Age estimation from median palatine suture using computed tomography reconstructed 3D images: a comparison of Northern and Southwestern Chinese populations.

To investigate the potential of computed tomography (CT) images of median palatine suture (MP) for adult age estimation in the Northern and Southwestern Chinese populations. A total of 1110 cranial CT scans from individuals aged 10-79 years, including 557 northern Chinese and 553 southwestern Chinese, were collected for analysis. After volume reformation and multiplanar reconstruction, a total of 20 slices of median palatine suture were selected from each individual. The closure of sutures was analyzed into four stages, and the cumulative scores of 20 slices were recorded as the suture closure score (SCS). The correlations between SCS and age were compared among the two Chinese populations residing in diverse geographic regions. Regression models were established for age estimation. The estimation accuracy was evaluated based on the test set. The mean absolute error (MAE) and the correlation between predicted age and chronological age were calculated to evaluate estimation accuracy. The SCS of MP exhibited a significant correlation with age (0.613, northern male; 0.678, southwestern male; 0.730, northern female; 0.704, Southwestern female; 0.662, total). Furthermore, there were statistically significant differences in SCS among different regions and sex groups (p < 0.001). The cubic regression model had the highest R2 value in all subjects, especially among Northern females and Southwestern males, while the power and quadratic regression models showed the highest R2 value in Northern males and Southwestern females, respectively. In the test set, the Northern cohort demonstrated a lower MAE (9.06 ± 7.32 years, males; 9.17 ± 5.28 years, females) compared to the Southwestern cohort (9.19 ± 7.49 years, male; 10.61 ± 6.83 years, female). Additionally, it was observed that males exhibited a lower MAE than females in both regional groups. This study demonstrated the potential utility of CT images of the MP for age estimation in Chinese populations, emphasizing the significance of incorporating regional and sex factors within this context.

Craniosynostosis: orofacial and oral health perspectives with masticatory insights.

BACKGROUND: Craniosynostosis (CS), premature fusion of one or more cranial sutures, leads to abnormal skull development, impacting both facial esthetics and oral function. This study aimed to evaluate the specific orofacial and oral health characteristics, including masticatory performance, in Thai patients with CS. METHODS: A comparative study was conducted with Thai CS patients aged 6-17 years and a control group of healthy individuals with similar age distribution. Assessments included craniofacial morphology, oral health status, and masticatory performance. Intergroup comparisons utilized appropriate statistical tests. RESULTS: The study included 24 CS patients with a mean age of 10.11 ± 2.98 years and 30 controls. CS patients exhibited a significantly higher prevalence of various oral conditions compared to controls: cleft palate (20.8%), anterior open bite (41.7%), anterior crossbite (54.2%), posterior crossbite (50%), combined anterior-posterior crossbite (45.8%), dental crowding in both maxilla and mandible (50% and 45.8% respectively), congenitally missing teeth (50%), supernumerary teeth (12.5%), and eruption failure (54.2%). Furthermore, CS patients exhibited significantly higher caries prevalence and susceptibility, alongside poorer oral hygiene, compared to controls. Regarding jaw relationships, CS patients exhibited a significantly higher proportion of Angle's Class III malocclusion (50%) compared to the control group, where Class I malocclusion was predominant (50%). Masticatory performance, assessed using the two-color gum mixing ability test, showed significantly higher hue variance in CS patients (0.12 ± 0.07) compared to the control group, indicating reduced chewing performance. CONCLUSION: This study underscores the significant orofacial and oral health challenges faced by children with CS, including a high prevalence of malocclusions, dental anomalies, elevated caries experience, and compromised masticatory function. These findings emphasize the importance of tailored interventions and comprehensive oral healthcare strategies to address the unique needs of this population and improve their overall quality of life.

Age Estimation by Machine Learning and CT-Multiplanar Reformation of Cranial Sutures in Northern Chinese Han Adults.

OBJECTIVES: To establish age estimation models of northern Chinese Han adults using cranial suture images obtained by CT and multiplanar reformation (MPR), and to explore the applicability of cranial suture closure rule in age estimation of northern Chinese Han population. METHODS: The head CT samples of 132 northern Chinese Han adults aged 29-80 years were retrospectively collected. Volume reconstruction (VR) and MPR were performed on the skull, and 160 cranial suture tomography images were generated for each sample. Then the MPR images of cranial sutures were scored according to the closure grading criteria, and the mean closure grades of sagittal suture, coronal sutures (both left and right) and lambdoid sutures (both left and right) were calculated respectively. Finally taking the above grades as independent variables, the linear regression model and four machine learning models for age estimation (gradient boosting regression, support vector regression, decision tree regression and Bayesian ridge regression) were established for northern Chinese Han adults age estimation. The accuracy of each model was evaluated. RESULTS: Each cranial suture closure grade was positively correlated with age and the correlation of sagittal suture was the highest. All four machine learning models had higher age estimation accuracy than linear regression model. The support vector regression model had the highest accuracy among the machine learning models with a mean absolute error of 9.542 years. CONCLUSIONS: The combination of skull CT-MPR and machine learning model can be used for age estimation in northern Chinese Han adults, but it is still necessary to combine with other adult age estimation indicators in forensic practice.

Onabotulinumtoxina (craniotomy scar combined with cranial suture line injections) for persistent post craniotomy headache: Case series with long-term follow-up.

BACKGROUND: There is no defined preventive treatment protocol for persistent post-craniotomy headache. In several small case series and individual case reports onabotulinumtoxinA injected into the craniotomy scar has shown possible efficacy. What is lacking is long term follow-up and if focusing on the cranial suture lines along with the craniotomy scar can enhance improvement and provide more sustained benefit. METHODS: Retrospective chart review with case series. RESULTS: Four patients (three women, one man) with ICHD-3 defined persistent post craniotomy headache were treated using a novel onabotulinumtoxinA injection protocol. All the patients presented with continuous head pain of moderate to severe intensity. All had severe allodynia on the side of their craniotomy. All had significant reduction in quality of life. Our application of onabotulinumtoxinA involved injection into both the surgical scar and the transected/irritated cranial suture lines noted on neuroimaging and physical examination. With treatment all patients demonstrated significant benefit including a reduction in daily pain intensity (75%-100%), developing periods of pain freedom (2-7 days per week) and having a dramatic improvement in quality of life (close to 100% in all). The benefit was sustained for at least five years of follow-up. CONCLUSION: From our case series it appears that injection not only along the painful craniotomy scar but into the involved cranial suture lines provides positive efficacy and sustained improvement in patients with persistent post craniotomy headache.

Three-dimensional morphometric analysis of cranial sutures - A novel approach to quantitative analysis.

Objective: Differences in complexity of cranial suture forms on the endocranial (i.e., deep) and ectocranial (i.e., superficial) skull surfaces have been noted in the literature, indicating through thickness three-dimensional (3D) suture variability depending on the chosen section and necessity for considering the complete 3D structure in many cases. This study aims to evaluate the variability of suture morphology through the skull thickness using a rat model, and to provide more robust metrics and methodologies to analyze suture morphology. Design: X-ray micro-computed tomographic (µCT) imaging methods were utilized in order to provide internal structure information. Methods were developed to isolate and analyze sutures widths and linear interdigitation index (LII) values on each adjacent offset transverse plane of the µCT datasets. LII was defined as the curved path length of the suture divided by the linear length between the ends of the region of interest. Scans were obtained on 15 female rats at ages of 16, 20, and 24 weeks (n = 5/age). Samples were imaged at 18 µm resolutions with 90 kV source voltage, 278 µA source amperage, and 0.7° increments. Suture widths and LII values were compared using a Kruskal-Wallis test. Results: 3D variability in local suture widths within individuals, as well as through thickness variabilities in planar widths and LII was observed. Kruskal-Wallis tests for bulk through thickness averaged suture widths and LII were found to be statistically insignificant, despite clear geometric differences through suture thicknesses. Conclusion: Although the bulk morphometric variability between age groups was found to be statistically insignificant, the 3D variability within individuals point to the importance of analyzing suture form using 3D metrics when studying suture development, response to functional activity, or morphometry in general.

Technical evolution of pediatric neurosurgery: craniosynostosis from 1972 to 2023 and beyond.

Very few clinical entities have undergone so many different treatment approaches over such a short period of time as craniosynostosis. Surgical treatments for this condition have ranged from simple linear craniectomies, accounting for the specific role of cranial sutures in assuring the normal growth of the skull, to more complex cranial vault reconstructions, based on the perceived role of the skull base in affecting the growth of the skull. While a great deal of evolution has occurred, there remains controversy regarding the ideal treatment including the best surgical technique, the optimal age for surgery, and the long-term morphological and neurodevelopmental outcomes. The evolution of the surgical management of craniosynostosis in the last 50 years has been affected by several factors. This includes the awareness of needing to operate on affected children during infancy to achieve the best results, the use of multistage operations, the availability of more sophisticated surgical tools, and improved perioperative care. In some forms of craniosynostosis, the operations can be carried out at a very young age with low morbidity, and with the postoperative use of a molding helmet, springs, or distractors, these operations prove to be as effective as traditional larger cranial reconstructions performed in older children. As a consequence, complex surgical operations have become progressively less utilized. A second relevant advance was the more recent advent of a molecular diagnosis, which allowed us to understand the pathogenesis of some associated malformations and neurodevelopmental issues that were observed in some children despite appropriate surgical treatment. Future research should focus on improving the analysis of longer-term outcomes and understanding the natural history of craniofacial conditions, including what issues persist despite optimal surgical correction. Progress in molecular investigations concerning the normal and pathological development of cranial sutures could be a further significant step in the management of craniosynostosis, possibly favoring a "medical" treatment in the near future. Artificial intelligence will likely have a role in establishing the diagnosis with less reliance on radiographic studies and in assisting with surgical planning. Overall, much progress has been made, but there remains much to do.

Sonographic guide for botulinum toxin injections for chronic migraine headache: EURO-MUSCULUS/USPRM approach.

Application of botulinum toxin A (BoNT-A) into the muscles of the head and neck area has become a widespread and reliable treatment modality for chronic migraine. The mechanism of action for BoNT-A is the inhibition of acetylcholine and local nociceptive peptide release at the terminal nerve endings. Cranial sutures have the highest concentration of nociceptive structures; therefore BoNT-A injection into the suture lines - as opposed to head and neck muscles - has been proposed for the treatment of chronic migraine. Nerve endings in sutures rapidly absorb BoNT-A and transfer it across the afferent nerve fibers in dura mater via orthodromic and antidromic transmission. In this article, ultrasound-guided BoNT-A application around the cranial sutures will be illustrated. It is noteworthy that suture injections would be safer and more efficient when applied with such guidance.

FGF signaling in cranial suture development and related diseases.

Suture mesenchymal stem cells (SMSCs) are a heterogeneous stem cell population with the ability to self-renew and differentiate into multiple cell lineages. The cranial suture provides a niche for SMSCs to maintain suture patency, allowing for cranial bone repair and regeneration. In addition, the cranial suture functions as an intramembranous bone growth site during craniofacial bone development. Defects in suture development have been implicated in various congenital diseases, such as sutural agenesis and craniosynostosis. However, it remains largely unknown how intricate signaling pathways orchestrate suture and SMSC function in craniofacial bone development, homeostasis, repair and diseases. Studies in patients with syndromic craniosynostosis identified fibroblast growth factor (FGF) signaling as an important signaling pathway that regulates cranial vault development. A series of in vitro and in vivo studies have since revealed the critical roles of FGF signaling in SMSCs, cranial suture and cranial skeleton development, and the pathogenesis of related diseases. Here, we summarize the characteristics of cranial sutures and SMSCs, and the important functions of the FGF signaling pathway in SMSC and cranial suture development as well as diseases caused by suture dysfunction. We also discuss emerging current and future studies of signaling regulation in SMSCs.

Analysis of skeletal stem cells by renal capsule transplantation and ex vivo culture systems.

Skeletal stem cells residing in the suture mesenchyme are responsible for proper development, homeostasis, and injury repair of the craniofacial skeleton. These naïve cells are programmed to differentiate into osteoblast cell types and mediate bone formation via an intramembranous ossification mechanism. The simplicity of this system also offers great advantages to studying osteoblastogenesis compared to the appendicular and axial skeletons. Recent studies utilizing genetically based cell tracing have led to the identification of skeletal stem cell populations in craniofacial and body skeletons. Although the genetic analysis indicates these cells behave like stem cells in vivo, not all of them have been thoroughly examined by stem cell isolation and stem cell-mediated tissue generation. As regeneration is an integral part of stem cell characteristics, it is necessary to further analyze their ability to generate tissue at the ectopic site. The establishment of an ex vivo culture system to maintain the stemness properties for extended periods without losing the regenerative ability is also pertinent to advance our knowledge base of skeletal stem cells and their clinical applications in regenerative medicine. The purpose of this review is to discuss our recent advancements in analyses of skeletal stem cells using renal capsule transplantation and sphere culture systems.

Computed Tomographic Evaluation of Cranial Suture Obliteration for Age Estimation in an Indian Population.

BACKGROUND: Age estimation is a critical aspect of human identification. Age is assessed using cranial suture obliteration, pelvic morphological changes, epiphyseal fusion of long bones, dental maturation, and other standard methods. METHODS: The present study investigated three-dimensional (3D) computed tomography (CT) scans of 263 individuals (183 males and 80 females) to assess the extent of ectocranial suture closure. The assessment of obliteration was done using a three-stage scoring method. Spearman's correlation coefficient (p < 0.05) was calculated to assess the relationship between cranial suture closure and chronological age. Simple and multiple linear regression models were developed using the cranial suture obliteration scores to estimate age. RESULTS: The standard errors of the estimate using multiple linear regression models developed for estimating age using obliteration scores of sagittal, coronal, and lambdoid sutures were 15.08 years in males, 13.27 years in females, and 14.74 years in the total study population. CONCLUSION: This study concludes that in the absence of additional skeletal age markers, this method can be used alone or in conjunction with other well-established methods of age assessment.

Control of craniofacial development by the collagen receptor, discoidin domain receptor 2.

Development of the craniofacial skeleton requires interactions between progenitor cells and the collagen-rich extracellular matrix (ECM). The mediators of these interactions are not well-defined. Mutations in the discoidin domain receptor 2 gene (DDR2), which encodes a non-integrin collagen receptor, are associated with human craniofacial abnormalities, such as midface hypoplasia and open fontanels. However, the exact role of this gene in craniofacial morphogenesis is not known. As will be shown, Ddr2-deficient mice exhibit defects in craniofacial bones including impaired calvarial growth and frontal suture formation, cranial base hypoplasia due to aberrant chondrogenesis and delayed ossification at growth plate synchondroses. These defects were associated with abnormal collagen fibril organization, chondrocyte proliferation and polarization. As established by localization and lineage-tracing studies, Ddr2 is expressed in progenitor cell-enriched craniofacial regions including sutures and synchondrosis resting zone cartilage, overlapping with GLI1 + cells, and contributing to chondrogenic and osteogenic lineages during skull growth. Tissue-specific knockouts further established the requirement for Ddr2 in GLI +skeletal progenitors and chondrocytes. These studies establish a cellular basis for regulation of craniofacial morphogenesis by this understudied collagen receptor and suggest that DDR2 is necessary for proper collagen organization, chondrocyte proliferation, and orientation.

We each have unique facial features that are key to our identities. These features are inherited, but the mechanisms are poorly understood. People with the genetic disease spondylo-meta-epiphyseal dysplasia, or SMED, have characteristic facial and skull abnormalities including a flattened face and shortened skull. SMED is associated with mutations that inactivate the gene encoding a protein called discoidin domain receptor 2 (DDR2), which is a receptor for collagen. Collagen is the major structural protein in the human body, supporting the structure of cells and tissues. It also controls cell behaviors including growth, migration and differentiation, and it helps form tissues such as cartilage or bone. At least some of the effects of collagen on cells depend on its interaction with DDR2. Since the facial and skull abnormalities in mice with mutations that stop DDR2 from working correctly resemble those of SMED patients, these mice can be used to understand the cellular basis for this disease, as well as the role of DDR2 in the embryonic development of the face and skull. Therefore, Mohamed et al. set out to understand how loss of DDR2 causes the characteristic facial and skull defects associated with SMED. Mohamed et al. used mice that had been genetically modified so that DDR2 could be inactivated in skeletal progenitor cells, cartilage cells and bone cells (osteoblasts). Examining these mice, they found that the shortened skulls and flat face characteristic of mice lacking DDR2 are due to bones at the skull base failing to elongate correctly due to defects in the growth centers that depend on cartilage. Mohamed et al. also discovered that the cells that normally produce DDR2 are the progenitors of cartilage and bone-forming cells, which partly explains why lacking this protein leads to issues in growth of these tissues. In addition to shedding light on the causes of SMED, Mohamed et al.'s results also provide general insights into the mechanisms controlling the formation of facial and skull bones that depend on interactions between cells and collagen. This information may help explain how other abnormalities in the face and skull emerge, and provide a basis for how the shape of the skull has changed during human evolution. In the future, it may be possible to manipulate the activity of DDR2 to correct skull defects.

Transmucosal pterygomaxillary disjunction using a piezoelectric device, in the context of the minimally invasive Le Fort I osteotomy protocol.

The aim of this study was to assess the accuracy and clinical implications of pterygomaxillary junction (PMJ) disjunction with a transmucosal PMJ osteotomy using a piezoelectric hand-piece device, in the context of Le Fort I osteotomy, by evaluating the level of PMJ disarticulation and the need for bone trimming around the pedicle. An ambidirectional 1-month follow-up cohort study was designed involving consecutive patients undergoing minimally invasive maxillary Le Fort I osteotomy through the twist technique. Two cohorts were defined according to whether or not the transmucosal PMJ osteotomy was performed. The site of PMJ disjunction was analysed radiographically. A total of 114 patients were included in the study, 57 in each group. The overall accuracy of the PMJ disjunction path was higher in the test group (43.9%) than in the control group (15.8%). Multiple logistic regression analysis identified the need for bone trimming (odds ratio 0.02; P < 0.001) and removal of the upper third molar (odds ratio 0.17; P < 0.001) as relevant factors. In conclusion, compared with the originally described twist technique, combination of the latter with the PMJ osteotomy increased its accuracy at the level of the PMJ. As a result, there is a decrease in resistance during down-fracture and decrease in the need for bone trimming around the pedicle, with preservation of the minimally invasive concept.

UCSQ Method Applied on 3D Photogrammetry: Non-Invasive Objective Differentiation Between Synostotic and Positional Plagiocephaly.

OBJECTIVE: Objective differentiation between unilateral coronal synostosis (UCS) and positional posterior plagiocephaly (PPP) based on 3D photogrammetry according to Utrecht Cranial Shape Quantificator (UCSQ). DESIGN: Retrospective study. SETTING: Primary craniofacial center. PATIENTS, PARTICIPANTS: Thirty-two unoperated patients (17 UCS; 15 PPP) (age < 1 year). INTERVENTIONS: Extraction of variables from sinusoid curves derived using UCSQ: asymmetry ratio forehead and occiput peak, ratio of gradient forehead and occiput peak, location forehead and occiput peak. MAIN OUTCOME MEASURE(S): Variables, derived using 3D photogrammetry, were analyzed for differentiation between UCS and PPP. RESULTS: Frontal peak was shifted to the right side of the head in left-sided UCS (mean x-value 207 [192-220]), and right-sided PPP (mean x-value 210 [200-216]), and to the left in right-sided UCS (mean x-value 161 [156-166]), and left-sided PPP (mean x-value 150 [144-154]). Occipital peak was significantly shifted to the right side of the head in left-sided PPP (mean x-value 338 [336-340]) and to the left in right-sided PPP (mean x-value 23 [14-32]). Mean x-value of occipital peak was 9 (354-30) in left- and 2 (350-12) in right-sided UCS. Calculated ratio of gradient of the frontal peak is, in combination with the calculated asymmetry ratio of the frontal peak, a distinctive finding. CONCLUSIONS: UCSQ objectively captures shape of synostotic and positional plagiocephaly using 3D photogrammetry, we therefore developed a suitable method to objectively differentiate UCS from PPP using radiation-free methods.

METTL5 regulates cranial suture fusion via Wnt signaling.

METTL5 is a methyltransferase that mediates eukaryotic 18S ribosomal RNA m6A modification, and its mutations lead to intellectual disability, microcephaly, and facial dysmorphism in patients. However, the role of METTL5 in craniofacial development remains poorly understood. This study demonstrates that Mettl5 knockout mice exhibit poor ossification, widened cranial sutures, and a cleidocranial dysplasia-like phenotype. Deletion of Mettl5 leads to increased proliferation and decreased osteogenic differentiation of suture mesenchymal stem cells. Mechanistically, we find that Wnt signaling is significantly downregulated after Mettl5 knockout. Overall, we reveal an essential role of METTL5 in craniofacial development and osteogenic differentiation of suture mesenchymal stem cells, making METTL5 a potential diagnostic and therapeutic target for craniofacial developmental diseases.

Cranial suture morphometry and mechanical response to loading: 2D vs. 3D assumptions and characterization.

Cranial sutures are complex soft tissue structures whose mechanics are often studied due to their link with bone growth in the skull. Researchers will often use a cross-sectional two-dimensional slice to define suture geometry when studying morphometry and/or mechanical response to loading. However, using a single cross section neglects the full suture complexity and may introduce significant errors when defining their form. This study aims to determine trends in suture path variability through skull thickness in a swine model and the implications of using a 'representative' cross section on mechanical modeling. To explore these questions, a mixture of quantitative analysis of computed tomography images and finite element models was used. The linear interdigitation and width of coronal and sagittal sutures were analyzed on offset transverse planes through the skull thickness. It was found that sagittal suture width and interdigitation were largely consistent through the skull thickness, whereas the coronal suture showed significant variation in both. The finite element study found that average values of displacement and strain were similar between the two-dimensionally variable and three-dimensionally variable models. Larger ranges and more complex distributions of strain were found in the three-dimensionally variable model. Outcomes of this study indicate that the appropriateness of using a representative cross section to describe suture morphometry and predict mechanical response should depend on specific research questions and goals. Two-dimensional approximations can be sufficient for less-interdigitated sutures and when bulk site mechanics are of interest, while taking the true three-dimensional geometry into account is necessary when considering spatial variability and local mechanical response.

A study of 285 cases of cranial vault suture closure in Chinese adults.

PURPOSE: To examine the presence and characteristics of cranial vault suture closure in Chinese adults and to explore whether craniosacral therapy (CST) manipulation is rational from the anatomical perspective. METHODS: Anthropological non-metric observation and craniometry were used to study 285 dry skull specimens of Chinese adults. RESULTS: A total of 91 specimens with closed extracranial sutures were observed, with an occurrence rate of 31.93%. Based on the mode of closure, there were 32 cases of single type closure, with sagittal suture closure predominating with 20 cases (21.98%); 59 cases of composite closure, with a partial closure of coronal suture + sagittal suture + lambdoid suture predominating with 26 cases (28.57%). In terms of the degree of closure, there were 13 cases (14.28%) of sagittal suture grade 0 closure and 78 cases (85.72%) of grade 1 - 4 closure; 34 cases (37.36%) of coronal suture grade 0 closure and 57 cases (62.64%) of grade 1 - 4 closure; 47 cases (51.65%) of lambdoid suture grade 0 closure and 44 cases (48.35%) of grade 1 - 4 closure. The segment and degree of coronal suture closure (46, 80.7%) and lambdoid suture (31, 70.45%) were mostly left-right symmetrical. The bone surfaces on either side of the cranial vault sutures are embedded in each other, forming a rough, complex and interlocking bone-suture-bone structure. CONCLUSION: This study observed the closure of the cranial vault suture, summarized its characteristics, and explored the irrationality of the CST manipulation. The anatomical characteristics of the cranial suture dictate that manipulation cannot push the cranial suture at will.

Embryonic requirements for Tcf12 in the development of the mouse coronal suture.

A major feature of Saethre-Chotzen syndrome is coronal craniosynostosis, the fusion of the frontal and parietal bones at the coronal suture. It is caused by heterozygous loss-of-function mutations in either of the bHLH transcription factors TWIST1 and TCF12. Although compound heterozygous Tcf12; Twist1 mice display severe coronal synostosis, the individual role of Tcf12 had remained unexplored. Here, we show that Tcf12 controls several key processes in calvarial development, including the rate of frontal and parietal bone growth, and the boundary between sutural and osteogenic cells. Genetic analysis supports an embryonic requirement for Tcf12 in suture formation, as combined deletion of Tcf12 in embryonic neural crest and mesoderm, but not in postnatal suture mesenchyme, disrupts the coronal suture. We also detected asymmetric distribution of mesenchymal cells on opposing sides of the wild-type frontal and parietal bones, which prefigures later bone overlap at the sutures. In Tcf12 mutants, reduced asymmetry is associated with bones meeting end-on-end, possibly contributing to synostosis. Our results support embryonic requirements of Tcf12 in proper formation of the overlapping coronal suture.

Spatial transcriptomics reveals a role for sensory nerves in preserving cranial suture patency through modulation of BMP/TGF-ß signaling.

The patterning and ossification of the mammalian skeleton requires the coordinated actions of both intrinsic bone morphogens and extrinsic neurovascular signals, which function in a temporal and spatial fashion to control mesenchymal progenitor cell (MPC) fate. Here, we show the genetic inhibition of tropomyosin receptor kinase A (TrkA) sensory nerve innervation of the developing cranium results in premature calvarial suture closure, associated with a decrease in suture MPC proliferation and increased mineralization. In vitro, axons from peripheral afferent neurons derived from dorsal root ganglions (DRGs) of wild-type mice induce MPC proliferation in a spatially restricted manner via a soluble factor when cocultured in microfluidic chambers. Comparative spatial transcriptomic analysis of the cranial sutures in vivo confirmed a positive association between sensory axons and proliferative MPCs. SpatialTime analysis across the developing suture revealed regional-specific alterations in bone morphogenetic protein (BMP) and TGF-ß signaling pathway transcripts in response to TrkA inhibition. RNA sequencing of DRG cell bodies, following direct, axonal coculture with MPCs, confirmed the alterations in BMP/TGF-ß signaling pathway transcripts. Among these, the BMP inhibitor follistatin-like 1 (FSTL1) replicated key features of the neural-to-bone influence, including mitogenic and anti-osteogenic effects via the inhibition of BMP/TGF-ß signaling. Taken together, our results demonstrate that sensory nerve-derived signals, including FSTL1, function to coordinate cranial bone patterning by regulating MPC proliferation and differentiation in the suture mesenchyme.

Identification of differentially expressed proteins between fused and open sutures in sagittal nonsyndromic craniosynostosis during suture development by quantitative proteomic analysis.

PURPOSE: Nonsyndromic craniosynostosis (NCS), the premature fusion of cranial sutures, results in an abnormal skull shape and is associated with a significant morbidity. Proteomics is a promising tool for disease characterization and biomarker discovery; we aimed to identify biologically relevant differentially expressed proteins for NCS. EXPERIMENTAL DESIGN: Label-based quantitative proteomic profiling using TMT was performed on protein extracted from mesenchymal stem cells, osteoblasts and bone tissue of five open and five fused sutures of sagittal NCS (sNCS) and analyzed using quantitative LC-MS/MS based bottom-up proteomics. Differential protein abundance between open and fused sutures was determined to identify biologically relevant proteins of interest. Proteins were validated in an independent sample set by western blot and immunohistochemistry. RESULTS: We observed 838 differentially expressed proteins between open and fused sutures of sNCS. Decorin, lumican, and asporin were significantly downregulated while COL4A1 and TGFß1|1 were upregulated in fused compared to open sutures. CONCLUSIONS AND CLINICAL RELEVANCE: The majority of significantly differentially expressed proteins between open and fused sutures were observed in the proteomes of osteoblasts suggesting that protein changes contributing to premature sagittal suture fusion occur predominantly at the osteoblast level. Our findings suggest a possible ineffective ECM deposition at the osteoblast cell stage.

Quantification of Severity of Unilateral Coronal Synostosis.

OBJECTIVES: Severity of unilateral coronal synostosis (UCS) can vary. Quantification is important for treatment, expectations of treatment and natural outcome, and education of the patient and parents. DESIGN: Retrospective study. SETTING: Primary craniofacial center. PATIENTS, PARTICIPANTS: Twenty-three preoperative patients with unilateral coronal craniosynostosis (age < 2 years). INTERVENTION: Utrecht Cranial Shape Quantifier (UCSQ) was used to quantify severity using the variables: asymmetry ratio of frontal peak and ratio of frontal peak gradient. MAIN OUTCOME MEASURES(S): The UCSQ variables were combined and related to visual score using Pearson correlation coefficient; UCSQ and visual score were additionally compared to Di Rocco classification by one-way analysis of variance or Kruskal-Wallis test. All measurements were made on computed tomography scans. RESULTS: Good correlation between UCSQ and visual score was found (r = 0.67). No statistically significant differences were found between group means of UCSQ in the 3 categories of Di Rocco classification (F2,20 = 0.047; P > .05). Kruskal-Wallis test showed no significant differences between group means of visual score in the 3 categories of Di Rocco classification (Kruskal-Wallis H (2) = 0.871; P > .05). CONCLUSIONS: Using UCSQ, we can quantify UCS according to severity using characteristics, it outperforms traditional methods and captures the whole skull shape. In future research, we can apply UCSQ to 3D-photogrammetry due to the utilization of external landmarks.