scholarly journals Establishment of a modified percutaneous CT-guided paraspinal intramuscular VX-2 squamous cell carcinoma dual tumor model in rabbits

PeerJ ◽  
2021 ◽  
Vol 9 ◽  
pp. e11536
Author(s):  
Liangliang Meng ◽  
Husheng Shan ◽  
Xiaofeng He ◽  
Jiantao Zhou ◽  
Jingxiang Huang ◽  
...  

Background The rabbit VX-2 tumor model is a commonly used transplanted tumor model and is widely used in surgical, radiological, and interventional studies. Most of the known tumor models for each site are single solid tumors. This study aimed to establish an accurate and stable intramuscular dual tumor model guided by computed tomography (CT). Methods In this study, we compared three different inoculation methods to select the most appropriate dual tumor model. Six New Zealand White rabbits were used as tumor-carrying rabbits for tumor harvesting. Thirty rabbits were divided into three groups as experimental rabbits. Group A applied the tumor cell suspension method, in which the suspension was injected into the designated location with a syringe under CT guidance. Groups B and C used tumor tissue strips obtained in vivo or under direct in vitro vision. The tumor tissue strips were implanted into the designated locations using a guide needle under CT guidance. The differences in tumorigenic rate, the size difference between bilateral tumors, and metastasis between the three methods were compared. Results It was found that group A obtained a 100% tumor survival rate, but the size of the tumor was more variable, and needle tract implantation metastasis occurred in 5 cases. In group B, tumor tissue strips were taken in vivo for implantation, in which one case failed to survive. Tumor tissue strips in group C were obtained in vitro under direct vision. The tumor tissue strips obtained in vitro by puncture using a biopsy needle in group C had a 100% tumorigenicity rate and stable tumor size. No significant needle tract implantation metastases were found in either group B or C. The variance of tumor size obtained in group A was significantly higher than in groups B and C. The variance of tumor size in group C was the smallest. Group C had high tumorigenicity and a more stable size and morphology of the formed tumors. Conclusion The results showed that the method of obtaining tumor tissue strips using in vitro direct vision puncture and implanting them into the muscle with CT guidance and guide needles can establish an accurate and stable dual tumor model. This dual tumor model can provide substantial support for relevant preclinical studies.

2018 ◽  
Vol 47 (1) ◽  
pp. 212-221 ◽  
Author(s):  
Cecilia Pascual-Garrido ◽  
Elizabeth A. Aisenbrey ◽  
Francisco Rodriguez-Fontan ◽  
Karin A. Payne ◽  
Stephanie J. Bryant ◽  
...  

Background: In this study, we investigate the in vitro and in vivo chondrogenic capacity of a novel photopolymerizable cartilage mimetic hydrogel, enhanced with extracellular matrix analogs, for cartilage regeneration. Purpose: To (1) determine whether mesenchymal stem cells (MSCs) embedded in a novel cartilage mimetic hydrogel support in vitro chondrogenesis, (2) demonstrate that the proposed hydrogel can be delivered in situ in a critical chondral defect in a rabbit model, and (3) determine whether the hydrogel with or without MSCs supports in vivo chondrogenesis in a critical chondral defect. Study Design: Controlled laboratory study. Methods: Rabbit bone marrow–derived MSCs were isolated, expanded, encapsulated in the hydrogel, and cultured in chondrogenic differentiation medium for 9 weeks. Compressive modulus was evaluated at day 1 and at weeks 3, 6, and 9. Chondrogenic differentiation was investigated via quantitative polymerase reaction, safranin-O staining, and immunofluorescence. In vivo, a 3 mm–wide × 2-mm-deep chondral defect was created bilaterally on the knee trochlea of 10 rabbits. Each animal had 1 defect randomly assigned to be treated with hydrogel with or without MSCs, and the contralateral knee was left untreated. Hence, each rabbit served as its own matched control. Three groups were established: group A, hydrogel (n = 5); group B, hydrogel with MSCs (n = 5); and group C, control (n = 10). Repair tissue was evaluated at 6 months after intervention. Results: In vitro, chondrogenesis and the degradable behavior of the hydrogel by MSCs were confirmed. In vivo, the hydrogel could be delivered intraoperatively in a sterile manner. Overall, the hydrogel group had the highest scores on the modified O’Driscoll scoring system (group A, 17.4 ± 4.7; group B, 13 ± 3; group C, 16.7 ± 2.9) ( P = .11) and showed higher safranin-O staining (group A, 49.4% ± 20%; group B, 25.8% ± 16.4%; group C, 36.9% ± 25.2%) ( P = .27), although significance was not detected for either parameter. Conclusion: This study provides the first evidence of the ability to photopolymerize this novel hydrogel in situ and assess its ability to provide chondrogenic cues for cartilage repair in a small animal model. In vitro chondrogenesis was evident when MSCs were encapsulated in the hydrogel. Clinical Relevance: Cartilage mimetic hydrogel may offer a tissue engineering approach for the treatment of osteochondral lesions.


1976 ◽  
Vol 85 (6_suppl) ◽  
pp. 2-32 ◽  
Author(s):  
Thomas R. Van De Water

An experiment was designed to answer the question as to whether or not the neural elements of the statoacoustic ganglion complex have a trophic effect upon the histodifferentiation of the sensory structures of the embryonic mouse inner ear anlage as it develops in vitro. The embryonic inner ear anlage with associated otic mesenchyme and statoacoustic ganglion complex was excised from 11, 12, and 13-day CBA/C57 mouse embryos. The inner ear explants of each gestational age group were further divided into two groups: the first group “A” (with) statoacoustic ganglion was explanted to the organ culture system without further surgical intervention; the second group “B” (without) statoacoustic ganglion underwent further surgical manipulation during which their statoacoustic ganglion complexes were dissected away prior to explantation to in vitro. The explanted embryonic inner ears were allowed to develop in organ culture until the equivalent of gestation day 21 in vivo was reached for each group; then all cultures were fixed and histologically processed and stained by a nerve fiber stain, in combination with a stain for glucoprotein membranes. Each specimen was code labeled and scored for histodifferentiation of sensory structures. Light microscopic observations confirmed that in group “A” cultures, statoacoustic ganglion neurons and their nerve fibers were present in association with the developed sensory structures; neither ganglion cell neurons nor their nerve fibers were found to be present in the sensory structures that developed in the group “B” organ culture specimens. Quantification revealed no consistent trend of greater occurrence of any sensory structure in the groups of explants analyzed. The presence of such a trend would have signified the probable existence of a trophic effect of the statoacoustic ganglion neural elements upon development of inner ear sensory structures in the group “A” explants of the 11, 12, and 13-day embryo inner ear organ culture specimens when compared to the aganglionic group “B” cultures. Microscopic comparison of the sensory structures and their sensory hair cells that developed in the organ cultures revealed no differences in the quality of the histodifferentiation of either group “A” or group “B” explants. A base to apex pattern of histodifferentiation of the organ of Corti sensory structures, which has been described to occur in vivo, was noted to occur in the in vitro developed cochlear ducts of all of the explanted inner ears without respect to whether neural elements were present (“A”) or absent (“B”) during development. It was concluded from the quantification of histodifferentiation data and the above observation on the pattern of differentiation of Corti's organ that no trophic effect of neural elements of the statoacoustic ganglion complex influencing the histodifferentiation of sensory structures of 11, 12, and 13-gestation day mouse embryo inner ear explants as they differentiate in vitro could be demonstrated.


Blood ◽  
1991 ◽  
Vol 77 (6) ◽  
pp. 1383-1388 ◽  
Author(s):  
LL Lenny ◽  
R Hurst ◽  
J Goldstein ◽  
LJ Benjamin ◽  
RL Jones

Abstract Full-unit transfusions of RBC enzymatically converted from group B to group O by treatment with alpha-galactosidase (ECO RBC) to group O and A normal healthy individuals exhibit excellent in vivo survival times (24-hour survival 95.1% +/- 2.3%, T50 36.9 +/- 4.6 days). These results confirm our earlier findings describing ECO RBC in vitro viability and normal in vivo survival time after small-volume infusions. No significant increase in pretransfusion anti-B titer or score is observed in either group O or A subjects provided that sufficient enzyme is used to treat the cells: Cells transfused to group O recipients require higher levels of enzyme (185 to 200 U/mL RBC) than those infused to group A (90 U/mL RBC). Two separate single-unit transfusions of ECO RBC to one group O recipient (4.5 months apart) also survived normally (24-hour survival 96% and 92%, T50 40 and 36 days) and did not increase preexisting anti-B levels in this subject. ECO RBC were not agglutinated or lysed by recipient sera before or after transfusion. Similarly, no antibody development to the alpha- galactosidase used in cell treatment (and washed from the product before transfusion) could be detected in any subject. The sustained increase in hemoglobin levels after transfusion of ECO RBC suggests that this product will be useful in treatment of acute and chronic anemia.


2019 ◽  
Vol 31 (1) ◽  
pp. 198
Author(s):  
F. Kotarski ◽  
B. Zimmer ◽  
C. Wrenzycki

The sex ratio of newborn calves and embryos produced in vivo is ~1:1. However, numerous studies on bovine in vitro-produced embryos suggest that the sex ratio may differ from 1:1 and that the rate of development may be influenced by the sex of the embryo under certain culture conditions. The duration of sperm-oocyte interaction and sperm pre-incubation also affect the sex ratio of bovine embryos produced in vitro. It is well documented that in vitro male embryos reach the more advanced stages earlier than do their female counterparts. Selection of developmentally more advanced embryos in anticipation that they have a greater developmental capacity may be one of the underlying causes of the disproportionate number of males among offspring born after transfer of in vitro-produced embryos. The aim of the present study is to test whether a pre-incubation of sperm before IVF might improve the developmental rates and also influence the sex ratio of the resulting embryos. Bovine cumulus-oocyte complexes were recovered from abattoir-derived ovaries by the slicing method. After 24h of maturation, fertilization was realised using a standard protocol. Prior to IVF, sperm cells from 2 different bulls were treated as follows: sperm within group A were pre-incubated in IVF medium for one hour. This step was omitted for sperm in group B (control). After 19h of co-culture of COC and sperm, presumptive zygotes were cultured in SOFaa for a period of 7 days. Cleavage and developmental rates were recorded at Day 3 and 7 (Day 0=IVF). Day 7 blastocysts from all groups were sexed using bovine and Y chromosome-specific primers. Data were analysed by ANOVA. As shown in Table 1, sperm pre-incubation did not affect the cleavage and developmental rates for the individual bull (P>0.05). On average, at Day 7 of development a higher number of blastocysts was determined when embryos had been produced from pre-incubated sperm (P ≤ 0.05). This held true for both bulls. The shift in favour of male embryos was detectable in all groups of embryos, with a drastic one for bull 1 after sperm pre-incubation. In conclusion, sperm pre-incubation accelerated embryo development and possibly enhanced the proportion of male embryos, which was already shifted toward males. Table 1.Developmental rates, developmental kinetics and sex ratio of embryos after sperm pre-incubation before IVF (mean±standard deviation)


Blood ◽  
1957 ◽  
Vol 12 (10) ◽  
pp. 883-906 ◽  
Author(s):  
WOLF W. ZUELZER ◽  
FLOSSIE COHEN ◽  
ABNER R. ROBINSON ◽  
KATHRYN BEATTIE

Abstract A "study in depth" is reported concerning the case of hemolytic disease of a group B infant born to a group O mother, whose group A twin was apparently unaffected. It was shown that the hemolytic disease of the affected infant was due to a specific anti-B antibody. The study included parallel examinations of the antibodies in the sera of the three individuals. The specificity of the B-anti-B reaction was demonstrated in vitro and in vivo. The powerful anti-B antibody of the mother had no effect on the group A twin, in whose serum anti-B was present in large amounts. In vitro, studied by the usual technics of cross absorption the maternal and the fetal anti-A and anti-B serum antibodies behaved as if strictly specific. By applying successive absorption, elution and neutralization techniques, however, it was possible to demonstrate additional cross-reacting antibodies in the maternal serum which could be separated from one another and from the specific anti-A and anti-B antibodies. From the erythrocytes of the normal group A twin such cross-reacting antibody could be eluted. The cross-reacting anti-B antibody, isolated in pure form in eluates, could be shown to be loosely attached to group A erythrocytes without producing visible agglutination reactions while after elution from A cells it did visibly agglutinate group B cells. It could be eluted from A cells, absorbed by fresh A cells and reeluted while retaining its anti-B effect. It was neutralized by group B saliva only. A separate anti-A antibody with similar properties was eluted from B cells and specifically neutralized by group A saliva. A partial affinity of these antibodies for heterologous erythrocytes but not for specific soluble substances was thus demonstrated. These findings support neither the linkage hypothesis of cross reactions between anti-A and anti-B nor the C-anti-C hypothesis of hemolytic disease. They are in keeping with the view that group O sera contain variable complexes of anti-A and anti-B antibodies, composed of multiple fractions with different partial specificities. It is suggested that the occurrence or non-occurrence of cross-reacting antibodies found in sera of group O mothers whose infants develop hemolytic disease is best explained on this basis. It is further stressed that the demonstration of an antibody in mother or child in ABO hemolytic disease does not necessarily indicate its pathologic significance.


2006 ◽  
Vol 91 (8) ◽  
pp. 3181-3188 ◽  
Author(s):  
Guneet Makkar ◽  
Ernest H. Y. Ng ◽  
William S. B. Yeung ◽  
P. C. Ho

Abstract Context: Impaired implantation in assisted reproduction cycles with high serum estradiol (E2) concentrations may be related to abnormal endometrial functions. Objective: The in vivo expression of T helper type 2 (Th2) cytokines in the periimplantation endometrium of infertile patients was compared between natural and stimulated cycles. Interventions and Main Outcome Measures: Uterine flushings and endometrial biopsies were collected 7 d after the LH surge in natural cycles or after human chorionic gonadotropin injection in stimulated cycles. Th2 cytokines were determined by immunolocalization and by ELISA. Natural cycles were in group A, whereas stimulated cycles with peak serum E2 of no more than 20,000 pmol/liter (moderate responders) and more than 20,000 pmol/liter (excessive responders) were classified as group B and group C, respectively. Results: Higher E2 had a negative effect on IL-11 and IL-6 expression in the endometrium and IL-11 concentration in the uterine flushing. In endometrial biopsies, a significantly lower immunostaining of stromal IL-11 (P < 0.001) and glandular IL-6 (P < 0.05) was detected in group C compared with that of groups A and B. IL-11 concentration by ELISA was significantly lower in group C (P < 0.05). Endometrial leukemia inhibitory factor and IL-4 expression was similar in the three groups. In uterine flushings, a significantly higher percentage of women in group C had undetectable IL-11 and a lower IL-11 concentration (P < 0.01) compared with group A, whereas no difference in IL-6 concentration was noted in the three groups. Conclusion: Reduced expression of IL-11 and IL-6 in periimplantation endometrium may account for lower implantation in excessive responders.


1978 ◽  
Vol 18 (94) ◽  
pp. 714 ◽  
Author(s):  
P Michell

In vivo and in vitro organic matter digestibilities (OMD's) were determined on cuts of Lolium perenne (perennial ryegrass cv. Tasmanian No. I ) , Lolium perenne (long rotation ryegrass cv. Grasslands ariki), Lolium perenne x L. multiflorum (short rotation ryegrass cv. Grasslands Manawa), Lolium multiflorum (tama ryegrass cv. Grasslands Tama) Dactylis glomerata (cocksfoot cv. Grasslands Apanui), Dactylis glomerata (cocksfoot cv. Currie), Festuca arundinacea (tall fescue cv. Demeter), Avena sativa (oats cv., Blythe), Phalaris tuberosa (phalaris cv. Australian), Trifoleum repens (white clover cv. Grasslands Huia) Medicago sativa (lucerne cv. Hunter River), Brassica napus (rape cv. Rangi) and Brassica oleracea (kale cv. Chou moellier). Linear regressions relating in vitro and in vivo OMD were calculated for the individual generic groups. There were no significant differences between the regressions of ryegrass, cocksfoot, phalaris, oats and lucerne (P > 0.05) (group A) or between clover and brassicas (P > 0.05) (group 6). However, the combined regression for group A (y = 0.78 x + 13.0) was significantly different from that of group B (y = 0.76 x + 19.1 ) (P < 0.01 ) and both were significantly different from the fescue regression (y = 1.23 x - 24.4) (P < 0.01 ). The in vitro technique underestimated the in vivo OMD of white clover and the brassicas by about 4.5 units and over-estimated that of fescue by about 3 units, relative to the other species. These differences were not caused by a modification of the in vitro method in which final residues were collected by filtering through sintered glass crucibles rather than by centrifuging. However, the filtering method gave in vitro results averaging 7.2 per cent higher than the centrifuging method.


1987 ◽  
Author(s):  
P E Makris ◽  
A Papadopoulos ◽  
D A Tsakiris

We aimed to investigate the changes of lipoxygenase products in platelets and the simultaneous behaviour of ‘in vivo’ or ‘in vitro’ aspirinised platelets, stimulated by two agonists, PAF and epinephrine (EPI). 12 healthy were included. 6 received 20mg of aspirin (ASA) per os for 7 days (group A), and in 6 (group B) platelets were aspirinised ‘in vitro’ (5 or lOmin incubation at 37°C with ASA 1M). In group A blood was drawn once at the beginning and once at the end of the trial, while in group B just ome. First, platelet aggregation was studied using two agonists simultaneously (0.6 ¼M EPI and 20 nM PAF). We incubated then all platelet samples with 0.5 M of the substance BW755C (kind offer of Dr Moncada) far 3 min at 37°C. Second we measured PL0 products according to Takayama et al (1980), in platelets with or without ASA, and in platelets with ASA and after treatment with BW755C, always after addition of both agonists. Our results showed: a) Irreversible aggregation was slightly enhanced by the simultaneous addition of PAF and EPI in both groups and in non-aspirinised platelets. After ASA treatment, each agonist alone did not induce irreversible aggregation, whereas their combination overcame this inhibition, a fact not noticed under BW755C (a known PLO inhibitor). b) PLO products were measured in nmol TBRS/10 platelets:Our results agree with Cerletti et al (1986) and confirm that the two agonists combined are capable of overcoming the inhibition caused by ASA, possibly by activating the PLO pathway (Cerletti et al, 1986). Respectively the quantitative determination of PLO products (about which we did not notice any other report insofar) confirm the above assumption, since inhibition by BW755C coincides with the steep fall of PLOlevels, which for group A is statisticallysignificant (p≺0.01, paired t-test) and for group B entirely significant (p ≺0.001).


2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Sarvesh Sabarathinam ◽  
Satish Kumar Rajappan Chandra ◽  
Vijayakumar Thangavel Mahalingam

AbstractMaha yogaraja guggulu (MYG) is a classical herbomineral polyherbal formulation being widely used since centuries. The aim of this study was to investigate the effect of MYG formulation and its major constituents E & Z guggulsterone on CYP3A4 mediated metabolism. In vitro inhibition of MYG and Guggulsterone isomers on CYP3A4 was evaluated by high throughput fluorometric assay. Eighteen Adult male Sprague–Dawley rats (200 ± 25 g body weight) were randomly divided into three groups. Group A, Group B and Group C were treated with placebo, MYG and Standard E & Z guggulsterone for 14 days respectively by oral route. On 15th day, midazolam (5 mg/kg) was administered orally to all rats in each group. Blood samples (0.3 mL) were collected from the retro orbital vein at 0.25, 0.5, 0.75, 1, 2, 4, 6, 12 and 24 h of each rat were collected. The findings from the in vitro & in vivo study proposed that the MYG tablets and its guggulsterone isomers have drug interaction potential when consumed along with conventional drugs which are CYP3A4 substrates. In vivo pharmacokinetic drug interaction study of midazolam pointed out that the MYG tablets and guggulsterone isomers showed an inhibitory activity towards CYP3A4 which may have leads to clinically significant interactions.


Blood ◽  
1991 ◽  
Vol 77 (6) ◽  
pp. 1383-1388
Author(s):  
LL Lenny ◽  
R Hurst ◽  
J Goldstein ◽  
LJ Benjamin ◽  
RL Jones

Full-unit transfusions of RBC enzymatically converted from group B to group O by treatment with alpha-galactosidase (ECO RBC) to group O and A normal healthy individuals exhibit excellent in vivo survival times (24-hour survival 95.1% +/- 2.3%, T50 36.9 +/- 4.6 days). These results confirm our earlier findings describing ECO RBC in vitro viability and normal in vivo survival time after small-volume infusions. No significant increase in pretransfusion anti-B titer or score is observed in either group O or A subjects provided that sufficient enzyme is used to treat the cells: Cells transfused to group O recipients require higher levels of enzyme (185 to 200 U/mL RBC) than those infused to group A (90 U/mL RBC). Two separate single-unit transfusions of ECO RBC to one group O recipient (4.5 months apart) also survived normally (24-hour survival 96% and 92%, T50 40 and 36 days) and did not increase preexisting anti-B levels in this subject. ECO RBC were not agglutinated or lysed by recipient sera before or after transfusion. Similarly, no antibody development to the alpha- galactosidase used in cell treatment (and washed from the product before transfusion) could be detected in any subject. The sustained increase in hemoglobin levels after transfusion of ECO RBC suggests that this product will be useful in treatment of acute and chronic anemia.


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