The objective of this study was to examine the fetal skeletons using both alizarin red stain and micro-computed tomography (CT) images; investigate differences, and to determine if the conclusions of the study were the same regardless of the examination method.
A candidate drug was given orally by gavage to pregnant New Zealand White rabbits on gestation day (GD) 7 to GD 19 (mating = GD 0) at doses of 0 (control), 0.02, 0.5, 5, and 15 mg/kg/day. Maternal toxicity was evident at ≥0.02 mg/kg/day. The 199 fetal skeletons (totaling 50,546 skeletal elements) obtained at cesarean delivery on GD29 were first stained with Alizarin Red S, then imaged by a Siemens Inveon micro-CT scanner. All fetal skeletons were examined by both methods, without knowledge of dose group, and the results were compared.
In total, 33 types of skeletal abnormalities were identified. There was 99.8% concordance of results comparing stain to micro-CT. Ossification of the middle phalanx of the forepaw digit 5 showed the greatest difference between the two methods.
Overall, micro-CT imaging is a realistic, and robust alternative to skeletal staining to examine fetal rabbit skeletons in developmental toxicity studies.

The current case study evaluated the effect of vital pulp therapy on a human dental pulp after a long-term period using micro-computed tomography (MCT) for the first time. In the presented report, the successful outcomes of full pulpotomy using calcium-enriched mixture (CEM) cement on an irreversible pulpitis case were documented clinically/radiographically over 5 years. Due to an unrestorable crown fracture at the 5-year recall, the tooth was extracted and evaluated by MCT; the images showed that CEM pulpotomy allowed the dental pulp to create complete dentinal bridges without pulp canal obliteration (PCO). These MCT results showed that CEM pulpotomy, as a bio-regenerative treatment, caused no negative consequence of PCO or calcific metamorphosis on dental pulp over the long term.

BACKGROUND: Melorheostosis (MEL) is an exceptionally rare sclerosing bone dysplasia with asymmetrically exuberant bone formation and soft tissue lesions in a segmental distribution. We aimed to summarize the clinical characteristics of Chinese MEL patients and identify their pathogenic cause.
METHODS: In total, 10 Chinese MEL patients were recruited, and clinical manifestations and radiological characteristics were recorded. Sanger sequencing of the LEMD3 gene was performed on peripheral blood samples of all patients, while the exome sequencing of matched peripheral blood, melorheostotic bone, and skin lesion samples was conducted on one patient who provided affected bone and skin tissues. Micro-computed tomography (micro-CT) was also used to scan the melorheostotic bone tissue.
RESULTS: We found the average age of the 10 MEL patients was 29.5 years (range 11-40 years), and the major symptoms were bone pain, restricted movement, and bone deformity. The lesions sites were mainly located in femur (8/10), tibia (8/10), fibula (6/10), and foot (7/10), the next was pelvis (4/10), and the last were patella (1/10), hand (1/10) and spine (1/10). Radiological examinations showed a mixture of hyperostosis consisting of classic \"dripping candle wax,\" \"osteoma-like,\" or \"myositis ossificans-like\" patterns in most patients. No germline pathogenic variants in the LEMD3 gene were found in all patients, but a disease-causing somatic variant of MAP2K1 (c.167A > C, p.Gln56Pro) was detected in melorheostotic bone from one patient. Moreover, the micro-CT analysis showed increased porosity in the melorheostotic bone with somatic MAP2K1 variant.
CONCLUSIONS: This is a summary of the clinical characteristics of Chinese MEL patients and we first identify the somatic MAP2K1 variant in Chinese patients. Our findings validate the molecular genetic mechanism of MEL and broaden its phenotype spectrum in the Chinese population.

UNASSIGNED: Mucopolysaccharidosis Type IVA (MPS IVA) or Morquio A Syndrome, is a rare metabolic disorder caused by compromised galactosamine-6 sulfatase (GALNS) encoded by GALNS gene (NM_000512.5), leading to keratin sulfate (KS), and chondroitin-6-sulfate accumulation in various organs. We present a 17-year-old woman with progressive bilateral hip pain and radiographic evidence of spondyloepiphyseal dysplasia.
UNASSIGNED: Diagnosis of MPS IVA was made based on whole-exome sequencing (WES) of blood samples collected from the patient and family members, high urinary glycosaminoglycan excretion, supportive clinical manifestations, radiographic examinations, including whole-body X-rays, cervical MRI, and pelvic CT. The patient underwent bilateral total hip arthroplasties sequentially, at a 1-month interval. Femoral heads were preserved for the micro-CT (μCT) analysis and the osteochondral histology examination.
UNASSIGNED: The patient presented with multiple skeletal deformities, including vertebras and long bone deformities. WES disclosed compound heterozygous variants at exon 11 (c.1156C>T) and exon 12 (c.1288C>G) of the GALNS (NM_000512.5). The μCT analysis revealed significant bone quantity loss and microarchitectural change in both weight-bearing area (WBA) and non-weight-bearing area (NWBA) of the femoral heads, while histological analysis showed structural abnormity of articular cartilage in the WBA of the femoral heads.
UNASSIGNED: We have found compound heterozygous variants of GALNS. This is also the first study to report the microarchitectural and histological changes of both subchondral bone and articular cartilage of the femoral head in a patient with MPS IVA.

This case study reports the advantages of micro-CT to aid the investigative process in a complex dismemberment case. Micro-CT was successfully implemented to scan all skeletal remains of a dismembered female. The digital models were utilized to (i) screen for any further injuries not related to the dismemberment, (ii) provide measurements from false starts non-destructively, and (iii) visually represent the evidence in a structured format in court to improve the understanding of the forensic evidence by the jury. Acquiring high-resolution scans in this manner improved the efficiency of the forensic investigation by screening the remains and provided complementary toolmark evidence to the investigating team and forensic pathologist. A total of 14 false starts were identified along with the directionality of each dismemberment cut. Furthermore, the visual 3D representation of the remains in court provided a powerful tool to communicate this important evidence to the jury and form a prosecution narrative. As a forensic radiological method, micro-CT provided valuable information both in the investigation and the court presentation.

OBJECTIVE: The aim of this study was to present the long-term course of a patient with nevus sebaceous syndrome (NSS). Recent genetic studies place the syndrome in the emerging group of so-called RASopathies. The focus of the report is on surgical treatment and morphological and genetic findings of the face and oral cavity.
METHODS: A female patient was treated for congenital alterations of facial skin and oral mucosa. The oral lesions were removed repeatedly. Eruption of teeth on the lesion sites was made easier by the measures taken. However, after repeated ablation of the affected gingiva, the periodontal papillomatous epithelium re-differentiated into the same reddish, conspicuous, hyperplastic epithelium. The teeth in the affected region showed noticeable changes in position, surface, and shape. A HRAS mutation was detected only in the regions of altered oral epithelia and not in adjacent soft tissues.
CONCLUSIONS: Reports on NSS rarely address oral manifestations. The recorded alterations of oral soft and hard tissues in NSS indicate a topographical relationship between the development of oral mucosa and teeth as well as the long-lasting impact of a sporadic mutation on organ development at this site.

Sea cucumber taxonomy and systematics has in the past heavily relied on gross external and internal anatomy, ossicle assemblage in different tissues, and molecular characterisation, with coloration, habitat, and geographical and bathymethric distribution also considered important parameters. In the present paper, we made these observations and techniques in detail and complemented them with the novel technique of micro-computed tomography of the calcareous ring. We investigated a single European species, the so-called gravel sea cucumber, Neopentadactylamixta (Östergren, 1898), using recently collected material from the Chausey Islands, Normandy, France. We redescribed the species, illustrated its ossicle assemblage through scanning electron microscopy, and visualised the calcareous ring through stacking photography and through micro-CT scanning. Additionally, a DNA fragment of 955 base pairs of the 18S ribosomal RNA gene was sequenced from one specimen, which showed a high similarity with the only sequence of N.mixta publicly available. We completed this integrative study by providing a detailed distribution of the occurrence of N.mixta based on published, verifiable accounts.

Introduction: Dental implant placement can be challenging when insufficient bone volume is present and bone augmentation procedures are indicated. The purpose was to assess clinically and histologically a specimen of 30%HA-60%β-TCP BCP 3D-printed scaffold, after 7-years. Case Description: The patient underwent bone regeneration of maxillary buccal plate with 3D-printed biphasic-HA block in 2013. After 7-years, a specimen of the regenerated bone was harvested and processed to perform microCT and histomorphometrical analyses. Results: The microarchitecture study performed by microCT in the test-biopsy showed that biomaterial volume decreased more than 23% and that newly-formed bone volume represented more than 57% of the overall mineralized tissue. Comparing with unloaded controls or peri-dental bone, Test-sample appeared much more mineralized and bulky. Histological evaluation showed complete integration of the scaffold and signs of particles degradation. The percentage of bone, biomaterials and soft tissues was, respectively, 59.2, 25.6, and 15.2%. Under polarized light microscopy, the biomaterial was surrounded by lamellar bone. These results indicate that, while unloaded jaws mimicked the typical osteoporotic microarchitecture after 1-year without loading, the BCP helped to preserve a correct microarchitecture after 7-years. Conclusions: BCP 3D-printed scaffolds represent a suitable solution for bone regeneration: they can lead to straightforward and less time-consuming surgery, and to bone preservation.

Spicules are mineral-based biocomposites skeletal structures that are widely distributed among phylogenetically distant groups of invertebrates (Porifera, Cnidaria, Mollusca, Echinodermata). Subepidermal spicules are formed under the ectodermal epithelium and are characterized for all groups except mollusks (Aplacophora, Polyplacophora, Bivalvia), their spicules are located on the surface of the body. However, one group of mollusks (Gastropoda: Heterobranchia) have unique subepidermal spicules that have never been detected above the ectodermal epithelium and similarly to those characterized for Porifera, Cnidaria and Echinodermata. Understanding subepidermal spicule formation in mollusks could help solve the question on the origin of spicules. Spicules in nudibranchs have been described for more than 150 years, yet ontogenetic dynamics of spicules have never been studied and the full mechanism of their formation remains unknown. Herein we investigate the spicule formation in different stages of postlarval development of the nudibranch Onchidoris muricata (O.F. Müller, 1776). For the first time, ontogenetic transformations of the spicule complex are described using experiments and different morphological methods. Our studies demonstrate that spicules of O. muricata form in the subepidermal space in early developmental stages immediately after veliger settlement. A single spicule forms inside a huge vacuole within a sclerocyte and remains there throughout the entire life of the specimen. Signs of spicule or sclerocyte migration under the epithelium in postlarval development was not found. Spicules only form during larval settlement, increasing only in size as development furthers. For the first time, spicule mineralization zones were detected at the tips of the spicules as well as the presence of collagen I in the overall composition of the spicules. Thus, our findings suggest that spicules form by an ectodermal cell that emerged under the ectodermal epithelium during the earliest stages of postlarval development.

To reconstruct the etiology of a perimortem injury observed on a Neolithic - Chalcolithic cranium (5060 - 4400 yrs cal. BP).
A cranium of an old adult male individual belonging to a collective burial from Cova Foradada site (Calafell, Tarragona, Spain).
The cranium was analyzed macroscopically and microscopically using micro-CT scan.
The fracture on the right parietal bone presents characteristics of perimortem trauma. The morphology of the point of impact allows for the interpretation of this fracture as a result of impact by an object with a straight and sharp edge. In addition, the same cranium presents two healed antemortem injuries.
The object that most likely caused the cranial fracture was a stone adze. The blow occurred from behind the individual, possibly by a right-handed attacker.
The potential to link cranial fractures with specific tools increases our understanding of interpersonal violence during the Neolithic.
It is not possible to infer if this cranial injury was related to a large-scale intergroup confrontation or an intragroup violent event.
To investigate additional similar sites in the northeastern Iberian Peninsula to better characterize the fracture pattern caused by stone axes and adzes as well as other objects used as weapons during the Neolithic - Chalcolithic ages.