Advancing rare cancer research by MALDI mass spectrometry imaging: Applications, challenges, and future perspectives in sarcoma.

MALDI mass spectrometry imaging (MALDI imaging) uniquely advances cancer research, by measuring spatial distribution of endogenous and exogenous molecules directly from tissue sections. These molecular maps provide valuable insights into basic and translational cancer research, including tumor biology, tumor microenvironment, biomarker identification, drug treatment, and patient stratification. Despite its advantages, MALDI imaging is underutilized in studying rare cancers. Sarcomas, a group of malignant mesenchymal tumors, pose unique challenges in medical research due to their complex heterogeneity and low incidence, resulting in understudied subtypes with suboptimal management and outcomes. In this review, we explore the applicability of MALDI imaging in sarcoma research, showcasing its value in understanding this highly heterogeneous and challenging rare cancer. We summarize all MALDI imaging studies in sarcoma to date, highlight their impact on key research fields, including molecular signatures, cancer heterogeneity, and drug studies. We address specific challenges encountered when employing MALDI imaging for sarcomas, and propose solutions, such as using formalin-fixed paraffin-embedded tissues, and multiplexed experiments, and considerations for multi-site studies and digital data sharing practices. Through this review, we aim to spark collaboration between MALDI imaging researchers and clinical colleagues, to deploy the unique capabilities of MALDI imaging in the context of sarcoma.

Genetic association and functional validation of ZFP36L2 in non-syndromic orofacial cleft subtypes.

BACKGROUND: Non-syndromic orofacial cleft (NSOC) is one of the most common craniofacial malformations with complex etiology. This study aimed to explore the role of specific SNPs in ZFP36L2 and its functional relevance in zebrafish models. METHODS: We analyzed genetic data of the Chinese Han population from two previous GWAS, comprising of 2512 cases and 2255 controls. Based on the Hardy-Weinberg Equilibrium (HWE) and minor allele frequency (MAF), SNPs in the ZFP36L2 were selected for association analysis. In addition, zebrafish models were used to clarify the in-situ expression pattern of zfp36l2 and the impact of its Morpholino-induced knockdown. RESULTS: Via association analysis, rs7933 in ZFP36L2 was significantly associated with various non-syndromic cleft lip-only subtypes, potentially conferring a protective effect. Zebrafish embryos showed elevated expression of zfp36l2 in the craniofacial region during critical stages of oral cavity formation. Furthermore, Morpholino-induced knockdown of zfp36l2 led to craniofacial abnormalities, including cleft lip, which was partially rescued by the addition of zfp36l2 mRNA. CONCLUSION: Our findings highlight the significance of ZFP36L2 in the etiology of NSOC, supported by both human genetic association data and functional studies in zebrafish. These results pave the way for further exploration of targeted interventions for craniofacial malformations.

Role of neuropeptides in orofacial pain: A literature review.

BACKGROUND: Neuropeptides play a critical role in regulating pain and inflammation. Despite accumulating evidence has further uncovered the novel functions and mechanisms of different neuropeptides in orofacial pain sensation and transmission, there is deficient systematic description of neuropeptides' pain modulation in the orofacial region, especially in the trigeminal system. OBJECTIVES: The present review aims to summarise several key neuropeptides and gain a better understanding of their major regulatory roles in orofacial inflammation and pain. METHODS: We review and summarise current studies related to calcitonin gene-related peptide (CGRP), substance P (SP), opioid peptide (OP), galanin (GAL) and other neuropeptides' functions and mechanisms as well as promising targets for orofacial pain control. RESULTS: A number of neuropeptides are clearly expressed in the trigeminal sensory system and have critical functions in the transduction and pathogenesis of orofacial pain. The functions, possible cellular and molecular mechanisms have been introduced and discussed. Neuropeptides and their agonists or antagonists which are widely studied to be potential treatment options of orofacial pain has been evaluated. CONCLUSIONS: Various neuropeptides play important but distinct (pro-nociceptive or analgesic) roles in orofacial pain with different mechanisms. In summary, CGRP, SP, NPY, NKA, HK-1, VIP mainly play proinflammatory and pro-nociceptive effects while OP, GAL, OXT, OrxA mainly have inhibitory effects on orofacial pain.

Changes in calpain-2 expression during glioblastoma progression predisposes tumor cells to temozolomide resistance by minimizing DNA damage and p53-dependent apoptosis.

BACKGROUND: Glioblastoma multiforme (GBM) is characterized by an unfavorable prognosis for patients affected. During standard-of-care chemotherapy using temozolomide (TMZ), tumors acquire resistance thereby causing tumor recurrence. Thus, deciphering essential molecular pathways causing TMZ resistance are of high therapeutic relevance. METHODS: Mass spectrometry based proteomics were used to study the GBM proteome. Immunohistochemistry staining of human GBM tissue for either calpain-1 or -2 was performed to locate expression of proteases. In vitro cell based assays were used to measure cell viability and survival of primary patient-derived GBM cells and established GBM cell lines after TMZ ± calpain inhibitor administration. shRNA expression knockdowns of either calpain-1 or calpain-2 were generated to study TMZ sensitivity of the specific subunits. The Comet assay and É£H2AX signal measurements were performed in order to assess the DNA damage amount and recognition. Finally, quantitative real-time PCR of target proteins was applied to differentiate between transcriptional and post-translational regulation. RESULTS: Calcium-dependent calpain proteases, in particular calpain-2, are more abundant in glioblastoma compared to normal brain and increased in patient-matched initial and recurrent glioblastomas. On the cellular level, pharmacological calpain inhibition increased the sensitivities of primary glioblastoma cells towards TMZ. A genetic knockdown of calpain-2 in U251 cells led to increased caspase-3 cleavage and sensitivity to neocarzinostatin, which rapidly induces DNA strand breakage. We hypothesize that calpain-2 causes desensitization of tumor cells against TMZ by preventing strong DNA damage and subsequent apoptosis via post-translational TP53 inhibition. Indeed, proteomic comparison of U251 control vs. U251 calpain-2 knockdown cells highlights perturbed levels of numerous proteins involved in DNA damage response and downstream pathways affecting TP53 and NF-κB signaling. TP53 showed increased protein abundance, but no transcriptional regulation. CONCLUSION: TMZ-induced cell death in the presence of calpain-2 expression appears to favor DNA repair and promote cell survival. We conclude from our experiments that calpain-2 expression represents a proteomic mode that is associated with higher resistance via "priming" GBM cells to TMZ chemotherapy. Thus, calpain-2 could serve as a prognostic factor for GBM outcome.

Dinuclear Nickel Complexes Using Hexadentate Benzothiazole-Based Diamine-Bisphenolate Ligands: Highly Active Catalysts for Copolymerization of Carbon Dioxide with Epoxides.

We reported for the first time the utilization of hexadentate benzothiazole-based diamine-bisphenolate ligands to synthesize structurally well-characterized dinickel dicarboxylate complexes and studied their catalysis for copolymerization of carbon dioxide with epoxides. Dinickel carboxylate complexes having a 1,3-diamine-bridged backbone were demonstrated to be high-performance catalysts for alternating copolymerization of CO2 and cyclohexene oxide (CHO) with high product selectivity. Particularly, acetate-supported nickel complex 2 enabled us to promote such CO2-copolymerization of this kind with a maximum turnover frequency of up to 2600 h-1 and gave good molecular weight controllability under high-pressure conditions. It is worth noting that bimetallic Ni catalyst 2 was also capable of mediating the catalytic CO2-polymerization of alicyclic epoxides at atmospheric pressure. Kinetic investigations of CO2/CHO copolymerization by 2 allowed us to determine the rate equation of -d[CHO]/dt = kp[2]1[CHO]1, and such catalysis exhibited a first-order dependence on both dinickel complex and CHO concentrations.

Response and regulatory mechanisms of heat resistance in pathogenic fungi.

Both the increasing environmental temperature in nature and the defensive body temperature response to pathogenic fungi during mammalian infection cause heat stress during the fungal existence, reproduction, and pathogenic infection. To adapt and respond to the changing environment, fungi initiate a series of actions through a perfect thermal response system, conservative signaling pathways, corresponding transcriptional regulatory system, corresponding physiological and biochemical processes, and phenotypic changes. However, until now, accurate response and regulatory mechanisms have remained a challenge. Additionally, at present, the latest research progress on the heat resistance mechanism of pathogenic fungi has not been summarized. In this review, recent research investigating temperature sensing, transcriptional regulation, and physiological, biochemical, and morphological responses of fungi in response to heat stress is discussed. Moreover, the specificity thermal adaptation mechanism of pathogenic fungi in vivo is highlighted. These data will provide valuable knowledge to further understand the fungal heat adaptation and response mechanism, especially in pathogenic heat-resistant fungi. KEY POINTS: • Mechanisms of fungal perception of heat pressure are reviewed. • The regulatory mechanism of fungal resistance to heat stress is discussed. • The thermal adaptation mechanism of pathogenic fungi in the human body is highlighted.

Studies on the radiographic visibility of the periodontal ligament in lower third molars: can the Olze method be used in the Chinese population?

The purpose of the present research is to study the radiographic visibility of the periodontal ligament in third molars from a northern Chinese population to determine if this methodology could be used for the 18-year threshold. A total of 1300 conventional orthopantomograms from 650 males and 650 females of northern Chinese subjects aged between 15 and 40 years were analyzed. The radiographic visibility of the periodontal membrane of fully mineralized third molars was assessed using the stages described by Olze et al. (2010). Stage 0 first appeared at the age of 17.05 years in males and at the age of 18.76 years in females. Stage 1 was first detected at 18.52 years in males and 19.59 years in females. Stage 2 was first achieved by males at the age of 22.33 years and by females at the age of 21.37 years. The earliest appearance of stage 3 was 26.85 years in males and 24.92 years in females. If stage 1 is determined, it is possible to prove that an individual has already attained the age of 18 years. Stages 2 and 3 can be used to determine that a person is over 21 years of age. However, it should also be noted that in 271 cases of tooth 38 and 255 cases of tooth 48, the visibility stage could not be reliably assessed because of fused roots or narrowed furcations. Therefore, this method seems to be of limited value for age estimation in the Chinese population.

Third molar maturity index (I3M) for assessing age of majority in northern Chinese population.

In many countries, assessment of legal age, also known as age of majority, has become increasingly important over the years. In China, individuals older than 18 years of age have full capacity regarding civil conduct and can be tried as an adult for criminal charges. Therefore, from a legal point of view, it is crucial to determine whether an individual is an adult. The developmental degree of the third molar is widely recognized as a suitable site for age estimation in late adolescence. This article uses the third molar maturity index (I3M) with a cutoff value of I3M = 0.08, which was established by Cameriere et al. in 2008, to distinguish whether an individual is a minor or an adult (≥ 18 years of age) in a northern Chinese population. A total of 840 digital orthopantomograms (OPTs) from 420 male and 420 female northern Chinese subjects aged 12 to 25 years were evaluated. It was found that an increase in I3M corresponded to a decrease in chronological age. In our study, I3M = 0.10 showed better accuracy in age discrimination in both men and women. This threshold also resulted in high sensitivity (0.929 and 0.809) and specificity (0.940 and 0.973) in males and females, respectively. The proportion of correctly classified subjects was 0.917 (95% CI, 0.898 to 0.935) in total, 0.938 (95% CI, 0.915 to 0.961) in male and 0.895 (95% CI, 0.866 to 0.925) in female subjects. Bayes post-test probabilities were 0.967 (95% CI, 0.947 to 0.986) in males and 0.983 (95% CI, 0.966 to 0.998) in females. These differences in threshold values between Chinese and Caucasian populations might be because the development of third molars is delayed in the Chinese population compared to the Caucasian population. In conclusion, I3M might be a useful method in legal and forensic practices to determine ages in late adolescence in northern Chinese individuals. However, a specific population should be tested before I3M is used for legal age estimation.

Increased m6A modification of BDNF mRNA via FTO promotes neuronal apoptosis following aluminum-induced oxidative stress.

N6-methyladenosine (m6A) RNA modification is a new epigenetic molecular mechanism involved in various biological or pathological processes. Exposure to aluminum (Al) has been considered to promote neuronal apoptosis resulting in cognitive dysfunction, yet whether m6A modification participates in the underlying mechanism remains largely unknown. Here, rats exposed to aluminum-maltolate [Al(mal)3] for 90 days showed impaired learning and memory function and elevated apoptosis, which were related to the increased m6A level and decreased fat mass and obesity-associated protein (FTO, an m6A demethylase) in the hippocampus. Accordingly, similar results presented in PC12 cells following Al(mal)3 treatment and FTO overexpression relieved the increased apoptosis and m6A level in vitro. Next, we identified brain-derived neurotrophic factor (BDNF) as the functional downstream target of FTO in a m6A-dependent manner. Furthermore, it was found that as the onset of aluminum neurotoxicity, oxidative stress may be the upstream regulator of FTO in aluminum-induced apoptosis. Taken together, these results suggest that increased m6A modification of BDNF mRNA via FTO promotes neuronal apoptosis following aluminum-induced oxidative stress.

Small extracellular vesicles derived from lipopolysaccharide-preconditioned dental follicle cells inhibit cell apoptosis and alveolar bone loss in periodontitis.

OBJECTIVE: This study aimed to explore the effects of small extracellular vesicles derived from lipopolysaccharide-preconditioned dental follicle cells (L-D-sEV) on periodontal ligament cells from periodontitis affected teeth (p-PDLCs) in vitro and experimental periodontitis in mice. DESIGN: In vitro, the biological function of p-PDLCs and the underlying molecular mechanism were investigated by flow cytometry, Western blot, and quantitative real-time PCR (qRT-PCR) analysis. Eighteen-eight-week-old male C57BL/6 mice were randomly divided into three groups: control (Con), periodontitis (Peri), and L-D-sEV groups. Mice periodontitis model was induced by placing the 5-0 silk thread (around the maxillary second molar) and P.gingivalis (1 ×107 CFUs per mouse). In vivo, the alveolar bone loss, osteoclast activity, and macrophage polarization were measured by micro-computed tomography and histological analysis. RESULTS: In vitro, the RANKL/OPG ratio and phosphorylation of JNK and P38 protein levels of p-PDLCs were significantly decreased after L-D-sEV administration. Besides, flow cytometry and qRT-PCR analysis showed that L-D-sEV reduced apoptosis of p-PDLCs, down-regulated apoptosis-related genes Caspase-3 and BCL-2-Associated X expression, and up-regulated B-cell lymphoma-2 gene levels. In vivo, L-D-sEV administration significantly reduced alveolar bone loss, inhibited osteoclast activity, and induced M2 polarization. The histological analysis showed that iNOS/CD206, RANKL/OPG, p-JNK/JNK, and p-P38/P38 ratios were significantly lower in the L-D-sEV group than in the Peri group. CONCLUSIONS: L-D-sEV administration alleviated alveolar bone loss by mediating RANKL/OPG-related osteoclast activity and M2 macrophage polarization, alleviating p-PDLCs apoptosis and proliferation via the JNK and P38 pathways.

SERBP1 interacts with PARP1 and is present in PARylation-dependent protein complexes regulating splicing, cell division, and ribosome biogenesis.

RNA binding proteins (RBPs) containing intrinsically disordered regions (IDRs) are present in diverse molecular complexes where they function as dynamic regulators. Their characteristics promote liquid-liquid phase separation (LLPS) and the formation of membraneless organelles such as stress granules and nucleoli. IDR-RBPs are particularly relevant in the nervous system and their dysfunction is associated with neurodegenerative diseases and brain tumor development. SERBP1 is a unique member of this group, being mostly disordered and lacking canonical RNA-binding domains. Using a proteomics approach followed by functional analysis, we defined SERBP1's interactome. We uncovered novel SERBP1 roles in splicing, cell division, and ribosomal biogenesis and showed its participation in pathological stress granules and Tau aggregates in Alzheimer's disease brains. SERBP1 preferentially interacts with other G-quadruplex (G4) binders, implicated in different stages of gene expression, suggesting that G4 binding is a critical component of SERBP1 function in different settings. Similarly, we identified important associations between SERBP1 and PARP1/polyADP-ribosylation (PARylation). SERBP1 interacts with PARP1 and its associated factors and influences PARylation. Moreover, protein complexes in which SERBP1 participates contain mostly PARylated proteins and PAR binders. Based on these results, we propose a feedback regulatory model in which SERBP1 influences PARP1 function and PARylation, while PARylation modulates SERBP1 functions and participation in regulatory complexes.

Can canines alone be used for age estimation in Chinese individuals when applying the Kvaal method?

Due to the secondary dentin formation, the dental pulp undergoes changes in shape throughout life. Based on this phenomenon, the Kvaal method has been applied to various populations for age estimation, and its usefulness has been verified. When applying the Kvaal method to Chinese subjects, we observed a relatively strong correlation between mandibular canines and age. This study notes the correlation between canines and chronological age and is the first to identify which canine is most closely related to chronological age. In addition, a new, simpler formula is determined based on canines according to Kvaal's methodology. The radiographs of 360 individuals from northern China were selected, from which the widths and lengths of the pulp from four canines were measured according to the Kvaal method. Next, inter- and intra-observer reliabilities were analyzed in order to assess the repeatability of these measurements. The correlation between measurements and age was examined, and Chinese-specific age estimation formulae were derived. The results revealed that the ratios from the left maxillary canine exhibited the strongest correlation with age compared to the other canines, whereas the left mandibular canine showed the weakest correlation, which may contribute to the overall poor correlation of mandibular canines with age. What's more, the formula derived from the left maxillary canine in this study displayed the highest coefficients of determination, and the formula derived from all canines showed the lowest residuals. Both of these formulae performed better than the Chinese-specific formula derived from six different types of teeth in our previous study, which had formerly possessed the highest coefficients of determination and the lowest residuals. Thus, we concluded that canines do play an important role in age estimation in the Chinese population, and the correlation between maxillary canines and chronological age is stronger than that of mandibular canines, although no distinct trend as to which side is better correlated with age was established. Going forward, we recommend the analysis of additional samples from different geographical regions and populations to further verify the importance of canines in age estimation.

Targeted Re-Sequencing of the 2p21 Locus Identifies Non-Syndromic Cleft Lip Only Novel Susceptibility Gene ZFP36L2.

rs7590268 present on the 2p21 locus was identified to be associated with non-syndromic cleft lip with or without cleft palate (NSCL/P) in several populations, including the Chinese Han population, indicating that 2p21 was a susceptibility locus for NSCL/P. However, previous studies have only identified common single-nucleotide polymorphism (SNP) within the THADA gene, neglecting the rare variants and other genes in 2p21; thus, this study was designed to investigate additional variants and novel susceptibility genes in 2p21. A total of 159 NSCL/P patients and 542 controls were recruited in the discovery phase, whereas 1830 NSCL/P patients and 2,436 controls were recruited in the replication phase. After targeted region sequencing, we performed association and burden analyses for the common and rare variants, respectively. Furthermore, RNA-seq, proliferation assay and cell cycle analysis were performed to clarify the possible function of the candidate gene ZFP36L2. Association analysis showed that four SNPs were specifically associated with non-syndromic cleft lip only (NSCLO) and two SNPs were associated with both NSCLO and NSCL/P. Burden analysis indicated that ZFP36L2 was associated with NSCLO (p = .0489, OR = 2.41, 95% CI: 0.98-5.90). Moreover, SNPs in the ZFP36L2 targeted gene JUP were also associated with NSCLO. ZFP36L2 also inhibited cell proliferation and induced G2 phase arrest in the GMSM-K cell line. Therefore, we proposed that ZFP36L2 is a novel susceptibility gene of NSCLO in the 2p21 locus, which could lead to NSCLO by modulating cell proliferation and cycle.

Treatment of inflammatory bone loss in periodontitis by stem cell-derived exosomes.

Periodontitis is the primary cause of tooth loss, but there is no effective treatment to repair inflammatory bone loss in periodontitis. Exosomes emerge as essential paracrine factors of mesenchymal stem cells (MSCs) that mediated tissue regeneration. Here, we investigated the potential of exosomes secreted by periodontal ligament stem cells (PDLSCs) as therapeutics for the bone defect in periodontitis. Exosomes secreted from PDLSCs derived from healthy periodontal ligaments (h-PDLSCs) and their function were evaluated on PDLSCs isolated from the inflammatory periodontal ligament of periodontitis patients (i-PDLSCs). Treatment of exosomes of h-PDLSCs led to an increase in the formation of mineralized nodules and the expressions of osteogenic genes and proteins in i-PDLSCs. Mechanistically, h-PDLSCs-exosomes suppressed the over-activation of canonical Wnt signaling to recover the osteogenic differentiation capacity of i-PDLSCs. To evaluate the therapeutic of exosomes on inflammatory bone loss, h-PDLSCs-exosomes loaded with Matrigel or ß-TCP were employed to repair bone defects in rat models of periodontitis. Compared to the vehicle-treated control group, h-PDLSCs-exosomes-treated rats resulted in more bone formation in the defect of alveolar bone. In conclusion, these results demonstrated that exosomes derived from healthy PDLSCs could rescue the osteogenesis capacity of endogenous stem cells under an inflammatory environment and promote regeneration of alveolar bone. Our findings suggest that MSCs-derived exosome is an effective and practical cell-free MSC therapeutic for the treatment of periodontitis. STATEMENT OF SIGNIFICANCE: There is no effective treatment to repair inflammatory bone loss in periodontitis. As essential paracrine factors of PDLSCs, exosomes might mediate tissue regeneration during stem cell therapy. Here, we reported that exosomes secreted from healthy PDLSCs promoted the osteogenic differentiation of PDLSCs derived from periodontitis tissue. Healthy PDLSCs-exosomes treatment resulted in accelerated bone formation in the defect of alveolar bone in rat models of periodontitis. Mechanistically, h-PDLSCs-exosomes suppressed the over-activation of canonical Wnt signaling to recover the osteogenic differentiation capacity of inflammatory PDLSCs. These findings suggest that MSCs-derived exosome is an effective and practical cell-free MSC therapeutic for the treatment of periodontitis.

Targeted re-sequencing on 1p22 among non-syndromic orofacial clefts from Han Chinese population.

Rs560426 at 1p22 was proved to be associated with NSCL/P (non-syndromic cleft lip with or without the palate) in several populations, including Han Chinese population. Here, we conducted a deep sequencing around rs560426 to locate more susceptibility variants in this region. In total, 2,293 NSCL/P cases and 3,235 normal controls were recruited. After sequencing, association analysis was performed. Western blot, RT-qPCR, HE, immunofluorescence staining, and RNA sequencing were conducted for functional analyses of the selected variants. Association analysis indicated that rs77179923 was the only SNP associated with NSCLP specifically (p = 4.70E-04, OR = 1.84), and rs12071152 was uniquely associated with LCLO (p = 4.00E-04, OR = 1.30, 95%CI: 1.12-1.51). Moreover, de novo harmful rare variant NM_004815.3, NP_004806.3; c.1652G>C, p.R551T in ARHGAP29 resulted in a decreased expression level of ARHGAP29, which in turn affected NSCL/P-related biological processes; however, no overt cleft palate (CP) phenotype was observed. In conclusion, rs12071152 was a new susceptible variant, which is specifically associated with LCLO among the Han Chinese population. Allele A of it could increase the risk of having a cleft baby. Rs77179923 and rare variant NM_004815.3, NP_004806.3; c.1652G>C, p.R551T at 1p22 were both associated with NSCLP among the Han Chinese population. However, this missense variation contributes to no overt CP phenotype due to dosage insufficiency or compensation from other genes.

Will different sagittal and vertical skeletal types relate the soft tissue thickness: A study in Chinese female adults.

Facial reconstruction is a classical technique in forensic anthropology to reestablish the contours of the soft tissues over the skull. The accurate facial soft tissue data plays an essential role in forensic facial reconstruction. However, according to previous studies, various skeletal types might relate to different thickness in facial soft tissue. Until now, there are few publications focusing on the relationship between facial soft tissue thickness (FSTT) and different skeletal types, and none of them analyze the FSTT according to various sagittal and vertical skeletal types. The aim of this study was to testify the possible existence of correlations between FSTT and different skeletal types. In order to exclude interference from age, sex, ethnicity and body mass index (BMI) factors, we collected lateral projection X-rays data of 270 Chinese female aged 19-26 years with nomal BMI and divided them into various skeletal groups. Soft tissue thickness measurements were mainly based on 10 anthropological landmarks of the skull and statistics were analyzed on the basis of different skeletal types. The greatest differences were observed in the upper lip region of maxilla and the mental region of mandible. The concave and hypodivergent skull types showed the thickest soft tissue in maxillary region, and the convex and hypedivergent skull types showed the thickest soft tissue in the mental region. This study provided a database for FSTT according to various skeletal types in Chinese female population, and our current studies demonstrated that considering various skeletal types will improve the accuracy of facial reconstruction.

A bone matrix-simulating scaffold to alleviate replicative senescence of mesenchymal stem cells during long-term expansion.

Replicative senescence during in vitro augmentation, which is mostly induced by the loss of physiological microenvironment, hinders the application of mesenchymal stem cells (MSCs) in the clinic. Here, we investigated whether MSCs senescence could be prevented by bio-scaffold mimicking the natural tissue matrix. Human umbilical cord mesenchymal stem cells (hUCMSCs) exhibited a senescent phenotype during a long-term passage in the conventional culture dish. To fabricate the bone matrix, a naturally based matrix composed of nano-hydroxyapatite/chitosan/poly lactide-co-glycolide (nHA/CS/PLGA) was produced. Long-term passage resulted in an obvious increase in the expression of senescence markers and a reduction in the expression of master genes involved in tissue regeneration. Functional assay confirmed that nHA/CS/PLGA scaffold preserved the proliferation and differentiation of hUCMSCs even after being passaged 27 times. Moreover, in vivo ectopic bone formation assay revealed that the bone formation of hUCMSCs cultured on the nano-scaffolds for the long term was as robust as the cells in the early passage. In summary, our results demonstrate that nHA/CS/PLGA scaffold effectively preserves the stemness and youth of hUCMSCs in the long-term passage. Taken advantage of its compatibility and bioactivity, nHA/CS/PLGA scaffold is of great potential in large-scale expansion of MSCs for stem cell therapy and tissue engineering.

Cone-beam computed tomography-guided three-dimensional evaluation of treatment effectiveness of the Frog appliance.

OBJECTIVE: To evaluate the effectiveness of the Frog appliance in three dimensions by using cone-beam computed tomography (CBCT) images. METHODS: Forty patients (21 boys and 19 girls), averaged 11.7 years old, with an Angle Class II division 1 malocclusion were included in our study. They had either late mixed dentition or early permanent dentition, and the maxillary second molars had not yet erupted. All patients underwent CBCT before and after the treatment for measuring changes in the maxillary first molars, second premolars, central incisors, and profile. Paired-samples t-test was used to compare the mean difference in each variable before treatment and after the first phase of treatment. RESULTS: The maxillary first molars were effectively distalized by 4.25 mm (p < 0.001) and 3.53 mm (p < 0.05) in the dental crown and root apex, respectively. The tipping increased by 2.25°, but the difference was not significant. Moreover the teeth moved buccally by 0.84 mm (p < 0.05) and 2.87 mm (p < 0.01) in the mesiobuccal and distobuccal cusps, respectively, whereas no significant changes occurred in the root apex. Regarding the anchorage parts, the angle of the maxillary central incisor's long axis to the sella-nasion plane increased by 2.76° (p < 0.05) and the distance from the upper lip to the esthetic plane decreased by 0.52 mm (p = 0.01). CONCLUSIONS: The Frog appliance effectively distalized the maxillary molars with an acceptable degree of tipping, distobuccal rotation, and buccal crown torque, with only slight anchorage loss. Furthermore, CBCT image demonstrated that it is a simple and reliable method for three-dimensional analysis.

Application of the Kvaal method for age estimation using digital panoramic radiography of Chinese individuals.

The Kvaal method as a non-invasive method for age estimation based on the changes of pulpal size caused by secondary dentin deposition has been applied to different ethnics. However, its results are often conflicting and its accuracy is relatively poor. This study aimed to verify the applicability of the Kvaal method to estimate the ages of the Chinese population using digital panoramic radiographs and to formulate specific regression equations for use in Chinese subjects. A sample of 360 digital panoramic radiographs from northern Chinese population were selected, from which the pulpal size and width of six tooth types (maxillary central incisor, lateral incisor, second premolar, mandibular lateral incisor, canine, and first premolar) were measured using Kvaal's method. Then, inter- and intra-observer reliabilities were calculated to evaluate the reproducibility of those measurements. The correlation between chronological age and measurements was also examined and the Chinese-specific equations for age estimation were conducted. After comparing the accuracy of the Chinese-specific equation with the original Kvaal equation, the age of the Chinese population was clearly overestimated by Kvaal's equation, and a Chinese-specific equation could ameliorate age prediction accuracy. In addition, the residuals of the Chinese-specific equation were significantly lower than that of the original Kvaal's method. Thus, we concluded that the original Kvaal equation cannot accurately estimate age in the Chinese population. Although accuracy improved using the Chinese-specific equation that was based on Kvaal's methodology, the standard deviation remained large. In view of the lack of accurate age estimation methods for adults, we propose finding more correlated variables or combining other age estimation methods to increase the accuracy for adult age estimation.