Regenerative muscles are required for swallowing and mastication, and are important for functional recovery from diseases involving oral muscular defects. Therefore, we generated three-layer hybrid sheets, similar to oral mucosal structures containing submucosal muscles, using rabbit oral mucosa epithelial, mesenchymal, and myoblastic progenitor cells, and examined the structural proteins. Each cell type was obtained from rabbit oral mucosa using enzymatic digestion. Isolated mesenchymal and myoblastic cells were multi-differentiated into osteoblasts, adipocytes, and chondrocytes or myotubes. Isolated epithelial cells were cultured on collagen gels containing isolated mesenchymal cells for 2 weeks, and these epithelial–mesenchymal cell sheets were laminated onto myoblastic cell sheets. The engineered hybrid sheets were multi-stratified in the epithelial and myoblastic layers in a time-dependent manner, expressing intermediate cytoskeletal filament proteins of epithelium and muscle. Hybrid sheets also expressed extracellular matrix basement membrane proteins. Immature cell markers for epithelial and myoblastic cells were observed continuously in hybrid sheet cultures. We established engineered three-dimensional rabbit oral mucosa hybrid sheets containing each immature cell type in vitro.

PURPOSE: We recently demonstrated the morphology of the anococcygeal ligament. As the anococcygeal ligament and raphe are often confused, the concept of the anococcygeal raphe needs to be re-examined from the perspective of fetal development, as well as in terms of adult morphology. MATERIALS AND METHODS: We examined the horizontal sections of 15 fetuses as well as adult histology. From cadavers, we obtained an almost cubic tissue mass containing the dorsal wall of the anorectum, the coccyx and the covering skin. Most sections were stained with hematoxylin and eosin or Masson-trichrome solution. RESULTS: The adult ligament contained both smooth and striated muscle fibers. A similar band-like structure was seen in fetuses, containing: 1) smooth muscle fibers originating from the longitudinal muscle coat of the anal canal and 2) striated muscle fibers from the external anal sphincter (EAS). However, in fetuses, the levator ani muscle did not attach to either the band or the coccyx. Along and around the anococcygeal ligament, we did not find any aponeurotic tissue with transversely oriented fibers connecting bilateral levator ani slings. Instead, in adults, a fibrous tissue mass was located at a gap between bilateral levator ani slings; this site corresponded to the dorsal side of the ligament and the EAS in the immediately deep side of the natal skin cleft. CONCLUSION: We hypothesize that a classically described raphe corresponds to the specific subcutaneous tissue on the superficial or dorsal side of the anococcygeal ligament.
Aged, 80 and over ; Anal Canal/*anatomy & histology/embryology ; *Cadaver ; Female ; *Fetus ; Humans ; Male ; Muscle, Smooth/*anatomy & histology/embryology ; Rectum/*anatomy & histology/embryology

Pacinian corpuscle-like structures were identified in the digital tendon sheaths and nail beds of hands obtained from eight of 12 human fetuses of gestational age 20–34 weeks (crown-rump length, 150–290 mm). The aberrant corpuscles were present in tight fibrous tissue connecting the flexor tendon sheath to the dorsal aponeurosis (138 corpuscles in the thumbs and all fingers of eight fetuses); loose fibrous tissue inside the sheath on the dorsal side of the tendon (37 corpuscles in the thumbs and all fingers of four fetuses); and the nail bed (10 clusters in the thumbs and second fingers of four smaller fetuses). The aberrant corpuscles in the tendon sheath were classified into two types: thin and short, with tightly packed lamellae, of diameter 20–40 µm and length 20–200 µm; and thick and long, with loosely packed lamellae, of diameter 70–150 µm and length 0.5–1.5 mm. The small corpuscles tended to form clusters, each containing 5–10 structures. Their similarity indicated that the tight and loose lamellae in these two types of corpuscles corresponded to typical immature and mature corpuscles, respectively, usually distributed along the palmar digital nerve. However, mature, large corpuscles were absent from the nail bed, and most aberrant corpuscles were smaller than typical corpuscles along the nerve. The aberrant corpuscles were apparently incorporated into the tendon sheath or nail bed during fetal vascular development, but they appeared to degenerate after birth due to mechanical stress from the tendon or nail.
Fetus* ; Fingers ; Gestational Age ; Hand ; Humans* ; Parturition ; Stress, Mechanical ; Tendons* ; Thumb

Background: The posterior transversus abdominis plane block (TAPB) and quadratus lumborum block (QLB) were developed for postoperative pain control after lower abdominal surgery. However, there is little data regarding their effects. Their analgesic effects and the distribution of the cutaneous sensory blockade were observed in patients undergoing laparoscopic gynecologic surgery. Methods: After an induction of general anesthesia, patients alternately received bilateral ultrasound-guided QLB type 2 (QLB2) or posterior TAPB using 20 ml of 0.375% levobupivacaine on each side. The measurements included visual analogue pain scores (VAS), cutaneous sensory blockade in each dermatome, demands for postoperative analgesics, and complications for up to 48 h after the block. Our primary endpoint was VAS at 24 h after the block. Results: Forty patients completed the study. The VAS at rest was significantly lower after QLB2 than that after TAPB at 48 h, but not at 24 h. Neither group differed in VAS when coughing at any point in time. Postoperative demands for fentanyl and other analgesics also did not differ for either block. The majority of injections produced a cutaneous sensory blockade in the T11 and T12 dermatomes in both groups. The median number of dermatomes blocked was limited to three dermatomes after either block. No severe complication related to either block was observed. Conclusions The analgesic effects of QLB2 and posterior TAPB did not differ in patients undergoing laparoscopic gynecologic surgery. The cutaneous sensory blockade produced was limited to three dermatomal levels in the majority of patients. However, these findings need to be confirmed through a larger comparative study.

In an attempt to clarify the function of the suboccipital muscles, we performed morphological observation of the suboccipital muscles for variations in the muscle belly and compared the morphology of their muscle fibers in terms of cross-sectional area by immunostaining with anti-myosin heavy chain antibodies. The cadavers of 25 Japanese individuals were used: 22 for morphological examinations and three for histological examinations. Among samples of the rectus capitis posterior major muscle (RCPma) and rectus capitis posterior minor muscle (RCPmi), 86.4% had a typical muscle appearance with a single belly, and 13.6% had an anomalous morphology. None of the samples of the obliquus capitis superior (OCS) or obliquus capitis inferior (OCI) muscles had an anomalous appearance. Measurement of cross-sectional area revealed that fast-twitch muscle fibers in the RCPma and OCI had a significantly greater cross-sectional area than those of the RCPmi and OCS. The cross-sectional area of intermediate muscle fibers was also significantly greater in the OCS than in the RCPma, RCPmi, and OCI. The cross-sectional area of slow-twitch muscle fibers was significantly greater in the OCS than in the RCPma, RCPmi, and OCI, and the RCPmi showed a significantly greater cross-sectional area for slow-twitch muscle fibers than did the RCPma, and OCI. Our findings indicate that the RCPmi and OCS exert a greater force than the RCPma and OCI, and act as anti-gravity agonist muscles of the head. Prolonged head extension in individuals with anomalous suboccipital muscle groups could result in dysfunction due to undue stress.
Antibodies ; Asian Continental Ancestry Group ; Cadaver ; Classification* ; Head ; Headache ; Humans ; Muscle Fibers, Fast-Twitch ; Muscle Fibers, Slow-Twitch ; Muscles

Although the pelvic autonomic plexus may be considered a mixture of sympathetic and parasympathetic nerves, little information on its composite fibers is available. Using 10 donated elderly cadavers, we investigated in detail the topohistology of nerve fibers in the posterior part of the periprostatic region in males and the infero-anterior part of the paracolpium in females. Neuronal nitric oxide synthase (nNOS) and vasoactive intestinal polypeptide (VIP) were used as parasympathetic nerve markers, and tyrosine hydroxylase (TH) was used as a marker of sympathetic nerves. In the region examined, nNOS-positive nerves (containing nNOS-positive fibers) were consistently predominant numerically. All fibers positive for these markers appeared to be thin, unmyelinated fibers. Accordingly, the pelvic plexus branches were classified into 5 types: triple-positive mixed nerves (nNOS+, VIP+, TH+, thick myelinated fibers + or -); double-positive mixed nerves (nNOS+, VIP-, TH+, thick myelinated fibers + or -); nerves in arterial walls (nNOS-, VIP+, TH+, thick myelinated fibers-); non-parasympathetic nerves (nNOS-, VIP-, TH+, thick myelinated fibers + or -); (although rare) pure sensory nerve candidates (nNOS-, VIP-, TH-, thick myelinated fibers+). Triple-positive nerves were 5-6 times more numerous in the paracolpium than in the periprostatic region. Usually, the parasympathetic nerve fibers did not occupy a specific site in a nerve, and were intermingled with sympathetic fibers. This morphology might be the result of an "incidentally" adopted nerve fiber route, rather than a target-specific pathway.
Adrenergic Fibers ; Aged* ; Cadaver* ; Female ; Humans ; Hypogastric Plexus* ; Male ; Myelin Sheath ; Nerve Fibers* ; Nitric Oxide Synthase Type I ; Tyrosine 3-Monooxygenase ; Vasoactive Intestinal Peptide

The mediobasal segment (S7) of the right lung has been considered to correspond to the cardiac lobe generally seen in mammals. To investigate fetal development of the right mediobasal segmental bronchus (B7), we examined paraffin-embedded serial sections of 15 embrynic and fetal lungs at 7-8 weeks (serial sections) as well as semiserial sections of 8 fetuses at 15-18 weeks (semiserial sections). All of the smaller specimens did not contain B7, but 2 of the 8 larger specimens carried B7: one was found in the immediately anterior side of the inferior pulmonary vein, while in the other, the subdivisions (B7a, B7b) were overriding the vein. Although the incidence might be underestimated because of observations using semiserial sections, the B7 was most likely to develop secondarily during a period from 8 to 15 weeks. Fetal topographical changes (mainly, the descent) of the middle lobe and the inferior pulmonary vein might relate with the secondarily budding of B7. The present result does not reduce a clinical relevance of B7 as a segmental bronchus of the lung segment system.
Bronchi ; Fetal Development* ; Fetus ; Incidence ; Lung* ; Mammals ; Pulmonary Veins ; Veins

There is little or no information about the distribution of elastic fibers in the human fetal head. We examined this issue in 15 late-stage fetuses (crown-rump length, 220-320 mm) using aldehyde-fuchsin and elastica-Masson staining, and we used the arterial wall elastic laminae and external ear cartilages as positive staining controls. The posterior pharyngeal wall, as well as the ligaments connecting the laryngeal cartilages, contained abundant elastic fibers. In contrast with the sphenomandibular ligament and the temporomandibular joint disk, in which elastic fibers were partly present, the discomalleolar ligament and the fascial structures around the pterygoid muscles did not have any elastic fibers. In addition, the posterior marginal fascia of the prestyloid space did contain such fibers. Notably, in the middle ear, elastic fibers accumulated along the tendons of the tensor tympani and stapedius muscles and in the joint capsules of the ear ossicle articulations. Elastic fibers were not seen in any other muscle tendons or vertebral facet capsules in the head and neck. Despite being composed of smooth muscle, the orbitalis muscle did not contain any elastic fibers. The elastic fibers in the sphenomandibular ligament seemed to correspond to an intermediate step of development between Meckel's cartilage and the final ligament. Overall, there seemed to be a mini-version of elastic fiber distribution compared to that in adults and a different specific developmental pattern of connective tissues. The latter morphology might be a result of an adaptation to hypoxic conditions during development.
Adult ; Capsules ; Cartilage ; Connective Tissue ; Ear Cartilage ; Ear Ossicles ; Ear, Middle ; Elastic Tissue ; Fascia ; Fetus ; Head ; Humans ; Joint Capsule ; Laryngeal Cartilages ; Ligaments ; Muscle, Smooth ; Muscles ; Neck ; Pterygoid Muscles ; Stapedius ; Temporomandibular Joint Disc ; Tendons ; Tensor Tympani

In the developing human musculoskeletal system, cell death with macrophage accumulation occurs in the thigh muscle and interdigital area. To comprehensively clarify the distribution of macrophages, we immunohistochemically examined 16 pairs of upper and lower extremities without the hip joint (left and right sides) obtained from 8 human fetuses at approximately 10-15 weeks of gestation. Rather than in muscles, CD68-positive macrophages were densely distributed in loose connective tissues of the flexor aspects of the extremities, especially in the wrist, hand and foot. In contrast, no or fewer macrophages were evident in the shoulder and the extensor aspects of the extremities. The macrophages were not concentrated at the enthesis of the tendon and ligament, but tended to be arranged along other connective tissue fibers. Deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick end-labeling revealed apoptosis in the hand lumbricalis muscles, but not in the area of macrophage accumulation. Likewise, podoplanin-positive lymphatic vessels were not localized to areas of macrophage accumulation. Re-organization of the connective tissue along and around the flexor tendons of the hand and foot, such as development of the bursa or tendon sheath at 10-15 weeks, might require the phagocytotic function of macrophages, although details of the mechanism remain unknown.
Apoptosis ; Cell Death ; Connective Tissue ; Deoxyuracil Nucleotides ; Deoxyuridine ; Extremities ; Fetus ; Foot ; Hand ; Hip Joint ; Humans ; Ligaments ; Lower Extremity ; Lymphatic Vessels ; Macrophages ; Muscles ; Musculoskeletal System ; Pregnancy ; Shoulder ; Tendons ; Thigh ; Wrist

We examined pharyngeal nerve courses in paraffin-embedded sagittal sections from 10 human fetuses, at 25-35 weeks of gestation, by using S100 protein immunohistochemical analysis. After diverging from the glossopharyngeal and vagus nerves at the level of the hyoid bone, the pharyngeal nerves entered the constrictor pharyngis medius muscle, then turned upward and ran superiorly and medially through the constrictor pharyngis superior muscle, to reach either the levator veli palatini muscle or the palatopharyngeus muscle. None of the nerves showed a tendency to run along the posterior surface of the pharyngeal muscles. Therefore, the pharyngeal nerve plexus in adults may become established by exposure of the fetal intramuscular nerves to the posterior aspect of the pharyngeal wall because of muscle degeneration and the subsequent rearrangement of the topographical relationship between the muscles that occurs after birth.
Adult ; Fetus ; Glossopharyngeal Nerve ; Humans ; Hyoid Bone ; Muscles ; Parturition ; Pharyngeal Muscles ; Pregnancy ; Vagus Nerve