Science topic

Wound Healing - Science topic

Restoration of integrity to traumatized tissue.
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Dear all,
I am currently searching for a topic to write my thesis on but to no avail. Preferably I would like the argument to be centered around plastics and reconstructive surgery since it is a specialty I would like to pursue after medical school.
The topics that interest me the most in plastic and reconstructive surgery are :
- Wound healing
- Vascularised Composite grafts
- Craniofacial surgery
Regarding these topics, I would like to know what within these arguments are trending questions.
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Vascularized composite grafts with active growth function in children.
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Dear all!
i encapsulated BSA-stabilized curcumin NP in the hydrogel matrix, and for antibacterial test i immersed it in 75% ethanol solution for 2 hours (referred in the lieratures), followed by washing with PBS buffer for 3 times. however, after alcohol sterilization i found alcohol color changed into light brown implying that BSA-Curcumin NP was extracted. actually, it is feasible i think.
So i am afraid this loss of encapsulated materials in the hydrogel will definitely affect its antibacterial, and further wound healing action. Then, which method is preferred for above type hydrogel? its nearly impossible to synthesize all polymer components aseptically even though hydrogel fabrication can be done aseptically. please let me get out from this delima. thanks!
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Alcohol does not sterilize at any concentration and you're very likely correct - it is extracting components. Spices are often treated with gamma to eliminate pathogens. Gamma to sterilize or heat to disinfect amight work - valiudate by comparing in vitro efficacy.
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Which kinds of these cells (HFF or HDF) are better for wound healing research ?
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(Human) dermal fibroblasts (HDF) originate from mesenchymal cells and are the main cell type present in skin connective tissue (dermis). They are located in particular, in the dermis and are the main actors of extracellular matrix (ECM) production and homeostasis. Fibroblasts interact with epidermal cells during hair development and in interfollicular skin. Moreover, they play an essential role during cutaneous wound healing and in bioengineering of skin.
In researches, Human Dermal Fibroblasts (NHDF) are isolated form the dermis of juvenile foreskin or adult skin from different locations like the face, the breasts, the abdomen, and the thighs. Those whoe are isolated from the foreskin are specifically labeled Human Foreskin Fibroblasts (HFF).
Such cells then undergo cryopreservation and can be used for wound healing studies and dermatological research to investigate diseases like scleroderma, fibrosarcoma, fibrosis, xeroderma pigmentosum, and histiocytoma. Moreover, fibroblasts are important for cancer research, tissue regeneration, and tissue engineering studies.
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I have data from different wound healing experiments (Epithelial cells) where each experiment produces data that is at a different range than the data from other experiments. The effect seems to be consistent between controls and corresponding treatments each day but cannot be compared to other days because the ranges are so wide.
Everything is the same protocol wise, the only source of change that is probably causing the difference is the wound size. I have to use a tool to scratch the surface of the plate but it's difficult to produce consistent wound sizes. Especially since I'm working on the um scale. For example, some experiments have a wound size of 300 um and other experiments might have a wound that is 600 um.
Within single experiments there seems to be some statistical difference but I don't know how to compare experiments from one day to the experiments of another day.
Does anybody know how to normalize this data and/or do statistical analysis?
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Hi Marjan,
To give you a little more context, we plate the cells and wait until they're confluent, then we do a scratch with a scraping tool. This is a 24 well plate that is then imaged under a microscope/incubator combo until the wound heals 100%, with images taken every 20 minutes.
What is abnormal is that sometimes the wound is very large and takes maybe 20 hours to heal and other times (on other days) the wound is small and takes maybe 12 hours to heal. Our treatment seems to be producing an effect in both scenarios when I plot the data but I don't know how to graph this data together or combine them as the timeframes are different.
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Post burn wound healing cosmotic appearance its relation direction of debridement of burned tissues and the the effect on body image of patient as well as quality if life so i am in need to understand the relation the debridement with post burn changes in cosmotic appearance of patient
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Thank you a lot for your valuable answer and so i mean that when we handle and manipulate the burned tissue and underneath in bizarre direction is this handling and removing the dead tissue affect positively on disfigurements of tissues post wound healing and thank you again
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I have loaded 2 compounds and a nanoparticle to a cream base. I need to study the drug release of the compounds and nanoparticles individually. Please suggest me a better technique/technology for carring out timely drug release.
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You can use Dialysis membrane or bag for the drug release study.
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I have been studying using PRP for chronic ulcer treatment but I was wondering if it could be used for chronic wound healing. I have several patients in the institution I work at that present chronic wounds that due to their advanced age do not heal. As PRP could promote re-epithelization, ¿could you help me in finding some articles that support the use of PRP in wound healing?
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I think you mean PRP for platelet rich plasma is this correct?
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hello. I am a student studying MTT assay and Wound healing.
Studying MTT and Wound healing, I learned that MTT measures cell viability and Wound healing measures cell proliferation rate and inhibition of migration.
However, I thought that it could be said that cell migration was inhibited by the inhibition of cell viability.
Can you tell me the exact difference between MTT and Wound healing?
Or do you have a paper to refer to when studying the difference between MTT and Wound healing?
thank you.
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Hello Eunji Han
The MTT assay is used to measure cellular metabolic activity as an indicator of cell viability, proliferation, and cytotoxicity. This colorimetric assay is based on the reduction of MTT to purple formazan crystals by metabolically active cells. The viable cells contain NAD(P)H-dependent oxidoreductase enzymes which reduces the MTT to formazan. The insoluble formazan crystals are dissolved using a solubilization solution and the resulting-colored solution is quantified spectrophotometrically. The darker the solution, the greater the number of viable, metabolically active cells.
On the other hand, Wound Healing assay is a standard in vitro technique for probing collective cell migration. In this assay, a cell-free area is created in a confluent monolayer by physical exclusion or by removing the cells from the area through mechanical, thermal, or chemical damage. The exposure to the cell-free area induces the cells to migrate into the gap. In this assay it is essential to inhibit proliferation of cells because closure of the wound should happen only due to cell migration and not by cell proliferation. For this reason, the cells need to be starved for at least 16 hours before the scratch (wound) is created. You may provide the cells with 0.5% serum containing media which may help prevent cell death but at the same time inhibit cell proliferation.
So, MTT assay measures cell viability and is mostly used to check the cytotoxic effects of compounds under study. Wound healing assay measures the basic cell migration parameters such as speed, persistence, and polarity. Cells at the edge of the created wound polarize and migrate into the wound space. This assay does not require the use of specific chemo attractants or gradient chambers, and it generates a strong directional migratory response. It is most reliably analyzed when performed using time-lapse imaging, which can also yield valuable cell morphology/protein localization information.
In short, MTT assay checks cell viability and Wound Healing assay checks cell migration in terms of speed, persistence and polarity.
Best.
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I have herbal extracts obtained by maceration in 80% ethanol that are desired to be evaluated for their wound healing activity using excision and incision wound model on albino rats.
Although the extracts are prepared using the same solvent they have different solubility profile in water (some form clear solution while others results in a homogeneous suspension).
We intend to formulate the extracts into a semisolid dermal preparation using either aqueous cream base or simple ointment base as they are the most frequently used bases with the least intrinsic efficacy.
Is there a suitable method to measure and compare the release of the extracts from the aqueous cream base and the simple ointment base in order to evaluate their suitability.
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Dear Jun,
yes, there are methods.Just read the attached article.
regards
Horst Liebl
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I will implement indirect co-culture by using transwell inserts. Macrophages will be in the inserts and lung cancer will be in the lower well. We will do a wound-healing assay for A549 during co-culture. What is the recommended pore size and number of inserts to do the assay? Are the characteristics of inserts that will be used for wound healing is same for western blot? if not what would be?
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ahmed how are you?
Do you happen to have a picture to clarify better? there is a classification regarding the wound and the evolution of healing, but it would help me to answer you. last year i studied something very similar.(I'm not very good at english)
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I went through few papers, in so,me they are saying that acidic pH help in wound healing and in some they are saying that neutral to alkaline is better for wound healing. So I am confused. It would be great if I can get some clarity on this.
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Acidic pH is helpful.
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While going through some literature I came across chemical signals such as Growth Factors (EGF/ FGF etc) released during wound healing. Are there any more such stimuli one can use to trigger drug release from a polymeric scaffold?
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In addition to chemical signals, other factors act in the healing process. Cell types and cytokines regulate each stage of the wound healing response. ECM, extracellular matrix; EGF, epidermal growth factor; FGF, fibroblast growth factor; G-CSF, granulocyte colony-stimulating factor; GM-CSF, granulocyte-macrophage colony-stimulating factor; HA, hyaluronic acid; IL, interleukin; MMP, matrix metalloproteinases; PDGF, platelet-derived growth factor; TGF, transforming growth factor; tPA, tissue plasminogen activator; uPA, urokinase-type plasminogen activator; VEGF, vascular endothelial growth factor.
I hope this helped you.
Regards,
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I use indirect MTT test to investigate the effect of the biomaterials I have developed. For this, I use the filtered extracts of my materials that have been kept in the medium in an incubator for 24 hours. As vehicle control, I also put the standard medium (containing 10% FBS, 1% Pen/Strep) into the incubator.
The vehicle control medium delays the closure in my scratch test for wound healing while increasing viability in MTT (significantly compared to the control medium that I did not keep in the incubator).
What factors in the medium can we connect the effect seen in these two tests? I attribute the delay in closing the scratch test to the serum, but I could not understand the increase in viability (we can say a kind of increase in mitochondrial activity) in MTT. Could it be pH-related?
Thank you very much.
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As you suggested, pH would be my first thought. For glutamine, there will be significant degradation in 24hr at 37°C which would be accelerated in the presence of 10% FBS, but I don't think the effect would be that drastic. Anyway, the cells are cultured in the same medium at 37°C for days. You can run an experiment with control medium warmed at 37°C for 24hr outside of the incubator (without CO2) and compare with the medium pre-incubated inside the incubator (assuming it is not in an air-tight vessel/container).
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I have prepared and hydrogel via freeze thaw method and then I performed its physical characterization like swelling, water content, moisture retention, gel content etc. Before conducting these experiments first I dried my hydrogels. I want to know it is the right way to perform the analysis? I have read the literature and it is mentioned that for these experiments hydrogels are dried but in few studies they have not mentioned whether they have processed the samples before these experiments.
I would appreciate the response
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Dear Noor,
Typically, the as-prepared hydrogels are dried before characterization (as you did). After the freeze-thaw process, it is also important to purify the hydrogel to remove non-crosslinked chains or other chemicals. The presence of liquid within the swollen hydrogel can lead you to the wrong interpretation of results collected from the experiments mentioned by you (swelling, water retention, etc.). Taking in mind that the drying process used by you (oven, vacuum, lyophilization, etc.) can also affect your results.
Regards, André
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in passage 0 the cells grown properly but after subculturing they did not adhere to the flask and showed cell death(round in shape and less number).Some of the cells that are frozen in -80 degree was revived in T75 flask the load found to be too low and no proper cell growth, cells are found to be round in shape and floating. What may be the reason? Kindly suggest me the possible reason for the above mentioned problem.
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Sam Augustine Kandathil NO sir, I didn't used coated plates
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I am working on a wound healing activity. I want to develop a wound healing formulation for which I am looking for a specific and standard formula. Can anybody help me for this with a standardized formula?
A helping hand would be greatly appreciated.
Thanking you
With Regards
Abhay
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The most neglected aspect of wound healing is cellular oxygenation and nutrition. Wound-healing fibroblasts need lots of oxygen and food, so that tissue perfusion and delivery of the glucose and oxygen to the cells is the key. Plus, hypercarbia is needed to release oxygen from the hemoglobin molecule via the Bohr Effect. CO2 enables every aspect of the mechanism of oxygen transport and delivery. This is basic physiology. Today’s doctors have totally forgotten the powerful therapeutic effects of Carbogen, which were well understood and accepted in the 1930’s. I was astonished when I learned of this during my investigation of anesthesia history. Read the attached paper that reviews the pathophysiology of carbon dioxide. Breathing Carbogen (a mixture of 5% carbon dioxide and 9% oxygen in a pressurized tank) is the most convenient and effective approach, but Big Pharma has wrapped its tentacles around everything that involves CO2 treatments, so that Carbogen must nowadays be special-ordered from the Airgas Corporation and costs $500.00 for a small tank. This is insane, because both oxygen and carbon dioxide are cheap. The price is obviously rigged to discourage utilization. Another approach would be to enclose the patient in a SCUBA “dry suit” and inject carbon dioxide inside the dry suit. The CO2 is absorbed directly through the skin, and will slightly exaggerate CO2 blood levels, which will release nitric oxide from the capillary endothelium, reduce microvascular flow resistance, improve tissue perfusion and enhance the release of oxygen from the hemoglobin molecule via the Bohr Effect so as to elevate tissue oxygenation and promote wound healing. There is no danger because CO2 stimulates respiratory drive and prevents excessive CO2 accumulation within the body.
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Dear All,
I am developing plant extract incorporated electrospun nanofibers for biological applications
Please discuss your ideas regrding which type of extract could I use for taking plant extract and criteria of polymer for the purpose
Please share your valuable experiences
Regards
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Dear all, I think the use of plants and plants extracts in wound healing are not a matter of discussion nowdays. It is an old science, art, and tradition. Please tell if you are working on a special system plant/polymer for electrospinning. My Regards
10.5772/intechopen.80215
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I have a set of data from wounds that are going through normal wound healing and those that have been treated to intentionally delay wound healing (n=3) in pigs. For normal/acute wounds, tissues were harvest at two timepoints: Day 5 and Day 10 from the point of wounding. For wounds with delayed healing, four timepoints were collected: Day 5, 10, 15 and 20 from the point of wounding. These wounds are all on the same pig and repeated twice on two other pigs. I would like to answer the question: Is wound healing significantly delayed in the treated wounds compared to acute wounds.
Initially, I just used an ANOVA to compare across all wounds and all timepoints but I'm not sure if this is the appropriate test to use. Should I be doing a pairwise comparison for each timepoint (eg. Acute D5 V Treated D5) instead of analyzing all the timepoints together (as acute does not have timepoints day 15 and day 20).
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What does the outcome variable look like; what scale is it?
Since you only have 2 groups (normal healing, delayed healing), t-test makes more sense.
You could indeed use a paired test to compare the difference between the two wounds in each pig.
The data from day 15 and 20 are only useful to test a hypothesis if you have something to test it against. For instance, you could test if day 15 with delayed healing is still 'less healed' than normal day 10.
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Can I find a recent update systematic review for growth factors in periodontal regeneration??!
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we are keen to evaluate certain molecules for their potential to accelerate wound healing process in in vitro set up.  Two cell culture models i.e 3T3 and hPBMCs we are using.  I would like to know if some other cell models that can support the testing.
thank you,
Asavari Joshi
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For wound healing, can we use THP-1 cell lines.
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We want to calculate the topical dose of a peptide drug which isn't used on wounded skin before but we have the systemic i.p dose
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For a new molecule, it is interesting to see first, a mathematical modeling for example with the equation of Pott and Guy (there are others but which do not provide more information) All the mathematical models are absolutely false, but they first allow us to verify whether the molecule has the potential to cross the skin barrier or if it is very limited. If your drug has a log P between 1 and 4 and a MW of less than 500, the amount absorbed may be sufficient. A peptide of high molecular weight will not cross the skin barrier, or so weakly that it will be of no benefit. You already have an idea if the molecule will be able to cross the barrier (the melting point, if it is a salified form, the formulation are other factors to take into consideration) If your molecule has a potential absorption, you can test in 2nd on "franz cells" human or porcine skin, otherwise it is a waste of time.
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I'm searching for a research article for cytokine study by ELISA for IL-6, TNF-a, IL-1B. Kindly recommend some good papers to proceed.
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Dear community,
I am doing wound healing experiments and found some substances that seem to enhance the proliferation of skin cells and might be even synergistic when applied together.
I would like to calculate it with CompuSyn by Ting-Chao Chou (a software making researchers lifes so much easier, thank you for that!!)
I cannot enter effects > 1.0, thats why I normalized the control to 0.75 so that all values are < 1.0.
Is this thought too simple? Does anybody have another suggestion?
Thanks in advance,
Lea
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No.
We do this simply by using GraphPad Prism.
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The lady 72-years old, 10 days after anterior resection because of rectal cancer (T3N0M0) developed anastomotic leak accompanied by surgical site infection. Since she has "frozen abdomen" we did not perform diverting ileostomy. We placed EndoSponge and abdominal vacuum assisted abdominal closure with quite good abdominal wound healing.
Is EndoSponge placed properly? It was not possible to place it deeper in the anastomotic cavity. The negative pressure works.
Do you usually allow the patients to consume liquid diet during EndoSponge treatment?
Thanks for your help
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I think will be a cause for complications, as greater opening in anastomosis, sepsis.
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Endogenous (self - burst) wound healing in patients with impaired metabolism, seeks for attention on many factors like tissue perfusion, peripheral neuropathy, tissue oxygenation, etc. whereas in induced diabetic models and excision wounds vascular and nerve supply are not compromised as same as in humans.
It is seen that other local and systemic factors like, hemoglobin, nutrition, limb activity, digestion etc. are not being focused in experimental studies.
In a nutshell, traumatic and atraumatic wounds, excision/incision and self burst wounds follows different healing methods.
The medicaments showing positive outcomes in excision models of diabetes induced Wistars should not be blindly followed without focusing on other healing factors.
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I agree 100% with Dr. Laborde especially with the first of 14 responses.
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hi everybody.
I'm using 75-85% deacetylated medium molecular weight chitosan.
and what if I want to load some wound healer medicine on it.
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Your question can be answered best by Prof Pierre Basmaji, a good friend of mine in Brazil
Kind regards
Horst Liebl
hl@el-cit. com
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Hello, I am new to the ImageJ and basic science world. I am working on a project that where we are measuring wound healing in B2-spectrin endothelial specific knockout mice. I have IHC images (below) where I have stained for CD-31 (red), and I would like to quantify the immunofluorescence of these images. I have found various tutorials online that discuss mostly quantizing individual cells. But I am more interested in the whole area, as this is essentially measuring the vascularity of the wound bed. The image attached is the combined channels but I can split the image into the respective channels for analysis.
So far, I have really only done the "analyze -> measure" feature and getting the mean gray value. I am I at least on the right track? Any tips? Thanks!
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Hi there,
yes, you're in the right track...
As usual, it depends on what you want to measure. However, measuring the whole image would be quite meaningless.
In this case, what you say is that you want to measure the whole area, and to do so you should first define your whole area (of the sample). One of your channels (usually the one that is a counter-staining) should be the right one to do so.
So, assuming the blue one is this one for DAPI (maybe I am wrong but still ok for the workflow):
-Over your RGB image (btw, when I opened your .png is a kind of 8-bit color image so I converted it to RGB)
1-Image>Color>Split Channels
-Take the image that better represents the whole area of the image (I took blue, however since the conversion 8-bit color to RGB it may contain artifacts but yours will not if you have the three separated channels to start with)
2-Image>Duplicate
3-Image>Adjust>Threshold
Adjust it manually to cover the whole area of the sample
4-Process>Binary>Close
5-Process>Binary>Fill holes
Here you get a binary image representing the whole area of your sample
Then:
6-Analyze>Analyze Particles...
Here you can try to filter by size and circularity (I used 1000-Inf for size, to get rid of the small particles). Check the Add to manager option as indicated in the workflow picture.
7-In the ROI manager: More>Save
This will save as a .zip file the ROIs corresponding to your binary mask derived from the image corresponding to the whole area of your image.
Then click over ("activate") your channel of interest (red channel in this case)
8-Analyze>Set measurements (check what you want to measure)
9-ROI manager>More>Multi measure
And you will get a list of the measurements in the different ROIs.
That's to measure the whole-area immunoreactivity of you red channel (picture attached). You can try to write a macro for all these steps to save some time for further analysis.
There are also plugins to measure vascularity (I haven't used them though), to do it manually you should define what's a vessel in your red image and then count the number of vessels in the whole area of your sample. To do so, I'd basically operate the red image as it was done with the blue channel in the example until you have particles that are vessels. You will have to optimize the thresholding and other binary operations may not be required. Then, again, you can use the ROI manager to filter by size and circularity (and play with the options) to get a count of the number of vessels and express it as number of vessels per area unit in each one of your conditions,
Cheers,
J
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Real time PCR primers given in the research article can be used for different species of rats for monitoring wound healing?
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Before doing the expressions studies, you will have to think,
Which genes?
Why these genes not others?
How many genes you will want?
Is there any relationship between these genes?
Are those genes part of the pathway?
Why that pathway?
Have those genes reported earlier?
If yes, collect references?
Have the primers for those genes reported earlier?
If no, design primers by yourself?
Record primers as much as possible?
Check these primers are for qPCR or simple PCR?
Select a reference gene?
Then perform expression analysis on the selected genes?
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I have been trying to do wound healing assay on Human Corneal Epithelial Cells with 2-well ibidi insert. However during media change with Mitomycin-C and washing with HBSS (after removing insert) steps some parts of HCEC monolayer keeps detaching away from the well plate.
My simplified protocol is:
Seed each insert with 70ul of cell suspension (density is 4x10^5 cells/ml)
Incubated around 12-16 hours.
Then changed media with Mitomycin-C and after 1 hour removed insert, and washed with HBSS. and added Media. Then take time lapsed images.
Anyone, who did work with HCEC have any suggestions about how keep HCEC monolayer intact?
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Hi Buuvee,
We assume that you have already tested for cell vitality after incubation with Mitomycin-C.
If the cells are only loosely adhering to the bottom of the dish and sensitive to media changes, two things can help:
- use a protein coating to increase cell attachment
- limit shear stress caused by fluid exchanges
- reduce wash steps (e.g. add drugs in higher concentration but lower volume (e.g. instead of 70 µL of solution with 10 µM substance x, remove 7 µl from well and add 7 µL of solution with 100 µM substance x)),
- prefer partial media exchange over a complete fluid exchange (e.g. for washing steps remove half volume, add back this volume, repeat),
- add and remove media extremely carefully/slowly.
We are happy to discuss your question in more detail. Please contact us at techsupport@ibidi.de.
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Can I use HT1080 (fibrosarcoma) cell line instead of the normal fibroblast cell line (Human dermal fibroblast, human keratinocytes) for wound healing studies?
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Hi, you cannot compare a fibrosarcoma cell line with normal fibroblasts. It depends on the aim of your study. First you need to identify if you want to work with primary cells or cell lines. Then if you choose cell lines, you need to select normal or cancer derived cell line. We would need more information to help you. Good luck
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Hi everyone,
Where can I find some information regarding sterility requirements for topical dosage forms?
To the best of my knowledge sterile topical drugs are: ophtalmic drugs, for wound healing and burns, gels used in endoscopy (anesthetic mostly) and certain dermoscometics?
Are there other type of drugs that should be manufactured in sterile dosage forms? (only topical).
Thanks very much!
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EVERYONE!! Or, at least, antiseptics.
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I have a Markov model from a colleague for wound healing. 4 stages open infcetd closed and deathd death. The paper used to inform the transition data has monthly healing rate over 6 months and an overall infection rate within those 6 months.
My colleague used the month 1 healing data for the transition probability and calculated a monthly probability for infection. I do not underttand why the firtst months healing data was suitable rather than calculating from using all of the healing data at months 1,2,3,4,5,and 6.
My colleague is no longer here and no-one else can explain. I am concerned our out come is wrong due to wrong data being used. we are using a time horison of 5 and then 10 years.
thanks for any advice
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You may be right. If you consider that healing is permanent and thus obviates the risk of subsequent infection, then his one month healing rate pertains. However, healing is not binary, because most wounds heal with a scar that is inferior in quality to native tissue. As a result, many wounds recur after appearing to be healed due to recurrent trauma or illness. Consider the fact that venous stasis ulcers of the ankle have a high relapse rate while an injury to the scalp heals rapidly after injury with minimal incidence of infection or recurrence. The main differentiating factor is the quality of blood flow and delivery of oxygen. Hypoxia is characteristic of the ankle due to its dependent position when we stand or sit.
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I will use 6 well plates. I'm studying the migration of the macrophages? Is it possible cell be migrate after it cell already differentiate monocytes into macrophages?
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A quick intuitive response to the question on whether PMA-differentiated THP-1 can migrate: No, as far as I know from imaging experiments. The cells become very large, spread out and don't look very active or motile. The differentiation primarily turns them to the gene expression profile needed to study macrophages, but that does not necessarily reconstitute the phenotype in vivo.
Questions about how to culture and differentiate THP-1 have been asked many times in this RG forum.
Briefly, the optimized dose is 5 ng/mL PMA. The exactly procedure can be found in this highly cited paper:
Good luck!
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I have been working with wound healing and I am facing problems as I am unable to find normal murine dermal fibroblast cell line to study the wound healing effect. The dermal fibroblast cell lines present in NCCS repository are melanoma cell lines. It would be kind, if I would get to know whether there are normal murine fibroblast cell lines in any of the repository concerned. The information regarding such cell lines will be highly appreciated.
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Thank you Fauzi Mb sir for the kind information
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Hello,
I am trying to measure the area of my scratch for wound healing using ImageJ. I have a large amount of images, so tracing the scratch manually is not a good option. I have tried MRI_Wound_healing_tool and I get many areas outside of the scratch selected.
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Great option!
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I am working with lymphatic endothelial cells and trying to test effect of few chemicals, on LECs by wound healing scratch assay. My confusion is, if I have 4 different chemicals cited in different literature to be used for cell culture studies, in different concentration let's say, if A is 10 um then B 25, C may be 100 and D 400um. What would be ideal concentration if I want to compare the effect of these individual chemical on one cell type. Should I be choosing one from literature individually or a standard concentration for all to be used, to make a comparison.
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Nimal Raveendran Thank you Nimal sir for helping me with your wisdom
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Aloe vera exhibits a range of therapeutic activities such as anti-microbial, antiviral, anti-cancer, anti-oxidant, anti-inflammation, skin protection, wound healing properties as well as the regulation of blood glucose and cholesterol. I want to know exactly what aloe vera medications are used for.
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antioxidant
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We have evaluated the wound healing potentials of different compounds at particular concentrations in laboratory animals. Now, we want to move in the direction of drug development for some topical products for wound healing. So, we require the complete guidelines which should be followed and fulfilled for further study in the direction of drug development for topical formulations for wound healing.
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refer Harry's Textbook of Cosmeticology
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I am to study the wound healing of infected wound in animal model. However I am not sure about the animal model to be used. Whether swiss albino mice or Wister albino rats or Sprague Dawley rats would be appropriate for the study that involves an infected wounds on the mice/rats. .
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Thank you Shaymaa Fadhel for the kind information
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I am looking for ways in which I can compare wound healing among different treatment groups
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Shaymaa Fadhel even with several measurements at different time points?
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Can anybody suggest me one/more plant names for doing project either in alone or combination therapy of diabetic mellitus, wound healing and anti oxidant property?
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Currently I use U-2OS for transient overexpression of genes of interest. Using Lipofectamine 3000 is quite efficient for overexpression and verified by WB. However, recently I tried to combine overexpression with scratch wound healing assay and found that groups transfected with empty vector (pCMV3) as well as expression vectors all grew slower than non-treated and Lipo-only groups. This rules out the toxicity of Lipofectamine 3000 to cells.
Does anyone have similar experience?
Transfection of simply plasmid hurts cells?
Should I reduce the amounts of DNA?
or merely lengthen time span of my observation? coz cells grow slower but not die out.
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Sure lipofection may induce cell death -among other things - due to membrane stress.
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We have evaluated the wound healing potentials of a compound at a particular concentration in ointment form in laboratory animals. Now, we are preparing different types of topical formulations (gel, ointment, spray etc) for wound healing studies on clinical wounds of domestic/livestock animals. Does it require to test each topical formulation again on laboratory animals before proceeding for study on clinical wounds?. Please, also tell me that what should be the other guidelines which I have to follow before going for study of our topical formulations on clinical wounds of livestock animals (sheep, goat, cattle etc) for the drug development process.
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I think yes every formulation should be tested because excipients will be different in each formulation and may lead to increased or decreased bioavailability.
Regards
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Alloxan induced diabetes in the rabbit experiment to perform wound-healing experiments.
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please look at this link , it will help you (rat model not rabbit)
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Hi! Can you please name some phytochemicals which prevent scar formation in the process of wound healing?
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Please go through the following RG links and PDF attachment.
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We have now tried a number of commercially available antibodies to detect the receptor of AGEs (RAGE, sometimes called AGER) in parrafin embedded mouse tissue with DAB staining. As a control we have used corresponding tissues of a RAGE-KO mouse. Whereas most of these antibodies worked well in lung tissue that accumulates RAGE to a high amount, we have faced significant problems when using tissues that express only minor amounts of RAGE. In particular we observed cross reactivity with muscle cells. If anybody has had good results using an anti RAGE antibody for IHC in mouse tissue please let us know!
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Anyone has recommendation for human Rage WT antibody?
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What are some biomarkers that can best describe the wound inflammatory phase during the wound-healing experiment in rats?
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Check this
Inflammation Biomarkers and Correlation to Wound Status After Full-Thickness Skin Grafting
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In our country, Iran , we use mummy to amen bone breakage and wound, traditionally.
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On the medical use of mummies, especially the powder called "Mumia vera" you can find a lot of information by googling "Mumia vera" o "Mummia vera" for instance: https://www.smithsonianmag.com/history/the-gruesome-history-of-eating-corpses-as-medicine-82360284/
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I need to know the process as well as a legit article/journal/literature. Any Ideas or suggestion would like to recommend. Thank you, kind sir and ma'am.
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Thank you so much.. it massively helps and supports our research study as well. Thank you Sir Vineet Sharma
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Hi,
I want some articles about the effect of bacteria on wound healing process in a positive way . preferably new ones (like last five years )
thank you in advance
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I'm working on breast cancer surgery, and I'm going to estimate wound healing process time in different groups after breast cancer surgery. Here I want to know the parameters that are required to estimate the time. Based on which parameters we can able to confirm the healing time as delayed.
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I guess you need to do a pilot study.
W.
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Phenytoin is known to increase the fibrotic growth of ginigiva when administered in therapeutic doses in human . Is it expected to facilitate fibrosis in wound area during healing
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Sounds like a bad idea. Phenytoin is a small molecule, and would presumably get into the bloodstream or lymph when added to an open wound. As such, it could cause fibrosis at distant sites.
Let the body do it's natural healing, or use a wound-healing treatment that's local.
Also, by what mechanism does phenytoin cause fibrosis?
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I am testing some compounds for wound healing in rats (rodent) and want to further extend the research for the drug development for wound healing in animals/human. So, please tell me the different parameters (as a toxicological data), which I should to do in rodent before proceeding to experimentation in large animal species. At present, I am using the compounds topically alone and in nanoformulation form.
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After documentation of the therapeutic effect on wound healing moddels
perfformed:
LD50
Local toxicological effects on the skin
effects on liver and renal functions
carcinogenic effect
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I have a collaborator who is looking to investigate wound healing in some tissue samples using IHC, and it's outside my area of expertise. Any common markers you can recommend?
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In general, the markers of wound healing that can be studied with IHC are Cytokeratin, laminin, collagen IV, vimentin, vinculin and fibronectin. Testing these is pretty straight-forward and commercially available.
There are other proteins and peptides which are more obscure and their testing wont be available in most labs, only advanced academic institutions perhaps.
Theoretically, matrix metallaproteinases and other enzymes produced in the remodelling phase of wound healing could also be used as markers and can be studied (for example by gelatin zymography) and the growth factors in the early stages such as VEGF produced during the angiogenesis phase can also serve the same purpose, and can be achieved by IHC and reverse-phase protein array. Not sure if these have been used specifically for the study of wound healing before and availability of testing may be more limited.
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Hello every body,
could any body kindly support me about the data of the subcutaneous injection of Sonic hedgehog to mice to improve wound healing
what is the recommended dose and the protocol used?
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thanks Subhash C. Juneja
your answer is very helpful to me
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i want to find swelling degree of cross linked films.i need to know whether any relation is there between swelling degree and biodegradability of films.Actually what is the real purpose of doing sweeling degree of films or water uptake of crosslinked polymer.what idea we get from doing this? i know for wound healing application sweeling degree of polymer should be between particular value. any other purpose is there to find sweeling degree of crosslinked fillms?plz answer this question?
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if i get swelling degree of films is above 100 means ...what information i will get from this?
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Glucose insulin platelets blood
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The causes are certainly many. The reduced oxygen supply is important but not the only one. It is necessary to speak of reduced blood supply and of altered regulation, in situ, of the blood supply. In the patient with type II diabetes rheological changes, microangiopathy and macroangiopathy are a deadly mix for the well-being of the tissues.
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Plasma jet in bio medicine
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Thank you sir for your help.
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I love to start working on finding a way to accelerate the wound healing process
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wound healing is a natural process of the body that is influenced by multifactors. modifying one factor might have an impact on another factor/pathway of healing. this certainly depends on our approach to accelerating wound healing. intrinsic factor or extrinsic factor
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so i want to evaluate the effect of a topical gel of a plant extract for wound healing, but still didnt know how to determine the frequency of dosing, because some article i read said its given daily, the other said every 2 day, an another said every 3 day
*files below are not mine
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thanks for your feedback sir, i want to test the extract on rats, and its must be safe and effective on the rats before i test it to human, so back to the primary question, how to determine the frequency and dose of the application?
thanks again sir Amit Kumar Srivastava
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Hi...
How to stain Myofibroblasts in myocardial infarcted hearts? What about Alpha- Smooth muscle actin in the detection of myofibroblasts? Is there any other specific markers for the detection?
Thank you.
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Dear Budharaju,
I have used CD140a (PDGFRa) for mouse fibroblasts and I am planning to work with CD90 in human fibroblasts. However this is not sufficient if you need to separate fibroblasts from myofibroblasts, I suppose.
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Hi,
I'm working within a project on the role that hyaluronic acid (HA) in tumor cell migration and invasion. I have started testing low molecular weight oligosaccharides (maximum 18 repeat units), but these compounds seem to have little effect in our models. For this reason, I would like to try some type of higher molecular weight HA (HMWHA), but I'm not sure about which MW size(s) should I try. Also, I don't know how these HMWHA would be used in an in vitro cell migration/invasion system (soluble? gelified? wound healing or transwell?), so any help in these will be really appreciated.
Many thanks,
Jordi
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Jordi Senserrich - Hyaluronic acid for research use is available here and manufacured in Europe - Contipro ship all over the world
Take a look at the catalog attached on page 42 for laboratory grade - Sodium hyaluronate - the SH MW 1250 - 1500 kDa would suit the MW you are looking for.
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Can you suggest few natural compounds having wound healing activity and can be used for in vitro wound healing activity ?
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Curcumin, Trigonella foenum-graecum, and Polygonum are useful natural compounds for wound healing.
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Hello! I want to prepare scaffolds for tissue engineering. For in vivo tests I need to preserve the wound dressing from drying. Can you provide some nice idea with coating or additional layer deposition for protection from drying? Any good practical experience?
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Introduce some additives e.g F127 into PCL fibers to increase its hydrophilic nature.
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Dear Ms Isgren,
I planning a study project on wound healing disorders in horses. Could you kindly let me know the median time untilwound healing was completed in your study?
Kindest regards,
Petra Weiermayer
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Hello,
Of which tissue it is !
Each tissue has particularity in terms of healing duration. Muscle healing period being important ( more than six month).
Mok
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I am interested to assess the skin permeation potential of some formulation prepared for topical use for wound healing or other problem of skin. For this study, I want to use franz diffusion cell assembly for assessing in vitro permeation of the drug present in formulation. So, please provide me the details of manufacturer/ company which provides good quality franz diffusion cell for research purposes.
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Hi.
Good luck!
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The silver nanoparticle has a high toxicity?
we know it has very good antibacterial effect but how it can good agent for wound healing despite its toxicity effect? has a negatively effect?
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Silver nanoparticle based wound dressing materials are shown as a more successful product in the market with the positive wound healing effect. Currently, many types of silver-based wound dressing materials are commercially available on the market (Aquacel Ag®, DynaGinate™ AG Silver Calcium Alginate Dressing, CuraFoam™ AG Silver Foam Dressing, DynaFoam™ AG Bordered Silver Foam Dressing, Biatain® Alginate Ag, and SilverIon®). Prof. Dr. Robert Burrell (Professor, University of Alberta, Alberta) developed the world’s first commercially available AgNP-based wound dressing (ACTICOAT®: Smith and Nephew, UK) which covers large areas of burn wounds, and accelerates the wound healing process and reduces scar formation.
Best wishes
DS Sundar
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Currently Importing Ob/Ob Mice is a challenge for me, I'm looking for a mice strains which mimics Diabetic foot ulcer. I'm researching on Mildly impaired Wound Healing and Mild Neuropathy.
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Thanks Marian Waheeb. I will look into it.
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I tried to use the institution database to find the article but unfortunately failed. The article needed for animal ethics approval form my institution. I would be grateful if you can provide me a copy. Thank you
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Thank you Beatrice. This is really helpful.
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I plan to prepare a natural wound healing cream using plant extracts as its active ingredients. A preliminary study showed that 300mg/kg (from in vivo study using rats weighing between between 180g to 220g) of plant extracts are very significant in healing the wound. Now, I am so confuse on how can I conduct a Total Microbial Count (TMC) by applying this 300mg/kg of extract.
Is that the concentration of extract from the preliminary study (300mg/kg) must be applied in development of wound healing cream? or the production of cream can use totally different concentrations/percentage ratios of extracts depending on the formulation?
Looking forward for experts particularly the ones in product development.
Thanks in advance.
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Hi Abdul,
You can treat the wounds in multiple ways. You could inject the extract in vehicle (e.g. sterile pbs) beneath the wounds to create a ‘bleb’ or you can mix the extract with cream and apply topically to the wounds. You can either apply the topical treatment daily or occlude the wounds with tegaderm. Also don’t forget your vehicle controls. For your experiment, to make sure you add 300mg extract per kg, you would have to weigh your animals to work out how many mg of extract to use per animal. If you were creating two wounds on the dorsum of the animals, I would split this amount between the two wounds. To me, the plant extract weight would not include the weight of the vehicle (e.g. Nivea), however, do you have more information about the previous study and what they did in their animal experiments? Or a reference?
Also, are you planning to work out total microbial count from the wounds as well? Or just look at wound healing via macroscopic images and/or histology?
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Dear researchers
I want to evaluate Antidiabetic and wound healing activities on the same time on rabbit. Can I do it ? What about ethic is it acceptable.
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The answer would be yes, you can do it. You need to establish experimental diabetes in rabbits first - I suggest using alloxan instead of streptozotocin. Then you can perform cutaneous wounds on the ventral side of rabbit ears using 6 mm punch devices - make sure to avoid removing cartilage. Finally, monitor wound closure and diabetic symptoms of treated vs non-treated. I think you might find the article linked below very useful. Concerning ethics, the single extra action would be preventing animals’ hypoglycemic shock by providing them plenty of food and water before and intermediately after alloxan administration. Consider using the streptozotocin-induced diabetes model in rats instead.
I wish the best for your research:-)
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gold nano can accelerate wound healing in rats?
how the nano gold in wound healing ?
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I know that nano silver particles are more effective!
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Patients affected by epidermolysis bullosa (EB) , a rare genetic connective tissue disorder with symptom of extremely fragile skin that blisters and tears from minor friction or trauma , are suffering from many problems . EB is always painful, often pervasive and debilitating, and is in some cases lethal before the age of 30 .Internal organs and bodily systems can also be seriously affected .
Daily wound care, pain management, and protective bandaging are the only options available for people with EB.
Although there is not a real cure for EB , I want to start researching in order to improve their quality of life and reduce their pain and problems .
So, I wounder if I could be guided by your answer what is most important issue ?
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Dear Hanie,
Maybe the following recent papers will help you:
Uitto J, Bruckner-Tuderman L, Christiano AM, McGrath JA, Has C, South AP, Kopelan B, Robinson EC. Progress toward Treatment and Cure of Epidermolysis Bullosa: Summary of the DEBRA International Research Symposium EB2015. J Invest Dermatol 2016;136(2):352-8. http://www.jidonline.org/article/S0022-202X(15)00051-2/pdf
Peking P, Koller U, Murauer EM. Functional therapies for cutaneous wound repair in epidermolysis bullosa. Adv Drug Deliv Rev 2017 Dec 15. pii: S0169-409X(17)30306-X. doi: 10.1016/j.addr.2017.12.003. [Epub ahead of print]. https://ac.els-cdn.com/S0169409X1730306X/1-s2.0-S0169409X1730306X-main.pdf?_tid=881f85db-003c-442b-af71-c7213b19c27a&acdnat=1520354699_9ce47bdfa891fa7aff060df1b77479a5
Parvizi MM, Lankarani KB, Handjani F, Ghahramani S, Parvizi Z, Rousta S. Health literacy in patients with epidermolysis bullosa in Iran. J Educ Health Promot 2017;6:105. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5747211/
Wally V, Hovnanian A, Ly J, Buckova H, Brunner V, Lettner T, et al. Diacerein Orphan Drug Development for Epidermolysis Bullosa Simplex: A Phase 2/3 Randomized, Placebo-Controlled, Double-Blind Clinical Trial. J Am Acad Dermatol. 2018 Feb 1. pii: S0190-9622(18)30130-0. doi: 10.1016/j.jaad.2018.01.019. [Epub ahead of print]. http://www.jaad.org/article/S0190-9622(18)30130-0/pdf
Dakiw Piaceski A, Larouche D, Ghani K, Bisson F, Cortez Ghio S, Larochelle S, Moulin MJ, Caruso M1, Germain L. Translating the combination of gene therapy and tissue engineering for treating recessive dystrophic epidermolysis bullosa. Eur Cell Mater 2018;35:73-86. http://www.ecmjournal.org/papers/vol035/pdf/v035a06.pdf
Best wishes from Germany,
Martin
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Hello there, the experiment I performed is a scratch wound assay on Hela43 cells transfected differently: a non-transfected control, emptyGFP, Cx43D3N, Cx43G138R, wildtype Cx43 and a non-transfected with Gap27 (positive control). the cells were photographed in 4 different time points, then the wound area was measured over time. I want to determine if there is a significant difference in the wound area between the different cell types, ie. if the mutants have any effects on the rate of wound healing at all.
ps if possible, please include as much information as you can about how to carry the test out on prism, I'm an undergraduate student and it's the first time I'm carrying out a statistical test like this!
thank you very much!!
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Sounds like you would perform a two way ANOVA, but I would still check with your supervisor and confirm that's the most accurate test for your data.
Here's a Prism tutorial:
Best of Luck
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Both cell proliferation and migration contribute to reepithelization of wounded skin. To observe the effects of any genes or compounds on cell migration, we usually use the scratch assay to assess the keratinocyte motility, and regard it as a measure for the cell migration in vivo. Normally, in the scratch assay we treat the cells with mitomycin c to block the cells' proliferation. Can we treat mice with any small molecular inhibitors to block the proliferation of keratinocytes so that we may assess the cell migration in vivo? If this is not possible, are there any other methods that can be applied for this purpose?
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I found an answer to the question. Dr. Ce´dric Blanpain lab applied 5-fluorouracil (5-FU) to block epidermal cell proliferation. Please refer to: DOI: 10.1038/ncomms14684
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