Bone - Science topic

Plasmacytoid cells can be one of the various types of myoepithelial cells seen in Pleomorphic adenoma. Plasmacytoid myoepithelial cells are seen usually when minor salivary glands of young people are affected whereas spindle morphology of myoepithelial cells is seen when major salivary glands are involved. Increased cellularity with fibrillary stroma and plasmacytoid cells on FNAC categorizes the tumor under category IVB (Salivary gland neoplasm of uncertain malignant potential (SUMP) in The Milan System for Reporting Salivary Gland Cytopathology (MSRSGC) for which management is surgical excision. After surgical excision, histopathology can help in establishing the benign nature of the tumor and differentiating whether the tumor is a Pleomorphic adenoma with plasmacytoid myoepithelial cells or a Plasmacytoid myoepithelioma (epithelial component is scant or absent). IHC can confirm the myoepithelial origin of tumor cells and differentiate it from melanoma, plasmacytoma etc.

According to the mechanism of infection, osteomyelitis can be divided into two categories: blood osteomyelitis; non-blood osteomyelitis, which can be caused by invasion of adjacent infection sites or infection of bone tissue directly. Live bone xenografts may will cause GVHD.

In our company have osteomyelitis mouse model，if you need more detail please send me mail.

Hi Ahmad Bakir

Thanks for your reply ; could you provide a bit more insight on the use of the calculator ?

I do have the value of my failure criteria stored in a state value for each integration point but since I would like to modify the mechanical properties of an element during the simulation if it exceeds the failure criteria and since, to my knowledge, the calculator is called at the end of the simulation, I still feel stuck on that issue.

I greatly appreciate your help

Primary salivary gland tumors arise from major or minor salivary glands. Bone is devoid of such glands but primary salivary glands can invade bone if present in vicinity esp in head and neck regions. They can also show osseus metaplasia.

Are you sure they are % values? Regardless of the instrument used, you can't have a total of 141%, that doesn't make sense. Are the values you have been given some sort of percentage of a standard value, rather than an absolute concentration in mg/kg?

Perhaps go back to the lab staff and ask what output the instrument provides. Check if they ran a comparison sample to see whether the instrument was calibrated properly to give accurate results.

Was the instrument calibrated for bone material or was it set up for soil or some other material? Did you do any surface preparation of the bone or was it analysed as is? Now I am wondering if the surface of the sample was not oriented correctly and scattered x-rays in all sorts of un-focused directions and you simply have been given poor-quality results.

Dear Zna

You can use ImageJ software to achieve this purpose (view YouTube) or contact Dr. Osman Sharif (Computer Science - KUST) for medical image processing using MatLab software.

Note: Both techniques need high-quality image resolution and sharpness with an accurate scale bar and unit of measurement.

Definition of joints is not exactly specific. Believe it or not, that is a complicated question. The answer depends upon what you mean by joint. For example:

Growing bones are divided into growth centers. This creates a growth joint where bone is connected to bone known as a metaphysis. If you count this as a joint then the answer depends upon how old the person is.

Some bones come together bone-to-bone, and then fuse later in life. This can be a natural process like with bone growth but occurs late in life. The sutures of the skull are an example. These joints are called synostosis.

A similar process can occur that is pathological. That can create bone spurs and new linkages, joints, among bones.

Some bones are linked together by ligaments, creating a joint known as a syndesmosis. If you count every instance where a ligament spans between two bones, and count every ligament as a separate joining, you would have an enormous number of joints.

Most of the time when we think of joints, we are thinking about the moveable joints known as synovial joints. There are a lot of these, and not all synovial joints are mobile and not all mobile joints are synovial.

In addition, there are a variable number of bones in the body. Sesamoids, small bones inside tendons, are common but variable. The number of linkages between sesamoids and more standard bone, such as the patellofemoral joint, would be impossible to count without a careful examination of the subject involved.

There are many other types of joints in the body that I am not mentioning, and there are even named joints that do not link bone-to-bone. So, again, I have to state that the answer to your question depends upon how you want to define joint. And, once you come up with a definition you like, you will always find someone who disagrees with you.

'A'ishah (A.S.) narrated that the Prophet (PBUH) said:

25 joints between the vertebrae.

72 joints between the vertebrae and the ribs.

2 joints between the bones of the sternum and the thoracic cage.

18 joints between the sternum and the ribs.

2 joints between the clavicle and the scapulae (shoulder blade).

2 joints between the scapulae and the thorax.

2 joints between the scapular bones.

6 joints between the elbows.

8 joints between the wrists.

70 joints between the hand bones.

2 hip joints.

6 joints between the knee bones.

6 joints between the ankles.

74 joints between the feet bones.

4 joints between the coccyx vertebrae.

6 joints between the bones acetabulm.

1 joint of the pubic sumphysis.

Check the attached files for details. A newborn has 275 to 300 bones, while most adults have 206. Here’s why: As a baby grows, their smaller bones join together to create larger bones. So, the bone count and bone joint both are variable but can result in approximately same as mentioned in the Hadees

The atrophy of the thyroid gland in Hashimoto's Thyroiditis can potentially lead to a decrease in the number of C cells present in the gland. As a result, there may be a reduction in the production of calcitonin by these C cells. Calcitonin is a hormone that helps regulate calcium levels in the body by promoting calcium deposition in bones and inhibiting calcium absorption in the intestines.

Therefore, if the thyroid gland is atrophied due to Hashimoto's Thyroiditis, there may be a decrease in calcitonin production, which could potentially impact calcium metabolism and bone health.

Thank you for the suggestions. I am little nervous to try the ultrasound to degas, could this cause the damage to the marrow structure. I will try the vaccum degassing to see if I can make that work.

The human cortical bone typically experiences maximum strain levels ranging from 0.7% to 2%. Beyond this range, the bone may reach its failure point, resulting in fracture. It's important to note that various factors such as bone density, mineral content, and structural integrity can influence the specific strain levels at which cortical bone may fracture. Additionally, the rate of loading and the direction of force applied also play significant roles in determining the strain threshold for bone fracture.

Late answer for other researchers who reach this page: You may also find valuable resources at https://www.embodi3d.com

Excellent question. I see that the loading protocols are facts from the time of non-osseointegrated implants. Therefore, we are dealing with it as if it were conventional implants, for example, such as just bone, Garbatio, endless thread, etc. We go back to the past regardless of an analysis of mechanical or sonic bone locking. In the past, there were no dietary restrictions. Today I understand that greater care must be taken with food that does not break the prosthesis, compromising mechanics or aesthetics. If this occurs, it will often lead to the need to remove it using counter torque (shearing), increasing the risk of failure. Colleagues often lose implants but this does not compromise momentarily and risks the long-term survival of the prosthesis. They end up reporting this as a success, transferring these problems to be repaired with the installation of new implants and prostheses later. I think that reality is that the numbers are masked by a statistical bias and that arrogant personality speaks louder.

I presume you know that these are two questions with totally different answers.

The first question (how to preserve ...) also can be defined in different aspects (temperature, wet or dried, environmental, PH, etc). I suggest reading a reference book named "Mechanical Testing of Bone and the Bone-Implant Interface".

Regarding the second question (prepare in cylindrical or cubic shape...), have you read about a device named "Microtome". It is like a lathe machine. Normally researchers use this device (not very complicated) for preparing the desired shape.

Good Luck

Hi Xudan. Oh I forgot to tell, but I'm using the Cryostat for bone slicing at 25 micrometers.

Hi Jordan Tzvetkov ,

Did you also consider using Kawamoto cryofilm tape to stick sections to slides? It will require different embedding though.

I'm very curious to hear whether you managed to solve this issue? Many thanks in advance for sharing any tips!

Best wishes,

Erinke

Depending on the throughput of samples it might not be worth optimizing all these assays (and identify a set of suitable reference genes). It might be easier and eventually cheaper to go for RNA-seq, even if only very little of the RNA-seq output will be used.

If using qrtPCR, I would not recommend TaqMan if you don't go for multiplexing (what comes with its own difficulties). The higher detection specificity of TaqMan probes as compared to dsDNA-specific dyes like SYBR Green makes it more difficult to confirm specific amplification what is important to ensure the interpretability of Ct values. If you use TaqMan probes you will need to run post-PCR gels to check what all was amplified.

Another scenario except multiplexing where using TaqMan probes would be advantagous is if you would not go for quantification but only for detection, possibly of single molecules, and if the amplification of the assay would not be sufficiently specific (e.g. amplifying primer-dimers).

Sorry, that I cannot answer this quetion, but this will not belong to my special work flow of damage analysis.

Hello Mansha Saxena

You may test the anti-proliferative activity of a natural plant extract compound in vitro against bone/osseous cancer on MG-63 osteosarcoma human cell line by using MTT assay at 570nm.

In MTT assay, try using various concentration of the extract say in the range of 25ug/ml to 500ug/ml for various time points (24, 48 and 72 hours).

You may follow the MTT protocol as provided below.

1. Plate the MG-63 cells in the 96-well plate at the desired cell density such that they are 80% confluent on the day of readout. If cells are overconfluent on the final day of the assay, you may end up with inaccurate results.

2. After 24 hours, when the cells have attached to the substratum, treat the cells with the extract by changing the media with the treatment media at various concentration of the extract as mentioned above.

3. Incubate the cells with the treatment media for 24, 48 and 72 hours.

4. After the treatment period, aspirate the treatment media and add MTT to a final concentration of 0.2 to 0.5mg/ml and incubate in the dark for 1 to 4 hours for the formation of formazan crystals.

5. After incubation, solubilize the crystals into a colored solution with the solubilizing solution which may include either dimethyl sulfoxide or acidified isopropanol solution, or a solution of 10% SDS in 0.01M hydrochloric acid.

6. Measure the absorbance at a wavelength of 570 nm.

You may also use live & dead assay, which is a quick two-color assay, used to determine the viability of cells based on plasma membrane integrity and esterase activity in live cells.

Also, you may use LDH assay which is a method used for the evaluation of cellular toxicity since LDH release is considered as an early event of necrosis.

For in vivo experiment, a xenograft for MG-63 may be created in nude mice and treated with the extract.

You may follow the protocol as provided below.

1. You may generate a human osteosarcoma tumor xenograft by subcutaneous (s.c.) in vivo injection of human MG-63 osteosarcoma cell line (10 -15×10^6 cells/flank/0.2 ml) in the right flank of adult athymic female nude mice.

2. After the tumor inoculation (say approximately 5- 7 days later) the animals may be randomly divided into four groups of say six mice each:

Group 1 (Negative control) receiving the vehicle control orally once daily for say 15 days.

Group 2 (positive control) receiving a standard drug for bone cancer orally once daily for 15 days.

Group 3 (Experimental group A) receiving a low dose of the extract dissolved in an appropriate solvent orally once daily for 15 days.

Group 4 (Experimental group B) receiving a high dose of the extract dissolved in an appropriate solvent orally once daily for 15 days.

The mice may be euthanized at the end of 15 days of treatment period and the tumors may be excised and weighed. The tumor volumes may be measured according to the formula given below:

Tumor volume = A × B^2 × 0.5

where A: length

B: width

You may also use the excised tumor fixed in formalin (37%) for histological analyses by a trained pathologist.

I hope this may be helpful.

Good Luck!

Follow these steps:

1. Import the Geometry: Load the pelvis and sacrum bone geometry file into a 3D modeling software or a CAD program capable of handling complex geometries. Ensure that the file format is compatible with the software you are using.

2. Duplicate the Bone Geometry: Create a duplicate copy of the original bone geometry to work on. This will allow you to preserve the original bone geometry while creating the cortical bone shell.

3. Scale the Duplicate Geometry: Scale up the duplicate bone geometry uniformly by 4mm in all directions. This will create a larger version of the bone geometry, which will serve as the outer boundary for the cortical bone shell.

4. Offset the Duplicate Geometry: In the CAD software, use the "offset" or "shell" feature to create a new surface that is 2mm away from the outer surface of the scaled duplicate bone geometry. This will generate the cortical bone shell with the desired thickness.

5. Boolean Operation: Perform a Boolean subtraction operation between the original bone geometry and the cortical bone shell geometry. This will remove the original bone geometry from the cortical bone shell, leaving behind the shell itself.

6. Clean and Refine the Geometry: After the Boolean operation, you may need to clean and refine the resulting geometry. Check for any overlapping or intersecting surfaces and make necessary adjustments to ensure a watertight and smooth cortical bone shell.

7. Create Holes for Screw Insertion: Identify the locations where you want to insert screws and create holes in the cortical bone shell geometry accordingly. The size and shape of the holes will depend on the specifications of the screws you intend to use.

8. Export the Final Geometry: Once you have completed the cortical bone shell and added the necessary holes, export the final geometry in a suitable file format (such as STL) that can be imported into a finite element analysis (FEA) software.

I am NOT a doctor, it should be used only for models.

Hope it helps: partial credit AI

I recommend to use another brand of cell culture plate and monitor the results. I guess some uncommon surface roughness is behind this event.

Here are some notable advancements along with their pros and cons:

1. Dual-Energy X-ray Absorptiometry (DXA):

- Pros: DXA is considered the gold standard for measuring bone mineral density (BMD) and diagnosing osteoporosis. It is widely available, relatively low-cost, and exposes patients to low radiation doses.

- Cons: DXA primarily measures BMD and may not fully capture bone quality and fracture risk. It also has limitations in distinguishing between trabecular and cortical bone.

2. Quantitative Computed Tomography (QCT):

- Pros: QCT provides volumetric BMD measurements and can differentiate between trabecular and cortical bone. It is useful for assessing bone strength and fracture risk prediction.

- Cons: QCT involves higher radiation doses compared to DXA. It is less widely available and more expensive. Additionally, QCT scans may have lower spatial resolution compared to DXA.

3. High-Resolution Peripheral Quantitative Computed Tomography (HR-pQCT):

- Pros: HR-pQCT provides detailed three-dimensional imaging of bone microarchitecture. It allows for assessment of trabecular and cortical bone compartments, and evaluation of bone strength and fracture risk.

- Cons: HR-pQCT is limited to peripheral skeletal sites (e.g., wrist or ankle) and is not suitable for central skeletal assessment. It is relatively expensive and less accessible than DXA.

4. Trabecular Bone Score (TBS):

- Pros: TBS is a texture analysis technique applied to DXA scans that provides information about bone microarchitecture. It complements BMD measurements in assessing fracture risk.

- Cons: TBS is an indirect measure of bone microarchitecture and relies on DXA scans. It may not be as informative in certain clinical conditions or when DXA scans are of low quality.

5. Biochemical Markers of Bone Turnover:

- Pros: Measurement of bone turnover markers (e.g., serum levels of osteocalcin or CTX) can provide insights into bone metabolism and response to treatment. They are useful for monitoring bone health and treatment efficacy.

- Cons: Bone turnover markers are influenced by various factors and may not provide a direct assessment of bone strength or fracture risk. Their clinical utility is more limited compared to imaging-based techniques.

Hope it helps: credit AI

I am NOT a doctor but Research with AI provided this info:

1. Identification of Vertebral Skeletal Stem Cells (VSSCs): The discovery and characterization of Vertebral Skeletal Stem Cells (VSSCs) in the context of bone metastasis could be a game-changer. VSSCs are a subset of skeletal stem cells specifically localized within the vertebral column. Understanding their unique properties, molecular signatures, and interactions with metastatic cancer cells could provide valuable insights into the mechanisms of bone metastasis.

2. Role of VSSCs in Bone Metastasis Progression: Investigating the role of VSSCs in bone metastasis could reveal their contribution to the initiation, establishment, and progression of metastatic cancer cells in the vertebral bone. Understanding the molecular signaling pathways and cellular interactions between VSSCs and cancer cells could unveil novel therapeutic targets for inhibiting bone metastasis.

3. Modulation of VSSC Behavior: Manipulating the behavior of VSSCs could offer therapeutic horizons in bone metastasis management. Identifying factors or compounds that regulate the activity, proliferation, or differentiation of VSSCs may provide avenues to control the growth and dissemination of metastatic cancer cells within the vertebral bone. Strategies targeting VSSCs could potentially disrupt the bone metastatic niche and impede cancer cell colonization.

4. Targeting the VSSC Microenvironment: The VSSC microenvironment plays a crucial role in bone metastasis. Investigating the cellular and molecular components of the VSSC niche, such as neighboring cells, extracellular matrix, and signaling molecules, could reveal new therapeutic targets. Modulating the VSSC microenvironment to create an inhospitable niche for cancer cells or to enhance the efficacy of existing therapies could revolutionize the treatment of bone metastasis.

5. Personalized Therapies for Bone Metastasis: Leveraging the knowledge of VSSCs and their interactions with metastatic cancer cells, it may be possible to develop personalized therapeutic approaches for patients with bone metastasis. By understanding the specific characteristics of VSSCs in individual patients, tailored treatment strategies could be designed, potentially leading to improved outcomes and reduced side effects.

6. Combination Therapies: Combining therapies targeting VSSCs with existing treatment modalities, such as chemotherapy, radiation therapy, or targeted therapies, could synergistically improve the management of bone metastasis. By simultaneously targeting cancer cells and the VSSC niche, it may be possible to disrupt the intricate interactions between metastatic cells and the bone microenvironment, leading to enhanced treatment efficacy.

7. Biomarkers for Bone Metastasis: Uncovering specific biomarkers associated with VSSCs or their interactions with metastatic cancer cells could aid in the early detection and monitoring of bone metastasis. Reliable biomarkers could facilitate the development of non-invasive diagnostic techniques, enabling timely intervention and improved patient outcomes.

Hope it helps.

I am not sure if all niches with dormant tumor cells are anti-metastatic. There can be a lack of critical growth factors, including oxygen and nutrients, as well as immune regulation of tumor cell numbers within disseminate micro-metastases. Here is another of a host of reviews looking into this.

Check out these articles.

Thanks

Atuoha:

It depends on what for do you need the views. For a quick radiological assessment of the distal radius a Posterior-to-Anterior and a lateral views are enough. Oblique views can separate the carpal bones and asses better some particular joints and bones.

Dynamic views such as ulnar deviation or radial deviation will demonstrate the alignment of the carpal bones under stress and can increase the likelihood of diagnosing some pathologies, especially scaphoid bone fractures.

For further reading I suggest the following paper

doi: 10.1016/j.hcl.2005.02.008.

Hand Clin. 2005 Aug;21(3):279-88.

Essential radiographic evaluation for distal radius fractures.

Robert J Medoff

From my view IdavLandmark is the better option

All the best for your work

Jiri Trousil Thanks a lot.

I'm glad I could help Gonzalo Javier Rodriguez Carpio :-) Let me know if you have any more questions about micro-excavation at the lab.

Best,

Maria

for high throughput screening, how about "Establishing cell line" using bone marrow-derived mesenchymal stem cells since they possess both myogenic and osteoblast potential?

I got the book, Thank you everyone

High definition radiograph will show the boundary line.

Hello Emilie; I'd recommend that you find a copy of.

Hall and Kelson. Mammals of North America. Two Volumes. The keys will provide detailed answers to your questions. The reference is old but has excellent descriptions that are timeless. Best regards, Jim Des Lauriers

Bone has notbeen found to have non-fibrillar colkagens. Even the minor Type V collagen that modulates fibril size is fibrillar.

You will get here: https://simtk.org/projects/openknee

Don't just consider large game, there were other important component in Neandertal diets. Of course, good evidence for plant foods are both harder to identify and often ignored.

2011 Henry, A. G., A. S. Brooks, and D. R. Piperno. Microfossils in calculus demonstrate consumption of plants and cooked foods in Neanderthal diets (Shanidar IIIm Iraq: Spy I and III, Belgium). Proceedings of the National Academy. of Sciences 108 (2): 486-491.

But in addition to the emphasis in research on larger body-sized animals, small game and invertebrates also should be considered, both for dietary and environmental reconstruction.

2014 Buck, L. T. and C. B. Stringer. Having the stomach for it:a contribution to Neanderthal diets? Quaternary Science Reviews 96: 161-167.

2000 Richards, M. P., P. B. Pettit, E. Trinkaus, F. H. Smith, M Paunovic, and I. Karavanic. Neanderthal diets at Vindija and Neanderthal predation: the evidence from stable isotopes. Proceedings of the National Academy. of Sciences 97 (13): 7663-7666.

2005 Bocherens, H., D. Drucker, D. Billion, M. Patou-Mathis, and B. Vendermeersch. Isotopic evidence for diet and subsistence pattern of the Saint-Cesaire I Neanderthal: review and use of multi-source mixing model. Journal of Human Evolution 49 (1): 71-87.

2006 Balter, V. and L. Simon. diet and behavior of the Saint-Cesaire Neanderthal inferred from biochemical data inversion. Journal of Human Evolution 51 (4) 329-338.

2008 Richards, M. P. and R. W, Schmitz. Isotope evidence for the diet of the Neanderthal type specimen. Antiquity 82 (317): 553-559.

2009 Richards, M. P. an E. Trinkaus. Isotopic evidence for the diets of European Neanderthals and early modern humans. Proceedings of the National Academy. of Sciences 106 (38): 16034-16039.

2009 Hublin J.-J., D Weston, P. Gunz, M. Richards, W. Roebrorks, J. Glimmerveen, and L. Anthonis. Out of the North Sea: the Zeeland Ridges Neanderthal. Journal of Human Evolution 57 (6): 775-785.

2011 Dusseldorp. G. L. Studying Pleistocene Neanderthal and eve hyena dietary habits: combining isotopic and archeozoological analyses. Journal of Archaeological Method and Theory 18 (3): 224-255.

2011 Cortez-Sanchez, M., A, Morales-Muniz, M. D. Simon-Vallejo, M. C. Lozano-Francisco, J. L. Vera-Pelaez, C. Finlayson, J. Rodriguez-Vidal, A Delgado-Huertas, F. J. Jimenez-Espejo, F. Martinez-Ruiz, and M. A. Martinez-Aguirre. Earliet known use of marine resources by Neanderthals. PLoS ONE 6 (9): e24026.

Klein, R. G. and D. W. Bird. Shellfishing and human evolution. Journal of World Prehistory 44: 198-205.

2005 Guthrie, R. D. The Nature of Paleolithic Art. Chicago university Press, Chicago. (mention of possible depictions of insects, the larvae of parasitic warble fly who commonly prey on reindeer and are used as food bty modern etnographically recoded groups).

1999 Hewitt, G.M. Post-glacial re-colonization of Eupean biota. Biological Journal of the Linnean Society 28 (4): 247-284

1994 Vaisanen, R. and K, Heliovaara. Hot spots of insect diversity on Northern Europe. Annales Zoologici Fennici 31 (1) 71-81.

2014 de Pablo, J. F.-L., E Badal, C. F. Garcia, A Martinez-Orti, and A. S. Serra. Land snails as a diet diversification proxy during the early upper Paleolithic in Europe. PLoS ONE 9 (8): e104898.

Dmitry Zinovkin ,

First of all, I’d propose you correct – at least - the title of your Query in RG to:

This might end up in more ‚attraction‘ to readers of RG and interested colleagues as well.

Think also on / about „full text search function“ in RG’s Q/A-archives.

Nobody will find your Question on >>SAFRANIN O stain<< easily, when you define it as „Saphranin“ or 'Sagranin O'.

OK. The issue / matter might be more complicated as initially thought. This due to not only several recipes / protocols available from (hand-)books, web sources, original articles, etc.

Your ‚working protocol‘ as stated in your initial QUESTION (see above) MIGHT be inaccurate or at least seems to be vague in particular regarding the steps one has to consider when performing such a complicated staining recipe / protocol. So it MIGHT be of benefit if you would let us know WHICH recipe / protocol you exactly are following (reference?).

You may compare then with a websource (found, e.g. at: http://www.newcomersupply.com/documents/staining/procedures/Safranin%20O%20Stain.1350.pdf ,

STAINING PROCEDURE: 1. Deparaffinize sections thoroughly in three changes of xylene, 3 minutes each. Hydrate through two changes each of 100% and 95% ethyl alcohols, 10 dips each. Wash well with distilled water.

a. See Procedure Notes #2 and #3. 2. Wash well in running tap water; rinse in distilled water. 3. Prepare fresh Weigert Iron Hematoxylin (1409); mix well. a. Solution A: Ferric Chloride, Acidified 20 ml b. Solution B: Hematoxylin 1%, Alcoholic 20 ml 4. Stain in fresh Weigert Iron Hematoxylin for 10 minutes. 5. Wash in running tap water for 10 minutes; rinse in distilled water. a. See Procedure Note #4. 6. Prepare 0.25% Fast Green Stain, Aqueous; combine and mix well. a. Fast Green Stain 2.5%, Aqueous (10852) 5 ml b. Distilled Water 45 ml 7. Stain in 0.25% Fast Green Stain, Aqueous for 5 minutes. 8. Rinse directly in Acetic Acid 1%, Aqueous (10012); 10-15 seconds. 9. Place directly in Safranin O Stain 1%, Aqueous for 5 minutes. 10. Dehydrate in two changes each of 95% and 100% ethyl alcohol.

Clear in three changes of xylene, 10 dips each; coverslip with compatible mounting medium.

RESULTS: - Cartilage: Red to orange - Mucin and mast cell granules: Red to orange - Bone, connective tissue and cytoplasm: Green - Nuclei: Black

Contrary to such a 'modern, & revised staining protocol' I'd like to offer you: SAWYEK (1940) A Standardized Technic for Safranin O, Stain Technology, 15:1, 3-7, DOI: 10.3109/10520294009110324, which in the SUMMARY reads:

>>ABSTRACT.-A method for control of staining with safranin O is described. The procedure is as follows:

As mentioned already above In addition, fixation matters too (NB: stated not only in THIS old article):

"Since staining affinity varies widely with the fixation employed, four representative fixing fluids were used: Petrunkevitch’s paranitrophenol-cupric-nitrate-nitric (Guyer, p. 34), Zenker’s bichromate sublimate-sodium-sulphatemetic (Lee, 1937, p. 46), Bouin’s picroformol-acetic (Lee, p. 58), and Petrunkevitch’s sublimate-nitric-acetic-alcohol (Lee, p. 45)."

So, IMHO, 'color deviations' from the "normal staining results" (as reported in any basic literature) may be a consequence of one to some to many alterations in parameters underlying your protocol: fixative, pH, (even sometimes osmolarity / ionic composition of solutions), dyes' quality, rinsing ('blueing': tap water quality, distilled water quality...), application time (dipping for seconds, incubating minutes or hours...etc., etc.).

So one has to go through "try and error" when faced with a problem "pink-violet" instead of "blue" stained bone (guessing it might have been the quality or application mode of the >Weigert's Iron Hematoxylin< stain and further treatment).

You can see (and perhaps the ‚professional‘ knows that for sure) that –especially with the staining sequence you use – not only dye quality is of interest, but starting with the fixation / fixative used, continuing with overall specimen processing AND treatments during staining itself will matter and yield different results in coloration (there are major differences in incubation / treatment steps for washing, clearing and last but not least, mounting of the sections too).

One (and I) cannot help with ‚trouble shooting‘, when not knowing - from scratch and, finally- the entire concept/protocol (i.e. what has been done from A-Z).

BERGMAN et al_2015: The Bone-Inflammation-Cartilage (BIC) Stain: A Novel Staining Method Combining Safranin O and Van Gieson’s Stains

described and „… established a novel staining method, the Bone-Inflammation-Cartilage (BIC) stain, in which Safranin O staining is combined with van Gieson’s staining to visualize and correctly assess bone erosions, the loss of articular cartilage, and inflammation in a single stain….“

(Free access, at least for me; cf.:

https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4804731/pdf/10.1369_0022155415591599.pdf )

You need to compare the steps you perform with those steps which are published in those many articles which can be found in literature searches or your own recipe / protocol collection.….

There are plenty of other / further staining protocols out in the world’s wild web

(e.g.: https://www.oarsijournal.com/article/S1063-4584(10)00240-2/pdf ,

or / and

https://eprints.nottingham.ac.uk/11979/1/PDF_of_final_thesis_submitted.pdf,

= Thesis for the degree of Doctor of Philosophy, S. ASHRAF July 2011: Contributions of Inflammation and Angiogenesis to Structural Damage and Pain in Osteoarthritis)

but one has to invest some time and ideas where to arrive with regard to the task you are dealing / have to deal with.

Elucidating / Correcting the ‚conundrum‘ regarding

German Isauro Garrido-Fariña 's hint at: „…en la diafanización a claramiento de Dowson….“

it might be interesting that in ENGLISH it may spell / correctly spells:

'Diaph o nisation (diaphonization)' and DAWSON, which can be found also by taking the Google ‚survey‘ for those terms (as one result out of some) e.g.:

http://uni-sz.bg/wp-content/uploads/vmf/file/04%20N_%20Tsandev.pdf

Bulgarian Journal of Veterinary Medicine, 2017, 20, Suppl. 1, 27–32 ISSN 1311-1477 (print); ISSN 1313-3543 (online):

by N. TSANDEV et al, 2017

Summary Tsandev, N., A. Atanasoff, G. Kostadinov, E. Petrova-Pavlova & I. Stefanov, 2017. Elaboration of transparent biological specimens for visualisation of developing cartilage and bone structures. Bulg. J. Vet. Med., 20, Suppl. 1, 27–32.

>>Several modifications of diaphonisation technique for preparation of transparent permanent preparations from fish, amphibians and reptiles are presented. It was demonstrated that the omission of some procedures, changed concentration of solutions, and duration of several diaphonization steps did not alter the quality of obtained permanent specimens. The prepared models could be used for monitoring of skeleton development, and later embedded in transparent polymers with respect to their use in museum collections and exhibitions. Key words: bone, clearing, diaphonisation, fish cartilage, morphology, transparent<< and, from the text:

„…Diaphonisation comprises consecutive fixation of biological specimens in formalin, bleaching with hydrogen peroxide, incomplete maceration in potassium hydroxide, staining with dyes and preservation in glycerol (Pramod et al., 2011). The technique is mainly used for fish, amphibians and reptiles due to their smaller body size and very delicate for dissection tissues (Taylor & Van Dyke, 1985)….“

Regarding the name „DAWSON“ (as mentioned in TSANDEV et al,2017):

cf.: Dawson, A., 1926. A note on the staining of the skeleton of cleared specimens with alizarin red S. Stain Technology, 1, 123–124.

If you haven’t found (Google) web-sources as for implementing 'literature searches' I mentioned yesterday, then I propose and recommend your doing a google search for keywords:

| Weigert hematoxylin Safranin O Fast Green Stain Bone Cartilage | =[5,860 results], or also

| Weigert hematoxylin Safranin O Fast Green Stain Bone Cartilage, Diaphonization or Diaphonisation |

IF you are not allowed or able to use e.g. Google search via unlimited Internet access, you might have problems to access and download the bibliographic sources I included in my post (nevertheless some may at least be accessible via Research Gate's database).

Since you have posted your Question in ResearchGate I think you have at least access to RG’s FULL TEXT RE-SEARCH in their ‚ARCHIVES‘….so take the time to access the SEARCH function (find within a rectangle in the upper menue bar with text: „Search for research, journals, people, etc.", insert your search term(s) / keyword(s), activate in browser and then choose from the menue popping up what seems to be appropriate to / for you: Research (Journals, Persons, Articles), Q/A,… you’ll wonder about the treasure of data saved already in RG. In case you are not served with (enough) results just vary or reduce the amount of (your) keyword(s).

E.g.: for term(s): | [safranin O] rats bones intra-articular fracture | :

by clicking the loupe-icon (automatically chooses RESEARCH=„publications“) will result in displaying an URL: https://www.researchgate.net/search.Search.html?query=%5Bsafranin+O%5D+++rats+bones++++intra-articular+fracture+&type=publication:

also displaying hits/results of your query: You’ll (might) have a lot of possible literature to evaluate.

Choosing the search option „QUESTIONS“ (with the former keywords) results in only two postings: your recent 'Question' and a question asked in 2014 on >decalcifying bone…<

Varying your search terms accordingly you’ll get more and more special references in RESEARCH or QUESTIONS which already have been posted and archived (e.g. Full text search: | safranin O stain bone intra-articular fracture | from QUESTIONS will yield a lot more information on the matter.

Hoping my elaboration helped you anyway…regards and best of luck...WHM

Michelle:

Janane's answer is a good one to use the maximum bending stress at the center of the tension surface to estimate parameters such as the elastic modulus and the stress at yielding or fracture, but in general measuring the elastic modulus by such mechanical techniques can be severely inaccurate as you're not measuring the strain accuarately enough (or even at all) and there is the situation mentioned by Janane with respect to the inhomogeneous nature of the structure of bone. The elastic modulus of bone is roughly 15 to 20 GPa but if you perform bend tests on mouse bone, you are like to get numbers as low as 3 GPa for these (and a few other) reasons.

The best way to measure the elastic modulus is any material is by measuring the propagation rate of a compression wave (or equivalent) but again even this is a problem in bone due to its inhomogeneous structure and geometry. If you are studying the cortical bone, the best way in my opinion is to use nanoindentation but to take many readings on a sinlge bone to compensate for the fact that each nanoindentation measure is examning only a very small volume.

I hope that this helps.

ROR

Afin de mieux comprendre votre problématique je conseille vivement la lecture de l article de Frost H.M. sur la théorie du Mécanostat. (Mécanisation déterminants of bonne modélisation.

Met. bone. Dis. Rel. Res.,4:217-229. 1982.

People who are overweight or have obesity appear to show a blunted response to vitamin D supplementation compared with normal weight individuals in a new analysis of a randomized trial.

Yes, there are recommended methods and procedures for lake sediment sampling for AMS radiocarbon dating. The following is a comprehensive guide on the recommended methods and procedures for lake sediment sampling for AMS radiocarbon dating. Lake sediment samples can be used for AMS radiocarbon dating to determine the age of the sediment and the lake's history. However, to obtain accurate results, it is essential to follow recommended methods and procedures for lake sediment sampling. The following are the recommended methods and procedures: 1. Sampling Location: Selecting the right location for sediment sampling is crucial. It is recommended to choose a location that is representative of the entire lake's sedimentation history. The location should be in a relatively undisturbed area of the lake, away from any inflows or outflows, and where sediment deposition is continuous. 2. Sampling Equipment: The equipment used for sediment sampling should be clean and free from any contaminants that could affect the radiocarbon dating results. Stainless steel corers are commonly used to collect sediment samples from lakes. 3. Sampling Depth: The depth of sediment sampling should be based on the lake's depth and the desired time resolution of the radiocarbon dating analysis. It is recommended to sample at least 10 cm intervals to capture changes in sedimentation rates accurately. 4. Sample Preservation: Once collected, it is essential to preserve the samples correctly to prevent any contamination or alteration of the sample material. Samples should be stored in airtight containers at low temperatures until they are ready for analysis. 5. Sample Processing: Before AMS radiocarbon dating analysis, it is necessary to process the samples properly. This includes drying, homogenizing, and removing any organic matter that could interfere with radiocarbon dating analysis. 6. Radiocarbon Dating Analysis: AMS radiocarbon dating analysis should be performed by an accredited laboratory using standard protocols and quality control. Following these recommended methods and procedures can ensure accurate and reliable radiocarbon dating results.

References: 1. "Guidelines for AMS radiocarbon analysis of lake sediments" by S. Juggins and A. J. Burridge, published in Quaternary Science Reviews. 2. "High-resolution AMS radiocarbon dating of annually laminated sediments from Lake Żabińskie, Poland: Varve chronology for the last 6000 years" by M. Blaauw et al., published in Quaternary Science Reviews. 3. "Radiocarbon dating of lake sediments" by P. Appleby and F. Oldfield, published in Philosophical Transactions of the Royal Society A: Mathematical, Physical and Engineering Sciences.

Hello!!

From my not very long experience, as you say, we can see the adypocites surrounding the bone marrow.

Commonly, the blue stands for collagen, which in the bone tissue normally is type 1, although at some levels of ossification we could see type 2 collagen.

I suppose that the cells within all the blue structures might be osteocytes.

Because it is bone marrow, the fibers that sorround it will be type 3 collagen, and would be positive with PAS because of the proteoglycans that sorround them or with silver staining.

I would love to answer you more specifically, but the image is not very close, so i can't identify any Megakaryocite or any blood-forming cell. If you have a bigger picture, please send it to me.

Wish you the best!!!

Hi Daniel

Thank you very much for the help, that is defintly a possibility, I will look more into it

Cheers

Jesper

Ziad Hazim Deleme Thank you for your nice and informative answer.

Hello, Sebastian Eberle . I have looked at your paper, and there was written that you considered convergence when the results did not change more than 5% in three subsequent refinement steps. May I know your base in defining such assumption? Actually I have seen other papers using this assumption as well, but maybe there is any mathematical evidence that proves this assumption..? Nonetheless, thank you very much for sharing your work. It's a meaningful guide for my own research. Best regards.

You may want to check my post elsewhere at this forum: https://www.researchgate.net/post/Alternative_methods_to_precipitate_collagen_proteins_with_neutral_salts

In Middle East countries

Dear Dr Desi Sandra Sari

Thank you very much Dr for your recommendation. I have contacted Dr. Fourier about that.

Best regards,

Maria

<reply by direct message>

:|

If you put a nice multimeter across the points of V1 and V2 do you see a change in voltage as you stretch the gauge?

I'd unhook the op-amp and Arduino for now, and just see if the strain gauge is happy: although that all looks fine. On its own the bridge ought to behave like a standard 'quarter bridge'.

In case some people may encounter this question in future, I add a reply. One way is that export the geometry as .obj or .stl. Then import in Meshmixer. In meshmixer using the make solids option or by inspecting the gaps they are filled. Aftar that the part can be exported as .stl and return to hypermesh or other software to proceed meshing and making .inp or etc.

There is no fixed amount like X/16. Optimal mesh size depends on the model geometry. It must assures that stress gradients are captured adequately

Growth factors and docking proteins necessary for biological joint replacement include:

-Transforming growth factor beta (TGF-β), which helps to regulate the production of both cartilage and bone.

-Bone morphogenetic proteins (BMPs), which induce the formation of bone and cartilage.

-Fibroblast growth factors (FGFs), which are involved in the differentiation of multipotent progenitor cells into osteoblasts and chondrocytes.

-Insulin-like growth factors (IGFs), which promote the proliferation of osteoblasts and chondrocytes.

-Vascular endothelial growth factor (VEGF), which stimulates the formation of new blood vessels.

-Adhesion proteins, such as integrins, which help to attach the newly formed cartilage and bone to the existing tissue. In terms of quantities, these proteins typically come in milligrams to grams, depending on the size of the joint replacement and the specific proteins needed.

In order to move this technology forward, research needs to be done to better understand the interactions between these proteins and other cells and molecules in the joint, as well as the effects of different growth factors on the formation of bone and cartilage. Additionally, further research is needed to develop ways to attach the newly formed cartilage and bone to the existing tissue, as well as to optimize the 3D printing process. Finally, clinical trials are needed to assess the safety and efficacy of this technology.

I think you will need to modify this example to do the segmentation (of sorts) you are trying to do, but I found this example of rendering color onto a volume: https://kitware.github.io/vtk-examples/site/Python/Rendering/Rainbow/

You may be looking for something different, I am not certain you can add color inherently to .stl files, you may need to use a different filetype like .obj.

Hi

Various types of HA-based materials such as pure HA, ion-doped HA, and HA/polymer composites, have been designed and used.

Please see this recently Review Article (2021)

Hydroxyapatite Based Materials for Bone Tissue Engineering: A Brief and Comprehensive Introduction

By Hui Shi, et.al.

https://doi.org/ 10.3390/cryst11020149

Thank you

Very good answer that submitted by the respected colleagues. I shall follow the next answers.

Kind regards!

One way analysis of variance (fixed effects model) for comparing all means and selected individual comparisons.. However from your results and with more treatment groups than biological replicates ,it does not look as if you will find any statistically significant differences. With multiple comparison adjustments this would become even less likely.

You could also analyse components of variance using a random effects model. This would compare between group variance with within group variance.

. For pure stochiometric mineral (hydroxyapatite) Ca/P ratio is 1.67. HA is Ca5(PO4)3(OH). So, we can see that for each 5 atoms of Ca it has 3 atoms of P. We can divide 5 by 3 (to get a Ca/P ratio), and result will be 1.666666666666667.

Recent research has stressed the importance of Vitamin D — not just for good bone health, but also for possibly preventing chronic disease when we are older. It has been linked to brain and heart health, obesity, mood, and, more recently, COVID-19. Yet, many children today are not getting enough Vitamin D.

Hello,

well, to me it looks like you did a segmentation of "air" and then created the pore-network for it. The problem is that your "air" contains not only the internal pores, but also the air surrounding your object. You have to get rid of that, to be able to calculate the pore-network only for the internal air.

You can do all necessary steps inside Avizo.

I suggest that you create a mask of your object (i.e. a binary label-field containing the filled shape of your object), and then use that mask to mask-out the external air, e.g. using "Mask" or "Arithmetic".

For creating the mask, you could start with your segmentation, invert it (so that it contains the bone material), and then apply a morphological "Closing" (maybe followed by some "Erosion".

For segmenting only the internal air, you could also use the module "Ambient Occlusion". Here is the link to a short introductory video for it: https://www.youtube.com/watch?v=pup_alqYxOY&list=PLoxdPzacxPYjDVMD4tPCaVbuQjxYizr_g&index=64

I hope that helps ...

Kind regards,

Andreas

Dear Dr. Liu Weidong,

you could try the freeware:

ImageJ / Fiji

A plugin for bone can be downloaded here:

https://www.liverpool.ac.uk/ageing-and-chronicdisease/bone-hist.

It is for instance described in:

van ’t Hof, R.J., Rose, L., Bassonga, E., and Daroszewska, A. (2017). Open source software for semi-automated histomorphometry of bone resorption and formation parameters. Bone 99, 69–79. https://doi.org/10.1016/j.bone.2017.03.051.

A good start would probably be to familarize yourself with the nomenclature used in bone histomorphometry described in the following excellent article:

Dempster, D.W., Compston, J.E., Drezner, M.K., Glorieux, F.H., Kanis, J.A., Malluche, H., Meunier, P.J., Ott, S.M., Recker, R.R., and Parfitt, A.M. (2013). Standardized nomenclature, symbols, and units for bone histomorphometry: a 2012 update of the report of the ASBMR Histomorphometry Nomenclature Committee. J. Bone Miner. Res. 28, 2–17. https://doi.org/10.1002/jbmr.1805.

Good luck

Stefan Tangl

Manarshhjot Singh

Hi,

Thank you for the reply. What you explained is right, assigning 100 properties is what I am doing right now using Mimics. based on the empirical relationship between the Hounsfield unit and material property (density), mimics automatically assigns 100 properties by dividing the bone region into 100 different parts. I cannot use the average value of bone properties, as the cortical region will have a higher value than the cancellous region, and the average value will not consider this difference.

right now I am using mechanical APDL, to export the bone as a .cdb file and then importing it into mimics. I just wanted to know if such a process is feasible with design modeller.

Hello Younas Masih

It is not the right choice. You need to work on blood cancer and bone cancer separately.

The bone is made up of compact bone, spongy bone, and bone marrow. Compact bone makes up the outer layer of the bone. Spongy bone is found mostly at the ends of bones and contains red marrow. Bone marrow is found in the center of most bones and is the soft, spongy tissue that has many blood vessels. There are two types of bone marrow: red and yellow. Red bone marrow contains blood stem cells that can become red blood cells, white blood cells, or platelets. Bone marrow makes nearly all the comp