Immunohistochemical Reaction of Calcitonin-gene-related Peptide and Type II Collagen and Morphological Changes of Cartilage Implants and Cultured Chondrocytes / 대한해부학회지

The purpose of this study was to investigate the immunohistochemical reaction of calcitonin-gene-related peptide (CGRP) and type II collagen and also morphological changes of cartilage implants and cultured chondrocytes isolated from the articular and costal cartilages. The chondrocytes were isolated from the head of the femur and the 11th costal cartilage of the 6 months old rabbits. De novo implants were prepared from the chondrocytes cultured on the perichondrium by culturing isolated articular chondrocytes. Cultured chondrocytes and implants were evaluated by immunohistochemical staining of CGRP and type II collagen and electron microscopy. Articular chondrocytes maintained the typical phenotype in the 1st and 2nd subcultures, but the costal chondrocytes were transformed into fibroblast-like cells. The articular chondrocytes cultured on the perichondrium were more flattened and formed the cartilage. Most chondrocytes were no loss of type II collagen immunostaining by culturing. Implants replaced by the cultured articular chondrocytes were generally increased CGRP and decreased type II collagen immunoreaction. Electron microscopically the cultured articular chondrocytes had a large euchromatic nucleus, a few granules, and abundant vesicles. During culture, the nucleus became atropy and the cytoplasm contained many large vacuoles. The chondrocytes cultured on the perichondrium showed a lot of segmented rough endoplasmic reticulum and fine short microvilli. During culture, articular chondrocytes maintained typical phenotype and type II collagen reaction. The cultured articular chondrocytes had some organelles and euchromatic nucleus with prominent nucleolus. The chondrocytes cultured on the perichondrium showed active secretion of the matrix with small vesicles and well developed endoplasmic reticulum. The implanted articular chondrocytes showed the decrease of their organelles after secretion of the marix and became increased CGRP and decreased type II collagen immunoreaction.

Morphometrical and Histological Changes of Glomeruli and Proximal tubules of Contralateral Kidney in Uninephrectomized Rat / 체질인류학회지

This study was to investigate the histomorphomeric and histological changes of glomeruli and proximal tubules of contralateral kidney in uninephrectomized rat. Rats were Sprague Dawley strain and female 1-year-old. Uninephrectomized rats were sacrificed after 20 days, 30 days, and 40 days. The results were as follows: 1. The mean areas and diameters of the glomeruli were markedly increased at the 30 days. 2. In the glomeruli, the mean nuclear areas of podocytes were markedly increased, but those of cells of the parietal layer were decreased. At the 20 days, those of the endothelial cells were slightly incerased. 3. The lumens of the endothelial cells were irregularly expanded and folded at the 20 days. The basal lamina of the 30 days was markedly thickened in the electron microscopic finding. 4. At the 30 days, the mean areas and mean thicknesses of proximal tubules were markedly increased, but those were not normal. At the 40 days, the mean thicknesses of proximal tubules were restored to the almost normal. 5. Light microscopic changes of the proximal tubules had no difference between the control and the experimental group. In electron microscopical findings, the cytoplasm of the proximal tubules were densely filled with the enlarged mitochondria at the 30 days. At the 40 days, some large vacuoles appeared in the cytoplasm.

Structural Changes of the Nucleus Pulposus Cells and Cartilage Cells of Age-Related Intervertebral Disc in Mongrel Dog / 대한해부학회지

The intervertebral discs of the dog from the newborn, the 1 year old, and the 5 years old were examined about the cell morphologies and their structural changes according to the advancement of age by histomorphometry and electron microscopy. The three regions of the disc - the nucleus pulposus, the inner and outer annulus fibrosus were considered. Each region was divided into the three zones - the peripheral, intermediate, and central zones. The cell morphology of the three regions were as follows : A. The nucleus pulposus 1. At the newborn, the cells of the nucleus pulposus had long cytoplasmic processes forming large extracellular vacuoles. In the peripheral and intermediate zones, the cells with euchromatic nucleus had fine cytoplasmic granule-like fibers and some of free ribosomes. In the central zone, the cells with heterochromatic nucleus had many cytoplasmic vacuoles filled with free ribosomes. 2. At the 1 years old, the cell-group composed of 4 to 8 cells contacted each other with their cytoplasmic processes. In the peripheral zone, the elliptical chondroblast-like cells with euchromatic nucleus had well-developed rough endoplasmic reticulum and mitochondria. In the intermediate zone, the foam-like large round cells with euchromaic nucleus had multiple cytoplasmic vacuoles filled with granular contents. In the central zone, the cells with heterochromatic nucleus had large cytoplasmic vacuoles, and seemed to be degenerated type of the outer zone-cells. 3. At the 5 years old, the cells in the periphery were rhomboid or polygonal with euchromatic nucleus, and had broad cytoplasmic contact surface with adjacent cells. They had small cytoplasmic vacuoles, well-developed Golgi apparatus and rough endoplasmic reticulum. The cells in the intermediate zone had heterochromatic nucleus and a lot of variable size of cytoplasmic vacuoles, and seemed to be degenerated form of the cells of the periphery. In the central zone, the star-like cells with heterochromatic nucleus had multiple sharp cytoplasmic processes and vacant cytoplasm because of large cytoplasmic vacuoles. B. The cartilaginous part and the inner annulus fibrosus 1. At the newborn, the outer zone of the cartilaginous part of the intervertebral disc showed the isogenous group with 2 to 3 chondrocytes within the cell nest. The cells were distributed mainly around the blood vessels, and had euchromatic nucleus. They had small cytoplasmic vacuoles, well-developed rough endoplasmic reticulum and abundant ribosomes. The cells of the inner zone showed heterochromatic nucleus, well-developed rough endoplasmic reticulum and expanding terminal cisternae. 2. At the 1 year old, the inner annulus fibrosus was divided into the fibrocartilaginous part and the hyaline cartilaginous part. The former, the outer zone showed the cell nest with 2 to 4 of elliptical chondrocytes which had euchromatic nucleus, small cytoplasmic vacuoles and scattered fine granules. The latter, the inner zone showed the chondrocytes within the dissociated cell nest with heterochromatic nucleus. They had disperse rough endoplasmic reticulum and microvilli-like cytoplasmic processes. 3. At the 5 years old, the annulus fibrosus was composed mainly of fibrocartilage. The chondrocyte showed heterochromatic nucleus, long cytoplasmic processes, large cytoplasmic vacuoles and densely packed abundant granules. C. The outer annulus fibrosus 1. At the 1 year old, the outer annulus fibrosus was composed of the palisading collagenous bundles. The chondrocytes with heterochromatic nucleus located in the cartilage cell nest. The cells had well-developed rough endoplasmic reticulum, mitochondria, and Golgi apparatus, and 4 to 5 processes of interterritorial matrix surrounded the collagenous bundles. 2. At the 5 years old, the collagenous bundles were invaded by the processes of interterritorial matrix, and made them incomplete ones. The cell had heterochromatic nucleus and scanty cytoplasm containing small mitochondria and poorly developed organelles. The Summary of The Above Mentioned Findings Are : 1. The cells of the nucleus pulposus were degenerated gradually from the periphery toward the center. 2. The cells of the nucleus pulposus were degenerated gradually according to the aging process. 3. With age, the cells of the annulus fibrosus were degenerated, the hyaline cartilage was replaced gradually by fibrocartilages, and fragmentation of the collagenous bundles appeared. 4. The older the age, the smaller the nucleus of the nucleus pulposus cells, and the larger the nucleus of the annulus fibrosus cells were encountered in the histomorphometric measurement.

A Study on Morphological and Immunohistochemical Changes of the Myometrium in the Pregnant and Postpartum Rat / 대한해부학회지

This experiment was performed to investigate morphological and immunohistochemical changes of the connective tissue, the nerve and the smooth muscle cell in the pregnant and postpartum rat myometrium. The materials were the non-pregnant control, the 2-, the 4-, the 10-day gestation and the 1-, the 2-, the 6-day birth of rat myometrium. The light microscopic findings were examed after van Gieson and malachite green stainings. The reaction of nitric oxide synthase activity, apoptosis, type IV collagen, and estrogen receptor were examined immunochemically. The results were as follows: A. Light microscopic findings: 1. In the van Gieson staining for collagen fibers and smooth muscles, it showed the increase of pinky collagen fibers and yellow smooth muscle cells from the 2-day gestation. At the 1-day birth, the yellow hypertrophic smooth muscle cells were in the form of the papilla. The collagen fibers of the 2-day birth were more abundant than that of the 1-day birth. 2. In the malachite green staining for nerve fibers, it showed the blue spots. The size of spots of the 4-day gestation enlarged and at the 10-day gestation the number of fine spots was increased. At the 1-day birth, it showed the decreased number and increased size of spots. B. Immunohistochemical findings : 1. The NOS activity showed the increased positive reaction at the 2-day gestation. At the 10-day gestation, the outer zone of myometrium showed prominent positive. At the 1-day birth, the endothelium of the vessels were weakly positive. 2. The apoptotic reaction was localized at the connective tissue cells between the outer and inner zones of myometrium. The positive reaction cells were a few at the 1-day birth, several at the 2-day birth, and many at the 6-day birth. 3. The type IV collagen reaction appeared at the perivascualr area and the comnnective tissues of the inner zone. The positive reaction showed weak at the control, moderate at the 2-day gestation, and strong at the 4-day gestation. At the 1- and 2-day birth, the connective tissue of inner zone was moderately positive. 4. The estrogen receptor reaction cells appeared at the papillary outer zone of the 10-day gestation. The reaction cells were several at the 1-day birth and a few at the 2-day birth. C. Electron microscopic findings : 1. At the 10-day gestation, one group of smooth muscle cells had well-developed sarcoplasmic reticulum and the other group with abundant mitochondria appeared. At the 2-day birth, one group smooth muscle cells had degenerating processes and the other group with the serrated processes showed. At the 6-day birth, smooth muscle cells were degenerated. 2. Mitochondria were abundant in the smooth muscle cell with many processes of the 10-day gestation. The onset of mitochondrial degeneration began at the 2-day birth. The mean section of mitochondria was increased by the 2-day birth and markedly decreased at the 6-day birth. 3. At the 1-day and 2-day birth, the electron-dense region of cell membrane was increased and submembranous caveolae were decreased. The mean section of caveolae was the mostly decreased at the 10-day gestation, nearly restored at the 1-day birth and enlarged at the 6-day birth.