Single-cell Transcriptomics Reveals Activation of Macrophages in All-trans Retinoic Acid (atRA)-induced Cleft Palate.

Cleft palate is among the most common birth defects with an impact on swallowing and speaking and is difficult to diagnose with ultrasound during pregnancy. In this study, we systematically capture the cellular composition of all-trans retinoic acid (atRA)-exposed and normal embryonic gestation 16.5 days mouse palate by the single-cell RNA sequencing technique. The authors identified 14 major cell types with the largest proportion of fibroblasts. The proportion of myeloid cells in atRA-exposed palate was markedly higher than those in the normal palate tissue, especially M1-like macrophages and monocytes. The upregulated genes of the different expression genes between atRA-exposed palate and normal palate tissue were linked to the biological processes of leukocyte chemotaxis and migration. Protein TLR2, CXCR4, THBS1, MRC1, transcription factor encoding genes Cebpb, Fos, Jun, Rela, and signaling pathway IL-17 and phagosome were found to be significantly involved in these processes. Subsequently, cellular communication network analysis suggested that myeloid-centered cell interactions SELL, SELPLG, MIF, CXCL, ANNEXIN, THBS, and NECTIN were significantly more activated in atRA-exposed palate. Overall, we delineate the single-cell landscape of atRA-induced cleft palate, revealing the effects of overexposure to atRA during palate tissue development and providing insights for the diagnosis of cleft palate.

A Concise and Accurate Method for Evaluating Alveolar Cleft ReconstructionDiet al Concise and Accurate Method for Evaluating.

Currently, there remain unresolved issues in the treatment of alveolar clefts, the resolution of which could greatly benefit many patients with cleft lip and palate. In alveolar cleft treatment research, a reliable tool for pre- and postoperative assessment is crucial. This study presents a concise and accurate method for postoperative evaluations of alveolar treatment, which can rapidly and accurately obtain the shape and volume of the newly formed bone. This study included helical computed tomography (CT) datasets of 20 patients who underwent alveolar bone grafting at our institute. Two observers independently measured the volume of newly formed bone in the patient's CT images one year postoperatively, with each observer performing the measurement twice. To acquire the volume of the newly formed bone at 1 year postoperatively, the model of the newly formed bone must be constructed first. The acquisition of this model involves Boolean operations on registered preoperative and postoperative cranial 3-dimensional (3D) images. The registration of the preoperative and postoperative models is performed in MIMICS software, and the registration results can be directly confirmed layer by layer on the CT images to ensure accuracy. The mean newly formed bone ratio in this study was 39.81%±17.96%, and the mean processing time was 05:11±01:41 minutes. The intraclass correlation coefficient for bone volume measurements between the two observers was 0.999, indicating high consistency and reproducibility. This method enhances accuracy, is time-efficient, and demonstrates high reliability in evaluating postoperative bone formation.

Enhancing Fat Graft Survival via Upregulating Autophagy of Adipocytes.

BACKGROUND: Autophagy is a cellular self-protection mechanism. The upregulation of adipose-derived stem cells' (ADSCs) autophagy can promote fat graft survival. However, the effect of interfering with adipocyte autophagy on graft survival is still unknown. In addition, autophagy is involved in adipocyte dedifferentiation. We investigated the effect of autophagy on adipocyte dedifferentiation and fat graft survival. METHODS: The classic autophagy regulatory drugs rapamycin (100 nM) and 3-methyladenine (3-MA; 10 mM) were used to treat adipocytes, adipocyte dedifferentiation was observed, and their effects on ADSCs were detected. In our experiments, 100 nM rapamycin, 10 mM 3-MA and saline were mixed with human adipose tissue and transplanted into nude mice. At 2, 4, 8 and 12 weeks postoperatively, the grafts were harvested for histological and immunohistochemical analysis. RESULTS: Rapamycin and 3-MA can promote and inhibit adipocyte dedifferentiation by regulating autophagy. Both drugs can inhibit ADSC proliferation, and 10 mM 3-MA can inhibit ADSC adipogenesis. At weeks 8 and 12, the volume retention rate of the rapamycin group (8 weeks, 64.77% ± 6.36%; 12 weeks, 56.13% ± 4.73%) was higher than the control group (8 weeks, 52.62% ± 4.04%; P < 0.05; 12 weeks, 43.17% ± 6.02%; P < 0.05) and the rapamycin group had more viable adipocytes and better vascularization. Compared with the control group, the volume retention rate, viable adipocytes and vascularization of the 3-MA group decreased. CONCLUSIONS: Rapamycin can promote adipocyte dedifferentiation by upregulating autophagy to promote fat graft survival. 3-MA can inhibit graft survival, but its mechanism includes the inhibition of adipocyte dedifferentiation and ADSC proliferation and adipogenesis. NO LEVEL ASSIGNED: This journal requires that authors assign a level of evidence to each submission to which Evidence-Based Medicine rankings are applicable. This excludes Review Articles, Book Reviews, and manuscripts that concern Basic Science, Animal Studies, Cadaver Studies, and Experimental Studies. For a full description of these Evidence-Based Medicine ratings, please refer to the Table of Contents or the online Instructions to Authors www.springer.com/00266 .

The Impact of Centrifugal Force on Isolation of Bone Marrow Mononuclear Cells Using Density Gradient Centrifugation.

BACKGROUND: Bone marrow mononuclear cells (BMMNCs) have great potential in bone regenerative therapy. The main method used today to obtain BMMNCs is Ficoll density gradient centrifugation. However, the centrifugal force for this isolation method is still suboptimal. OBJECTIVES: To determine the optimal centrifugal force in Ficoll density gradient centrifugation of bone marrow (BM) to achieve high stem/progenitor cell content BMMNCs for regenerative therapy. METHODS: BM was aspirated from nine minipigs and divided into three groups according to different centrifugal forces (200 g, 300 g and 400 g). Immediately after BMMNCs were obtained from each group by Ficoll density gradient centrifugation, residual red blood cell (RBC) level, nucleated cell counting, viability and flow cytometric analyses of apoptosis and reactive oxygen species (ROS) generation were measured. The phenotypic CD90 and colony formation analyses of BMMNCs of each group were performed as well. Bone marrow-derived mesenchymal stem cells (BMSCs) were harvested at passage 2, then morphology, cell phenotype, proliferation, adipogenic, chondrogenic and osteogenic lineage differentiation potential of BMSCs from each group were compared. RESULTS: The 300 g centrifugal force was able to isolate BMMNCs from BM with the same efficiency as 400 g and provided significantly higher yields of CD90+ BMSCs and fibroblastic colony-forming units of BMSC (CFU-f(BMSC)), which is more crucial for the regenerative efficacy of BMMNCs. Meanwhile, 200 g hosted the most RBC contamination and minimum CFU-f (BMSC) yield, which will be disadvantageous for BMMNC-based cell therapy. As for in vitro cultured BMSCs which were isolated from BMMNCs by different centrifugal forces, no significant differences were found on morphology, cell proliferation rate, phenotypic marker, adipogenic, chondrogenic and osteogenic differentiation potential. CONCLUSIONS: 300 g may be the optimal centrifugal force when using Ficoll density gradient centrifugation to isolate BMMNCs for bone regenerative therapy. NO LEVEL ASSIGNED: This journal requires that authors assign a level of evidence to each submission to which Evidence-Based Medicine rankings are applicable. This excludes Review Articles, Book Reviews and manuscripts that concern Basic Science, Animal Studies, Cadaver Studies and Experimental Studies. For a full description of these Evidence-Based Medicine ratings, please refer to the Table of Contents or the online Instructions to Authors www.springer.com/00266.

Identification of a Novel TP63 Variant in a Chinese Patient with Orofacial Clefts and Ectrodactyly: Case Report and Literature Review.

The TP63 gene is essential for epithelial proliferation, differentiation, and maintenance during embryogenesis. Despite considerable clinical variability, TP63-related symptoms are characterized by ectodermal dysplasia, distal limb malformations, and orofacial clefts. We identified a novel TP63 variant (c.619A > G, p.K207E) in a seven-month-old Chinese patient with orofacial clefts and ectrodactyly but no evident signs of ectodermal dysplasia. This phenotype was rarely reported before. We summarized the presence of the three main TP63-related manifestations in the literature and noted different distributions of CP- and CL/P-related variants regarding p63 structural domains.

Using a New Deep Learning Method for 3D Cephalometry in Patients With Hemifacial Microsomia.

ABSTRACT: Deep learning algorithms based on automatic 3D cephalometric marking points about people without craniomaxillofacial deformities have achieved good results. However, there has been no previous report about hemifacial microsomia (HFM). The purpose of this study is to apply a new deep learning method based on a 3D point cloud graph convolutional neural network to predict and locate landmarks in patients with HFM based on the relationships between points. The authors used a PointNet++ model to investigate the automatic 3D cephalometry. And the mean distance error (MDE) of the center coordinate position and the success detection rate (SDR) were used to evaluate the accuracy of systematic labeling. A total of 135 patients were enrolled. The MDE for all 32 landmarks was 1.46 ± 1.308 mm, and 10 landmarks showed SDRs at 2 mm over 90%, and only 4 landmarks showed SDRs at 2 mm under 60%. Compared with the manual reproducibility, the standard distance deviation and coefficient of variation values for the MDE of the artificial intelligence system was 0.67 and 0.43, respectively. In summary, our training sets were derived from HFM computed tomography to achieve accurate results. The 3D cephalometry system based on the graph convolutional network algorithm may be suitable for the 3D cephalometry system in HFM cases. More accurate results may be obtained if the HFM training set is expanded in the future.

Heterogeneity of Orofacial Clefts and Associated Anomalies in China.

Despite the large number of affected individuals in China, information on the descriptive epidemiology of orofacial clefts in the Chinese population remains limited. Therefore, the authors aimed to report a detailed clinical classification of orofacial clefts, including microform cleft lip (CL), and associated malformations in patients from a major cleft surgical unit in China. The authors reviewed the medical records of 718 patients who underwent primary cleft repair surgery at their center between December 2016 and April 2023. The sex distribution varied with cleft type and extent, and males had a higher proportion of clefts with increased severity than females. In patients with a unilateral CL, the ratio of left-to-right-sided clefts was 1.85:1. The frequency of associated congenital malformations in patients with microform CL only, overt CL only, CL and palate, and cleft palate only was 8.0%, 7.6%, 14.9%, and 30.9%, respectively. A total of 98 malformations were observed in 69 patients with congenital anomalies of unknown origin. Among these, cardiovascular anomalies were the most common, identified in 27 patients (39.1%), followed by head and neck and musculoskeletal anomalies. The high risk of heart defects highlights the importance of routine echocardiography. Microform CL exhibited increased prevalence and a similar frequency of associated malformations with overt CL, suggesting that this mild phenotype requires further attention by clinicians.

A 3-Dimensional Measurements of Bone and Airway Variables After Le Fort I Distraction Osteogenesis in Patients With Cleft Lip and/or Palate-Induced Midface Hypoplasia: A Retrospective Study.

The authors retrospectively analyzed the effects of Le Fort I advancement with distraction osteogenesis on skeletal and airway variables in patients with midfacial hypoplasia induced by cleft lip and/or palate using 3-dimensional computed tomography reconstructions. The authors enrolled 23 subjects with moderate-to-severe midface hypoplasia induced by cleft lip and palate who were treated with Le Fort I distraction osteogenesis (mean age, 19.22±3.48 y; male/female ratio, 20/3); computed tomography images (1 before distraction and another at completion of distraction) were acquired. A 3-dimensional craniometric findings and airway volumes for the nasal cavity, nasopharynx, velopharynx, and upper and lower oropharynx were compared before and after distraction. The relationships between craniofacial morphology and changes in airway volume were also assessed ( P <0.05 was considered significant). Significant increases were observed in airway volumes for the nasal cavity (13.85%), nasopharynx (50.82%), velopharynx (29.57%), and upper oropharynx (36.92%) ( P =0.007, P <0.001, P =0.023, and P <0.001, respectively), whereas no significant changes were observed for the lower oropharynx ( P =0.117). Maxillary horizontal advancement was positively correlated with the airway volumes of the nasopharynx and upper oropharynx after distraction osteogenesis ( rs =+0.451, P =0.031; rs =+0.548, P =0.007); however, no significant correlations were observed for the nasal cavity and velopharynx. The authors' finding indicate that despite rotation of the mandible along with the maxilla, this change does not impact airway volume at the mandibular level. Le Fort I distraction osteogenesis can be feasible for patients with cleft lip and palate-induced midface hypoplasia, with satisfactory appearance and occlusion. Long-term detailed follow-up of the patients postdistraction osteogenesis is warranted.

A Novel Neurovascular Protection Method In Nasolabial Muscle Biomechanical Bionic Surgery For Secondary Unilateral Cleft Lip Repair: A Three-Dimensional Evaluation Of Short And Long-Term Results.

BACKGROUND: Neurovascular protection within the upper lip muscles is essential in muscle tension reconstruction and functional recovery during surgery. This study aimed to investigate a novel neurovascular protection method based on the intramuscular neurovascular partitions of the upper lip applied to nasolabial muscle biomechanical bionic surgery for secondary unilateral cleft lip repair and to evaluate postoperative outcomes. METHODS: From November 30, 2019 to October 31, 2020, 50 consecutive patients with secondary unilateral cleft lips who underwent the modified biomechanical bionic surgery were reviewed retrospectively. Three-dimensional (3D) photographs of patients were collected preoperatively, 7-day postoperatively, and during long-term follow-up (at least 6 months). The Global Aesthetic Improvement Scale was used to evaluate surgical subjective outcomes, and statistical analysis of nasolabial measurements on 3D photographs was used to evaluate objective outcomes before and after surgery. RESULTS: The Global Aesthetic Improvement Scale showed that 94% of patients had improved upper lip morphology and 92% had improved nasal morphology 7-day postoperatively. In all, 84% maintained favorable nasolabial morphology during long-term follow-up. Statistical results showed that the postoperative alar width, philtral depth, columellar angle, and nasal base inclination angle were significantly improved compared with preoperatively (P<0.01). The differences between the above 4 indexes were not statistically significant (P >0.05) between 7-day postoperatively and long-term follow-up, indicating that the postoperative lasting outcomes were satisfactory. CONCLUSIONS: The novel method of neurovascular protection in nasolabial muscle biomechanical bionic surgery can achieve a long-term improvement of labial-nasal morphology and function in patients with secondary unilateral cleft lip.

Using a New Deep Learning Method for 3D Cephalometry in Patients With Cleft Lip and Palate.

Deep learning algorithms based on automatic 3-dimensional (D) cephalometric marking points about people without craniomaxillofacial deformities has achieved good results. However, there has been no previous report about cleft lip and palate. The purpose of this study is to apply a new deep learning method based on a 3D point cloud graph convolutional neural network to predict and locate landmarks in patients with cleft lip and palate based on the relationships between points. The authors used the PointNet++ model to investigate the automatic 3D cephalometric marking points. And the mean distance error of the center coordinate position and the success detection rate (SDR) were used to evaluate the accuracy of systematic labeling. A total of 150 patients were enrolled. The mean distance error for all 27 landmarks was 1.33 mm, and 9 landmarks (30%) showed SDRs at 2 mm over 90%, and 3 landmarks (35%) showed SDRs at 2 mm under 70%. The automatic 3D cephalometric marking points take 16 seconds per dataset. In summary, our training sets were derived from the cleft lip with/without palate computed tomography to achieve accurate results. The 3D cephalometry system based on the graph convolutional neural network algorithm may be suitable for 3D cephalometry system in cleft lip and palate cases. More accurate results may be obtained if the cleft lip and palate training set is expanded in the future.

Three-dimensional Anatomy of the Velopharyngeal Muscles in the Cleft Palate.

OBJECTIVE: We have used micro-computed tomography (CT) to elucidate the relationship between the muscle fibers in specimens with cleft palate. These findings could be useful for muscle reconstruction in cleft palate repair and to better understand cleft palate speech. DESIGN: Cadaveric anatomical study. PARTICIPANTS: This study included three specimens with cleft palate. INTERVENTION: The specimens were stained with phosphomolybdic acid and scanned by Micro-CT. MAIN OUTCOME MEASURE(S): The anatomy of the muscles. RESULTS: Using 2D projection images and 3D reconstruction models, subtle anatomical structures could be observed in the muscles. The attachment of the levator veli palatini (LVP) was not at the posterior edge of the hard palate or palatine aponeurosis (PA), but at the anterior 21.71-44.2% of the cleft edge. The palatopharyngeal (PP) was composed of two bundles: inferior and superior heads, which clasped the LVP. The uvularis was unevenly distributed, and located on both sides of the cleft edge, originating at the edge. The palatoglossus, superior constrictor of pharynx and anatomical structure around the pterygoid hamulus, were normal. The PA, PP and LVP were attached to the cleft edge from front to back, in that order. The position of the uvularis was not fixed. CONCLUSIONS: With the help of Micro-CT technology, detailed anatomical features and the relationship between muscles could be visualized. In the specimen with cleft palate, muscles in the soft palate were associated with the pharyngeal muscles, which formed the 3D "velopharyngeal muscles complex." These findings provide anatomical evidence for muscle reconstruction in cleft palate repair.

Cadaveric Study and Micro-Computed Tomography of the Anatomy of Palatine Aponeurosis and its Link to the Soft Palate Muscles and Pharyngeal Muscles.

OBJECTIVE: There have been few studies on the anatomy of palatine aponeurosis (PA). Herein, we elucidated the relationship between the PA and soft palate muscles and pharyngeal muscles. DESIGN: Two cadaveric specimens were dissected to observe the gross anatomy of the PA. Six cadaveric specimens were processed and scanned by micro-computed tomography to determine the elaborate anatomy. Images were exported to Mimics software to reconstruct a three-dimensional model. RESULTS: The PA covered the anterior (32.1%-38.8%) of the soft palate, extending from the tensor veli palatini (TVP) and connecting to 3 muscles: palatopharyngeus (PP), uvula muscle, and superior pharyngeal constrictor (SC). The SC and PP are attached to the PA on the medial side of the pterygoid hamulus. SC muscle fibers were attached to the hamulus, forming a distinct gap between the hamulus. Some muscle fibers of the PP and uvula originated from the PA. The PA extended from the TVP to the midline and the posterior edge of the hard palate. The PA was not uniformly distributed, which was complementary to the attached muscles in thickness. CONCLUSIONS: PA, as a flexible fibrous membrane, maintains the shape of the soft palate. It extends from the TVP and covers anteriorly about one-third of the soft palate. The PA provides a platform for the soft palate muscles and pharyngeal muscles, connecting to the PP, uvula muscle, and SC. These muscles are important for palatopharyngeal closure and middle-ear function. It is necessary to minimize the damage to the PA during surgical interventions.

Increased Fat Graft Survival by Promoting Adipocyte Dedifferentiation.

BACKGROUND: Some adipocytes undergo dedifferentiation after fat transplantation, and this may affect the survival of fat grafts. However, this effect has not been adequately studied. OBJECTIVES: This study aimed to clarify the effect of promoting the dedifferentiation of mature adipocytes on the survival of fat grafts. METHODS: Mature adipocytes and adipose stem cells (ASCs) were treated with OSI-906 (a specific inhibitor of insulin receptor and insulin-like growth factor-1 receptor) in vitro, and then the dedifferentiation of mature adipocytes and the proliferation of ASCs were evaluated. In the in vivo experiment, human lipoaspirates mixed with phosphate-buffered saline (Group A) or OSI-906 (Group B) were compared in nude mice. Grafts were harvested at 2, 8, and 12 weeks, and volume retention rate, histologic, and immunohistochemical analyses were conducted. RESULTS: OSI-906 can promote the dedifferentiation of mature adipocytes and inhibit the proliferation of ASCs. At 12 weeks, Group B showed a better volume retention rate (mean [standard deviation, SD], 62.3% [7.61%]) than group A (47.75% [6.11%]) (P < .05). Moreover, viable adipocytes and vascularization showed greater improvement in Group B than in Group A. CONCLUSIONS: This study suggests that promoting the dedifferentiation of mature adipocytes can improve the survival rate and quality of fat grafts.

Muscle Flap Reconstruction Based on Muscle Tension Line Groups to Repair the Philtrum of Patients With Microform Cleft Lip or Secondary Cleft Lip.

ABSTRACT: The muscle flap reconstruction technique was developed based on the concept of muscle tension line groups, which elucidates how nasolabial muscle tension helps maintain the shape of the philtrum. To investigate the operative effect, we reviewed 43 patients with microform cleft lip and 102 patients with secondary cleft lip treated with muscle flap reconstruction between January 2018 and June 2020. The patients were scanned using the digital three-dimensional stereophotogrammetry face system pre- and post-operatively, and comparative analysis of three-dimensional (3D) images was used to highlight variations of the philtrum. Visual analog scales were used to assess surgical outcomes. More than 6 months after the surgery, comparative 3D images of 37 patients (86.04%) with microform cleft lip and 86 patients (84.31%) with secondary cleft lip showed visible improvement in the prominence of the affected column. In addition, visual analog scale scores showed that 38 microform cleft lip patients (88.37%) and 89 secondary cleft lip patients (87.25%) had a good appearance. The postoperative prominence of the philtral column in both groups improved significantly compared to before surgery (P < 0.001 and P < 0.001, respectively). There was no significant difference in scores for philtrum prominence pre- and post-operatively between the 2 groups (P > 0.05). Muscle flap reconstruction is an effective means to create the 3D configuration of the philtrum. The biomechanical properties of muscles play a vital role in the morphological maintenance of the philtrum.

An Auxiliary Cranioparietal Anchorage System to Improve the Stability of the Rigid External Distraction Device in Children With Crouzon Syndrome.

ABSTRACT: Rigid external distraction (RED) device instability because of thin skulls in syndromic craniosynostosis patients remains challenging. The authors propose the use of an auxiliary cranioparietal anchorage system to improve RED device stability in patients with Crouzon syndrome. Five consecutive patients with Crouzon syndrome underwent Le Fort III osteotomy with midface advancement using a modified external device. Cephalometric analysis and three-dimensional computed tomography images were evaluated preoperatively and after device removal. Mechanical analyses of the RED device with and without the auxiliary cranioparietal anchorage system were performed. Both postoperative photographs and follow-up computed tomographs showed obvious midfacial advancement with no intracranial pin perforation or external frame migration. Mechanical analysis showed that the new system reduced the possibility of postoperative external frame migration. The auxiliary cranioparietal anchorage system might be considered an adjunct to the RED device in patients with Crouzon syndrome to avoid postoperative complications.

Three-Dimensional Measurement of the Zygomatic-Maxillary Complex in Unilateral Cleft Lip and Palate Patients With Maxillary Retrusion.

ABSTRACT: The purpose of this study was to investigate the anatomical features of the zygomatic-maxillary complex in unilateral cleft lip and palate (UCLP) patients with maxillary retrusion. Individuals were grouped and craniofacial measurements were carried out for 34 individuals in the UCLP with maxillary retrusion group (UMRG) and 50 from a control group (CG). The authors measured the length, width, and height of the maxilla and zygoma and also measured predetermined regions on the midface in each group. Independent sample group t tests were performed to determine differences between groups (with significance set at P < 0.05). Multiple points on the midface (the most posterior point on the contour of the maxillary alveolar process, lowest point of the zygomaticomaxillary suture, furthest point to the zygomatic self-base plane, superior point in the infraorbital foramen, and lowest point of the inferior margin of the orbit) to the coronal plane were smaller in the UMRG than in the CG (all results P < 0.05). In addition, the maxillary length was significantly reduced in the UMRG than in the CG (P < 0.05). In summary, for the UCLP patients with maxillary retrusion, the deficiency in the midface gradually reduced going upward, with the deficiency in the maxillary alveolar level the most serious. The zygoma was influenced to a lesser extent.

Transposition Mucosal Flap Combined With Autologous Fat Transplantation to Repair Lip Defects Caused by Sclerotherapy of Hemangioma.

ABSTRACT: Infantile hemangiomas are the most common benign childhood tumors and can occur on any part of the human body. Sclerosing agents are used in the early stage of treatment of infantile hemangioma. Sometimes a lip defect remains after sclerosing agent treatment. We developed a simple technique to repair lip defects. The authors performed transposition mucosal flap and autologous fat transplantation surgery on patients who had lip defects caused by sclerosing agents. The flap was transposed 90° from the intraoral labial mucosa to the vermilion defect. Autologous fat was transplanted to the white lip defect. If necessary, a secondary fat transplantation may be performed every half year. All patients were followed up to evaluate the effect of the operation. Patients were asked to rate their satisfaction with the surgery between 1 and 10. Digital three-dimensional evaluation was performed. Sixteen patients underwent the surgery successfully, and the flaps were all viable. No complications occurred after surgery (5 males, 11 females; age range, 5-27 years; 12 upper lip, 3 lower lip, and 1 median lip). The patients were satisfied with the aesthetic outcome of surgery (mean score, 9). Seven patients wanted to undergo a second fat transplantation, whereas 9 patients felt it was unnecessary to transplant fat again. Transposition mucosal flap combined with autologous fat transplantation is reliable and minimally invasive. It is an effective method for repairing moderate lip defects mainly involving vermilion caused by a sclerosing agent.

Vascular anatomy of the velopharyngeal muscles and its clinical implications: A fresh cadaveric study based on micro-computed tomography.

Poor speech improvement after levator veli palatini (LVP) reconstruction may be related to intraoperative vascular injury. We aimed to examine the vascular anatomy of the velopharyngeal muscles to provide a guide for arterial protection in cleft palate repair. Fresh adult cadaveric heads were injected with gelatin/lead oxide. The velopharyngeal specimens were stained with iodine and scanned using micro-computed tomography. Three-dimensional reconstruction models were obtained using a computer-aided design software. The ascending palatine artery (APaA), especially the posterior branch, is the main artery supplying the velopharyngeal muscles. The posterior branch of the APaA reaches the dorsal part of the musculus uvulae in the posterior one third of the soft palate (SP) and lies 1.75 mm (standard deviation, 0.06) under the nasal mucosa; the anterior branch penetrates the anterolateral side of the LVP to reach the anterior one third of the SP and lies 7.09 mm (0.03) under the oral mucosa. The posterior APaA, anterior ApaA, and ApaA trunk had mean diameters of 0.41 mm (0.04), 0.46 mm (0.06), and 0.65 mm (0.04) at 0.5, 1, and 1.5 cm distance from the palatal midline, respectively. To minimize vascular injury, mobilization of muscles during intravelar veloplasty should be performed within a distance of 1 cm from the palatal midline, and dissection of the oral submucosa should be reduced in the anterior one third of the SP, while wide dissection of the nasal submucosal should be avoided in the posterior one third of the SP.

Washing Lipoaspirate Improves Fat Graft Survival in Nude Mice.

BACKGROUND: The optimal fat processing technique of fat grafting has not been determined. We have proved the importance of washing lipoaspirate to remove blood, but the necessity of washing when there is no obvious bleeding during liposuction is not clear. OBJECTIVES: The purpose of this study is to further investigate the effect of washing on fat graft survival and the underlying mechanisms, from the perspective of inflammation, oxidative stress and apoptosis. METHODS: To exclude the influence of blood, de-erythrocyte infranatant (dEI) isolated from lipoaspirate was obtained. Purified fat processed by cotton pad filtration mixed with dEIs after sedimentation (sedimentation group), washing (washing group) or phosphate buffer solution (control group) was transplanted to nude mice subcutaneously. Samples were harvested at 1 day and 1, 3, 8 weeks after transplantation. Volume and weight retention, histologic examination, immunostaining of perilipin-1, CD31, CD45 and Ly6g, mRNA expression of PPAR-γ, C/EBPα, VEGF, bFGF, IL-6, IL10, TNF-α, TGF-ß, Bax and Bcl-2, and protein contents of 8-iso-PGF2α, IL-6, IL10, TNF-α and TGF-ß were all compared among groups. RESULTS: After transplantation, volume and weight retention, histologic scores, viable adipocytes and vascularization were all improved in the washing group, with increased expression of adipogenic and angiogenic genes. Compared with the sedimentation group, the washing group had milder inflammation, lower levels of oxidative stress and apoptosis. CONCLUSIONS: Washing lipoaspirate to eliminate mixed components can improve fat graft survival and promote adipogenesis and angiogenesis, possibly by relieving inflammation, reducing oxidative stress injury and inhibiting apoptosis. NO LEVEL ASSIGNED: This journal requires that authors assign a level of evidence to each submission to which Evidence-Based Medicine rankings are applicable. This excludes Review Articles, Book Reviews, and manuscripts that concern Basic Science, Animal Studies, Cadaver Studies, and Experimental Studies. For a full description of 47 these Evidence-Based Medicine ratings, please refer to the Table of Contents or the online Instructions to Authors http://www.springer.com/00266 .

Micro-Computed Tomography-Based Three-Dimensional Anatomical Structure of the Region Around the Pterygoid Hamulus.

OBJECTIVE: Palatoplasty would involve the structures around the pterygoid hamulus. However, clinicians hold different opinions on the optimal approach for the muscles and palatine aponeurosis around the pterygoid hamulus. The absence of a consensus regarding this point can be attributed to the lack of investigations on the exact anatomy of this region. Therefore, we used micro-computed tomography to examine the anatomical structure of the region surrounding the pterygoid hamulus. DESIGN: Cadaveric specimens were stained with iodine-potassium iodide and scanned by micro-computed tomography to study the structures of the tissues, particularly the muscle fibers. We imported Digital Imaging and Communications in Medicine images to Mimics to reconstruct a 3-dimensional model and simplified the model. RESULTS: Three muscles were present around the pterygoid hamulus, namely the palatopharyngeus (PP), superior constrictor (SC), and tensor veli palatini (TVP). The hamulus connects these muscles as a key pivot. The TVP extended to the palatine aponeurosis, which bypassed the pterygoid hamulus, and linked the PP and SC. Some muscle fibers of the SC originated from the hamulus, the aponeurosis of which was wrapped around the hamulus. There was a distinct gap between the pterygoid hamulus and the palatine aponeurosis. This formed a pulley-like structure around the pterygoid hamulus. CONCLUSIONS: Transection or fracture of the palatine aponeurosis or pterygoid hamulus, respectively, may have detrimental effects on the muscles around the pterygoid hamulus, which play essential roles in the velopharyngeal function and middle ear ventilation. Currently, cleft palate repair has limited treatment options with proven successful outcomes.