PURPOSE: This study attempts to evaluate the effect of lens cortex and nucleus remnants on posterior capsular opacification with method of the cell culture according to in vitro capsular bag model. METHODS: After bovine lens were isolated, we performed continuous curvilinear capsulorhexis and hydrodissectioin of the lens fiber mass. At this stage a tension ring was implanted and then the preparations placed in organ culture for up to 6 weeks. Lens cortex and nucleus material was added at the culture media in group 2, 3, 4, 5, 6 with amount of 1/16, 1/32, 1/64, 1/96, 1/128 of one lens volume. Group 1 was control group that was not added lens materials. Cell coverage of the posterior lens capsule was recorded and the capsules were examined, both pre-and post-coverage, for proliferative activity. RESULTS: After a lag period outgrowth was observed across the posterior capsule. The proliferative activity was greater at the groups that were added more amount of the lens cortex and nucleus material. CONCLUSIONS: it is important that we should not remain any lens cortex material remnant at cataract surgery.
Capsules ; Capsulorhexis ; Cataract ; Cell Culture Techniques ; Culture Media ; Epithelial Cells* ; Organ Culture Techniques

BACKGROUND: Careful manipulation of hair grafts is essential for a good yield of transplanted hair. OBJECTIVE: The aim of this study was to evaluate some of the factors responsible for poor graft yield, such as dehydration of the graft and the temperature and duration of preservation. METHODS: First, for the dehydration study, isolated single hair follicles were left on dry gauze for 0, 5, 10, 20, and 30 minutes at room temperature. Secondly, to evaluate the effect of preservation temperature and time on the hair graft, follicles were preserved in saline for 5 minutes as a control, then for 6, 24, and 48 hours both at room temperature and at 4℃, respectively. Viability of preserved follicles was judged based on organ culture. RESULTS: Elongation of hair folliciles was seen in 96%, 94%, 94%, 83%, and 68% for 0-, 5-, 10-, 20-, and 30-minute air-exposed groups, respectively. Survival was seen in 95%, 92%, 40% and 34% at room temperature and 96%, 94%, 76% and 50% at 4℃ for follicles preserved in saline for 5 min (control), then for 6, 24, and 48 hours, respectively. CONCLUSION: We suggest that, alone with careful manipulation of hair units, high survival can be achieved with the avoidance of graft dehydration and preservation of the grafts at low temperatures if the operation time extends for more than 6 hours.
Dehydration* ; Hair Follicle ; Hair* ; Organ Culture Techniques ; Transplants*

BACKGROUND: Skin organ culture is widely used as a tool to investigate skin biology or skin disease. OBJECTIVE: The objective of the present study was to develop an ideal skin organ culture model for evaluation of melanin pigmentation. METHODS: An air-liquid interface and submerged method were used. The histology of the cultured skin was studied with H&E stain. To examine the epidermal pigmentation, Fontana-Masson stain and NKI/beteb stain were performed. Pigment modifiers (arbutin, LY294002) were applied to the culture medium for 3 days as an air-liquid interface culture. RESULTS: The general architecture of the skin was well maintained for 5 days. The melanin pigment decreased during culture without change of the number of melanocytes. As expected from previous reports, the effect of pigment modifiers (arbutin, LY294002) on cultured skin was demonstrated. CONCLUSION: The results indicate that this skin organ culture model is useful in evaluating the melanin pigmentation
Biology ; Melanins* ; Melanocytes ; Organ Culture Techniques* ; Pigmentation* ; Skin Diseases ; Skin*

OBJECTIVEStudies have showed that L type calcium channel plays an important role in dentin calcification and affects tooth development and tooth reparation after injury. The objective of this article is to study the effects of nimodipine, blocking agent of L type calcium channel, on human dentinogenesis using human tooth slice organ culture in vitro.

METHODSYoung healthy human premolars were collected, and cut into 2 mm-thick transverse slices by low speed diamond saw. Agarose beads dipped in nimodipine solution and PBS weresy minetrically placed on tooth slices, and the slices were then embedded in a semisolid agarose-based medium and cultured with organ culture method for 1 week. Fluorescent band of tetracycline, Von-Kossa staining, immunohistochemical staining of the slices and transmission electron microscopy (TEM) of odontoblasts were observed to evaluate dentinogenesis changes of the slices.

RESULTSTooth slices were successfully cultured in vitro for 1 week and the odontoblasts could maintain their original morphology. After treatment with nimodipine, the fluorescent band of tetracycline was narrow and weak, and globular calcification in predentine was decreased compared with the control. TEM showed that secretory vesicles in odontoblast were somewhat increased, hut iminunohistochemical staining for collagen I showed no difference between the two groups.

CONCLUSIONNimodipine can influence the calcification of dentine, but has no obvious influence on the synthesis and secretion of dentine matrix. The results show that L type calcium channel is important in dentin calcification.

Dentin ; Dentinogenesis ; Humans ; Nimodipine ; Odontoblasts ; Organ Culture Techniques

PURPOSE: To assess the effect of the remnants after lens extraction on posterior capsular opacification with lens epithelial cell culture through in vitro capsular bag model. METHODS: After isolating porcine lens capsules, sterile non-toxic PMMA (polymethyl- mathacrylate) tension ring was inserted into the capsule. These were placed in organ culture medium up to 6 weeks. The grade of cell coverage of the posterior lens capsule was recorded to check the proliferative activity. RESULTS: In the process of cell culture, outgrowth of the epithelial cells was observed across the posterior capsule after a lag period. The rate of cell coverage was dependent upon the added factors. The proliferative activity was the greatest in the group where lens cortical and nuclear materials were added, and other groups showed no difference from a control group. CONCLUSIONS: To reduce posterior capsular opacification, it is important that we should not leave the lens cortical material behind during cataract surgery.
Capsules ; Cataract Extraction* ; Cataract* ; Cell Culture Techniques ; Epithelial Cells* ; Organ Culture Techniques ; Polymethyl Methacrylate

OBJECTIVEThe purpose of this study was to establish a palatal organ culture method and to investigate the palatogenesis in vitro.

METHODS20 pregnant 14-day mice were killed, embryos were separated ascetically, and palatal shelves were dissected and placed on a modified Trowell's system. All explants were cultured 24 h and 48 h respectively. Finally, all explants were embedded and stained by Hematoxylin and Eosin.

RESULTSAll explants grew healthy. After incubation for 24 h, medial edge epithelium maintained, whereas after 48 h, medial edge epithelium disappeared, bilateral mesenchymal cells contacted, palates fused.

CONCLUSIONThis method provides an effective way for investigating the etiology of cleft palate in vitro.

Animals ; Cleft Palate ; Epithelium ; Female ; In Vitro Techniques ; Mice ; Organ Culture Techniques ; Palate ; cytology ; Pregnancy

BACKGROUND AND OBJECTIVES: Organotypic culture of organ of Corti maintains the basic organization of the spiral lamina and can conserve several factors responsible for the neuronal growth of the nervous components. The explant culture technique has been widely used in organ culture system, however, the floating drop method using collagen gel was also developed as a simple and reliable method. In order to study the effect of growth factors on the regenerative and protective ability of cochlear hair cells, we first had to establish an in vitro model of the inner ear. MATERIALS AND METHODS: Organ of Corti was obtained from newborn rats and cultured with the floating drop method using collagen gel. Immunohistochemical staining was used to visualize the stereocilia and scanning electron microscopic study was also carried out. RESULTS: Explants were maintained up to 10 days without contamination. Morphologically, immunofluorescent staining with phalloidin showed well preserved outer and inner hair cells with stereocilia on the second day of culture. On the tenth day of culture, the staining result showed inner and outer hair cells, although the stereocilia were poorly stained. In scanning electron microscopic examination, an explant on the tenth day of culture showed preserved outer and inner hair cells and stereocilia, although damaged hair cells and stereocilia were also observed. CONCLUSION: The floating drop method was an appropriate method for maintaining the organ of Corti in vitro with the advantage being the easiness in its manual manipulation.
Animals ; Collagen ; Culture Techniques ; Ear, Inner ; Hair ; Humans ; Infant, Newborn* ; Intercellular Signaling Peptides and Proteins ; Neurons ; Organ Culture Techniques ; Organ of Corti* ; Phalloidine ; Rats* ; Spiral Lamina ; Stereocilia

Laser photocoagulation was applied in vitro to organ culture exoplants of porcine retinal pigment epithelium(RPE) attached to Bruch's membrane, choroid, and sclera using the Del Priore method for de-monstrating the proliferation of RPE cells and characterizing the response with respect to power level of treatment. Six-millimetor-round buttons of eyewall were treated with 20-30 spots from the argon bluegreen laser using a 500 micrometer spot size, 0.1s duration, and variable powers(100mW, 300mW, and 500mW). Group 1 is untreated control group and group 2(100mW) and group 4(500mW) showed less active proliferation of RPE than group 3(300mW). In group 3, all RPE cells, Bruch's membrane, and a part of choroid were disrupted and lifted off three hours after laser photocoagulation and then RPE cells began to proliferate actively at third day. The treated area became completely covered with several layers of RPE cells. The proliferation of RPE cells turned out to be larger when the power was moderate(300mW) as compared to the case when the power was too high(500mW) or too low(100mW).
Argon ; Bruch Membrane ; Choroid ; Light Coagulation* ; Organ Culture Techniques* ; Retinal Pigment Epithelium* ; Retinaldehyde* ; Sclera

Since adult human skin can be grown in chernically defined medium without serum, the skin organ culture has gained a great interest as a method for studies concerning skin biology, pharmacology and toxicology. however, serum supplementation has extensively been used to improve the viahility of tissue culture. This study was undertaken to evaluate the effect of serum on the histologic changes ohserved during the organ culture of the normal human skin. The general architecture of the skin was well maintained for 6 days with or without seru. After then, fetal calf serum or autologous human serum was found to enhance the viability of the epidermis. A confluent layer of necrotic spinous ceils was ovserved earlier and more widespread without serum. The addition of serum had an impressive effect on epibolization. In the absenee of serum, the formation of the epibolus was not only minimal, but also, susceptible to degeneration, and no epibolus remained at 10 days rif incubation. No difference can be found between fetal calf serm and autologous human serum in the formation of the epibolus. There was no favorable effect of serum on the formation of new stratum corneum. The thickness of new straturn corneum increased in parallel with the number of parakeratatic cells, increasing most rapidly between 6 and 8 days of incubation. Parakeratosis was more prominent in the presence of serurn.
Adult ; Biology ; Epidermis ; Humans* ; Organ Culture Techniques* ; Parakeratosis ; Pharmacology ; Selective Estrogen Receptor Modulators ; Skin* ; Toxicology

OBJECTIVETo compare the changes of nucleus pulposus after in vitro culture of rabbit whole intervertebral disc and spinal motion segment.

METHODSTwenty-one New Zealand white rabbits which were randomly divided into organ group with 8 rabbits and segment group with 13 rabbits. Fifty intervertebral discs and 50 spinal motion segments were harvested respectively under aseptic conditions from two groups. These specimens were maintained in organ culture with hyperosmotic media (410 mOsm/kg), then 10 discs of the two groups were observed respectively by HE staining, immunohistochemistry of collagen type III, proteoglycan content and cells viability of nucleus pulposus before culture and at 3, 7, 14, 21 days after culture.

RESULTSHE staining showed the intervertebral disc tissue structure remained intact after culture of 21 days organ group and 14 days segment group,but there was severely degenerated of 21 days segment group. The intensity value of type II collagen immunohistochemical staining in the nucleus pulposus were not changed significantly between 21 days organ group and 14 days segment group (P > 0.05), but the staining of segment group at 21 days became shallower, there was significant difference compared with before each time points and organ group at 21 days (P < 0.05). PAS/AB staining of proteoglycan of nucleus pulposus showed that there were not decrease of tinting strength of two groups within 7 days, but the strength weakened slightly of two groups at 14 days, and the tinting strength became weaker at 21 days segment group, the change is more obvious than the organ group. The intensity value of fluorescence staining of nucleus pulposus cells was not changed significantly within 7 days of two groups (P > 0.05), the intensity value decreased slightly at 21 days organ group and 14 days segment group, but there were no significant difference compared with before time points (P > 0.05) however at 21 days segment group the intensity decreased as cells viability of nucleus pulposus decreased,and there was a significant difference compared with before each time points and organ group at 21 days (P < 0.05).

CONCLUSIONIt is not obviously degenerated of the discs of organ group cultured within 21 days and segment group cultured within 14 days, but there was significant degeneration of the intervertebral disc of segment group after cultured 21 days, so the rabbit spinal motion segment can be used on research about the biomechanics of intervertebral disc as a vitro experimental model within 14 days.

Animals ; Collagen Type II ; analysis ; Female ; Immunohistochemistry ; Intervertebral Disc ; chemistry ; pathology ; Male ; Organ Culture Techniques ; Rabbits