Archives of oral biology最新文献_第3页

Intriguing astrocyte responses in CA1 to reduced and rehabilitated masticatory function: Dorsal and ventral distinct perspectives in adult mice CA1中星形胶质细胞对咀嚼功能减退和恢复的反应耐人寻味：成年小鼠背侧和腹侧的不同视角

IF 2.2 4区医学 Q2 DENTISTRY, ORAL SURGERY & MEDICINE

Archives of oral biology

Pub Date : 2024-10-05 DOI: 10.1016/j.archoralbio.2024.106097

Micaele Maria Lopes Castro , Fabio Leite do Amaral Junior , Fabíola de Carvalho Chaves de Siqueira Mendes , Daniel Clive Anthony , Dora Maria Tuna de Oliveira Brites , Cristovam Wanderley Picanço Diniz , Marcia Consentino Kronka Sosthenes

Objective

We sought to investigate the plasticity of diet-induced changes in astrocyte morphology of stratum lacunosum-moleculare (SLM) in CA1.

Design

Three diet regimes were adopted in 15 mice, from the 21st postnatal day to 6 months. The first diet regimen was pellet feed, called Hard Diet (HD). The second, with reduced masticatory, received a pellet-diet followed by a powdered-diet, and it was identified as Hard Diet/Soft Diet (HD/SD). Finally, the group with rehabilitated masticatory was named Hard Diet/Soft Diet/Hard Diet (HD/SD/HD). In the end, euthanasia and brain histological processing were performed, in which astrocytic immunoreactivity to glial-fibrillary-acidic-protein (GFAP) was tested. In reconstructed astrocytes, morphometric analysis was performed.

Results

Astrocyte morphometric revealed that changes in masticatory regimens impact astrocyte morphology. In the dorsal CA1, switching from a hard diet to a soft diet led to reductions in most variables, whereas in the ventral, fewer variables were affected, highlighting regional differences in astrocyte responses. Cluster analysis further showed that diet-induced changes in astrocyte morphology were reversible in the dorsal region, but not in the ventral region, indicating a persistent impact on astrocyte diversity and complexity in the ventral even after rehabilitation. Correlation tests between astrocyte morphology and behavioral performance demonstrated disrupted relationships under masticatory stress, with effects persisting after rehabilitation.

Conclusion

Changes in the diet result in significant alterations in astrocyte morphology, suggesting a direct link between dietary modulation and cellular structure. Morphometric analyses revealed distinct alterations in astrocyte morphology in response to changes in the masticatory regimen, with both dorsal/ventral regions displaying notable changes. Moreover, the regional differential effects on astrocytes underscore the complexity of mastication on neuroplasticity and cognitive function.

目的我们试图研究饮食诱导的 CA1 腔隙层（SLM）星形胶质细胞形态变化的可塑性。设计我们对 15 只小鼠从出生后第 21 天到 6 个月采用了三种饮食方案。第一种饮食方案是颗粒饲料，称为硬质饮食（HD）。第二组小鼠咀嚼功能减退，先吃颗粒饲料，再吃粉末饲料，称为硬质饮食/软质饮食（HD/SD）。最后，咀嚼功能得到恢复的一组被命名为硬质饮食/软质饮食/硬质饮食（HD/SD/HD）。最后，进行安乐死和脑组织学处理，检测星形胶质细胞对神经纤维酸性蛋白（GFAP）的免疫反应。结果星形胶质细胞形态计量学显示，咀嚼方式的改变会影响星形胶质细胞的形态。在背侧 CA1 中，从硬质饮食转为软质饮食会导致大多数变量的减少，而在腹侧，受影响的变量较少，这突显了星形胶质细胞反应的区域差异。聚类分析进一步表明，饮食诱导的星形胶质细胞形态变化在背侧区域是可逆的，但在腹侧区域则不可逆，这表明即使在康复后，星形胶质细胞的多样性和复杂性在腹侧仍会受到持续影响。星形胶质细胞形态与行为表现之间的相关性测试表明，在咀嚼应激下，两者之间的关系被破坏，康复后影响依然存在。形态计量学分析表明，星形胶质细胞形态随着咀嚼方式的改变而发生明显变化，背侧/腹侧区域均有显著变化。此外，对星形胶质细胞的区域性不同影响凸显了咀嚼对神经可塑性和认知功能的复杂性。

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引用次数: 0

Familial hypodontia in bronze age Northwest China (1046–771BC) 中国西北青铜时代（公元前 1046-771 年）的家族性牙齿发育不全。

IF 2.2 4区医学 Q2 DENTISTRY, ORAL SURGERY & MEDICINE

Archives of oral biology

Pub Date : 2024-10-02 DOI: 10.1016/j.archoralbio.2024.106104

Yaohan Wu , Qiang Ma , Baiwei Han , Yuanyuan Shen , Shaoqing Wen

Objective

This research aimed to report hypodontia cases in a Middle Bronze Age high-tier cemetery in China and test the possible hereditary behind hypodontia by performing kinship tests on those individuals.

Design

In this study, dental anomalies were observed on a human skeletal sample (n = 45) uncovered from Yaoheyuan, China. Ancient DNA analysis was subsequently employed on a subsample of the Yaoheyuan individuals (n = 15), including individuals observed hypodontia and individuals randomly sampled from the cemetery for preliminary investigation on the cemetery demography. Kinship estimation tests (READ, TKGWV2, KIN, and F3 test) were subsequently employed.

Results

The Yaoheyuan elite population had a prevalence (n = 7, 15 %) of tooth agenesis in either the maxilla or mandible, with one to two teeth missing. All missing teeth were incisors, except for one individual missing maxillary second molar. Preliminary ancient DNA results indicate that several kinship groups existed among interred individuals, including those with hypodontia, indicating the hereditary origin of these cases.

Conclusions

The prevalence of hypodontia observed on site is high compared to that in both modern East Asian populations and archaeological samples in the Chinese population. The preliminary kinship analysis suggests a case of familial hypodontia. Ancient DNA analysis should be thoroughly conducted in future studies to understand the genetic markers contributing to those hypodontia cases among the Yaoheyuan individuals.

研究目的本研究旨在报告中国青铜时代中期高级墓地中的牙列不齐病例，并通过对这些个体进行亲缘关系测试，检验牙列不齐背后可能存在的遗传性：本研究在中国姚河源出土的人类骨骼样本（n = 45）中观察到牙齿异常。随后，对姚河源的一个子样本（n = 15）进行了古DNA分析，其中包括观察到的牙齿畸形个体，以及为初步调查墓地人口结构而从墓地随机取样的个体。随后采用了亲缘关系估计检验（READ、TKGWV2、KIN和F3检验）：姚河源精英人群的上颌或下颌普遍存在牙齿缺失（n = 7，15%），缺失牙齿数量在 1 到 2 颗之间。除了一颗上颌第二臼齿缺失外，所有缺失的牙齿都是门牙。初步的古 DNA 检测结果表明，被埋葬者（包括牙齿缺失者）之间存在多个亲属群体，这表明这些病例具有遗传性：结论：与现代东亚人群和中国人群的考古样本相比，现场观察到的齿列不齐发生率较高。初步的亲缘关系分析表明这是一例家族性牙齿发育不全。在今后的研究中，应该对古DNA进行深入分析，以了解导致姚河源人牙齿发育不全的遗传标记。

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引用次数: 0

Metabolomic insights into idiopathic xerostomia: The central role of caffeine metabolism in salivary biochemistry 代谢组学对特发性口腔干燥症的启示：咖啡因代谢在唾液生物化学中的核心作用

IF 2.2 4区医学 Q2 DENTISTRY, ORAL SURGERY & MEDICINE

Archives of oral biology

Pub Date : 2024-10-02 DOI: 10.1016/j.archoralbio.2024.106102

Feifei Hou , Yuan Cui , Lu Ye , Fangman Chen , Chuanji Wu , Yang Meng , Peiyang Yuan , Xuemei Qiu , Xiaodong Feng , Lu Jiang

Objective

This study aims to delineate the salivary metabolomic profile of patients with idiopathic xerostomia using untargeted metabolomics techniques, with the goal of addressing the lack of clear diagnostic markers and providing insights into the pathophysiological mechanisms of the condition.

Design

In this observational, cross-sectional study, saliva samples from 33 patients with idiopathic xerostomia and 34 healthy controls were analyzed using Ultra Performance Liquid Chromatography Quadrupole Time of Flight Mass Spectrometry (UPLC-QTOF MS). Metabolomic profiling was complemented by multivariate statistical analysis to differentiate between affected individuals and controls.

Results

Metabolomic analysis delineated a pronounced differentiation between patients with idiopathic xerostomia and healthy controls. A total of 195 metabolites displayed significant differential expression, each with a variable importance in projection (VIP) greater than 1 and a P-value less than 0.05. Pathway enrichment analysis, according to the Kyoto Encyclopedia of Genes and Genomes (KEGG), identified 22 metabolites that participated in 18 distinct metabolic pathways. Among these, the caffeine metabolism pathway, characterized by notable alterations in impact values (VIP, P-value, Log2-fold change, Rich factor), emerged as the most significantly disrupted, underscoring its potential role in the pathophysiology of idiopathic xerostomia (P = 0.0000395).

Conclusions

The salivary metabolomic profiling revealed distinct alterations in idiopathic xerostomia, with a significant reduction in caffeine metabolism pathways, underscoring potential neuropathic involvement. This study advances our understanding of the metabolic alterations in xerostomia, suggesting that salivary metabolomics may offer viable biomarkers for diagnosing and understanding the etiology of idiopathic xerostomia. Future research should focus on therapeutic targeting of these metabolic disturbances and evaluating their reversibility with treatment.

目的本研究旨在利用非靶向代谢组学技术描述特发性口腔干燥症患者的唾液代谢组学特征，以解决缺乏明确诊断标志物的问题，并深入了解该病症的病理生理机制。设计在这项观察性横断面研究中，使用超高效液相色谱-四极杆飞行时间质谱（UPLC-QTOF MS）分析了33名特发性口干症患者和34名健康对照者的唾液样本。代谢组分析辅以多元统计分析，以区分患者和对照组。共有 195 种代谢物显示出显著的差异表达，每种代谢物在预测中的可变重要性（VIP）均大于 1 且 P 值小于 0.05。根据《京都基因和基因组百科全书》（KEGG）进行的通路富集分析发现，有 22 种代谢物参与了 18 条不同的代谢通路。结论唾液代谢组学分析揭示了特发性口腔干燥症的明显改变，其中咖啡因代谢途径显著减少，突显了潜在的神经病理性参与。这项研究加深了我们对口腔干燥症代谢改变的了解，表明唾液代谢组学可为诊断和了解特发性口腔干燥症的病因提供可行的生物标记物。未来的研究应侧重于针对这些代谢紊乱的治疗，并评估其治疗后的可逆性。

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引用次数: 0

Effects of preservatives on the activities of salivary enzymes 防腐剂对唾液酶活性的影响。

IF 2.2 4区医学 Q2 DENTISTRY, ORAL SURGERY & MEDICINE

Archives of oral biology

Pub Date : 2024-10-01 DOI: 10.1016/j.archoralbio.2024.106098

Moon-Jong Kim , Ji-Youn Chang , Yoon-Young Kim , Jae Wook Lee , Hong-Seop Kho

Objectives

To investigate the effects of common preservatives used in oral health care products on the enzymatic activities of lysozyme, peroxidase, and α-amylase in-solution and on-hydroxyapatite surface phases.

Design

The preservatives used in this study were sodium benzoate, methylparaben, propylparaben, and benzalkonium chloride. Hen egg-white lysozyme, bovine lactoperoxidase, and α-amylase from Bacillus sp. served as sources of purified enzymes. Human unstimulated whole saliva was used as a source of salivary enzymes. Hydroxyapatite beads were used as the surface phase. The preservatives were incubated with purified enzymes or saliva samples in-solution or on-hydroxyapatite surface phases, respectively. Enzymatic activities of lysozyme, peroxidase, and α-amylase were measured by hydrolysis of fluorescein-labelled Micrococcus lysodeikticus, oxidation of fluorogenic 2′,7′-dichlorofluorescin, and hydrolysis of fluorogenic starch, respectively.

Results

The effects of the preservatives on the enzymatic activities of lysozyme and peroxidase were more distinct in the saliva samples than purified substances, and in the in-solution phase than on-hydroxyapatite surface phase, and the opposite was true for α-amylase. The most significant result was apparent decrease in peroxidase activities caused by the parabens in the in-solution phase (P<0.05). Sodium benzoate and parabens inhibited lysozyme activity in the in-solution phase, but differently for the purified and salivary lysozymes. Parabens and benzalkonium chloride inhibited the enzymatic activity of α-amylase from Bacillus sp., not saliva samples, only on-hydroxyapatite surface (P<0.05).

Conclusions

Each preservative affected the enzymatic activities of lysozyme, peroxidase, and α-amylase differently. Based on the effects on salivary enzymes, sodium benzoate or benzalkonium chloride was recommended as preservatives rather than parabens.

研究目的研究口腔保健产品中常用的防腐剂对溶液和羟基磷灰石表面溶液中溶菌酶、过氧化物酶和α-淀粉酶酶活性的影响：本研究使用的防腐剂为苯甲酸钠、苯甲酸甲酯、苯甲酸丙酯和苯扎氯铵。母鸡蛋清溶菌酶、牛乳过氧化物酶和芽孢杆菌α-淀粉酶是纯化酶的来源。人类未经刺激的全唾液被用作唾液酶的来源。羟基磷灰石珠被用作表面相。防腐剂分别与纯化酶或唾液样品在溶液中或羟基磷灰石表面相上进行孵育。溶菌酶、过氧化物酶和α-淀粉酶的酶活性分别通过荧光素标记的溶菌水解、荧光素 2',7'-二氯荧光素的氧化和荧光素淀粉的水解进行测定：防腐剂对溶菌酶和过氧化物酶活性的影响在唾液样本中比在纯化物质中更明显，在溶液相中比在羟基磷灰石表面相中更明显，而对α-淀粉酶的影响则相反。最重要的结果是对羟基苯甲酸酯类物质在溶液相中导致过氧化物酶活性明显降低（PConclusions：每种防腐剂对溶菌酶、过氧化物酶和α-淀粉酶的酶活性都有不同的影响。根据对唾液酶的影响，建议使用苯甲酸钠或苯扎氯铵而不是对羟基苯甲酸酯类防腐剂。

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引用次数: 0

Impact of temporomandibular disorder on food preference behavior in adult male rats 颞下颌关节紊乱对成年雄性大鼠食物偏好行为的影响

IF 2.2 4区医学 Q2 DENTISTRY, ORAL SURGERY & MEDICINE

Archives of oral biology

Pub Date : 2024-09-28 DOI: 10.1016/j.archoralbio.2024.106100

Saeid Ghorbanizadeh , Mehdi Abbasnejad , Mahnaz Zamyad , Saeed Esmaeili-Mahani , Reza Kheirandish , Mojdeh Mansoori

Objective

Temporomandibular disorders (TMD) are a group of conditions affecting the jaw, surrounding muscles and associated structures. Researches indicate that TMD has implications for changes in nutritional behavior. This study investigates the impact of TMD on the food preference behavior of rats, examining various food models with differing caloric content, taste, and texture.

Design

Forty-two male Wistar rats (200–250 g) were divided into six groups, comprising three control and three pain or TMD groups. Induction of TMD by injection of complete Freund adjuvant (CFA) into the left temporomandibular joint (TMJ) of rat serves as a model for studying TMD. Twenty-four hours post-TMD induction, the preference of animals to foods with differences in calories, taste and texture was evaluated with the help of an automatic preference measuring device.

Results

In terms of caloric preferences, the pain group exhibited a distinct shift towards high-calorie food. Taste preferences were marked by an increased preference for sweet food in the pain group. Texture preferences were altered, with the pain group displaying a preference for low-texture food.

Conclusion

TMD in rats leads to increased preferences for high-calorie and sweet foods, as well as altered preferences for food textures. These findings highlight the influence of TMJ pain on food preference behavior in rats.

目的颞下颌关节紊乱症（TMD）是一组影响颌骨、周围肌肉和相关结构的疾病。研究表明，TMD 会影响营养行为的变化。本研究调查了 TMD 对大鼠食物偏好行为的影响，研究了各种不同热量、口味和质地的食物模型。设计将 42 只雄性 Wistar 大鼠（200-250 克）分为 6 组，其中包括 3 个对照组和 3 个疼痛或 TMD 组。在大鼠左侧颞下颌关节（TMJ）注射全氟佐剂（CFA）诱导 TMD，作为研究 TMD 的模型。在诱导 TMD 24 小时后，利用自动偏好测量仪评估了动物对热量、味道和质地不同的食物的偏好。口味偏好方面，疼痛组对甜食的偏好明显增加。结论颞下颌关节紊乱导致大鼠对高热量和甜味食物的偏好增加，并改变了对食物质地的偏好。这些发现突显了颞下颌关节疼痛对大鼠食物偏好行为的影响。

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引用次数: 0

Effects of chronic unpredictable mild stress-induced depression on bitter taste receptor expression in mice 慢性不可预测的轻度应激诱导抑郁对小鼠苦味受体表达的影响

IF 2.2 4区医学 Q2 DENTISTRY, ORAL SURGERY & MEDICINE

Archives of oral biology

Pub Date : 2024-09-27 DOI: 10.1016/j.archoralbio.2024.106099

Zhaoxiang Wang , Min Zhang , Meng Huang , Luyue Zhang , Gencheng Han , Ge Li , Junkai Cao

Objective

With the rapid increase in the pace of life, people are facing increasing pressures of all kinds, and depression has gradually become a serious psychological disorder in human society, strongly affecting normal social and physiological activities. Depression can disrupt an individual's taste perception and potentially result in taste disorders by affecting and altering taste receptors. This disruption can consequently impact their food preferences and overall eating experiences.

Design

In this study, we used the chronic unpredictable mild stress (CUMS) method to establish a depression model in male C57BL/6 J mice and explored the changes in taste receptor expression in the lingual circumvallate papillae (CP) to elucidate the effects of depression on taste. After 6 weeks of CUMS, behavioral performance evaluations, such as forced swim, open field, and elevated plus maze tests, were conducted in depression model mice. A further two-bottle choice test was subsequently performed to determine the effect of depression on bitter taste, and the expression of bitter taste receptors in the lingual CP was detected via immunofluorescence staining.

Results

In this study, we found for the first time that mice with CUMS-induced depression had decreased bitter taste sensitivity through a two-bottle choice test and demonstrated that the expression of T2r5, a receptor related to bitter taste perception, and the expression of secondary taste signaling proteins in the lingual CP were significantly decreased in mice exposed to CUMS, as determined via qRTPCR and immunofluorescence staining.

Conclusions

Our study highlights how CUMS influences the perception of bitterness in the peripheral taste system, potentially elucidating stress-induced changes in eating habits.

目的随着生活节奏的加快，人们面临的各种压力越来越大，抑郁症逐渐成为人类社会的一种严重心理障碍，强烈影响着人们正常的社交和生理活动。抑郁症会影响和改变味觉感受器，从而扰乱个体的味觉感知，并可能导致味觉失调。本研究采用慢性不可预知轻度应激（CUMS）法在雄性C57BL/6 J小鼠中建立抑郁模型，并探讨舌周乳头（CP）味觉受体表达的变化，以阐明抑郁对味觉的影响。服用 CUMS 6 周后，对抑郁模型小鼠进行了行为表现评估，如强迫游泳、开阔地和高架加迷宫测试。随后再进行双瓶选择测试，以确定抑郁对苦味的影响，并通过免疫荧光染色检测舌CP中苦味受体的表达。结果在这项研究中，我们首次发现由 CUMS 引起的抑郁小鼠通过双瓶选择试验降低了对苦味的敏感性，并通过 qRTPCR 和免疫荧光染色检测证明，与苦味感知相关的受体 T2r5 的表达以及舌 CP 中次级味觉信号蛋白的表达在暴露于 CUMS 的小鼠中显著下降。结论我们的研究强调了 CUMS 如何影响外周味觉系统对苦味的感知，从而有可能阐明压力引起的饮食习惯变化。

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引用次数: 0

An intronic variant in LAMB3 contributes to junctional epidermolysis bullosa and enamel hypoplasia via translational attenuation LAMB3的内含子变异通过翻译衰减导致交界性表皮松解症和釉质发育不全。

IF 2.2 4区医学 Q2 DENTISTRY, ORAL SURGERY & MEDICINE

Archives of oral biology

Pub Date : 2024-09-27 DOI: 10.1016/j.archoralbio.2024.106101

Yi Yang , Yao Wang , Man Qin , Yuming Zhao , Cristina Has , Xin Wang

Objectives

This study aimed to investigate the genetic etiology of a family affected by junctional epidermolysis bullosa (JEB) and generalized enamel hypoplasia, and to explore how an intronic variant influenced the 5’ untranslated region (5’UTR), thereby affecting LAMB3 expression and contributing to the pathogenesis of the disease.

Design

Whole-exome and whole-genome sequencing were used to screen for genetic defects in the patient. Mutational consequences were characterized through luciferase assays, splice assay, in silico analyses, and verification using the patient's gingival sample.

Results

A nonsense variant (c.2983 C>T; p.Gln995*) and an intronic variant (c.–38+2 T>C) of LAMB3 were identified. In vitro assays demonstrated that the intronic variant activated a cryptic splice site, resulting in a 120 bp intronic inclusion. This splicing alteration significantly reduced the translation efficiency of the downstream coding sequence, while overall mRNA expression remained unaffected. Bioinformatic analysis unveiled the creation of three upstream AUG codons, leading to the presence of two upstream open reading frames (uORFs) and one overlapping ORF. The longer uORF's AUG exhibited a moderate Kozak strength similar to that of the main ORF's AUG. Structural analysis of the mutant 5’UTR sequence revealed a more complex secondary structure, characterized by a large branch loop and a stem-loop preceding the coding sequence's start codon.

Conclusion

This study suggests that variants affecting the 5’UTR may contribute to the genetic etiology of JEB. These findings could help enhance the diagnostic accuracy and efficiency in JEB patients.

研究目的本研究旨在调查一个家族交界性表皮松解症（JEB）和全身釉质发育不全的遗传病因，并探讨一个内含子变异如何影响5'非翻译区（5'UTR），从而影响LAMB3的表达并导致该病的发病机制：设计：利用全外显子组和全基因组测序筛选患者的基因缺陷。通过荧光素酶测定、剪接测定、硅学分析以及使用患者牙龈样本进行验证，确定了突变后果的特征：结果：发现了LAMB3的一个无义变体（c.2983 C>T；p.Gln995*）和一个内含子变体（c.-38+2 T>C）。体外试验表明，内含子变异体激活了一个隐性剪接位点，导致一个 120 bp 的内含子。这种剪接改变大大降低了下游编码序列的翻译效率，而整个 mRNA 的表达不受影响。生物信息学分析揭示了三个上游 AUG 密码子的产生，导致了两个上游开放阅读框（uORF）和一个重叠 ORF 的存在。较长的 uORF 的 AUG 表现出中等的科扎克强度，与主 ORF 的 AUG 相似。对突变体 5'UTR 序列的结构分析表明，其二级结构更为复杂，编码序列起始密码子之前有一个大的分支环和一个茎环：本研究表明，影响5'UTR的变异可能是JEB的遗传病因之一。这些发现有助于提高 JEB 患者的诊断准确性和效率。

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引用次数: 0

Roles of the histone methyltransferase SET domain bifurcated 1 in epithelial cells during tooth development 组蛋白甲基转移酶 SET 域分叉 1 在牙齿发育过程中上皮细胞中的作用

IF 2.2 4区医学 Q2 DENTISTRY, ORAL SURGERY & MEDICINE

Archives of oral biology

Pub Date : 2024-09-26 DOI: 10.1016/j.archoralbio.2024.106094

Yuri Takagiwa, Norihisa Higashihori, Sakurako Kano, Keiji Moriyama

Objective

This study aimed to reveal the effects of SET domain bifurcated 1 (SETDB1) on epithelial cells during tooth development.

Design

We generated conditional knockout mice (Keratin 14-Cre;Setdb1^f/f mice), in which Setdb1 was deleted only in epithelial cells. At embryonic day 14.5 (E14.5), immunofluorescence staining was performed to confirm the absence of SETDB1 within the epithelium of tooth embryos from Keratin 14-Cre;Setdb1^f/f mice. Mouse embryos were harvested after reaching embryonic day 13.5 (E13.5), and sections were prepared for histological analysis. To observe tooth morphology in detail, electron microscopy and micro-CT analysis were performed at postnatal months 1 (P1M) and 6 (P6M). Tooth embryos were harvested from postnatal day 7 (P7) mice, and the epithelial components of the tooth embryos were isolated and examined using quantitative RT-PCR for the expression of genes involved in tooth development.

Results

Keratin 14-Cre;Setdb1^f/f mice exhibited enamel hypoplasia, brittle and fragile dentition, and significant abrasion. Coronal sections displayed abnormal ameloblast development, including immature polarization, and a thin enamel layer that detached from the dentinoenamel junction at P7. Electron microscopic analysis revealed characteristic findings such as an uneven surface and the absence of an enamel prism. The expression of Msx2, Amelogenin (Amelx), Ameloblastin (Ambn), and Enamelin (Enam) was significantly downregulated in the epithelial components of tooth germs in Keratin 14-Cre;Setdb1^f/f mice.

Conclusions

These results indicate that SETDB1 in epithelial cells is important for tooth development and clarify the relationship between the epigenetic regulation of SETDB1 and amelogenesis imperfecta for the first time.

目的本研究旨在揭示 SET domain bifurcated 1（SETDB1）在牙齿发育过程中对上皮细胞的影响。设计我们产生了条件性基因敲除小鼠（Keratin 14-Cre;Setdb1f/f mice），其中 Setdb1 仅在上皮细胞中被删除。在胚胎第 14.5 天（E14.5），进行免疫荧光染色以确认 Keratin 14-Cre;Setdb1f/f 小鼠牙胚上皮细胞中 SETDB1 的缺失。小鼠胚胎在发育到胚胎第 13.5 天（E13.5）后收获，并制备切片进行组织学分析。为了详细观察牙齿形态，在出生后第 1 个月（P1M）和第 6 个月（P6M）进行了电子显微镜和显微 CT 分析。结果角蛋白 14-Cre;Setdb1f/f 小鼠表现出釉质发育不全、牙齿脆而脆弱以及明显的磨损。冠状切片显示釉母细胞发育异常，包括极化不成熟，釉质层很薄，在P7时从牙本质釉质交界处脱落。电子显微镜分析显示了一些特征性结果，如表面不平整和缺乏釉质棱镜。在角蛋白14-Cre;Setdb1f/f小鼠的牙胚上皮成分中，Msx2、釉质素（Amelogenin，Amelx）、釉母细胞素（Ameloblastin，Ambn）和釉质素（Enam）的表达显著下调。

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引用次数: 0

Modulation of regenerative responses by retinoic and ascorbic acids in human apical papilla cells 视黄酸和抗坏血酸对人类顶端乳头细胞再生反应的调节作用。

IF 2.2 4区医学 Q2 DENTISTRY, ORAL SURGERY & MEDICINE

Archives of oral biology

Pub Date : 2024-09-24 DOI: 10.1016/j.archoralbio.2024.106095

Lídia de Oliveira Fernandes , Igor Paulino Mendes Soares , Rafael Antonio de Oliveira Ribeiro , Victória Peruchi , Maria Luiza Barucci Araujo Pires , Caroline Anselmi , Maria Luisa Leite , Carlos Alberto de Souza Costa , Josimeri Hebling

Objective

This study investigated the bioactive effects of retinoic acid and ascorbic acid on hSCAPs in vitro.

Design

Cells were obtained from human third molars (n=4) and characterized for mesenchymal stem cell markers by flow cytometry. The experimental groups: control (α-MEM); vehicle control group (α-MEM + 0.17 % DMSO); retinoic acid 0.1, 1, and 10 µM; and ascorbic acid 3, 30, and 300 µM (n=8) were tested for cell viability (alamarBlue; 1, 3, and 7 days), total collagen synthesis (Sirius Red; 1 and 7 days), mineralized matrix formation (Alizarin red; 14 days), and the regulation of gene expression related to mineralization (ALPL and DSPP), cell migration (ITGAV and CXCL12) angiogenesis (VEGFA) and collagen synthesis (COL1A1 and COL3A1; RT-qPCR) on 1 and 7 days. ACTB and GAPDH were used as reference genes. Data were analyzed by ANOVA and complementary tests at a 5 % significance level.

Results

Ascorbic acid 300 µM increased viability, and retinoic acid reduced it dose-dependently. Retinoic acid 0.1 µM and ascorbic acid 30 and 300 µM increased mineralized matrix formation and total collagen synthesis, and retinoic acid 10 µM decreased. On day 1, 0.1 µM retinoic acid upregulated the gene expression of COL1A1, COL3A1, VEGFA, CXCL12, ALPL, DSPP e ITGAV, and 300 µM ascorbic acid upregulated COL1A1, COL3A1 and DSPP. However, on day 7, retinoic acid downregulated ALPL, COL3A1, CXCL12, and VEGFA and downregulated ITGAV and VEGFA.

Conclusion

Retinoic acid 0.1 µM and ascorbic acid 300 µM biostimulated hSCAPs to differentiate into pro-regenerative phenotypes with potential application for REPs.

研究目的本研究探讨了维甲酸和抗坏血酸对体外 hSCAPs 的生物活性作用：细胞取自人类第三磨牙（n=4），通过流式细胞术鉴定间充质干细胞标记。实验组：对照组（α-MEM）；载体对照组（α-MEM + 0.17 % DMSO）；视黄酸 0.1、1 和 10 µM；抗坏血酸 3、30 和 300 µM（n=8），检测细胞存活率（茜草蓝；1、3 和 7 天）、总胶原合成（天狼星红；1 和 7 天）、矿化基质形成（茜素红；14 天），以及 1 天和 7 天内与矿化（ALPL 和 DSPP）、细胞迁移（ITGAV 和 CXCL12）、血管生成（VEGFA）和胶原合成（COL1A1 和 COL3A1；RT-qPCR）相关的基因表达调控。参考基因为 ACTB 和 GAPDH。数据采用方差分析和显著性水平为 5% 的互补检验进行分析：结果：抗坏血酸 300 µM 可提高存活率，而维甲酸可降低存活率，两者呈剂量依赖性。维甲酸 0.1 µM、抗坏血酸 30 µM 和 300 µM 增加了矿化基质的形成和胶原蛋白的总合成，而维甲酸 10 µM 则减少了矿化基质的形成和胶原蛋白的总合成。第1天，0.1 µM维甲酸上调COL1A1、COL3A1、VEGFA、CXCL12、ALPL、DSPP e ITGAV的基因表达，300 µM抗坏血酸上调COL1A1、COL3A1和DSPP的基因表达。然而，在第 7 天，维甲酸下调 ALPL、COL3A1、CXCL12 和 VEGFA，下调 ITGAV 和 VEGFA：视黄酸 0.1 µM 和抗坏血酸 300 µM 可生物刺激 hSCAPs 分化为促再生表型，有望应用于 REPs。

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引用次数: 0

Influence of bone morphogenetic protein (BMP) signaling and masticatory load on morphological alterations of the mouse mandible during postnatal development 骨形态发生蛋白（BMP）信号传导和咀嚼负荷对小鼠出生后发育过程中下颌骨形态改变的影响

IF 2.2 4区医学 Q2 DENTISTRY, ORAL SURGERY & MEDICINE

Archives of oral biology

Pub Date : 2024-09-24 DOI: 10.1016/j.archoralbio.2024.106096

Amber Uptegrove , Coral Chen , Madison Sahagun-Bisson , Anshul K. Kulkarni , Ke’ale W. Louie , Hiroki Ueharu , Yuji Mishina , Maiko Omi-Sugihara

Objective

Bone homeostasis relies on several contributing factors, encompassing growth factors and mechanical stimuli. While bone morphogenetic protein (BMP) signaling is acknowledged for its essential role in skeletal development, its specific impact on mandibular morphogenesis remains unexplored. Here, we investigated the involvement of BMP signaling and mechanical loading through mastication in postnatal mandibular morphogenesis.

Design

We employed conditional deletion of Bmpr1a in osteoblasts and chondrocytes via Osterix-Cre. Cre activity was induced at birth for the 3-week group and at three weeks for the 9-week and 12-week groups, respectively. The conditional knockout (cKO) and control mice were given either a regular diet (hard diet, HD) or a powdered diet (soft diet, SD) from 3 weeks until sample collection, followed by micro-CT and histological analysis.

Results

The cKO mice exhibited shorter anterior lengths and a posteriorly inclined ramus across all age groups compared to the control mice. The cKO mice displayed an enlarged hypertrophic cartilage area along with fewer osteoclast numbers in the subchondral bone of the condyle compared to the control group at three weeks, followed by a reduction in the cartilage area in the posterior region at twelve weeks. Superimposed imaging and histomorphometrical analysis of the condyle revealed that BMP signaling primarily affects the posterior part of the condyle, while mastication affects the anterior part.

Conclusions

Using 3D landmark-based geometric morphometrics and histological assessments of the mandible, we demonstrated that BMP signaling and mechanical loading reciprocally contribute to the morphological alterations of the mandible and condyle during postnatal development.

目的骨骼的稳态依赖于多种因素，包括生长因子和机械刺激。尽管骨形态发生蛋白（BMP）信号在骨骼发育中的重要作用已得到公认，但其对下颌骨形态发生的具体影响仍未得到研究。设计我们通过 Osterix-Cre 有条件地缺失成骨细胞和软骨细胞中的 Bmpr1a。3周组在出生时诱导Cre活性，9周组和12周组分别在3周时诱导Cre活性。条件性基因敲除（cKO）小鼠和对照组小鼠从 3 周开始食用普通食物（硬质食物，HD）或粉状食物（软质食物，SD），直到样本采集完毕，然后进行显微 CT 和组织学分析。与对照组相比，cKO 小鼠在三周时髁状突软骨下的肥厚软骨面积增大，破骨细胞数量减少，十二周时后部软骨面积缩小。结论通过对下颌骨进行基于三维地标的几何形态计量学和组织学评估，我们证明了BMP信号传导和机械负荷对下颌骨和髁状突在出生后发育过程中的形态改变具有互为因果的作用。

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引用次数: 0