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1.
J. appl. oral sci ; 31: e20220489, 2023. graf
Article in English | LILACS-Express | LILACS | ID: biblio-1430629

ABSTRACT

Abstract Objective: This study aimed to evaluate neuronal markers in stromal cells from human exfoliated deciduous teeth (SHED) and standardize the isolation and characterization of those cells. Methodology: Healthy primary teeth were collected from children. The cells were isolated by enzymatic digestion with collagenase. By following the International Society for Cell and Gene Therapy (ISCT) guidelines, SHED were characterized by flow cytometry and differentiated into osteogenic, adipogenic, and chondrogenic lineages. Colony-forming unit-fibroblasts (CFU-F) were performed to assess these cells' potential and efficiency. To clarify the neuronal potential of SHED, the expression of nestin and βIII-tubulin were examined by immunofluorescence and SOX1, SOX2, GFAP, and doublecortin (DCX), nestin, CD56, and CD146 by flow cytometry. Results: SHED showed mesenchymal stromal cells characteristics, such as adhesion to plastic, positive immunophenotypic profile for CD29, CD44, CD73, CD90, CD105, and CD166 markers, reduced expression for CD14, CD19, CD34, CD45, HLA-DR, and differentiation in three lineages confirmed by staining and gene expression for adipogenic differentiation. The average efficiency of colony formation was 16.69%. SHED expressed the neuronal markers nestin and βIII-tubulin; the fluorescent signal intensity was significantly higher in βIII-tubulin (p<0.0001) compared to nestin. Moreover, SHED expressed DCX, GFAP, nestin, SOX1, SOX2, CD56, CD146, and CD271. Therefore, SHED had a potential for neuronal lineage even without induction with culture medium and specific factors. Conclusion: SHEDs may be a new therapeutic strategy for regenerating and repairing neuronal cells and tissues.

2.
Braz. dent. sci ; 25(2): 1-12, 2022. ilus, tab
Article in English | LILACS, BBO | ID: biblio-1363062

ABSTRACT

Objective: 1) To critically review the published literature on applications of dental stem cells in the regeneration of intraoral tissues. 2) To provide an evidence-based level on research regarding application of dental stem cells in intraoral tissues regeneration. Methodology: This systematic review is conducted as per the JBI guidelines and reported as per the PRISMA. An initial literature search of papers published between 2004 and 2018 yielded 421 manuscripts. Nineteen studies satisfied the inclusion / exclusion criteria and were included for qualitative synthesis. Studies were categorized as animal (11) and human (8) trials. Five independent reviewers critically assessed the included studies. Risk of bias was assessed using SYstematic Review Centre for Laboratory animal Experimentation (SYRCLE) bias risk tool, robins-I tool for non-randomised clinical trial and Cochrane Collaboration's Tool for randomised clinical trial. Evidence levels were assessed based on JBI Criteria. Results: Animal trials mainly focused on periodontal regeneration. A high or unclear Risk of bias was more commonly found amongst animal studies. Laboratory, clinical and radiographic evaluation were used to assess the outcome. A total of Eight Human studies were conducted on a total samples size of 153 upon a wide age ranging from seven years to 60 years. Nearly 70% of the human studies used DPSC for regenerating alveolar bone defects. Conclusion: Appropriate well designed double-blind randomized clinical trials of longer duration are yet to be performed. Evidence for the included studies were 1C and 1D as per the JBI Criteria. Stem cell therapy demonstrated promising results in Periodontal tissue and alveolar bone regeneration. However, the number of studies to claim such a benefit are very limited (AU)


Objetivo: 1) Revisar criticamente a literatura publicada sobre aplicações de células-tronco dentárias na regeneração de tecidos intraorais. 2) Fornecer um nível baseado em evidências sobre pesquisas relacionadas à aplicação de células-tronco dentárias na regeneração de tecidos intraorais. Metodologia: Esta revisão sistemática é conduzida de acordo com as diretrizes do JBI e relatada de acordo com o PRISMA. Uma pesquisa bibliográfica inicial de artigos publicados entre 2004 e 2018 resultou em 421 manuscritos. Dezenove estudos satisfizeram os critérios de inclusão / exclusão e foram incluídos para síntese qualitativa. Os estudos foram categorizados como ensaios em animais (11) e humanos (8). Cinco revisores independentes avaliaram criticamente os estudos incluídos. O risco de viés foi avaliado usando a ferramenta de risco de viés do Centro de Revisão Sistemática para Experimentação com Animais de Laboratório (SYRCLE), a ferramenta robins-I para ensaios clínicos não randomizados e a Ferramenta da Colaboração Cochrane para ensaios clínicos randomizados. Os níveis de evidência foram avaliados com base nos critérios JBI. Resultados: Os ensaios em animais focaram principalmente na regeneração periodontal. Um risco alto ou pouco claro de viés foi mais comumente encontrado entre os estudos com animais. Avaliações laboratorial, clínica e radiográfica foram utilizadas para avaliar o resultado. Um total de oito estudos em humanos foram conduzidos em um tamanho total de amostras de 153 com ampla faixa etária, variando de sete a 60 anos. Quase 70% dos estudos em humanos usaram DPSC para regeneração de defeitos ósseos alveolares. Conclusão: Ensaios clínicos randomizados duplo-cegos apropriados e bem elaborados de maior duração ainda precisam ser realizados. As evidências para os estudos incluídos foram 1C e 1D de acordo com os critérios JBI. A terapia com células-tronco demonstrou resultados promissores na regeneração do tecido periodontal e do osso alveolar. No entanto, o número de estudos para reivindicar tal benefício é muito limitado (AU)


Subject(s)
Humans , Animals , Stem Cells , Tooth, Deciduous , Guided Tissue Regeneration, Periodontal , Dental Pulp
3.
Article | IMSEAR | ID: sea-210929

ABSTRACT

Milking behaviour of dairy cows has serious impacts on their production efficiency. A number of genetic and environmental factors controls and influences milking behaviour of dairy cow. The aim of present study was to investigate the influence of housing comfort on expressivity of milking behaviours of cows in parlour, milk yield and compositions. Forty Jersey crossbred cows of similar production levels were selected and divided into 2 groups based on age, production and parity. Subsequently, cows were kept in two different types of loose house; each containing 20 animals. Two types of housing patterns were compared - (i) Traditional shed (T0) and (ii) Thermo-comfortable shed (T1). Impact of housing comfort significantly transformed the expressions of dairy cows behaviour even in milking parlour. Milking temperament scores and stepping during milking were significantly lower in cows kept in T1 compared to that of T0. Cows of thermo-comfortable shed showed more docile, calm and less nervous behaviour than those kept in traditional shed. Housing patterns significantly influenced daily milk yield (kg) being 2.86% more in T1 as compared to T0. Similarly milk compositions were better and significantly higher in T1 group of cows than that of T0. It was concluded that staying comfort of living by resignificantly modulated the expression of dairy cows behaviours even in milking parlour, demonstrated favourable milking temperament, reduced nervousness, enhanced milk yield and showed better milk compositions in Jersey crossbred cows

4.
Clinics ; 75: e1656, 2020. graf
Article in English | LILACS | ID: biblio-1133444

ABSTRACT

OBJECTIVES: Mesenchymal stem cells (MSCs) are potentially ideal for type 2 diabetes treatment, owing to their multidirectional differentiation ability and immunomodulatory properties. Here we investigated whether the stem cells from human exfoliated deciduous teeth (SHED) in combination with hyperbaric oxygen (HBO) could treat type 2 diabetic rats, and explored the underlying mechanism. METHODS: SD rats were used to generate a type 2 diabetes model, which received stem cell therapy, HBO therapy, or both together. Before and after treatment, body weight, blood glucose, and serum insulin, blood lipid, pro-inflammatory cytokines (tumor necrosis factor-alpha and interleukin-6), and urinary proteins were measured and compared. After 6 weeks, rats were sacrificed and their organs were subjected to hematoxylin and eosin staining and immunofluorescence staining for insulin and glucagon; apoptosis and proliferation were analyzed in islet cells. Structural changes in islets were observed under an electron microscope. Expression levels of Pdx1, Ngn3, and Pax4 mRNAs in the pancreas were assessed by real-time quantitative polymerase chain reaction (RT-qPCR). RESULTS: In comparison with diabetic mice, those treated with the combination or SHE therapy showed decreased blood glucose, insulin resistance, serum lipids, and pro-inflammatory cytokines and increased body weight and serum insulin. The morphology and structure of pancreatic islets improved, as evident from an increase in insulin-positive cells and a decrease in glucagon-positive cells. Terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) staining of islet cells revealed the decreased apoptosis index, while Ki67 and proliferating cell nuclear antigen staining showed increased proliferation index. Pancreatic expression of Pdx1, Ngn3, and Pax4 was upregulated. CONCLUSION: SHED combined with HBO therapy was effective for treating type 2 diabetic rats. The underlying mechanism may involve SHED-mediated increase in the proliferation and trans-differentiation of islet β-cells and decrease in pro-inflammatory cytokines and apoptosis of islets.


Subject(s)
Humans , Animals , Male , Mice , Rats , Mesenchymal Stem Cell Transplantation , Diabetes Mellitus, Experimental/therapy , Diabetes Mellitus, Type 2/therapy , Insulin-Secreting Cells , Hyperbaric Oxygenation/methods , Stem Cells , Tooth, Deciduous , China , Rats, Sprague-Dawley , Diabetes Mellitus, Type 2/chemically induced , Mesenchymal Stem Cells , Insulin
5.
Article | IMSEAR | ID: sea-187764

ABSTRACT

Background: Stem cells from Human Exfoliated Deciduous teeth (SHED) were identified by Miura in 2003. SHED have been described as a suitable, accessible and potential source for regenerative medicine and therapeutic applications. However, the best group of deciduous teeth for the obtention of stem cells (SCs) has not been established. Therefore, this research aimed to determine the dental organs group from which SHED can be obtained with higher potentiality, considering their biomolecular features. Methodology: Deciduous teeth from 64 healthy children were collected and divided into two groups: anterior and posteriors. Dental pulp tissue was removed to determine their genetic, phenotypic, and spectroscopic profiles by RT-qPCR, immunofluorescence, and Fourier Transform Infrared (FTIR) spectroscopy respectively. Results: The results showed a higher gene (CD73 and NANOG) and protein (NANOG and SOX2) expression of mesenchymal and pluripotent markers in anterior SHED. CD146 gene expression between the two groups shows no statistical significant difference. Furthermore, the analysis of deciduous dental pulps by FTIR spectroscopy showed spectral bands related to biological samples, indicating the higher state of potentiality in anterior deciduous dental pulps. Conclusion: The deciduous dental pulp harbor a heterogenous population of SCs with different potentiality; however, the expression of multipotent and pluripotent markers was higher in the pulps from anterior deciduous teeth respect to posterior deciduous teeth. The storage and obtention of SHED from anterior teeth is more recommended respect to posterior teeth. However, it is necessary to analyze more stem cell markers and to study the differentiation capability of SHED.

6.
Journal of Practical Stomatology ; (6): 425-431, 2015.
Article in Chinese | WPRIM | ID: wpr-463572

ABSTRACT

Dental stem cells(DSCs)possess the characteristics of stem cells and can be effectively obtained from iatro-waste products (such as impacted wisdom tooth and the extracted teeth for orthodontic reason).It has been proved that DSCs are the important sources of stem cells for tissue engineering and regenerative medicine research.Research of these stem cells will create broader space for tissue engi-neering and regenerative medicine and will have important values in translational research.This review gives an overview of the research pro-gress of dental stem cells,and presents some new findings of several common dental stem cells as well as the application in tissue regenera-tion.

7.
Indian J Exp Biol ; 2014 Oct; 52(10): 1011-1016
Article in English | IMSEAR | ID: sea-153801

ABSTRACT

DNA from molted feathers is being increasingly used for genetic studies on birds. However, the DNA obtained from such non-invasive sources is often not of enough quantity and quality for isolation of new microsatellite markers. The present study examined the potential of shed feathers of near threatened Painted Stork as a source of its DNA for cross-species amplification of microsatellites. Thirty-one shed feathers of varying conditions (‘good’ and ‘deteriorated’) and sizes (‘large’, ‘intermediate’ and ‘small’) collected in a north Indian population were used to isolate DNA by a standard isopropanol method and 11 microsatellite markers already developed in the Wood Stork were screened for amplification. Nine plucked feathers from two dead Painted Storks were also used to compare the DNA yield and amplification success. The DNA yield of feathers varied significantly in relation to the calamus size and condition. Among molted feathers, ‘good’ and ‘large’ samples provided more DNA than ‘deteriorated’ and ‘small’ ones, respectively. ‘Large’ plucked feathers yielded more DNA than ‘large’ molted feathers. DNA was almost degraded in all the samples and ratio of absorbance at 260/280 nm varied from 1.0 to 1.8, indicating impurity in many samples. Independent of DNA yields, all microsatellites were cross-amplified in all kinds of feathers, with >80% success in different feather categories. It is concluded that the shed feathers can be successfully used to isolate DNA in the Painted Stork and for cross-species amplification of microsatellites.


Subject(s)
Animals , Birds/genetics , DNA/genetics , Feathers/chemistry , Genetics, Population/methods , Microsatellite Repeats , Species Specificity
8.
Journal of the Korean Ophthalmological Society ; : 1030-1038, 2014.
Article in Korean | WPRIM | ID: wpr-89991

ABSTRACT

PURPOSE: To investigate the influence of water-shed zone (WSZ) and nocturnal dip (ND) on the progression of the glaucomatous visual field (V/F) defects in open-angle glaucoma (OAG) patients when the intraocular pressure (IOP) was maintained under the target pressure. METHODS: We performed fluorescence angiography (FAG), 24-hour ambulatory blood pressure monitoring (24-hr ABPM), and V/F tests. We examined the relationships among WSZ in early-FAG, ND over 10% (dip), and the progression of the glaucomatous V/F defects using chi-square, Fisher's exact, and multivariate logistic regression tests. A p-value < 0.05 was considered statistically significant. RESULTS: When considering the correlation between WSZ and dip, statistical significance was found in OAG (p = 0.024, odds ratio (OR) = 3.308) and normal tension glaucoma (NTG) (p = 0.029, OR = 4.364) patients. In patients with dip, glaucomatous V/F defects significantly progressed (OAG: p = 0.003, OR = 5.938, NTG: p = 0.005, OR = 13.929). In patients with WSZ, the glaucomatous V/F defects progressed in all groups (OAG: p = 0.002, OR = 5.156, NTG: p = 0.024, OR = 4.750, primary open angle glaucoma (POAG): p = 0.021, OR = 8.750). In the patients with WSZ involving optic nerve head, the glaucomatous V/F defects had progressed in OAG (p = 0.004, OR = 5.958) and NTG (p = 0.009, OR = 8.333) groups. Based on binary logistic regression analysis, dip (p = 0.010, OR = 6.227) significantly affected V/F progression only in OAG patients. CONCLUSIONS: In the OAG and NTG groups, ND over 10% influenced the progression of the glaucomatous V/F defects. The patients with WSZ tended to have ND over 10% in OAG and NTG groups and glaucomatous V/F defects progressed in all patients. Therefore, performing early FAG and 24-hr ambulatory blood pressure monitoring may be helpful for glaucoma patients with progressing glaucomatous V/F defects even when the IOP was maintained under the target pressure.


Subject(s)
Humans , Blood Pressure Monitoring, Ambulatory , Fluorescein Angiography , Glaucoma , Glaucoma, Open-Angle , Intraocular Pressure , Logistic Models , Low Tension Glaucoma , Odds Ratio , Optic Disk , Visual Fields
9.
Indian J Exp Biol ; 2013 Mar; 51(3): 235-240
Article in English | IMSEAR | ID: sea-147587

ABSTRACT

The snake shed skin though considered as biological waste products have been mentioned in folk and traditional medicine for treatment of ailments like skin disorders, parturition problems etc. Shedded skin extract (5 mg.kg-1, sc) did not produce any change in the estrous cycle of normal cycling female mice. However in 10 mg.kg-1, sc dose, the extract caused a temporary cessation of the estrous cycle at diestrous phase in normal cycling female mice for 10 days. SSAE (10 mg.kg-1, sc) caused a significant change in the level of LH, FSH, progesterone, estradiol, IL-1β, IL-6 and TNF-α. Histopathology of uterus and ovary showed structural disorientation in both. The results substantiate the influence of snake shed skin in mice reproductive cycle.


Subject(s)
Animals , Cytokines/metabolism , Elapidae , Estradiol/metabolism , Estrous Cycle/drug effects , Estrous Cycle/metabolism , Female , Fertility/drug effects , Gene Expression Regulation/drug effects , Hormones/metabolism , Mice , Ovary/metabolism , Ovary/pathology , Progesterone/metabolism , Reproduction , Skin/chemistry , Uterus/metabolism , Uterus/pathology
10.
The International Medical Journal Malaysia ; (2): 19-26, 2013.
Article in English | WPRIM | ID: wpr-627298

ABSTRACT

Stem cells from human exfoliated deciduous teeth (SHED) are highly proliferative, clonogenic cells capable of differentiating into osteoblasts and inducing bone formation. It is a potential alternative for stem cell bone regeneration therapy. However, stem cell therapy carries the risk of immune rejection mediated by inflammatory cytokines of the human defense system. Objective: This preliminary research studies the interaction between SHED and the immune system by determining the inflammatory cytokines profile and osteogenic potential of SHED. Methods: Human fetal osteoblasts (hFOb) cell line and isolated SHED were cultured and total RNA was extracted, followed by reverse transcription cDNA synthesis. Semi-quantitative reverse transcription PCR and Multiplex PCR were performed to detect the expression levels of OPG/RANKL and TNF-α, IL-1β, IL-6, IL-8 and TGF-β in both cell types. Results: Analysis showed that SHED expressed significantly lower amounts of IL-1β, IL-6, and IL-8 compared to hFOB. IL-1β is a potent bone-resorbing factor, while IL-6 and IL-8 induce osteoclastogenesis and osteolysis respectively. SHED did not express TNF-α which stimulates osteoclastic activity. SHED demonstrated high OPG/RANKL ratio, in contrast with that of marrow stem cells described in previous studies. Our findings suggest that SHED may have improved immunomodulatory profile in terms of promoting relatively lower inflammatory reaction during transplant and enhancing bone regeneration. Conclusion: SHED has a potential to be a good source of osteoblasts for bone regeneration therapy. Further studies on the immunomodulatory properties of SHED-derived osteoblasts are necessary to enable stem cell therapy in immunocompetent hosts.

11.
Article in English | IMSEAR | ID: sea-144143

ABSTRACT

Stem cells of the dental pulp are a population of postnatal stem cells with multilineage differentiation potential. These cells are derived from the neural ectomesenchyme, similar to most craniofacial tissues, and specific niches in the pulp have been identified. Since the isolation of dental pulp stem cells (DPSC) and stem cells from exfoliating deciduous teeth (SHED), numerous studies have attempted to define and characterize these cells, and embryonic stem cell features have been reported in both DPSC and SHED. These cells have a vast repertoire of differentiation - osteogenic, odontogenic, myogenic, adipogenic, neurogenic, and melanocytic, and have even demonstrated transdifferentiation to corneal cells and islet cells of pancreas. The combined advantages of multipotency/pluripotency and the relative ease of access of pulp tissue for autologous use render DPSC/ SHED attractive options in regenerative dentistry and medicine. This review gives a bird's eye view of current knowledge with respect to stem cells from the dental pulp.


Subject(s)
Dental Pulp , Humans , Multipotent Stem Cells/cytology , Pluripotent Stem Cells/cytology , Stem Cells , Tooth, Deciduous/cytology
12.
Archives of Orofacial Sciences ; : 15-20, 2011.
Article in English | WPRIM | ID: wpr-627421

ABSTRACT

The aim of this study was to determine the genotoxicity of a locally produced nanocomposite by Universiti Sains Malaysia, Malaysia using Comet assay. Stem cells from human exfoliated deciduous teeth (SHED) were treated with the nanocomposite at five different concentrations (0.006, 0.0125, 0.025, 0.05, and 0.1 mg/ml) along with concurrent negative (medium alone) and positive control (zinc sulfate heptahydrate) and incubated at 37°C for 24 hours in an incubator at 5% CO2. The tail moment was used to assess the extent of DNA damage. The tail moment for the group of SHED treated with nanocomposite (for all the five different concentrations) was not statistically significant as compared to the negative control, suggesting that the locally produced dental nanocomposite did not induce any DNA damage. Hence, it can be concluded that the locally produced nanocomposite is non-genotoxic on stem cells from human exfoliated deciduous teeth.

13.
RBCF, Rev. bras. ciênc. farm. (Impr.) ; 44(2): 233-248, abr.-jun. 2008. graf, tab
Article in Portuguese | LILACS | ID: lil-488720

ABSTRACT

A rutina é empregada como antioxidante e na prevenção da fragilidade capilar. Estudos de penetração in vitro através da pele humana seria a situação ideal, entretanto, há dificuldades de sua obtenção e manutenção de sua viabilidade. Entre os demais modelos de membrana, a muda de pele de cobra se apresenta como estrato córneo puro, fornecendo barreira similar ao humano e é obtida sem a morte do animal. Os objetivos desta pesquisa foram desenvolver e avaliar a estabilidade de uma emulsão cosmética contendo rutina e, como promotor de penetração cutânea, o propilenoglicol; e avaliar a penetração e a retenção cutânea in vitro da referida substância ativa da formulação, empregando um modelo de biomembrana alternativo. A emulsão foi desenvolvida com rutina e propilenoglicol, ambos a 5,0 por cento p/p. Quantificou-se a rutina das emulsões por espectrofotometria a 361,0 nm, método previamente validado. A penetração e retenção cutânea in vitro foram realizadas em células de difusão vertical com muda de pele de cobra de Crotalus durissus, como modelo de biomembrana alternativo, e água destilada e álcool etílico absoluto 99,5 por cento (1:1), como fluido receptor. O experimento foi conduzido em um período de seis horas, a 37,0 ± 0,5 ºC e agitação constante de 300 rpm. Empregou-se o método espectrofotométrico validado a 410,0 nm para a quantificação da rutina após penetração e retenção cutânea. A emulsão não promoveu a penetração cutânea da rutina através da muda de pele de C. durissus, retendo 0,931 ± 0,0391 mg de rutina/mg de muda de pele de cobra. Nas condições de armazenamento a 25,0 ± 2,0 ºC; 5,0 ± 0,5 ºC e 45,0 ± 0,5 ºC, a emulsão apresentou-se quimicamente estável durante 30 dias. De acordo com os resultados, a emulsão não favoreceu a penetração cutânea da rutina, mas apenas sua retenção no estrato córneo de C. durissus, condição considerada estável no período de 30 dias.


Rutin is employed as antioxidant and to prevent the capillary fragility and, when incorporated in cosmetic emulsions, it must target the action site. In vitro cutaneous penetration studies through human skin is the ideal situation, however, there are difficulties to obtain and to maintain this tissue viability. Among the membrane models, shed snake skin presents itself as pure stratum corneum, providing barrier function similar to human and it is obtained without the animal sacrifice. The objectives of this research were the development and stability evaluation of a cosmetic emulsion containing rutin and propylene glycol (penetration enhancer) and the evaluation of rutin in vitro cutaneous penetration and retention from the emulsion, employing an alternative model biomembrane. Emulsion was developed with rutin and propylene glycol, both at 5.0 percent w/w. Active substance presented on the formulation was quantified by a validated spectrophotometric method at 361.0 nm. Rutin cutaneous penetration and retention was performed in vertical diffusion cells with shed snake skin of Crotalus durissus, as alternative model biomembrane, and distilled water and ethanol 99.5 percent (1:1), as receptor fluid. The experiment was conducted for six hours, at 37.0 ± 0.5 ºC with constant stirring of 300 rpm. Spectrophotometry at 410.0 nm, previously validated, determined the active substance after cutaneous penetration/retention. Emulsion did not promote rutin cutaneous penetration through C. durissus skin, retaining 0.931 ± 0.0391 mg rutin/mg shed snake skin. The referred formulation was chemically stable for 30 days after stored at 25.0 ± 2.0 ºC, 5.0 ± 0.5 ºC and 45.0 ± 0.5 ºC. In conclusion, it has not been verified the active cutaneous penetration through the model biomembrane, but only its retention on the Crotalus durissus stratum corneum, condition considered stable for 30 days.


Subject(s)
Cosmetic Stability , Emulsions , Propylene Glycol , Rutin/metabolism , Skin Absorption
14.
Korean Journal of Obstetrics and Gynecology ; : 82-86, 2000.
Article in Korean | WPRIM | ID: wpr-204495

ABSTRACT

OBJECTIVE: To evaluate the viability and the characteristics of shed endometrial tissues obtained from menstrual fluid during in-vitro culture. METHODS: The menstrual fluids were collected using Wallace catheter from uterine cavity in 10 women with regular menstruation. The menstrual fluids were washed twice, and the pellets, containing blood cells and shed endometrium, were collected and diluted fivefold with Ham's F-10 medium containing 10% fetal bovine serum. The cell suspension was placed on culture dishes, and cultured for 7 days in an incubator. To evaluate the characteristics of the cultured endometrial cells, immunohistochemical (IHC) staining was performed using anti-cytokeratin and anti-vimentin antibody. RESULTS: The mean volume of menstrual fluids and pellets were 0.7ml and 0.3ml, respectively. Only 15% of the shed endometrial tissues were attached and proliferated in culture dishes, which was considered to have viability. Initially, endometrial epithelial cells and fibroblasts were attached and proliferated, and the area of these cells was increased according to prolong the culture time. Stromal cell colonys were located and proliferated on the epithelial cells. IHC staining showed strongly positive for cytokeratin in epithelial cells and for vimentin in stromal cells. In the confocal microscopic observation of 3-dimensional structure of cultured endometrium, cytokeratin-positive cells (epithelial cells) were located in the pheriphery and cytokeratin-negative cells (stromal cells) inside of the structure. CONCLUSION: From our study, shed endometrial tissues in menstrual fluid showed meaningful viability and closed relationship between epithelial cells and stromal cells during in-vitro culture. Thus, we suggest that the in-vitro culture system of shed endometrium is a suitable model for researches of endometriosis.


Subject(s)
Female , Humans , Blood Cells , Catheters , Endometriosis , Endometrium , Epithelial Cells , Fibroblasts , Incubators , Keratins , Menstruation , Stromal Cells , Vimentin
15.
The Journal of the Korean Orthopaedic Association ; : 702-706, 1998.
Article in Korean | WPRIM | ID: wpr-644464

ABSTRACT

In the orthopaedic field, some elective surgeries such as joint replacement, spinal surgery and limb salvage procedures for musculoskeletal tumors frequently need various amounts of blood transfusions. However, homologous transfusion occasionally results in various side effects, such as allergic reaction, febrile reaction, and the transmission of infectious diseases such as syphilis, hepatitis and AIDS, ctc. Recently, these complications especially in elective surgery might result in medicolegal or social problems. Risks from transfusions in elective surgery can be minimized with prebanked autologous transfusion. To evaluate the necessity of prehanked autogenous transfusion, fifty five patients who had unilateral hybrid total knee arthroplasty (noncemented at the femoral side and cemented at the tibial and patellar sides) were operated on by the same surgeon from April 199S to July 1997 and had autogenous shed blood transfusion were evaluated for postoperative blood loss, amount of autogenous shed blood, amount of transfusion, hemoglobin and hematocrit. The results were as follows: 1. The distribution of preoperative hemoglobin was from 9.6g/dL to 16.5g/dL (average: 1.8g/dL). 2. The distribution of the amount of blood loss for three days postoperatively was from 156ml to 2001 ml (average: 798ml). 3. The distrihution of the amount of transfusion of autogenous shed blood was from 30ml to 600ml (average: 448ml). 4. There were two patients who had febrile reactions above 38 after transfusion of autogenous shed blood. 5. Forty-six patient(84%) had a homologous transfusion and the average amount of transfusion was 1.9 pint. 6. Total amount of homologous transfusion was decreased according to the increased amount of hemoglobin and the amount of transfusion was statistically decreased above the level of I 3g/dL(Students t-test, P=0.0005). 7. There were no significant differences in the amount of homologous transfusion between age, sex, type of disease, type of implants. In conclusion, most of our patients(84%) needed homologous blood transfusion in unilateral hyhrid total knee arthroplasty and the amount of transfusion decreased in patients who had hemoglobin above 13.0g/dL. So we recommend preparing banked autogenous hlood preoperatively in patients who have a lower hemoglobin level in unilateral hyhrid total knee arthroplasty.


Subject(s)
Humans , Arthroplasty , Blood Transfusion , Communicable Diseases , Hematocrit , Hepatitis , Hypersensitivity , Joints , Knee , Limb Salvage , Postoperative Hemorrhage , Social Problems , Syphilis
16.
The Journal of the Korean Orthopaedic Association ; : 651-657, 1995.
Article in Korean | WPRIM | ID: wpr-769672

ABSTRACT

The major purpose for the use of autotransfusion is to prevent the transmission of blood borne infectious agents, such as human immunodeficiency virus and non-A & non-B hepatitis virus. To evaluate the efficacy and quality of autolgous shed blood for autotransfusion, eighty patients who had total knee arthroplasty from Dec. 1992 to Mar. 1994, were included in one of two groups: Group I, who received the autotransfusion, or Group II, who did not. Each group included 20 patients of unilateral TKR and 20 patients of bilateral TKR. The Orth-evac system(Deknatel, USA) was used to salvage drained blood in the first six hours after the operation. All of the patients were evaluated for the postoperative blood loss, transfusion requirements, hemoglobin, hematocrit, platelet count, blood pressure and body temperatures. l. In bilateral TKR, the reinfusion of shed blood reduced the requirements for homologous blood by 41.4%(1.2 pints in group I versus 2.9 pints in group II). In unilateral TKR, it was decreased to 36.4%(0.4 pint in group I vs 1.1 pint in group II). 2. In bilateral TKR, the requirements for homologous transfusion was decreased from 95% of patients in control group to 55% in group I .In unilateral TKR, it was decreased from 60% to 20%. 3. There were four patients who had high fever above 39℃ after autotransfusion. 4. At the immediate postoperative period there were two patients who had hypovolemic shock in group I patients who had bilateral TKR. 5. There was no clotting abnormality, no transfusion reaction and no thromboembolic disease in group I patients. In conclusion, the reinfusion of autologous shed blood after TKR is an acceptable alternative to the homologous transfusion without untoward effect.


Subject(s)
Humans , Arthroplasty, Replacement, Knee , Blood Transfusion, Autologous , Body Temperature , Fever , Hematocrit , Hepatitis Viruses , HIV , Knee , Platelet Count , Postoperative Hemorrhage , Postoperative Period , Shock , Transfusion Reaction
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