OBJECTIVE: The objective of this study was to determine the photodynamic therapeutic response of U-87 human glioma cell in vitro as well as in the nude rat xenograft model using photofrin as photosensitizer. MATERIAL AND METHOD: U-87 cells were cultured on 96-well culture plates, photofrin(Quadralogic Technologies Inc., Vancouver, Canada) was added into the cell culture medium at concentration of 1ng/ml, 2.5ng/ml, 5ng/ml, 10ng/ml and 20ng/ml. 24 hour after drug treatment, cells were treated with optical(632nm) irradiation of 100mJ/cm2, 200mJ/cm2 and 400mJ/cm2. Photofrin(12.5mg/kg, i.p.) was administered to 28 nude rats containing intracerebral U-87 human glioma as well as 26 normal nude rats. 48 hours after administration, animals were treated with optical irradiation(632nm) of 35J/cm2, 140J/cm2 and 280J/cm2 to exposed tumor and normal brain. The photofrin concen-tration was measured in tumor and normal brain in a separate population of animals. RESULTS: By MTT assay, there was 100% cytotoxicity at any dose of photofrin with optical irradiation of 200mJ/cm2 and 400mJ/cm2. But at the optical irradiation of 100mJ/cm2 cells were killed in dose dependent manner 28.5%, 49.1%, 54.4%, 78.2%, and 84.6% at concentration of 1ng/ml, 2.5ng/ml, 5ng/ml, 10ng/ml and 20ng/ml, respectively. Dose dependent PDT lesions in both tumor and normal brain were observed. In the tumor lesion, only superficial tissue damage was found with optical irradiation of 35J/cm2. However, in the optical irradiation group of 140J/cm2 and 280J/cm2 the volume of lesions was measured of 7.2mm3 and 14.0mm3 for treatment at 140J/cm2 and 280J/cm2, respectively. The U-87 bearing rats showed a photofrin concentration in tumor tissue of 6.53+/-2.16ng/g, 23 times higher than that found in the contralateral hemisphere of 0.28+/-0.15ng/g. CONCLUSION: Our data indicate that the U-87 human glioma in vitro and in the xenografted rats is responsive to PDT. At these doses, a reproducible injury can be delivered to human glioma in this model. Strategies to spare the normal brain collateral damage are being studied.
Animals ; Brain ; Brain Neoplasms ; Cell Culture Techniques ; Dihematoporphyrin Ether* ; Glioma* ; Heterografts ; Humans* ; Photochemotherapy ; Rats ; Rats, Nude

In the development of radio frequency (RF) coils for better quality of the mini-type permanent magnetic resonance imager for using in the small animal imaging, the solenoid RF coil has a special advantage for permanent magnetic system based on analyses of various types.of RF coils. However, it is not satisfied for imaging if the RF coils are directly used. By theoretical analyses of the magnetic field properties produced from the solenoid coil, the research direction was determined by careful studies to raise further the uniformity of the magnetic field coil, receiving coil sensitivity for signals and signal-to-noise ratio (SNR). The method had certain advantages and avoided some shortcomings of the other different coil types, such as, birdcage coil, saddle shaped coil and phased array coil by using the alloy materials (from our own patent). The RF coils were designed, developed and made for keeled applicable to permanent magnet-type magnetic resonance imager, multi-coil combination-type, single-channel overall RF receiving coil, and applied for a patent. Mounted on three instruments (25 mm aperture, with main magnetic field strength of 0.5 T or 1.5 T, and 50 mm aperture, with main magnetic field strength of 0.48 T), we performed experiments with mice, rats, and nude mice bearing tumors. The experimental results indicated that the RF receiving coil was fully applicable to the permanent magnet-type imaging system.
Animals ; Magnetic Fields ; Magnetic Resonance Imaging ; instrumentation ; Magnets ; Mice ; Mice, Nude ; Phantoms, Imaging ; Radio Waves ; Rats

Proliferative scarring in the form of keloids and hypertrophic scars continues to be a clinical problem for some patients. The lack of an animal model for such scarring has been an obstacle to studying the biology and effective therapy of these entities. Consequently we created an accurate reproductive animal model to systematically study them. Human proliferative scars were explanted into flaps based on isolated vascular pedicles in congenitally rats. We compared the procollagen type III peptide levels of proliferative scar tissue before and after explanting. The procollagen type III peptide levels of explanted proliferative scar tissue remained increased as before explanting. Histological analysis of the explanted proliferative scar tissue revealed that all explants retained their original histotypic character even after 1 year. We could also retain the volume of implanted proliferative scar for 1 year and studied in vitro cellular proliferation. Fibroblast cultures from explanted scars demonstrated less aggressive growth characteristic than those from original surgical specimens. The advantages of this animal model are as follows: 1. The explants retain their histotypical character for a long period. 2. Placement of the explants outside the dorsum of a nude rat makes serial observation and measurement easier. 3. Agents under test can be injected into the explants through a catheter inserted into a single pedicle of island flap without the possibility of spreading systematically.
Animals* ; Biology ; Catheters ; Cell Proliferation ; Cicatrix* ; Cicatrix, Hypertrophic ; Collagen Type III ; Fibroblasts ; Humans ; Keloid ; Models, Animal* ; Rats ; Rats, Nude

Diet has long been recognized as a strong factor in prostate carcinogenesis, with nutrients participating in either the development or the prevention of cancer. In this review, we concentrate on the role of dietary factors in prostate cancer development. The most significant dietary factors in prostate carcinogenesis are energy, total fat, animal fat, milk, calcium and red meat. However; evidence from case-control, epidemiological and laboratory studies does not support the causative role of any single nutritional component in prostate cancer development, and many questions remain to be further studied about the association of dietary factors with prostate cancer.
Animals ; Diet ; Dietary Fats ; adverse effects ; Humans ; Male ; Mice ; Mice, Nude ; Prostatic Neoplasms ; epidemiology ; etiology ; Rats ; Rats, Wistar ; Risk Factors

Objective: To explore the effects of porcine acellular dermal matrix (ADM) combined with human epidermal stem cells (ESCs) on wound healing of full-thickness skin defect in nude mice. Methods: The morphology of porcine ADM was analyzed by photograph of digital camera, the cell residues in porcine ADM were observed by hematoxylin-eosin (HE) staining, the surface structure of porcine ADM was observed by scanning electron microscope, the secondary structure of porcine ADM was analyzed by infrared spectrometer, the porcine ADM particle size was analyzed by dynamic light scattering particle size analyzer, and the porcine ADM potential was analyzed by nano-particle size potentiometer. The morphology of porcine ADM was observed by inverted fluorescence microscope when it was placed in culture medium for 30 min, 1 d, and 5 d (n=2). The porcine ADM was divided into 5 min group, 10 min group, 20 min group, 30 min group, 60 min group, and 120 min group according to the random number table (the same grouping method below) in static state at normal temperature for the corresponding time to calculate the water absorption by weighing method (n=3). Swiss white mouse embryonic fibroblasts (Fbs) were divided into blank control group (culture medium only), and 50.0 g/L ADM extract group, 37.5 g/L ADM extract group, 25.0 g/L ADM extract group, 12.5 g/L ADM extract group, and 6.5 g/L ADM extract group which were added with the corresponding final concentrations of ADM extract respectively. At post culture hour (PCH) 24, 48, and 72, the cell survival rate was detected by cell counting kit 8 and the cytotoxicity was graded (n=5). The erythrocytes of a 6-week-old male Sprague-Dawley male rat were divided into normal saline group, ultra-pure water group, and 5 mg/mL ADM extract group, 10 mg/mL ADM extract group, and 15 mg/mL ADM extract group which were treated with the corresponding final concentrations of porcine ADM extract respectively. After reaction for 3 h, the absorbance value of hemoglobin was detected by microplate reader to represent the blood compatibility of porcine ADM (n=3). ESCs were isolated and cultured from the discarded prepuce of a 6-year-old healthy boy who was treated in the Department of Urology of the First Affiliated Hospital of Army Medical University (the Third Military Medical University) in July 2020, and then identified by flow cytometry. The porcine ADM particles of composite ESC (hereinafter referred to as ESC/ADM) were constructed by mixed culture. After 3 days of culture, the composite effect of ESC/ADM was observed by HE staining and laser scanning confocal microscope. Thirty-six 7-8-week-old male non-thymic nude mice were divided into phosphate buffer solution (PBS) alone group, ADM alone group, ESC alone group, and ESC/ADM group, with 9 mice in each group, and the wound model of full-thickness skin defect was established. Immediately after injury, the wounds were treated with the corresponding reagents at one time. On post injury day (PID) 1, 7, 11, and 15, the wound healing was observed and the wound healing rate was counted (n=3). On PID 7, the epithelialization of wounds was observed by HE staining and the length of un-epithelialized wound was measured (with this and the following sample numbers of 4). On PID 11, the dermal area and collagen deposition of wounds were observed by Masson staining and the dermal area of wound section was calculated, the number of cells expressing CD49f, a specific marker of ESC, was calculated with immunofluorescence staining, the mRNA expression of glyceraldehyde-3-phosphate dehydrogenase (GAPDH) in ESC after wound transplantation was detected by real-time fluorescence quantitative reverse transcription polymerase chain reaction. Data were statistically analyzed with independent sample t test, one-way analysis of variance, analysis of variance for repeated measurement, and least significant difference t test. Results: The porcine ADM was white particles and composed of reticular structure, with no cells inside, disordered structure, and rough surface. The absorption peak of porcine ADM appeared at the wave numbers of 1 659, 1 549, and 1 239 cm^-1, respectively. The main particle size distribution of porcine ADM in solution was 500 to 700 nm, with negative charge on the surface. The morphology of porcine ADM in static state at 30 min and on 1 and 5 d was relatively stable. The water absorption of porcine ADM remained relatively high level in static state from 30 min to 120 min. The cytotoxicity of mouse embryonic Fbs in 6.5 g/L ADM extract group, 12.5 g/L ADM extract group, and 25.0 g/L ADM extract group was grade 1 at PCH 24, and the cytotoxicity of the other groups was 0 grade at each time point. After reaction for 3 h, the absorbance value of hemoglobin of erythrocytes in ultra-pure water group was significantly higher than the values in normal saline group and 15 mg/mL ADM extract group (with t values of 8.14 and 7.96, respectively, P<0.01). After 3 days of culture, the cells of the fourth passage showed pebble-like morphology, with low expression of CD71 and high expression of CD49f, which were identified as ESCs. There was ESC attachment and growth on porcine ADM particles. On PID 1, the wound sizes of nude mice were almost the same in PBS alone group, ADM alone group, ESC alone group, and ESC/ADM group. On PID 7, 11, and 15, the wound contraction of nude mice in each group was observed, especially in ADM alone group, ESC alone group, and ESC/ADM group. On PID 7, the wound healing rates of nude mice in ESC alone group and ESC/ADM group were significantly higher than the rate in PBS alone group (with t values of 2.83 and 4.72 respectively, P<0.05 or P<0.01). On PID 11, the wound healing rate of nude mice in ESC/ADM group was significantly higher than that in PBS alone group (t=4.86, P<0.01). On PID 15, the wound healing rates of nude mice in ADM alone group, ESC alone group, and ESC/ADM group were significantly higher than the rate in PBS alone group (with t values of 2.71, 2.90, and 3.23 respectively, P<0.05). On PID 7, the length of un-epithelialized wound of nude mice in ADM alone group, ESC alone group, and ESC/ADM group was (816±85), (635±66), and (163±32) μm, respectively, which were significantly shorter than (1 199±43) μm in PBS alone group (with t values of 5.69, 10.19, and 27.54 respectively, P<0.01). On PID 11, the dermal areas of wound section of nude mice in ADM alone group, ESC alone group, and ESC/ADM group were significantly larger than the area in PBS alone group (with t values of 27.14, 5.29, and 15.90 respectively, P<0.01); the collagen production of nude mice in ADM alone group and ESC/ADM group was more obvious than that in PBS alone group, and the collagen production of nude mice in ESC alone group and PBS alone group was similar. On PID 11, in the wounds of nude mice in ESC alone group and ESC/ADM group, the cells with positive expression of CD49f were respectively 135±7 and 185±15, and the mRNA expressions of GAPDH were positive; while there were no expressions of CD49f nor mRNA of GAPDH in the wounds of nude mice in PBS alone group and ADM alone group. Conclusions: ESC/ADM particles can promote the wound healing of full-thickness skin defects in nude mice, which may be related to the improved survival rate of ESCs after transplantation and the promotion of dermal structure rearrangement and angiogenesis by ADM.
Acellular Dermis ; Animals ; Fibroblasts ; Humans ; Male ; Mice ; Mice, Nude ; Rats ; Rats, Sprague-Dawley ; Stem Cells ; Swine ; Wound Healing

In many tissue engineering application, highly open porous scaffolds are required for efficient cell seeding and culture. Synthetic biodegradable polymers such as poly (L-lactic acid)(PLLA) and its copolymers with D-lactic and glycolic acids(PLGA) are widely used as a porous scaffold. The suitable biodegradability and dimensional stability of porous scaffolds during in vivo implantation play an important role in tissue engineering application. In this study, we investigated in vivo biodegradation and dimensional stability of acellular porous polymer scaffolds prepared by using a gas foaming technique with non-toxic effervescent mixture. In addition, we have engineered cartilage tissue 3D cultured on PLGA scaffolds in nude mouse in order to compare with degradation and deformation on acellular porous polymer scaffolds and to form tissue-engineered cartilage tissue. Sodium bicarbonate and citric acid crystals were used as an effervescent mixture. These particles were milled and sieved to yield various range of sizes(50 - 100, 100 - 300, and > 300 micrometer). After polymer scaffolds fabricated, biodegradation test was performed in subcutaneous tissue of male rats during 12 weeks. Degradability of polymer scaffolds were evaluated by weight difference, gel permeation chromatography(GPC), and SEM as each period. Tissue-engineered cartilage by transplanting 3D cultured chondrocytes onto PLGA 85:15 scaffolds in nude mouse was also made and compared with acellular scaffolds. In conclusion, highly open porous biodegradable scaffolds are prepared by gas foaming method using sodium bicarbonate and citric acid as a non-toxic effervescent mixture. Furthermore, tissue-engineered cartilage formation by in vivo 3D culture onto modified PLGA scaffolds in nude mouse was significantly improved as compared to controls.
Animals ; Cartilage ; Chondrocytes ; Citric Acid ; Humans ; Male ; Mice ; Mice, Nude ; Polymers ; Porifera* ; Rats ; Sodium Bicarbonate ; Subcutaneous Tissue ; Tissue Engineering

A major obstacles to evaluation of newly-developed treatment strategy for human lung cancer has been the lack of appropriate experimental animal models. We describe a new experimental model of orthotopically-developed non-small cell lung cancer in nude rat, involving inoculation of tumor cell suspension by thoracotomy. Over 40 direct implantation to the periphery of the lung has been performed to date, each requiring less than 1 hour for completion. This model has been used to perform a series of experiments to investigate whether the rat lung and surrounding structures trapped tumor cells with 2 different non-small cell lung cancer cell lines(NCI-H460 and NCI-H1299). Every animal showed development of tumor masses, which were loculated at the periphery of the lung paren- chyma and identified also by radiography. After 3 weeks of the inoculation, tumor develop ment at the mediastinal strutures were identified. The life expectancies of the victims were different between the cell lines, but were approximately 5 weeks when NCI-H460 cell line was used. This new orthotopic lung cancer model may be facilitate future studies of the new therapeutics of localized non-small cell lung cancer .
Animals* ; Carcinoma, Non-Small-Cell Lung* ; Cell Line ; Humans ; Life Expectancy ; Lung ; Lung Neoplasms ; Models, Animal* ; Models, Theoretical ; Radiography ; Rats ; Rats, Nude* ; Thoracotomy

BACKGROUND: Demineralized bone matrix (DBM) is used for bone healing due to its osteoinductivity, but it requires a carrier for clinical application. Here, we report the effects on the osteoinductivity of DBM by use of a poloxamer 407-based hydrogel as the carrier, compared to sterile water. METHODS: DBM-W and DBM-H represent 27 wt% of DBM with sterile water and DBM with a poloxamer 407-based hydrogel, respectively. Both of the compositions were applied to human mesenchymal stem cell (MSC) cultures, and monitored for alkaline phosphatase (ALP) staining and ALP activity. Six 10-week-old athymic nude rats were used for abdominal muscle grafting with either DBM-W or DBM-H, and were tested by plane radiography, microfocus X-ray computed tomography (CT), and decalcified histology to evaluate ectopic bone formation. RESULTS: The DBM-W group showed stronger ALP staining at 7, 14, and 21 days of treatment, and significantly higher ALP activity at 7 and 14 days of treatment, compared to the DBM-H group. Plane radiography could not confirm the radio-opaque lesions in the rat ectopic bone formulation model. However, ectopic bone formation was observed in both groups by micro-CT. Compared to the DBM-H group, the DBM-W group showed higher bone volume, percent bone volume and trabecular number, and the difference in percent bone volume was statistically significant. Decalcified histology found bony tissue with lamellation in both groups. CONCLUSIONS: Our results suggest that poloxamer 407-based hydrogel has efficacy as a DBM carrier since it shows ectopic bone formation, but its effects on the quality and quantity of osteoblastic differentiation in rat abdominal ectopic bone and MSC are considered negative.
Animals ; Bone Matrix/*physiology ; Cell Culture Techniques ; Decalcification Technique ; Excipients/*pharmacology ; Hydrogels/pharmacology ; Male ; Mesenchymal Stromal Cells/*drug effects ; Osteogenesis/*drug effects ; Poloxamer/*pharmacology ; Rats ; Rats, Nude

Vitiligo is a pigmentary disorder induced by a loss of melanocytes. In addition to replacement of pure melanocytes, cocultures of melanocytes with keratinocytes have been used to improve the repigmentation outcome in vitiligo treatment. We previously identified by in vitro studies, that adipose-derived stem cells (ADSCs) could be a potential substitute for keratinocytes in cocultures with melanocytes. In this study, the efficacy of pigmentation including durability of grafted melanocytes and short-term safety was examined in the nude mouse and Sprague-Dawley rat after grafting of primary cultured human melanocytes, with or without different ratios of primary cultured human ADSCs. Simultaneous grafting of melanocytes and ADSCs, which were separately cultured and mixed on grafting at the ratios of 1:1, 1:2, or 1:3, showed better efficacy than that of pure melanocytes. Grafting of melanocytes cocultured with ADSCs resulted in a similar outcome as the grafting of cell mixtures. Skin pigmentation by melanocytes : ADSCs at the ratios of 1:1 and 1:2 was better than at 1:3. No significant difference was observed between the 1-week and 2-week durations in coculturing. Time-course microscopic examination showed that the grafted melanocytes remained a little longer than 6-week post-grafting. No inflammatory cell infiltration was observed in the grafted skin and no melanocytes were detectable in other organs. Collectively, grafting of melanocytes and ADSCs was equally safe and more effective than grafting of melanocytes alone. Despite the absence of significant differences in efficacy between the group of 1:1 and that of 1:2 ratio, 1:2 ratio for 1-week coculturing may be better for clinical use from the cost-benefit viewpoint.
Animals ; Coculture Techniques ; Humans ; Keratinocytes ; Melanocytes* ; Mice ; Mice, Nude ; Pigmentation ; Rats ; Rats, Sprague-Dawley ; Skin Pigmentation ; Skin* ; Stem Cells* ; Transplants ; Vitiligo

PURPOSE: Dialysis and renal transplantation, the current therapies for end-stage renal disease (ESRD), have limitations including severe complications, donor organ shortage, and allograft failure. The present study investigated the possibility of using a tissue engineering technique for renal reconstruction as a new method to replace the current suboptimal treatments for ESRD. We reconstituted renal units in vivo by transplanting isolated renal segments on three-dimensional, biodegradable polymer scaffolds. METHODS: Renal segments were freshly isolated from Sprague-Dawley rat kidneys and seeded onto porous mesh matrices fabricated from polyglycolic acid, a biodegradable synthetic polymer. The renal segment-seeded scaffolds were implanted into subcutaneous spaces of athymic mice for two and four weeks. Retrieved specimens were examined by histological analyses. RESULTS: The tubular structures with hollow centers and vascular tufts of glomerulus-like structures were identified by histological analyses of the 2 and 4 week specimens. In contrast, no renal-like structures were observed from unseeded polymer implants (negative controls). CONCLUSION: These results suggest a possibility of reconstituting the renal structures by transplanting renal segments on polymer scaffolds and could be applid for partial or full replacement of kidney function in the treatment of ESRD.
Allografts ; Animals ; Dialysis ; Humans ; Kidney ; Kidney Failure, Chronic ; Kidney Transplantation ; Mice ; Mice, Nude ; Polyglycolic Acid ; Polymers* ; Rats ; Rats, Sprague-Dawley ; Tissue Donors ; Tissue Engineering