• Journal of International Academy of Physical Therapy Research
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• v.3 no.1
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• pp.345-355
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• 2012

This study purposed to examine the effect of low power laser on pain response and axonal regeneration. In order to prepare peripheral nerve injury models, we crushed the sciatic nerve of Sprague-Dawley rats and treated them with low power laser for 21 days. The rats were divided into 4 groups: normal group(n=10); control group(n=10) without any treatment after the induction of sciatic nerve crush injury; experimental group I(n=10) treated with low power laser(0.21$mJ/mm^2$) after the induction of sciatic nerve crush injury; and experimental group II(n=10) treated with low power laser(5.25$mJ/mm^2$) after the induction of sciatic nerve crush injury. We measured spontaneous pain behavior(paw withdrawal latency test) and mechanical allodynia(von Frey filament test) for evaluating pain behavioral response, and measured the sciatic function index for evaluating the functional recovery of peripheral nerve before the induction of sciatic nerve crush injury and on day 1, 7, 14 and 21 after the induction. After the experiment was completed, changes in the H & E stain and toluidine blue stain were examined histopathologically, and changes in MAG(myelin associated glycoprotein) and c-fos were examined immunohistologically. According to the results of this study, when low power laser was applied to rat models with sciatic nerve crush injury for 21 days and the results were examined through pain behavior evaluation and neurobehavioral, histopathological and immunohistological analyses, low power laser was found to affect pain response and axonal regeneration in both experimental group I and experimental group II. Moreover, the effect on pain response and axonal regeneration was more positive in experimental group I to which output 0.21$mJ/mm^2$ was applied than in experimental group II to which 5.25$mJ/mm^2$ was applied.

• The korean journal of orthodontics
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• v.21 no.1 s.33
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• pp.53-76
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• 1991

The study was designed to analyse the reorganization of the rabbit TMJ meniscus which was anteriorly displaced by surgery. The author compared the anteriorly displaced groups with control group. After surgical opening of the left rabbit TMJ space, cut the posterior attachment of the meniscus, and pushed it under the undercut area of the condyle head. Experimental groups were sacrificed by 1, 2, 4, 8 weeks after surgery. The samples were analysed with light microscope under T-B stain and electron microscope. The results were as follows: 1) The rabbit TMJ meniscus consisted of thick anterior and posterior band running different way, and comparative thin intermediate band runining antero-posteriorly. 2) Round oval shape chondrocyte-like cells were imbeded between the collagen fiber bundles and composed of proteoglycan granules, that showed metachromasia with toluidine blue, around the cell matrix. 3) Type II collagen fiber bundles in experimental group occured degenerative changes in organic patterns at 8 weeks, but those of type I collagen fiber bundles sustained longer, 4) The typical fibrocartilage of the rabbit TMJ meniscus was changed into fibrotic mode in process of time and showed the degenerative changes, which contained hyperplasia, calcification, resorption and hyalinization in the connective tissue. 5) The hyperplastic change of the synovial membrane in 4 week group and transitional change from fibrocyte to chondrocyte in cell type in 8 week group were observed. 6) The diameters of collagen fibers were diminished with the degenerative changes, the shape of the fibers became wavier and more nonorganic in running pattern and fiber bundle spaces widened.

• The korean journal of orthodontics
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• v.14 no.1
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• pp.53-65
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• 1984

The purpose of this study was to investigate the histopathological change and adaptation process of the mandibular joint of the rat by muscle alteration. For this study, one hundred and twenty eight rats of 25 - and 60 - day old of age were used. Unilateral and bilateral detachment, with anterior positioning of the Masseter muscle, was performed under anesthesia. The animal was sacrified 10, 20, 50, 80 days postoperatively. This alteration in muscle function led to change in neuromuscular activity and demonstrated the adaptive nature of the condyle cartilage to functional demand. The results were as follows : 1. In the right muscle detached group, operated at 25 days of age, marked decrease on the chondroblastic zone was found in the condyle head on the right side of animals examined 10 days postoperatively. Comparing with the control group, no difference was found on the chondroblastic zone in the condylar head of animals examined 20, 50 and 80 days postoperatively. 2. In the bilateral muscle detached group, operated at 25 days of age, the chondroblastic zone was slightly decreased in the anterior parts of condylar head of animals examined 10 days postoperatively. 3. In the unilateral and bilateral muscle detached group, operated at 60 days of age, no significant change was found in the mandibular joint regardless of the post operative experimental periods. 4. Under Toluidine blue staining, slightly decreased metachromasia was found in the condyle head on the right side of unilateral experimental animals, operated at 25 days of age and examined 10 days postoperatively. 5. Under Masson's trichrome staining, increased metachromasia was found in the condyle head on the right side of unilateral experimental animals, operated at 25 days of age and examined 10 days postoperatively. In summary, the condyle of the rat could respond to changes in neuromuscular activity depend on the level of maturation of the tissue, because the endochondral bone formation of the condyle of the rat was almost ended within 3 months.

• Journal of Periodontal and Implant Science
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• v.25 no.2
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• pp.341-356
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• 1995

The purpose of this study was to evaluate the effect of demineralized freeze dried bone and demineralized bone gel with guided tissue regeneration treatment around titanium implants with dehisced bony defects and also evaluate space maintaining capacity of demineralized bone gel type and DFDB powder type under e-PTFE membrane. In 3 Beagle dogs, mandibular premolar was extracted and four peri-implant osteotomies were formed for dehiscence. After insertion of implants, the four peri-implant defects were treated as follows. 1) In control group. no graft material and barrier membrane were applied. 2) In experimental group.1, the site was covered only with the e-PTFE membrane. 3) In experimental group 2,received DFDB powder and covered by the e-PTFE membrane. 4) In experimental group 3, demineralized bone gel and e-PTFE membrane were used. By random selection, animals were sacrificed at 4, 8, 12 weeks. The block sectioned specimens were prepared for decalcified histologic evaluation(hematoxylin and eosin staining) and undecalcified histologic evahiation(Von Kossa's and toluidine blue staining) with light microscopy. The results of this study were as follows. 1) In control group, there was a little new bone formation and connective tissue was completely filled in the defect area. 2) Experimental group 1 showed lesser quantity of bone formation as compared to the bone grafted group. Thin vertical growth of new bone formation around implant fixture was shown. 3) Experimental group 2 showed thick bucco-lingual growth of new bone formation and grafted bone particles were almost resorbed in 12 week group. 4) In experimental group 3, most grafted bone particles were not resorbed in 12 week group and thick bucco-lingual bone formation was shown in dehisced defect base area. 5) There was no remarkable differences in space making capacity and new bone formation procedure between demineralized freeze-dried bone powder type and demineralized bone gel type.

• Journal of Periodontal and Implant Science
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• v.34 no.1
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• pp.93-100
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• 2004

많은 연구들에서 hMSC를 얻기 위해 centrifugation, fluoroscence activated cell sorter(FACS), magnetic activated cell sorter(MACS)가 이용되어져 왔다. 그러나 centrifugation만을 이용한 경우 순도가 떨어지며 FACS나 MACS의 경우에는 비용, 시간이 많이 드는 단점이 있다. 따라서 이 연구에서는 antibody cocktail을 이용하여 hMSC를 좀더 쉽게 얻어내는 방법에 대해 알아보았다. 사람의 골반에서 12G의 바늘을 이용하여 골수를 흡입한 후 heparin이 들어있는 시험관에 넣고 처리과정을 시행하기 전에 냉장고에 보관하며 가능한 한 빨리 처리 과정을 실시한다. 얻은 골수에 적당량의 RosetteSep( Stemcell Technologies)을 첨가한 후 실온에서 20분간 반응시킨다. 그 후 적당량의 Ficoll-paque위에 골수와 RosetteSep의 혼합물을 섞이지 않게 올리고 원심분리를 이용하여 원하는 세포층을 얻어낸다. 이 세포층을 따로 분리한 뒤 배양한다. 배양 시 세포가 80%이상 차기 전에 계속 passage를 시행하며 배양한다. 이는 세포가 밀도가 높아져 원치 않는 세포로 분화되는 것을 막기 위함이다. 배양된 세포가 다양한 분화능력을 가지고 있는지 알아보기 위해 세 가지로 분화를 유도하였다. 적절한 배지와 적절한 환경에서 배양함으로써 얻어진 세포를 osteoblast, chondroblast, adipocyte로 분화를 유도하였다. 분화된 세포가 원하는 형질의 세포로 분화되었는지를 확인하기 위하여 osteoblast의 경우 alizarin red staining, alkaline phosphatase activity, chondroblast의 경우 toluidine blue staining, adipocyte의 경우 Oil-Red-O staining으로 염색하여 분화를 확인하였다. 분리해낸 세포는 각각 세 가지 세포로 분화가 되었으며 이는 RosetteSep이 hMSC를 성공적으로 분리해냈다는 것을 보여준다. 그러나 모든 세포가 분화를 보이지는 않았으며 따라서 hMSC의 순도를 높이기 위한 연구가 더 필요하다. RosetteSep을 이용하면 다른 방법들 보다 쉽게 hMSC를 얻을 수 있으나 기존의 방법과 순도의 측면에서 더 비교할 필요가 있다.

• Biomedical Science Letters
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• v.16 no.4
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• pp.349-357
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• 2010

We have examined the histological changes of skin during hair follicle growth after depilation in C57BL/6N mice. We first studied on histological changes of number of mast cells and thickness of skin during hair follicle growth periods (telogen, 1 day, 3 day, 5 day, 10 day, 14 day, 17 day and 21 day after depilation) by toluidine blue, Giemsa and H&E staining methods. We second studied immunoreactive density of cytokines and Brdu labeled cells in skin during hair follicle growth periods after depilation in C57BL/6N mice by immunohistochemical methods. The histological changes on skin thickness was increased from telogen to 14 day. The number of mast cells was decreased in 3,5 and 10 day and increased in 14, 17 and 20 day after depilation. Immunoreactive density of cytokines [protein kinase C-${\alpha}$ (PKC-${\alpha}$), c-kit, and vascular endothelial growth factor (VEGF)] in 1, 3, 5, 10, and 14 day after depilation was mildly stained in bulge and cutaneous trunci m., but immunoreactive density of cytokines in 17 and 21 day was heavily stained in epidermis, bulge, outer root sheath (ORS), inner root sheath (IRS) and cutaneous trunci m.. Immunoreactive density of Brdu labeled cells in skin in 1 and 3 day was heavily stained in bulge, epidermis and connective tissue under the cutaneous trunci m.. In all periods, immunoreactive density of Brdu labeled cells in skin was heavily stained in bulge, subcutaneous tissue, cutaneous trunci m, ORS and IRS. These experiments suggest that histological changes related to hair follicle growth elevated mast cell counts, skin thickness and epidermis thickness and heavily stained immunoreactive density of cytokines and Brdu labeled cutaneous trunci m. and connective tissue under the cutaneous trunci m. after depilation in C57BL/6N mice.

• Biomedical Science Letters
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• v.16 no.4
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• pp.365-376
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• 2010

The purpose of this study was to clarify the effect of light emitting diode (LED) irradiation on healing of impaired wound and alteration of mast cells in experimental diabetic rats. Twenty-four male Sprague-Dawley rats were divided into four groups: excision (Ex), excision-LED irradiation (Ex-LED), diabetes + excision (DM) and diabetes + excision + LED irradiation (DM-LED). Diabetes was induced in rats by streptozotocin (STZ) injection (70 mg/kg, single dose) and 6 mm punch excision wounds were created on the back after shaving hair. The LED-irradiated rats were treated to a daily dose of $5\;J/cm^2$ LED (630 nm) light for 11 days after surgery, and were killed at day 1, 3, 7 and 11. The lesion and adjacent skin tissues were excised, fixed with 10% buffered formalin and embedded with paraffin. For evaluation of wound healing, hematoxylin-eosin (HE) and Masson trichrome staining were performed. Mast cells (MCs) were stained with toluidine blue (pH 0.5) and quantified using a computerized image analysis system. The proliferation activity of keratinocyte in skin tissues was analyzed on sections immunostained with proliferative cell nuclear antigen (PCNA). The results showed that wound healing rate, collagen density and neo-epidermis length, number of PCNA-positive cells, fibroblasts and mast cells were significantly higher in the LED-irradiated rats than in the DM and Ex rats throughout the periods of experiment. Exceptionally, the number of MCs was significantly lower at day 11 compared with day 7 after surgery in the all groups. These findings suggest that the LED irradiation may promote the tissue repair process by accelerating keratinocyte and fibroblast proliferation and collagen production in normal rats as well as in diabetic rats, and MCs may play an important role at an early stage of skin wound healing in normal and diabetic rats.

• The Korean Journal of Malacology
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• v.4 no.1
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• pp.1-16
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• 1988

The authors observed histochemical and ultrastructural characters on the osphradium of Rapana venosa Valenciennes using light microscope, scanning and transmission electron microscpes. The results were as follows:1)The basic structure of osphradium was bipectinated shape, which consisted of a septum situating in the center of osphradium and numerous osphradial leaflets. On the other hand, Epidermis of ospradial leaflets formed the structure of pseudostratified ciliated columnar epithelium which was composed of an epithelial cell layer, a basal cel layer and a neuropile. 2) Ciliated dpithelial cells:A large number of these cells were observed on the lateral and ventral regions but a small number of them were observed on the dorsal region. These cells had cylindrical microvilli, slender mitochondria and serve fibers.3) Supporting cells: These cells had cylindrical microvilli, spongy layer, electron dense granules, mitochondria and nerve fibers4) Four types secretory epothelial cells: Four distinct types of secretory epithelial cells were recognized and were arbitrily designated as Type I, Type II, Type III and Type IV.cell type I: These cells contained electron denwe granules(diameter, 0.94-1.56${\mu}{\textrm}{m}$), well developed Golgi apparatus and rough endoplasmic reticula, cell type II: These cills contained two types of granules of the different electron density. One was high electron density granules which were 0.4-1.0${\mu}{\textrm}{m}$ in diameter, The other was low electron density granules which were 0.75-1.2${\mu}{\textrm}{m}$ in diameter.cell type III:These cells had fibrous secretory materials and exhibited strongly positive reaction with Toluidine blue.cell type IV:A large number of this type of cells were observed on the ventral region of ospgradial leaflets and positively reacted with periodic acid Schiff reagent. 5)Dark cells contained several electron dense cillaty rootlets and unmerous granules but cellular organelles were not observed.6) Four types basal cells: Four distinci types of basal cells were recognized and arbitrarily designated as Type I, Type II, Type III and Type IV.Cell type I(light cell): These cells exhibited low electuon density and contained short smooth endoplasmic reticula, several vacuoles and granules.

• Korean Journal of Clinical Laboratory Science
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• v.44 no.4
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• pp.229-238
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• 2012

Cyclooxygenase(COX-2) is an inducible enzyme that catalyzes the synthesis of prostaglandins (PGs) from arachidonic acid. Over-expression of COX-2 has been reported to be associated with progressive hepatic fibrosis in chronic hepatic C infection and rat liver fibrosis induced by carbon tetrachloride($CCl_4$). Recently, it is well known that mast cell products can stimulate the proliferation of hepatic stellate cells and key players in liver fibrosis. But little is known regarding their role in $CCl_4$-induced liver fibrosis in rat. Our aim was to investigate the relation between COX-2 expression and mast cells during liver fibrosis after $CCl_4$ treatment. Thirty Wistar rats were divided into five groups (non-treated 0, 2, 4, 6 and 8-week after $CCl_4$-treatment). Reverse transcription polymerase chain reaction (RT-PCR) and immunohistochemistry were used to assess the expression of ${\alpha}$-smooth muscle actin (${\alpha}$-SMA), collagen-1 and COX-2 in liver tissue from $CCl_4$-treated rats. The density of collagen and mast cells were determined using a computerized image analysis system in liver sections stained with picrosirius red and toluidine blue, respectively. The expression levels of ${\alpha}$-SMA, collagen-1 and COX-2 mRNA were significantly higher at 2 wk in $CCl_4$-treated groups than non-treated group. The number of mast cells in liver tissues increased gradually from 2 wk to 6 wk depending on the fibrosis severity but decreased abruptly at 8 wk. The significant increase of collagen-1 and ${\alpha}$-SMA mRNA expression in $CCl_4$-treated rats was continued until 6 wk while the COX-2 mRNA was significantly decreased at 8 wk. These results suggest that increased mast cells are closely associated with COX-2 over-expression during hepatic fibrogenesis of $CCl_4$-treated rats.

• Physical Therapy Korea
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• v.12 no.1
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• pp.80-90
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• 2005

The purpose of this study was to evaluate the effects of pulsed ultrasound on wound healing and observe during the wound healing process the distribution of mast cells according to histopathologic findings. Eighty Sprague Dawley rats which were divided into 4 groups received full thickness skin wounds on the back. Each of the 5 animals was sacrificed immediately and then sacrificed again 1, 3, 6, and 12 days after injury. Specimens from the wounds were removed during healing and routinely processed with a hematoxylin-eosin stain and a toluidine blue stain. The authors then observed the distribution of mast cells under a light microscope. The results of this study were as follows: The rate of wound healing and the length of the wounds of the pulsed ultrasound group II was significantly faster than group I on day 6 and day 12 (p<.001). Group III showed the most significant effect after12 days (p<.001). Group IV also showed a significant effect at 12 days (p<.01). A low-intensity ultrasound .5 $W/cm^2$ resulted in a fast healing rate. During the wound healing process mast cells had a tendency to decrease in the acute inflammatory phase. During the wound healing process mast cells were thought to contribute to the healing of the wound.