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BACKGROUND@#Large-scale muscle tissue engineering remains a major challenge. An axial vascular pedicle and perfusion bioreactor are necessary for the development and maintenance of large-scale engineered muscle to ensure circulation within the construct. We aimed to develop a novel experimental model of a large-scale engineered muscle flap from an existing rat groin fat flap.@*METHODS@#A fat flap based on the superficial inferior epigastric vascular pedicle was excised from rats and placed into a perfusion bioreactor. The flaps were kept in the bioreactor for up to 7 weeks, and transdifferentiation of adipose to muscle tissue could have taken place. This system enabled myogenic-differentiation medium flow through the bioreactor at constant pH and oxygen concentration. Assessment of viability was performed by an immunofluorescence assay, histological staining, a calcein-based live/dead test, and through determination of RNA quantity and quality after 1, 3, 5, and 7 weeks.@*RESULTS@#Immunofluorescence staining showed that smooth muscle around vessels was still intact without signs of necrosis or atrophy. The visual assessment of viability by the calcein-based live/dead test revealed viability of the rat adipose tissue preserved in the bioreactor system with permanent perfusion. RNA samples from different experimental conditions were quantified by spectrophotometry, and intact bands of 18S and 28S rRNA were detected by gel electrophoresis, indicating that degradation of RNA was minimal.@*CONCLUSIONS@#Flow perfusion maintains the long-term viability of a rat groin engineered muscle flap in vitro, and a large-scale vascularized muscle could be engineered in a perfusion bioreactor.
Subject(s)
Animals , Male , Rats , Bioreactors , Groin , Perfusion , RNA , Rats, Inbred Lew , Surgical Flaps , Tissue EngineeringABSTRACT
<p><b>BACKGROUND</b>Large-scale muscle tissue engineering remains a major challenge. An axial vascular pedicle and perfusion bioreactor are necessary for the development and maintenance of large-scale engineered muscle to ensure circulation within the construct. We aimed to develop a novel experimental model of a large-scale engineered muscle flap from an existing rat groin fat flap.</p><p><b>METHODS</b>A fat flap based on the superficial inferior epigastric vascular pedicle was excised from rats and placed into a perfusion bioreactor. The flaps were kept in the bioreactor for up to 7 weeks, and transdifferentiation of adipose to muscle tissue could have taken place. This system enabled myogenic-differentiation medium flow through the bioreactor at constant pH and oxygen concentration. Assessment of viability was performed by an immunofluorescence assay, histological staining, a calcein-based live/dead test, and through determination of RNA quantity and quality after 1, 3, 5, and 7 weeks.</p><p><b>RESULTS</b>Immunofluorescence staining showed that smooth muscle around vessels was still intact without signs of necrosis or atrophy. The visual assessment of viability by the calcein-based live/dead test revealed viability of the rat adipose tissue preserved in the bioreactor system with permanent perfusion. RNA samples from different experimental conditions were quantified by spectrophotometry, and intact bands of 18S and 28S rRNA were detected by gel electrophoresis, indicating that degradation of RNA was minimal.</p><p><b>CONCLUSIONS</b>Flow perfusion maintains the long-term viability of a rat groin engineered muscle flap in vitro, and a large-scale vascularized muscle could be engineered in a perfusion bioreactor.</p>
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<p><b>Background</b>Fat grafting technologies are popularly used in plastic and reconstructive surgery. Due to its size limitation, it is hard to directly inject untreated fat tissue into the dermal layer. Nanofat, which was introduced by Tonnard, solves this problem by mechanically emulsifying fat tissue. However, the viability of the cells was greatly destroyed. In this study, we reported a new method by "gently" digesting the fat tissue to produce viable adipocytes, progenitors, and stromal stem cells using collagenase I digestion and centrifugation. This was named "Vivo nanofat".</p><p><b>Methods</b>Human liposuction aspirates were obtained from five healthy female donors with mean age of 28.7 ± 5.6 years. Colony-forming assay, flow cytometry analysis, and adipogenic and osteogenic induction of the adherent cells from the Vivo nanofat were used to characterize the adipose mesenchymal stem cells (MSCs). To investigate in vivo survival, we respectively injected Vivo nanofat and nanofat subcutaneously to the back of 8-week-old male BALB/c nude mice. Samples were harvested 2 days, 2 weeks, and 4 weeks postinjection for measurement, hematoxylin and eosin staining, and immunostaining.</p><p><b>Results</b>Our results showed that the Vivo nanofat contained a large number of colony-forming cells. These cells expressed MSC markers and had multi-differentiative potential. In vivo transplantation showed that the Vivo nanofat had lower resorption ratio than that of nanofat. The size of the transplanted nanofat was obviously smaller than that of Vivo nanofat 4 weeks postinjection (0.50 ± 0.17 cm vs. 0.81 ± 0.07 cm, t = -5783, P = 0.01).</p><p><b>Conclusion</b>Vivo nanofat may serve as a cell fraction injectable through a fine needle; this could be used for cosmetic applications.</p>
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The cystine knot (CK) motif comprises an internal ring formed by two disulfide bonds and their connecting backbone segments which is threaded by a third disulfide bond .It is present in peptides and proteins of a variety of species ,in-cluding fungi ,plants ,marine molluscs ,insects and spiders .CK polypeptide is one of the ideal model molecules for drug design and molecular engineering research because of its stable structure and variety of bioactivities .Here we summarized the main structural features of both inhibitor cystine knot (ICK) peptide and cyclic cystine knot (CCK) peptide ,including primary se-quence ,topology ,permutation ,synthesis and folding characteristics ,as well as its applications on drug design and molecular engineering .
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<p><b>BACKGROUND</b>Many studies on periostin have focused on its role in tumors and vascular reconstruction. However, the effect of periostin on stem cell function remains unclear. The aim of this study was to enhance vitality in adipose-derived stem cells (ADSCs), the effect of periostin on the function of ADSCs was observed.</p><p><b>METHODS</b>Human ADSCs (hADSCs) were isolated from human adipose tissue by collagenase I digestion and collected in multi-periods for in vitro culture. CD29, CD34, CD44, CD45 and CD105 were detected by flow cytometry. In addition, directed differentiation of hADSCs was induced using adipogenic, osteogenic and chondrogenic induction mediums. The induced morphological changes were observed using oil red O, Alizarin red and alcian blue staining. Periostin was administered to hADSCs in an acidic environment. The treatments of cells were divided into three groups: a periostin group (P); an acidic control group (A); a normal group (N). Then the resulting cell proliferation and migration were detected using a Cell Counting Kit-8 (CCK-8) and a transwell chamber assay, respectively.</p><p><b>RESULTS</b>The detection rates of CD29, CD44, CD105, CD34 and CD45 were 98.89%, 93.73%, 86.99%, 0.19% and 0.16%. The specific staining of cells was positive after induction culture. The mean absorbance of the cells in group P and A at 12 hours were 16.67% and 22.22% greater than group N, respectively (P < 0.01). The mean absorbance of cells from group P was 20.00% greater than that of group A at 48 hours (P < 0.05). The mean number of migratory cells per visual field in group A was 50.38% lower than that in group N (P < 0.05). The migratory cell number in group P was 119.98% greater than that in group A (P < 0.05).</p><p><b>CONCLUSIONS</b>The acidic environment impacted hADSC proliferation and inhibited cell migration. However, periostin was able to promote the proliferation and migration of hADSCs despite the acidic environment.</p>
Subject(s)
Adult , Female , Humans , Adipose Tissue , Cell Biology , Antigens, Surface , Cell Adhesion Molecules , Pharmacology , Cell Differentiation , Cell Movement , Cell Proliferation , Cells, Cultured , Stem Cells , PhysiologyABSTRACT
OBJECTIVE@#To explore effect of calcium citrate on bone integration in a rabbit femur defect model, and to compare the bone formation with different sizes by radiological and histological study.@*METHODS@#Twenty-four male Japanese white rabbits were randomly divided into three groups (Group A, B, C) in this study. Under anesthesia, defects of four sizes (1.2, 1.5, 2.0 and 2.5 mm) were created in each of the rabbits. Commercially pure calcium citrate powder was placed inside the medullary compartment of the femur (Experimental), while in the contralateral femur (Control) nothing was implanted. The defects were analyzed using radiography and histological analysis by using Imagepro-Plus 6.0 software after animal was sacrificed at 4th(Group A), 6th(Group B) and 8th(Group C) weeks postoperatively. Four samples were analyzed for each size of defect and each healing period.@*RESULTS@#The histological and the radiologic evaluation were performed after sacrification of all rabbits on postoperative 4th and 6th weeks, It showed significant difference between the experimental group and the control group when these defects were less than or equal to 2.0 mm. No statistical difference was observed when these defects were larger than 2.0 mm at all healing periods except at the 4th week.@*CONCLUSIONS@#Calcium citrate affects the early periods of bone defects healing mechanism in Japanese white rabbits positively, especially when the defect is not too large. We suggest further studies on calcium citrate to determine the effects of various dosages, administration ways and the experimental time on the bone defects.
Subject(s)
Animals , Male , Rabbits , Bone Density Conservation Agents , Pharmacology , Bone Regeneration , Calcium Citrate , Pharmacology , Femur , Diagnostic Imaging , Radiography , Random Allocation , Wound HealingABSTRACT
Objective: To separate the nematocyst venom (NV) and tentacle-only extract (TOE) from the jellyfish Cyanea capillata and to analyze the difference between their haemolytic activities and the influencing factors. Methods: NV and TOE were separated by autolysis and centrifugation. The influence of concentration, temperature and pH value on the haemolytic activities of NV and TOE were observed. Results: NV and TOE were successfully separated. The concentration-associated haemolytic curves were "S" shaped for both NV and TOE. The HU50 of NV and TOE were 8 μg/ml and 67 μg/ml, respectively; and the haemolysis of NV was about 8.4 times of that of TOE. Temperature had great influence on haemolysis of both and the highest haemolysis were both at 40°C. pH value also had influence on haemolysis. The strongest haemolysis activity was found both at pH 8 and TOE was more sensitive. Conclusion: NV and TOE both have haemolysis activity, with the activity of NV stronger than that of the latter. The haemolysis activity is influenced by concentration, temperature and pH value.
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AIM To study the therapeutic effect of bone marrow transplantation on mustard gas-poisoned mice. METHODS Mustard-poisoned mice model was constructed by injected mustard gas 20 mg·kg~(-1) subcutaneously. At 4 h, 1 d and 2 d after poisoned by mustard gas, the suspended bone marrow cells (106 cells per time) were injected into mice respectively. After poisoned 3, 5, 7 and 10 d, the numbers of leukocyte and marrow nucleated cells were recorded periodically. At the same time, after poisoned 4 h and 1 d by mustard gas, mice in another group were injected granulocyte colony stimulating factor (G-CSF) 74 μg·kg~(-1) combined with transplantation and changes of the cell number were observed. RESULTS Compared with poisoned group without transplantion, leukocyte number in bone marrow transplanted group presented ascending trend, but there was no statistical significance until the 10 d after poisoned. However, the number of marrow nucleated cell increased significantly from the 5th day. After mice injected bone marrow cells combined with G-CSF, leukocyte increased markedly in earlier stage compared with transplantation only. CONCLUTION The bone marrow transplantation has significant effects on improvement of marrow hematogenic system inhibited by mustard gas.
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<p><b>OBJECTIVE</b>To probe into the mechanism of fibrillin 1 in pathologic scar, by examining the expressions of fibrillin 1 and TGF-beta1 as well as their correlations in the tissues of keloid, hypertrophic scar and normal skin.</p><p><b>METHODS</b>The tissues of keloid, hypertrophic scar and normal skin were tested. RT-PCR was used to assess the mRNA expression levels of the aimed genes. The distribution of fibrillin 1 in scars and normal skin was examined by immunohistochemistry staining.</p><p><b>RESULTS</b>The mRNA level of fibrillin 1 in keloid (0.802 +/- 0.116) was increased by 218.25% (P < 0.01) than that in normal skin (0.252 +/- 0.067). The expression of the gene in hypertrophic scar (0.628 +/- 0.144) was higher by 149.21% (while, P > 0.05) than that in normal skin. The expression of TGF-beta1 in keloid and hypertrophic scar were more than that in normal skin. The expression of fibrillin 1 was related to that of TGF-beta1 positively (r = 0.820, P < 0.01). Fibrillin 1 protein was stained positively in basic membranes, endothelial cells, fibroblasts and extracellular matrix of skin tissues. In dermal, the protein levels of fibrillin 1 in keloid (0.117 +/- 0.042) was decreased than those in normal skin (0.185 +/- 0.043) and hypertrophic scar (0.181 +/- 0.048), the inhibition rates were 36.76%, 35.36% respectively (both P < 0.01).</p><p><b>CONCLUSIONS</b>The expression of fibrillin 1 in keloid was changed and related to the expression of TGF-beta1 positively, which appears that fibrillin 1 was a cicatrix specific gene. Fibrillin 1 might play an important role in the formation of keloid.</p>
Subject(s)
Humans , Cicatrix, Hypertrophic , Metabolism , Pathology , Fibrillin-1 , Fibrillins , Keloid , Metabolism , Pathology , Microfilament Proteins , Metabolism , RNA, Messenger , Genetics , Transforming Growth Factor beta1 , Genetics , MetabolismABSTRACT
<p><b>OBJECTIVE</b>To probe into periostin's role in the pathological mechanism of hyperplasic scars, by examining the expression of periostin in hyperplasic scar tissues. To investigate the correlations between periostin and TGF-beta1, TGF-beta R I, TGF-beta R II.</p><p><b>METHODS</b>RT-PCR was used to assess the mRNA expression levels of TGF-beta1, TGF-beta R I, TGF-beta R II in three kinds of tissues, which are keloid (K), hypertrophic scar (HS) and normal skin (SK). The protein expression of periostin was measured with Western blotting.</p><p><b>RESULTS</b>The mRNA level of periostin in K was higher than that in SK. The mRNA expression of TGF-beta1 in K was higher than that in HS and SK. The mRNA level of TGF-beta R I in K was higher than that in HS and SK. The significances above all was at P < 0.01. The protein expression level of periostin in HS increased, compared with that in SK (P < 0.05). Periostin was related to TGF-beta1 positively (P <0.01).</p><p><b>CONCLUSIONS</b>The periostin's expression is increased in keloids. Periostin is a cicatrix specific gene. Periostin appears to play an important role in the formation of keloids, which is related to TGF-beta1 closely.</p>
Subject(s)
Adult , Female , Humans , Male , Cell Adhesion Molecules , Metabolism , Cicatrix, Hypertrophic , Metabolism , Pathology , Keloid , Metabolism , Pathology , Receptors, Transforming Growth Factor beta , Metabolism , Transforming Growth Factor beta1 , MetabolismABSTRACT
OBJECTIVE@#To explore the risk factors of gastric cancer in the rural area of Henan province.@*METHODS@#Three hundred and twenty-five families with gastric cancer and 325 control families (1010 persons in each group) were selected among the rural residents in 4 counties of Henan province. Totally 2020 people were surveyed and assessed using population-based case-control family study.@*RESULTS@#Gastric cancer was related to stomach upset, irregular dietary, hobby for salty taste, residual food, and history of mental stimulus.@*CONCLUSION@#Stomach upset, irregular dietary, hobby for salty taste, residual food, and history of mental stimulus are the risk factors of gastric cancer.
Subject(s)
Humans , Case-Control Studies , China , Epidemiology , Feeding Behavior , Risk Factors , Rural Population , Stomach Neoplasms , Genetics , Surveys and QuestionnairesABSTRACT
This paper introduces several novel HPC-based monitoring devices for community medicine. They support net transmission and have superiorities of portability, small size, good mobility, easy use and strong adaptivity.
Subject(s)
Humans , Blood Pressure Monitoring, Ambulatory , Community Health Services , Computers, Handheld , Electrocardiography, Ambulatory , Equipment Design , Monitoring, Physiologic , TelemedicineABSTRACT
We constructed the AFP promotor, suicide gene and EGFP eukaryotic expression vector recombinant plasmid, and this plasmid DNA was encapsulated by biodegradable, biocompatible polymer PLGA to prepare nanoparticles. The results demonstrate that the mean diameter of DNA-PLGA-NP is 68 nm, the encapsulation ratio reaches to 80%, and the PLGA nanoparticles can protect plasmid DNA from digestion by deoxyribonuclease I (DNaseI) and sonication-induced shearing in vitro.