东亚中更新世古人类下颔第二臼齿釉质-齿质连接面三维形状和釉质厚度分布

东亚中更新世古人类在头骨、下颌骨、牙齿等解剖部位表现出不同程度的形态多样性,中期成员代表为直立人,而晚期成员的演化地位具有较大争议。为进一步了解东亚中更新世古人类内部的形态变异特点和为东亚中更新世晚期古人类分类提供依据,本文使用微分同胚的表面匹配（Diffeomorphic Surface Matching, DSM）和形态测量图（Morphometric map）对下颌第二臼齿（M₂）釉质-齿质连接面的形状和齿冠侧面釉质厚度分布模式进行了量化分析。结果显示：1）东亚中更新世古人类与晚期人属成员（尼安德特人和现代人）存在较明显的形态差别;2）该时段晚期的东亚古人类相对中期直立人在侧面釉质厚度分布规律上具有独特性,并在釉质-齿质连接面的三维形状上与晚期人属成员更加接近。本文在以往对东亚中更新世古人类牙齿内外结构单个性状研究的基础上,使用三维形态测量方法进一步量化了M₂釉质-齿质连接面三维形状和侧面釉质厚度分布模式两项重要特征的变异特点,这对未来该时段同类型牙齿的形态鉴定以及解决东亚中更新世晚期古人类的分类地位具有一定意义。     

A three-phase liquid chromatographic method for δC analysis of amino acids from biological protein hydrolysates using liquid chromatography-isotope ratio mass spectrometry

We report a three-phase chromatographic method for the separation and analysis of δ¹³C values of underivatized amino acids from biological proteins (keratin, collagen, and casein) using liquid chromatography-isotope ratio mass spectrometry (LC-IRMS). Both precision and accuracy of δ¹³C values for standard amino acid mixtures over the range of approximately 8 to 1320 ng of carbon per amino acid on the column were assessed. The precision of δ¹³C values of amino acids was found to be better at higher concentrations, whereas accuracy improved at lower concentrations. The optimal performance for this method was achieved with between 80 and 660 ng of carbon of each amino acid on the column. At amino acid amounts lower than 20 ng of carbon on the column, precision and accuracy may become compromised. The application of this new three-phase chromatographic technique will allow the analysis of δ¹³C of amino acids to be carried out as a routine method and benefit fields of research such as biomedicine, forensics, ecology, nutrition, and palaeodiet reconstruction in archaeology.     

Preliminary observations on increasing root length during the eruptive phase of tooth development in modern humans and great apes

Ground sections of incisors, canines, and molars were selected that showed clear incremental markings in root dentine. The sample comprised 98 Homo sapiens, 53 Pan troglodytes, and a more limited combined sample of 51 Gorilla and Pongo sections. Daily rates of root dentine formation, together with the orientation of incremental markings in roots close to the cement-dentine junction (CDJ), were used to calculate root extension rates for the first 10mm of root formed beyond the buccal enamel cervix. Modern human anterior tooth roots showed a more regular pattern of increase in root length than those in great apes. In Pan, root growth rose quickly to higher rates but then flattened off. The fastest extension rates in modern humans were in incisor roots (10-12 microm per day), followed by canines (8-9 microm per day). Extension rates in Pan rose to slightly greater values in canines ( approximately 12-14 microm per day) than in incisors ( approximately 10-11 microm per day). Molar tooth roots in both modern humans and great apes grew in a nonlinear manner. Peak rates in molars reduced from M1 to M3 (8, 7, and 6 microm per day, respectively). Like humans, root growth in Pan peaked earlier in M1s at rates of between 8 and 9 microm per day, and later in M3s at rates of 7 to 8 microm per day. The more limited data set for Gorilla and Pongo molars suggests that extension rates were generally higher than in Pan by approximately 1.0-1.5 microm per day. There were greater differences in peak extension rates, with Gorilla and Pongo extension rates being between 2.5 and 4.5 microm per day higher than those in Pan. These findings highlight for the first time that root growth rates differ between tooth types in both pattern and rate and between taxa. They provide the basis with which to explore further the potential comparative relationships between root growth, jaw growth, and the eruption process.     

Ultrastructure of developing tooth plates in the Australian lungfish, Neoceratodus forsteri (Osteichthyes: Dipnoi)

While the lungfish dentition is partially understood as far as morphology and light microscopic structure is concerned, the ultrastructure is not. Each tooth plate is associated with a dental lamina that develops from the inner layer of endodermal cells that form the oral epithelium. Dentines, bone and cartilage of the jaws differentiate from mesenchyme cells aggregating beneath the oral endothelium. Enamel, in the developing and in the mature form, has similarities to that of other early vertebrates, but unusual characters appear as development proceeds. Ameloblasts are capable of secreting enamel, and, with mononuclear osteoclasts, of remodelling the bone below the tooth plate. The forms of dentine, all based largely on an extracellular matrix of collagen and mineralised with biological apatite, differ from each other and from the underlying bone in the ultrastructure of associated cells and in the mineralised extracellular matrices produced. Cell processes emerging from the odontoblasts and from the osteoblasts vary in length, degree of branching and of anastomoses between the processes, although all of the cell types have large amounts of rough endoplasmic reticulum. Mineralisation of the extracellular matrices varies among the enamel, dentines and bone in the tooth plate. In addition, the development of the hard tissues of the tooth plates indicates that many of the similarities in fine structure of the dentition in lungfish, to tissues in other fish and amphibia, apparent early in development, disappear as the dentition matures.     

Odontoblasts induced from mesenchymal cells of murine dental papillae in three-dimensional cell culture

In an organ culture system under a three-dimensional microenvironment that provides the conditions needed for odontoblast differentiation, a row of odontoblasts can be induced (Kikuchi et al. 1996, 2001). Therefore, in a newly designed three-dimensional cell culture system that fulfils the conditions necessary for odontoblast differentiation (Kikuchi et al. 2002), we examined whether dental papilla cells in rat mandibular incisors could differentiate into tubular dentine-forming cells. In our previously established organ culture system, CM-Dil-labeled cells that were microinjected into isolated dental papillae were replaced by a row of odontoblasts. In a three-dimensional cell culture system, which consists of two kinds of type I collagen in the upper layer over multi-layered cells seeded onto collagen containing Matrigel in the lower layer and which acts as a structural meshwork, dental papilla cells were incubated as multi-layered cells in an artificial extracellular matrix (ECM). The cells aggregated to form a cell mass and invaginated as a cell mass into the ECM. The cells also extended fine fibrillar processes into the ECM. With regard to invagination, the proteolytic activities of matrix metalloproteinase-2 (MMP-2)/membrane type 1-matrix metalloproteinase (MT 1-MMP) were observed on the outer multi-layers of cells within a cell mass adjacent to the ECM. The cell mass progressively shrank to about one-half to one-third of its original diameter and was organized as a tissue surrounded by a newly secreted ECM, like dental pulp-dentine. The cells adjacent to the secreted ECM were constructed as a row of polarized columnar cells. They extended slender processes into the new ECM, which is characteristic of tubular matrix. Dentine sialophosphoprotein (DSPP) and dentine matrix protein 1 (DMP 1) genes, which are specific for odontoblast differentiation, were expressed in an aggregated cell mass where tubular matrix-forming cells were induced. Furthermore, the tubular matrix became mineralized under prolonged culture. These results imply that the putative progenitor cells/stem cells residing in dental papillae can differentiate into odontoblasts under appropriate conditions in vitro.     

How the microstructure of dentine can contribute to reconstructing developing dentitions and the lives of hominoids and hominins

Accounts of dentine microstructure are less well established in the primate life history literature than those of enamel microstructure. The aim of this paper is to draw some basic comparisons between enamel and dentine, but at the same time to show how dentine microstructure can make a major but different contribution to reconstructing past lives than enamel can. Dentine has both an organic and an inorganic component. The organic component contains growth factors, stable isotopes and DNA that survive long after death. The mineral component contains trace elements and preserves an incremental record of tooth growth. These can be used to put a time scale to many past events when the chemistry or microstructure of dentine has become altered during tooth growth. Dentine microstructure allows us to reconstruct tooth root growth in the past and has contributed to a fuller understanding of the modular nature of developing dentitions among hominoids and hominins.     

Untersuchungen zur Mineralisierungsdichte im Hartgewebe mit Protein-Polysaccharid bzw. mit Kollagen als Hauptbestandteil der Matrix

Zusammenfassung Ultradünnschnitte von schmelznahem Dentin sowie von Osteopetroseknochen-Partien zeigten im Elektronenmikroskop, daß die proteinpolysaccharid-reichen Zonen des peritubulären Dentins sowie der sog. dichten Zonen des Osteopetrose-Knochens punktartige Calzium-Phosphat-Keime bzw. Ketten von Punktkeimen in dichter Zusammenlagerung besitzen, während in den kollagenreichen Zonen neben diesen Ketten und Nadeln größere Blättchen-Kristallite auftreten. Durch Entmineralisierung-Kontrastierung der Ultradünnschnitte mit Phosphorwolframsäure wurde gezeigt, daß im peritubulären Dentin und den dichten Partien des Osteopetroseknochens elektronenmikroskopisch keine Kollagenfasern mit Querstreifen vorkommen; diese sind aber im intertubulären Dentin und den normalen Partien des Osteopetroseknochens verbreitet. Durch Registrierung der Calzium- und Phosphor-Röntgenimpulse mit der elektronenmikroskopischen Mikrosonde (Cameca) konnte — bei Annahme angenähert gleicher Schnitt-Dicke in benachbarten Zonen — gefunden werden, daß der Calzium-Gehalt pro Volumeneinheit im Mittel von 100 Einzelmessungen im peritubulären Dentin um etwa 40 % höher liegt als im kollagenreichen intertubulären Dentin und in den proteinpolysaccharid-reichen Zonen der Osteopetrose-Proben um 65 % höher als in den normalen kollagenreichen Bezirken. Da entsprechende Werte für Phosphor gefunden wurden, bedeuten diese Prozentangaben zugleich dieselben Unterschiede in der Mineralisierungsdichte.

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Investigations of mineralization in hard tissues with protein-polysaccharides or collagen as main constituents of the matrix

Summary Ultrathin sections of mature dentine in the vicinity of enamel and sections of osteopetrotic bone were examined. In the protein-polysaccharide-rich regions of the peritubular dentine, as well as in the so-called dense zones of the osteopetrotic bone, dot-like nuclei, or short chains of them, in close packing were found, whereas in the intertubular collagen-rich dentine zones, as well as the normal regions of osteopetrotic bone, many platelike crystals were found among the chains of dots. By demineralization-staining experiments with phosphotungstic acid, it was found that in the peritubular dentine as well as the so-called dense zones of the osteopetrotic bone there were no collagen fibres visible, while they dominated in the intertubular dentine as well as the normal regions of the osteopetrotic bone. The calcium and phosphorous concentrations in the sections were examined in the Cameca electronmicroscopical probe. Assuming that neighbouring regions were of nearly equal thickness, the mean calcium content per unit volume from 100 single point measurements for each region was found to be about 40 % higher in the peritubular region than in the collagen-rich intertubular region, and about 65 % higher in the so-called dense regions of the osteopetrose bone than in the normal collagen-rich zones. The same was found for phosphorous.

Wir danken Frl. Ch. Lippert, Frl. I. Schütte und Herrn J. Schreiber für wertvolle Hilfe bei der technischen Durchführung der Untersuchungen. Der Deutschen Forschungsgemeinschaft danken wir für die Unterstützung unserer Arbeit.     

Investigation of the early mineralisation on collagen in dentine of rat incisors by quantitative electron spectroscopic diffraction (ESD)

The earliest crystallites in dentine appear as chains of dots in ultra-thin sections viewed by transmission electron microscopy. These dots rapidly coalesce along the longitudinal directions of the collagen microfibrils to form needle-like structures that coalesce preferentially in lateral directions to form ribbon-like or plate-like crystallites. This morphological interpretation is supported by line-scans of the corresponding zero-loss filtered electron spectroscopic diffraction patterns, which demonstrate the crystalline structure of the dentine mineral (apatite). The intensity ratio of the Debye-Scherrer rings of the characteristic Bragg-reflections (002 to 300, together with 1 or 2 unresolved reflections) shows a maximum in the region of early chain-like and needle-like crystallites, decreasing with maturation of the dentine mineral to the ribbon-plate-like crystallites. Detailed investigations using line-scans of the zero-loss filtered electron spectroscopic diffraction patterns through the dentine zone show that the intensity ratio found near the mineralisation front is repeated 3–5 times at distances of about 10–20 m. This may represent a circadian pattern of mineralisation corresponding to light microscopically visible incremental lines in dentine.     

鳜牙齿形态结构及与不同种属间比较观察

牙齿是肉食性鱼类重要的摄食器官。为探究鳜(Siniperca chuatsi)牙齿形态结构,采用解剖镜观察了鳜牙齿分布、形态与数量。并比较其与大眼鳜(S. kneri)、斑鳜(S. scherzeri)及中国少鳞鳜(Coreoperca whiteheadi)牙齿差异。采用茜素红染色、组织切片、扫描电镜、X射线能谱及红外光谱观察并检测了鳜牙齿结构、元素组成和化学成分。结果显示,鳜牙齿具上颌齿、下颌齿、犁齿、腭齿和咽齿。其中,上颌齿前端有犬齿,其余为绒毛状齿;下颌齿最内列有犬齿,其余为绒毛状齿;犁齿、腭齿、咽齿均为绒毛状齿。鳜和大眼鳜下颌齿列数为3列,斑鳜和中国少鳞鳜为4列。茜素红染色显示,犬齿和绒毛状齿外层均为透明的牙釉质,内层为包裹髓腔的牙本质;组织切片显示,牙本质结构疏松,内有许多孔隙;牙尖部分形成牙釉质帽;中央为髓腔,内有牙髓组织。扫描电镜显示,犬齿和绒毛状齿表面均由釉柱组成,釉质层有釉柱横纹。X射线能谱显示,犬齿和绒毛状齿主要元素都为碳(C)、氧(O)、氮(N)、钙(Ca)和磷(P)。红外光谱显示,犬齿和绒毛状齿主要无机成分为碳酸羟基磷灰石。结果表明,鳜牙齿较发达,上、下颌齿有犬齿,犬齿和绒毛状齿结构与组成基本相似。