Hypoplasia of medial pterygoid process in sphenoid bone relates to decreased mesenchymal cell proliferation in the Runx2-haploinsufficient cleidocranial dysplasia mouse model.

OBJECTIVE: Hypoplasia of the medial pterygoid process of the sphenoid bone is a distinct skeletal phenotype in runt-related transcription factor 2 (Runx2) heterozygous mice and patients with cleidocranial dysplasia. The aim of this study was to investigate the involvement of Runx2 in hypoplasia by regulating cell proliferation in the mesenchymal cell condensation region. DESIGN: A total of thirty mouse embryos were used. The medial pterygoid process region in the Runx2+/+, Runx2+/-, and Runx2-/- mouse embryos were histologically investigated. Immunohistochemistry for Runx2 and proliferating cell nuclear antigen (PCNA) was carried out. RESULTS: In embryonic day 14.5, mesenchymal cell condensation appeared at the future medial pterygoid process in Runx2+/+ mice, but was obscure in Runx2+/- mice. In these areas, cells showed a dual expression of Runx2 and PCNA in both Runx2+/+ and Runx2+/- mice. However, the number of Runx2- and PCNA-positive cells was decreased in Runx2+/- mice. In Runx2-/- mice, mesenchymal cell condensation appeared on embryonic day 18.5 at the medial pterygoid process region, associated with a few PCNA-positive cells. Moreover, the PCNA-positive cell rate in the medial pterygoid process was significantly lower in Runx2-/- mice than in Runx2+/+ and Runx2+/- mice. On embryonic day 18.5, Runx2+/- and Runx2-/- mice showed significantly shorter axial length of medial pterygoid process compared to that in Runx2+/+ mice. CONCLUSIONS: The present study demonstrates that Runx2 is involved in cell proliferation in the mesenchymal cell condensation region of the medial pterygoid process during mouse embryonic development.

Changes of CD90 expression and immunoreactive cell localisation in rat dental pulp after cavity preparation.

CD90 expression and immunoreactive cell localisation in rat dental pulp cells after cavity preparation was investigated. Cavity preparation was performed on the maxillary first molar of 8-week-old Wistar rats (n = 36), and immunohistochemistry and quantitative real-time PCR were performed. CD90-immunoreactivity was observed among subodontoblastic cells in the control group. One day after cavity preparation, the CD90-immunoreactivity disappeared under the cavity area. While CD90-immunoreactivity was faint after 3 days, the re-arrangement of odontoblasts was detected in contact with dentine. After 5 days, the odontoblasts were observed beneath the dentine, and CD90-immunoreactive cells were localised under the odontoblast layer. Immunofluorescence showed co-localisation of CD90 and nestin was detected after 3 days. After 5 days, CD90-immunoreactivity increased at the subodontoblastic layer. mRNA expression of CD90 and DSPP decreased after cavity preparation, and gradually recovered (P < 0.01). These results suggest that CD90-immunoreactive cells in the subodontoblastic layer contribute to regeneration of odontoblast and subodontoblastic layers following cavity preparation.

Sphenoid bone hypoplasia is a skeletal phenotype of cleidocranial dysplasia in a mouse model and patients.

Cleidocranial dysplasia (CCD) is an autosomal dominant disorder caused by heterozygous mutations in RUNX2. Affected individuals exhibit delayed maturation or hypoplasia in various bones, mainly including those formed by intramembranous ossification. Although several reports described deformation of the sphenoid bone in CCD patients, details of the associated changes have not been well documented. Most parts of the sphenoid bone are formed by endochondral ossification; however, the medial pterygoid process is formed by intramembranous ossification associated with secondary cartilage. We first investigated histological changes in the medial pterygoid process during different developmental stages in Runx2+/+ and Runx2+/- mice, finding that mesenchymal cell condensation of the anlage of this structure was delayed in Runx2+/- mice as compared with that in Runx2+/+ mice. Additionally, in Runx2+/+ mice, Osterix-positive osteoblastic cells appeared at the upper region of the anlage of the medial pterygoid process, and bone trabeculae appeared to associate with subsequent secondary cartilage formation. By contrast, few Osterix-positive osteoblastic cells appeared at the upper region of the anlage of the medial pterygoid process, and no bone trabeculae appeared thereafter in Runx2+/- mice. At more advanced embryonic stages, endochondral ossification occurred at the lower part of the medial pterygoid process in both Runx2+/+ and Runx2+/- mice. After birth, well-developed bone trabeculae occupied two-thirds of the cranial side of the medial pterygoid process, and cartilage appeared beneath these bones in Runx2+/+ mice, whereas thin trabecular bone appeared at the center of the cartilage of the medial pterygoid process in Runx2+/- mice. In adult mice, the body and medial pterygoid processes of the sphenoid bone comprised mature bones in both Runx2+/+ and Runx2+/- mice, although the axial length of the medial pterygoid processes was apparently lower in Runx2+/-mice as compared with that in Runx2+/+mice based on histological and micro-computed tomography (CT) examinations. Moreover, medical-CT examination revealed that in CCD patients, the medial pterygoid process of sphenoid bone was significantly shorter relative to that in healthy young adults. These results demonstrated that the medial pterygoid process of the sphenoid bone specifically exhibited hypoplasia in CCD.

Tree of Vater-Pacinian corpuscles in the human finger and thumb: a comparison between the late fetal stage and old age.

Using histological sections of 12 hands from 12 human fetuses at 20-34 weeks of gestation (150-290 mm) and 14 fingers (index and small) from seven donated cadavers of elderly individuals (aged 78-95 years), we compared the features of Vater-Pacinian corpuscles between these two stages of life. Corpuscles with thin, tightly packed lamellae appeared to undergo a change to thick, loosely packed lamellae at 23-32 weeks. The typical fetal corpuscle had two parts: (1) a rod-like proximal part (0.2-0.6 mm in length) extending along the proximodistal axis of the finger, and (2) a distal end (0.1 mm) after acute bending of the proximal part. Corpuscles were associated with palmar digital nerves in the fingers, but were also present along the dorsal nerves in the thumb. A flower bouquet- or tree-like arrangement including 5-10 corpuscles extended to the dermis of the skin along a perforating artery. Serial sections of the thumb and fifth finger revealed approximately 80-180 corpuscles in the distal phalangeal segment. In elderly individuals, the corpuscles were distributed along the palmar digital nerve, but (1) their density was much lower than in fetuses and (2) a bouquet- or tree-like arrangement was rarely seen. In the distal segment, there were fewer than 40 adult corpuscles, being 0.2-0.5 mm thick and 1.0-2.5 mm long. Wavy or coiled corpuscles were evident. Because of the considerable differences in the distribution and number of corpuscles between the fetus and adult, they appear to undergo considerable depletion with age, especially along thin, superficial nerve branches.

Morphology and relationships of the biceps brachii and brachialis with the musculocutaneous nerve.

INTRODUCTION: Major anatomical textbooks generally state that the biceps brachii muscle (BB) is composed of long and short heads, whereas the brachialis muscle (BR) consists of a single head. However, the numbers of heads comprising the BB and the BR are very variable. The purpose of this study was to investigate how the branching patterns of the musculocutaneous nerve (MC) influence the number of heads of the BB and the BR. MATERIALS AND METHODS: Morphological examinations of the BB and MC were conducted using cadavers of 22 Japanese individuals, and morphological examinations of the BR and the MC were conducted in 9 of those 22 individuals. RESULTS: A three-headed BB was observed in 7 of the 22 specimens (31.8%). Most of these specimens showed a Type III branch pattern (after penetrating the long head or the short head, the MC innervated the supernumerary head or communicated with the main root again). The number of BR heads was categorized into three types: Type A, two heads (superficial and deep heads, 22.2%); Type B, three or four heads (two or three superficial heads and one deep head, 44.4%); and Type C, multiple heads (33.3%). Among these categories, branches of the MC in Type A specimens were most simple. CONCLUSION: A supernumerary head of the BB seemed to be present if the MC penetrates it. The BR basically consists of superficial and deep heads, and the number of superficial heads is affected by branches of the MC.

Developmental mechanism of muscle-tendon-bone complex in the fetal soft palate.

OBJECTIVE: This study was performed to investigate how the palatine aponeurosis, medial pterygoid process (MPP) of the sphenoid bone, and tensor veli palatini (TVP) muscle form the pulley: muscle-tendon-bone complex. DESIGN: Mice at embryonic day (ED) 14-17 were used as sample in this study. Azan staining was performed to observe the morphology, and immunohistochemical staining of desmin was performed to closely observe the development of the myotendinous junction. To confirm the bone formation process, immunohistochemical staining of type II collagen (col II), tartrate-resistant acid phosphatase (TRAP), and alkaline phosphatase (ALP) staining were performed. Furthermore, to objectively evaluate bone formation, the major axis and width of the MPP were measured, and osteoclasts that appeared in the MPP were counted. RESULTS: At ED 14 and 14.5, ALP showed a reaction throughout the MPP. The col II-positive area expanded until ED 16.5, but it was markedly reduced at ED 17. The TVP initially contacted with the palatine aponeurosis at ED 16.5. The major axis and width of the MPP and the number of TRAP-positive osteoclasts were significantly increased as the TVP and palatine aponeurosis joined. CONCLUSIONS: Therefore, in addition to the tissue units: muscle, tendon, and bone, the interaction in organogenesis promotes rapid growth of the pulley: muscle-tendon-bone complex.

[Clinical Investigation of the Effects of Filgrastim BS1 on Neutropenia Following Oral Cancer Chemotherapy (TPF Therapy)].

The time for the neutrophil count to recover after subcutaneous injection of filgrastim BS1 or lenograstim was studied in patients suffering from neutropenia following preoperative combined chemotherapy using docetaxel, nedaplatin, or cisplatin (in divided doses for 5 days)and 5-fluorouracil for oral cancer. 1. There was no significant difference in the minimum leukocyte and neutrophil counts after chemotherapy. 2. There was no significant difference in the maximum leukocyte and neutrophil counts after chemotherapy. 3. Time for leukocytes to recover from their minimum count(>4,000/mm3)or for neutrophils to recover from their minimum count(>2,000/mm3)and the number of days on which treatment was administered tended to be shorter in the filgrastim BS1 group. Thus, it was concluded that filgrastim BS1 is just as effective as other prior G-CSF agents in treating patients suffering from neutropenia following chemotherapy(TPF therapy).