp Stem Cells Derived from Elderly Patients and Osteo-Induced by a Helioxanthin Derivative.

Human dental pulp stem cells (DPSCs) have high clonogenic and proliferative potential. We previously reported that a helioxanthin derivative (4-(4-methoxyphenyl)pyrido[40,30:4,5]thieno[2-b]pyridine-2-carboxamide (TH)) enhances osteogenic differentiation of DPSCs derived from young patients. However, in the clinical field, elderly patients more frequently require bone regenerative therapy than young patients. In this study, we examined and compared the osteogenic differentiation potential of TH-induced DPSCs from elderly patients and young patients to explore the potential clinical use of DPSCs for elderly patients. DPSCs were obtained from young and elderly patients and cultured in osteogenic medium with or without TH. We assessed the characteristics and osteogenic differentiation by means of specific staining and gene expression analyses. Moreover, DPSC sheets were transplanted into mouse calvarial defects to investigate osteogenesis of TH-induced DPSCs by performing micro-computed tomography (micro-CT). We demonstrated that osteogenic conditions with TH enhance the osteogenic differentiation marker of DPSCs from elderly patients as well as young patients in vitro. In vivo examination showed increased osteogenesis of DPSCs treated with TH from both elderly patients and young patients. Our results suggest that the osteogenic differentiation potential of DPSCs from elderly patients is as high as that of DPSCs from young patients. Moreover, TH-induced DPSCs showed increased osteogenic differentiation potential, and are thus a potentially useful cell source for bone regenerative therapy for elderly patients.

Effects of Helioxanthin Derivative-Treated Human Dental Pulp Stem Cells on Fracture Healing.

Bone defects affect patients functionally and psychologically and can decrease quality of life. To resolve these problems, a simple and efficient method of bone regeneration is required. Human dental pulp stem cells (DPSCs) have high proliferative ability and multilineage differentiation potential. In our previous study, we reported a highly efficient method to induce osteogenic differentiation using DPSC sheets treated with a helioxanthin derivative (4-(4-methoxyphenyl)pyrido[40,30:4,5]thieno[2,3-b]pyridine-2-carboxamide (TH)) in a mouse calvarial defect model. However, the localization of the DPSCs after transplantation remains unknown. Therefore, in this study, we investigated the localization of transplanted DPSCs in a mouse fracture model. DPSCs were collected from six healthy patients aged 18-29 years, cultured in normal medium (NM), osteogenic medium (OM), or OM with TH, and fabricated them into cell sheets. To evaluate the efficacy of fracture healing using DPSCs treated with OM+TH, and to clarify the localization of the transplanted DPSC sheets in vivo, we transplanted OM+TH-treated DPSC sheets labeled with PKH26 into mouse tibiae fractures. We demonstrated that transplanted OM+TH-treated DPSCs sheets were localized to the fracture site and facilitated bone formation. These results indicated that transplanted OM+TH-treated DPSCs were localized at fracture sites and directly promoted fracture healing.

Follicular lymphoid hyperplasia of the posterior maxillary site presenting as uncommon entity: a case report and review of the literature.

BACKGROUND: Follicular lymphoid hyperplasia (FLH) is characterized by an increased number and size of lymphoid follicles. In some cases, the etiology of FLH is unclear. FLH in the oral and maxillofacial region is an uncommon benign entity which may resemble malignant lymphoma clinically and histologically. CASE PRESENTATION: We report the case of a 51-year-old woman who presented with an asymptomatic firm mass in the left posterior maxillary site. Computed tomography scan of her head and neck showed a clear circumscribed solid mass measuring 28 × 23 mm in size. There was no evidence of bone involvement. Incisional biopsy demonstrated benign lymphoid tissue. The patient underwent complete surgical resection. Histologically, the resected specimen showed scattered lymphoid follicles with germinal centers and predominant small lymphocytes in the interfollicular areas. Immunohistochemically, the lymphoid follicles were positive for CD20, CD79a, CD10, CD21, and Bcl6. The germinal centers were negative for Bcl2. Based on these findings, a diagnosis of benign FLH was made. There was no recurrence at 1 year postoperatively. CONCLUSIONS: We diagnosed an extremely rare case of FLH arising from an unusual site and whose onset of entity is unknown. Careful clinical and histopathological evaluations are essential in making a differential diagnosis from a neoplastic lymphoid proliferation with a nodular growth pattern.

Simple Technique for Reducing the Buccal Fat Pad During Mandibular Orthognathic Surgery.

Sagittal split ramus osteotomy and intraoral vertical ramus osteotomy are commonly performed for the correction of jaw deformities. However, during mandibular orthognathic surgeries such as sagittal split ramus osteotomy and intraoral vertical ramus osteotomy , the authors sometimes encounter exposure of the buccal fat pad (BFP), which decreases the surgical field. The exposed BFP makes it difficult to perform these operations, may result in unexpected complications, and may increase the operation time. Therefore, the authors herein describe a simple, safe, and convenient technique for reducing the volume of the exposed BFP during mandibular orthognathic surgery using an electric knife in the coagulation mode.

Usefulness of Early Plate Removal in Patients With Occlusal Discrepancies After Sagittal Split Ramus Osteotomy.

The aim of this study was to evaluate the effects of early removal of fixed plates in patients with occlusal discrepancies after sagittal split ramus osteotomy (SSRO) with a focus on the positional relationship of the temporomandibular joint (TMJ). Sagittal split ramus osteotomy with/without Le Fort I osteotomy was performed on 98 patients with mandibular prognathism. Of these 98 patients, 15 with occlusal discrepancies and/or TMJ symptoms underwent early plate removal after SSRO. Finally, 12 consecutive patients were evaluated in this study: 7 underwent bilateral SSRO, 1 underwent unilateral SSRO, and 4 underwent bilateral SSRO with maxillary advancement. The axiolateral TMJ Schuller method was used to evaluate the TMJ position. The authors measured 3 spaces (anterior, superior, and posterior joint spaces) between the condyle and glenoid fossa in the sagittal plane. The anterior and superior joint spaces were significantly larger immediately after the operation than before the operation. After early plate removal, these spaces significantly decreased in size. The posterior joint space increased, but with no significant difference. Three months after SSRO, the size of each of the 3 spaces was closely related to its preoperative size. In conclusion, these results suggest that early removal of fixed plates is 1 treatment option for postoperative occlusal discrepancies after SSRO.

Intravenous Dexamethasone Administration Before Orthognathic Surgery Reduces the Postoperative Edema of the Masseter Muscle: A Randomized Controlled Trial.

PURPOSE: Dexamethasone seems to suppress postoperative swelling. However, the standard administration dose of dexamethasone for bilateral sagittal split osteotomy (BSSO) has not been reported. This study focused on clarifying the most effective dose of dexamethasone for BSSO. MATERIALS AND METHODS: This research was planned as a prospective, randomized controlled, double-blind study. Patients undergoing BSSO were randomly assigned to receive intravenous preoperative dexamethasone under 3 different dose conditions: 16 mg, 8 mg, and 0 mg (control). The endpoints of this study were 1) postoperative changes in masseter muscle thickness and buccal soft tissue thickness; 2) postoperative changes in maximum incisal opening; 3) postoperative changes in sensation of the chin and lower lip region; 4) postoperative changes in blood examination findings (white blood cell count, neutrophil count, C-reactive protein level, and lymphocyte count); and 5) types of complications. Data were recorded at 2 to 4 time intervals: before surgery, postoperative day 1, postoperative day 2, and postoperative day 3. Average age, gender, average body mass index, average surgery time, and average blood loss also were examined. Data were analyzed by 1-way analysis of variance (Bonferroni multiple-comparisons test) after the Bartlett test. RESULTS: We enrolled 24 patients, including 5 men and 19 women, in this study. The rate of increase in the thickness of the masseter muscle 24 hours after BSSO was 38.4% in the 16-mg group (n = 8), 57.7% in the 8-mg group (n = 8), and 56.1% in the 0-mg group (n = 8). The rate of increase in masseter muscle thickness in the 16-mg group was significantly lower than that in the 0-mg group (P < .05). Regarding the number of lymphocytes after surgery, the 16-mg and 8-mg groups maintained preoperative levels whereas there was a reduced number of lymphocytes in the control group. No statistically significant results were obtained for the following study endpoints: postoperative changes in maximum incisal opening and postoperative changes in sensation of the chin and lower lip region. CONCLUSIONS: This investigation showed that the most effective dose of dexamethasone for BSSO is 16 mg.

Short Lingual Osteotomy Using a Piezosurgery Ultrasonic Bone-Cutting Device During Sagittal Split Ramus Osteotomy.

Short lingual osteotomy is a useful method for the performance of sagittal split ramus osteotomy involving interference between the proximal and distal bone fragments when lateral differences exist in the setback distance. However, this procedure occasionally results in abnormal fracture and nerve injury; expert surgical skill is thus required. We herein describe a novel technique involving the use of an ultrasonic bone-cutting device (Piezosurgery; Mectron Medical Technology, Carasco, Italy) for vertical osteotomy posterior to the mandibular foramen. Successful short lingual osteotomy was performed using this technique with avoidance of abnormal fracture and neurovascular bundle damage.

Orthognathic Surgery in Patients With Von Willebrand's Disease: A Report of Four Cases and Literature Review.

Von Willebrand's disease (VWD), characterized by quantitatively or qualitatively abnormal von Willebrand factor (VWF), is the most common inherited bleeding disorder. There is limited evidence of treatment using orthognathic surgery in patients with VWD. This report focuses on four patients with VWD who underwent orthognathic surgery and received Factor VIII/VWF concentrates (Confact F) preoperatively. One patient with type 3 (severe) VWD underwent delayed extubation owing to laryngeal edema and exhibited epistaxis thereafter. No perioperative complications were observed in any of the other patients. Two of the four patients were diagnosed with VWD during preoperative screening. Most young adults do not experience general anesthesia and, therefore, may not have undergone blood tests at a hospital. Thus, preoperative screening and adoption of a multidisciplinary approach to orthognathic surgery is important in patients with bleeding disorders such as VWD. Close communication between anesthetists, surgeons, and hematologists is essential to ensure effective management during the perioperative period.

Squamous cell carcinoma of mandibular gingiva producing both parathyroid hormone-related protein and granulocyte colony-stimulating factor: a case report.

We describe a case of mandibular gingival carcinoma with hypercalcaemia and leukocytosis caused by tumour-derived parathyroid hormone-related protein (PTHrP) and granulocyte colony-stimulating factor (G-CSF). A 54-year-old man presented to our Department of Oral and Maxillofacial Surgery with a chief complaint of a left-sided mandibular gingival ulcer. A 42 mm × 20 mm sized ulcer was found on the left lower molar gingiva. Squamous cell carcinoma was pathologically diagnosed. The patient underwent a hemimandibulectomy, left-sided radical neck dissection, plate reconstruction, pectoralis major musculocutaneous flap reconstruction, and tracheostomy under general anaesthesia. Pathologically, two metastatic lymph nodes were identified. Residual tumour was suspected at the resection margins. Eight weeks after surgery, the patient started postoperative concurrent chemoradiotherapy (CCRT). Two weeks after CCRT, the patient developed hypercalcaemia. Serum levels of PTHrP and G-CSF increased in parallel with the progression of hypercalcaemia and leukocytosis. Immunohistochemical analysis of the surgical specimen showed positivity for G-CSF. Based on these clinical and pathological findings, the patient was diagnosed with hypercalcaemia and leukocytosis associated with malignancy and was treated with denosumab. Irradiation was terminated at 50 Gy because CT showed rapid disease progression. Chemotherapy was initiated, however, four weeks after the start of chemotherapy, a CT scan showed increased metastases and pleural dissemination. Therefore, chemotherapy was discontinued. One week after the chemotherapy was discontinued, the patient died of respiratory failure.

A novel technique for preventing skin pressure sores using a rubber tube during surgical treatment of mandibular condyle fractures.

Craniofacial surgery occasionally results in sores and necrosis of the facial skin because of pressure from surgical instruments. During surgical treatment of mandibular condylar process fractures, the main mandibular fragment is routinely retracted downward using a wire to achieve a satisfactory anatomic reduction. This procedure may injure the facial skin. This potential complication is easily overlooked by medical staff, but it is easily preventable. We herein describe a method of using a rubber tube to avoid causing pressure sores of the facial skin during surgical treatment of mandibular condylar process fractures.

Platelet-Rich Fibrin-Conditioned Medium as an Alternative to Fetal Bovine Serum Promotes Osteogenesis of Human Dental Pulp Stem Cells.

Human dental pulp stem cells (DPSCs) exhibit multilineage differentiation capabilities and superior clonogenic and proliferative properties. However, the use of animal-derived components such as FBS raises concerns regarding the clinical application of stem-cell-based therapies. Platelet-rich fibrin (PRF) derived from human blood is rich in fibrin, platelets, and growth factors and acts as a bioactive scaffold for grafting with biomaterials. In this study, we assessed the efficacy of PRF-conditioned medium (CM) in promoting DPSCs proliferation and osteogenic differentiation compared with the standard culture medium supplemented with FBS. A comparison of DPSCs cultured in FBS and PRF-CM revealed no differences in characteristics or morphology. However, cells cultured with PRF-CM exhibited inferior proliferation rates and cell numbers during passage in comparison with those cultured with FBS. In contrast, DPSCs cultured in PRF-CM showed significantly higher levels of calcification, and RT-PCR confirmed that the gene expression levels of markers associated with osteoblast differentiation were significantly increased. The PRF-CM approach offers a convenient, straightforward, and advantageous method for culturing DPSCs, without relying on animal-derived components. In summary, this study introduces a novel application of PRF-CM for enhancing the osteogenesis of DPSCs, which provides an alternative to FBS culture medium and addresses concerns associated with the use of animal-derived components in clinical settings.

VCAM-1 and GFPT-2: Predictive markers of osteoblast differentiation in human dental pulp stem cells.

INTRODUCTION: Dental pulp stem cells (DPSCs) have high proliferative and multilineage differentiation potential in mesenchymal stem cells. However, several studies have indicated that there are individual differences in the potential for osteogenic differentiation of DPSCs, and the factors determining these differences are unknown. OBJECTIVE: To identify the genes responsible for the individual differences in the osteogenic differentiation ability of DPSCs. METHODS: We divided DPSCs into high and low osteogenic differentiation ability groups (HG or LG) with ALP and von Kossa stain, and compared the gene expression patterns using RNA-seq. In addition, genes that may affect osteogenic differentiation were knocked down using small interfering RNA (siRNA) and their effects were investigated. RESULTS: The RNA-seq patterns revealed that VCAM1 and GFPT2 were significantly expressed at higher levels in the HG than in the LG. The results of siRNA analysis showed that VCAM1 and GFPT2 knockdown significantly reduced the expression of osteogenic markers. Furthermore, we analyzed the involvement of these two genes in cell signaling in DPSC differentiation. The results indicated that the VCAM1-mediated Ras-MEK-Erk and PI3K/Akt pathways are involved in the osteogenic differentiation of DPSCs, and that GFPT2-mediated HBP signaling influences the osteogenic differentiation of DPSCs. CONCLUSIONS: These findings indicate that DPSCs that highly express VCAM1 and GFPT2 have a high capacity for osteogenic differentiation. Evaluation of VCAM1 and GFPT2 expression in undifferentiated DPSCs may predict the outcome of bone regenerative therapy using DPSCs. Moreover, the expression levels of VCAM1 and GFPT2 in DPSCs may be useful in setting criteria for selecting donors for allogeneic cell transplantation for bone regeneration.

Computed tomography evaluation of risk factors for an undesirable buccal split during sagittal split ramus osteotomy.

Sagittal split ramus osteotomy (SSRO) sometimes induces an irregular split pattern referred to as a bad split. We investigated the risk factors for bad splits in the buccal plate of the ramus during SSRO. Ramus morphology and bad splits in the buccal plate of the ramus were assessed using preoperative and postoperative computed tomography images. Of the 53 rami analyzed, 45 had a successful split, and 8 had a bad split in the buccal plate. Horizontal images at the height of the mandibular foramen showed that there were significant differences in the ratio of the forward thickness to the backward thickness of the ramus between patients with a successful split and those with a bad split. In addition, the distal region of the cortical bone tended to be thicker and the curve of the lateral region of the cortical bone tended to be smaller in the bad split group than in the good split group. These results indicated that a ramus shape in which the width becomes thinner towards the back frequently induces bad splits in the buccal plate of the ramus during SSRO, and more attention should be paid to patients who have rami of these shapes in future surgeries.

RNAase-III enzyme Dicer maintains signaling pathways for differentiation and survival in mouse cortical neural stem cells.

An important function of the RNAase-III enzyme Dicer is to process microRNA precursors into ~22-nucleotide non-coding small RNAs. But little is known about the role of Dicer in mammalian brain formation and neural stem cell (NSC) development. Here we show that Dicer plays a crucial role in controlling mouse cortical NSC development. We found that Dicer function is essential for expanding cortical neural progenitors and NSCs. We have identified a population of Dicer-deficient NSCs that can self-renew, and that display normal karyotype and heterochromatin protein expression levels but show enlarged nuclei. Dicer-deficient NSCs display abnormal differentiation and undergo cell death when mitogens are withdrawn. Dicer deletion affects the levels of many proteins, as revealed by a mass spectrometry proteomic approach. We have found that an increase of anti-survival and/or pro-apoptosis proteins and a decrease of pro-survival and/or anti-apoptosis proteins contribute to the cell death of Dicer-deficient NSCs, implying a general role for Dicer in protecting cells from apoptosis. Our results demonstrate important functions for Dicer in regulating NSC development by maintaining proper signaling pathways related to cell survival and differentiation.

A Case of Orthognathic Surgery for Jaw Deformity in a Patient with Spinocerebellar Ataxia.

Spinocerebellar ataxia (SCA) is a progressive neurodegenerative disease that can cause various ataxia symptoms. Here we report a patient with spinocerebellar ataxia who underwent orthognathic surgery to correct a mandibular protrusion with facial asymmetry. A 33-year-old woman was admitted to our hospital for orthognathic surgery. She started preoperative orthodontic treatment after a diagnosis of mandibular protrusion with facial asymmetry. Two and a half years later, after completing preoperative orthodontic treatment, she returned to our hospital after being diagnosed with spinocerebellar ataxia. After discussing the risk of surgery with the anesthesiologist and neurologist, we elected to perform orthognathic surgery after the patient provided informed consent. Sagittal split ramus osteotomy and intraoral vertical ramus osteotomy were performed under general anesthesia, but no remarkable perioperative complications occurred. After a 3-year follow-up, the occlusion has remained stable, and no postoperative relapse occurred. Whether we should provide surgical treatment for SCA patients is controversial. However, when long-term predictions were considered, altering an occlusion could improve a patient's quality of life in the present case.

Transcription factor Lmo4 defines the shape of functional areas in developing cortices and regulates sensorimotor control.

Proper formation of the shape and size of cortical functional areas is essential for complex brain function, including sensory perception and motor control. Our previous work identified the transcription factor Lim domain only 4 (Lmo4), a regulator in calcium-dependent gene transcription, that has unique, region-specific expression in postnatal mouse cortices with high expression anteriorly and posteriorly but very low expression in between. Here we report that Lmo4 expression coincides with the timing of the development of the somatosensory barrel field. Lmo4 cortical deletion causes changes in expression patterns of cortical regional markers and results in rostro-medial shrinkage but not rostral or caudal shift of the somatosensory barrel subfield. Fine regulation of accurate shape of the barrel subfield by Lmo4, as well as Lmo4-mediated calcium-dependent gene expression, is critical for normal brain functions, as Lmo4-deficient mice display impaired sensorimotor performance. Moreover, even though Lmo4 has broad expression in the central nervous system, it plays a subtle role in the development of non-cortical regions. Our results reveal a new mechanism of cortical area formation and normal sensorimotor control that is regulated by genes with region-specific expression in the developing cortex.

Different timings of Dicer deletion affect neurogenesis and gliogenesis in the developing mouse central nervous system.

MicroRNAs, processed by the RNAase III enzyme Dicer, are approximately 22 nucleotide endogenous noncoding small RNAs. The function of Dicer in the mouse central nervous system (CNS) development is not well understood. Here, we show that specifically deleting Dicer expression in the CNS and in the cerebral cortex using two Cre lines results in reduced progenitor numbers, abnormal neuronal differentiation, and thinner cortical wall. Incomplete Dicer deletion during early embryonic stages contributes to normal development of early-born neurons in the cortex and motor neurons in the spinal cord. However, at late embryonic stages when Dicer is completely ablated in the CNS, the migration of late-born neurons in the cortex and oligodendrocyte precursor expansion and differentiation in the spinal cord are greatly affected. Our studies of different timings of Dicer deletion demonstrate the importance of the Dicer-mediated microRNA pathway in regulating distinct phases of neurogenesis and gliogenesis during the CNS development.

Identification of neurospheres generated from human dental pulp stem cells in xeno-/serum-free conditions.

INTRODUCTION: Cell-based therapies require an emerging alternative treatment using easily harvested cell sources. Neural stem cells derived from various tissues, including brain, bone marrow, skin and retina can give rise to both neurons and glial cells. Recently, human dental pulp stem cells (DPSCs) and stem cells from human exfoliated deciduous teeth (SHED) were demonstrated to have mesenchymal stem cell-like abilities such as self-renewal and multi-lineage differentiation, including neuron and glial cells. Moreover, DPSCs and SHED show a higher proliferation rate and a higher number of population doublings compared with adult bone marrow stromal stem cells. Therefore, DPSCs are a useful source that can be applied in cell replacement therapy for various neurological disorders. Generally, the conventional culture methods for DPSCs have used serum, therefore the undefined components in culture medium may complicate investigations of the molecular mechanisms that control the self-renewal and differentiation of DPSCs. However, neural stem cells proliferate to form 'neurospheres' in suspension in vitro in the presence of epidermal growth factor (EGF) and basic fibroblast growth factor (bFGF). No study to date has obtained neurospheres from DPSCs in serum-free conditions in primary culture. Thus, the aim of this study was to establish a method for the proliferation and neural differentiation of DPSCs in xeno- and serum-free conditions in primary culture. METHODS: DPSCs were obtained from the dental pulp of wisdom teeth from healthy individuals (18-41 years old) and cultured in conventional medium containing 15% fetal bovine serum and xeno-/serum-free medium. We evaluated the proliferation of DPSCs, neurosphere generation, and neural differentiation under xeno-/serum-free conditions by flow cytometry, immunohistochemistry, and real-time polymerase chain reaction. RESULTS: In proliferation medium without xeno/serum, DPSCs can proliferate and generate neurospheres, however, the neurospheres had limited self-renewal ability. Under differentiation conditions, class III ß-tubulin (TUBB3) and microtubule-associated protein (MAP2) were more significantly expressed in neurospheres derived from DPSCs in xeno-/serum-free culture conditions than in DPSCs in conventional culture conditions. CONCLUSIONS: Our result demonstrated that neurosphere generation from DPSCs in xeno-/serum-free culture may be an accessible source for clinical cell replacement therapies for neuronal degenerative diseases.