This document discusses bone biopsy and decalcification. Bone biopsy is described as a procedure to remove bone samples for examination under a microscope to diagnose bone disorders like cancer or infection. It can be done through closed or open methods. Bone decalcification is defined as removing calcium from bone to make it flexible for pathological examination. Methods of decalcification discussed include acids, chelating agents like EDTA, and tests for completion like x-ray or ammonia testing.

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BONE BIOPSY
AND
DECALCIFICATION
MUHAMMAD SHOAIB
M.PHIL. Biochemistry

2. HENRY MOORE
NO ONE EVER KNOWS WHAT
EACH DAY IS GOING TO
BRING. THE IMPORTANT
THING IS TO BE OPEN AND
READY FOR IT.

3. SHIFA
TAMEERE MILLAT
UNIVERSITY
FOR
APPLIEDSCIENCES STUDIES

4. Calcium ions found in the bones are responsible for its
rigid posture and without it, bones may be flexible and
most of time unable to carry their body weight.
Diseases such as osteomalacia and rickets are as a
result of lack of calcium in the bones of those areas.
De-calcification of bones as our topic is about the
removal of these calcium from the bones. As such, we
are going to look at the following about the topic:
•The definition of decalcification of bones
•The purpose of decalcification of bones
•The principle of bone decalcification
•The method of decalcification of bones
How to test for the completion of decalcification

5. INTRODUCTION
BIOPSY
A biopsy is a procedure performed to remove tissue or
cells from the body for examination under a
microscope.
BONE BIOPSY
A bone biopsy is a procedure in which a small sample of
bone is taken from the body and looked at under a
microscope for cancer, infection, or other bone
disorders.

6. TYPES OF BIOPSY
Bone biopsy can be done by following
two methods
• Closed or needle biopsy
• Open biopsy

7. Closed or needle biopsy
• A drill biopsy is generally used to obtain a small specimen
• clean the skin over the bone where the biopsy sample will be taken
and local anesthesia in skin is given.
• The radiologist or surgeon puts a long, thin needle through the
skin into the bone
• Doctor may make a small cut in your skin before putting the needle
in so the needle passes easily
• Take out a small amount of bone through the needle.
• After a closed or needle biopsy, a small bandage is placed over
the area and pressure is put on the area to stop any bleeding

8. Open biopsy
• An open biopsy takes 30 to 60 minutes
• Before an open biopsy, general anesthesia is given to
the patients
• Before making a skin cut the area need to be shaved
and clean
• The surgeon makes a cut to see the bone and take out a
small piece.
• After an open biopsy, the cut is cleaned and closed
with stitches (sutures). A bandage is put on the area.
The stitches are taken out about 14 days after the
biopsy.

9. Purpose of bone biopsy
• Confirm the diagnosis of a bone disorder
• determine if a bone tumor is malignant (cancerous)
or benign
• evaluate bone pain or tenderness .
• determine the cause of an unexplained infection or
inflammation
• Find the cause of ongoing bone pain.
• Check bone problems seen on an X-ray.

10. Risks after bone biopsy
• bruising and discomfort at the biopsy site
• bone fracture
• prolonged bleeding from the biopsy site
• infection near the biopsy site or in the
bone

11. Bone biopsy results
Normal The biopsy sample shows normal bone tissue.
Abnormal
Bone tissue may show signs of infection, cancer, or
another bone disorder (including Paget's disease,
osteomyelitis, a bone cyst, or a benign bone
growth called an osteoma). The bone tissue may
also show osteoporosis or osteomalacia, which
means the bones are weak.
Most cancer of the bone spreads to the bone from
another part of the body, such as the breast,
lungs, prostate, or other organs. But bone cancer
can also start in the bone itself (such as
osteosarcoma or multiple myeloma

12. BONE
DECALCIFICATION

13. INTRODUCTION
Decalcification
Loss of calcium salts from a bone or tooth.
OR
The process of removing calcareous matter.
Bone decalcification
Removal of calcium ions from the bone through
histological process thereby making the bone
flexible and easy for pathological investigation.

14. PURPOSE OF BONE DECALCIFICATION
The purpose is to make the bone flexible
and easy for pathological investigation.
This is necessary in order to obtain soft
sections of the bone using the microtome
Failure to decalcification results in torn,
ragged sections and damage to the
cutting edge of the microtome knife

15. PRINCIPLE OF DECALCIFICATION
Insoluble calcium salt are converted into
soluble calcium salts by the action of
decalcifying agent so that the tissue
become soft.
Chelating agent binds to calcium ions
present in the bone and decalcification is
carried out.

16. Methods of decalcification
• Acid decalcifying agents.
• Ion exchange resins with acid and
decalcifying fluids.
• Electrolytic decalcification.
• Chelating agents.

17. Acid decalcifying agent
• The commonest method of
decalcification is dissolving calcium
salts in an acid solution.
• Some of the acid decalcifying agents
are..
 Nitric acid.
 Formic acid.
 Trichloroacetic acid

18. Nitric acid
MEHTOD
Thin slices of fixed tissue are placed in a
freshly prepared 5-10% solution of nitric
acid in distilled water.
Decalcification through this should not
extends beyond 48 hours.
Formalin is added to nitric acid to protect
the tissue against maceration and
swelling.

19. Formic acid
• Formic acid is widely used as a decalcifying
agent.
• For routine 10% of formic acid in distilled
water is recommended and higher
concentration gives more rapid
decalcification.
• A large volume of fluid is used and renewed
every 48 hours.

20. Ion exchange resins with acid
decalcifying fluids
The removal of calcium ions from the
decalcifying fluid by the resins leads to
quicker and more efficient decalcification.
Electrolytic Decalcification
It is the speedier decalcification without damage
to cytological features and staining.
Drawback:
Heat produced in the process may cause the
charring of specimen in the process

21. Chelating Agents
• EDTA is a chelating agent, it is a white
crystalline powder soluble in distilled
water to about 20%.
• As a decalcifying agent it combines with
calcium ions to form soluble, non ionized
compounds.
• The volume of solution for decalcifying
should be 150 times that of the tissue.
• The solution should be renewed every
5 to 7 days during decalcification.

22. Advantages of EDTA
• Deposits of iron and other metals may also be
removed by EDTA.
• Tissue is not hardened after decalcification.
• It can be good for Bone ,Teeth and any calcified
tissue.
• This is also the preferred solution for
decalcifying bone material for transmission
electron microscopy .

23. Test for completion of Decalcification
There are two method for determining the
completion of progress of decalcification.
 By X-ray examination.
 By Ammonia method.

24. X-ray Examination
• The most reliable method for
determining decalcification.
• But as this facility is not presented in all
the laboratories

25. Ammonia Method
• In this method ammonia is added drop by
drop in the decalcifying solution
cloudiness indicate the presence of
calcium.
• The specimen is then placed in a fresh
solution of decalcifying fluid and test is
repeated after a suitable interval of time.

26. SUMMARY

27. BONE BIOPSY
It is a procedure in which bone samples are removed (with a
special biopsy needle or during surgery) to determine if cancer
or other abnormal cells are present.
Done by two methods
• closed method
• Open method
BONE DECALCIFICATION
Removal of calcium ions from the bone through histological
process thereby making the bone flexible and easy for
pathological investigation.

28. QUOTE :
THE ONLY WAY TO DISCOVER THE LIMIT OF
THE POSSIBLE IS TO GO BEYOND THEM
INTO THE IMPOSSIBLE.