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BOTOX®     (BOTOX® COSMETIC) Wrinkle free Solutions

Dr. Hari Muraleedharan Msc,PhD,MA

Microbiologist

                                It is the hottest craze. With one minute injection, within minutes your face can be youthful, devoid of any unsightly wrinkles. BOTOX® clinics are popping up all over the country, helping clients venture back a few years into their past. A hub for beauty, unblemished skin, and eternal youth. Now a day’s most of the senior film stars were using BOTOX® to keep their porcelain skin wrinkle free. Although BOTOX® has been used for other conditions in the past such as neck spasms, cranial nerve disorders and eye spasms, when the FDA finally approved BOTOX® to be used to treat wrinkles, the popularity of the toxin rose tremendously.

                  BOTOX® is currently the only form of botulisum toxin approved for such cosmetic purposes. It is a nonsurgical, physician-administered treatment that can temporarily reduce moderate to severe frown lines between the brows in patients 18 to 65 years of age. It is a mixture of several protoplasmic proteins that are released from cells after autolysis. BOTOX® is the brand name for a special formulation of botulinum toxin A manufactured by Allergan Inc., of Irvine, CA, U.S.A.,. BOTOX® is used to generate lines or wrinkles between the eyebrows look better. Initially, the toxin was used for eye muscle problems.

What is BOTOX® COSMETIC?

BOTOX® COSMETIC (Botulinum Toxin Type A) Purified Neurotoxin Complex is a sterile, vacuum-dried purified botulinum toxin type A, produced from fermentation of Hall strain Clostridium botulinum type A grown in a medium containing casein hydrolysate, glucose and yeast extract. It is purified from the culture solution by dialysis and a series of acid precipitations to a complex consisting of the neurotoxin, and several accessory proteins. The complex is dissolved in sterile sodium chloride solution containing albumin human and is sterile filtered (0.2 microns) prior to filling and vacuum-drying. The specific activity of BOTOX® COSMETIC is approximately 20 units/nanogram of neurotoxin protein complex. Each vial of BOTOX® COSMETIC contains 100 units (U) of Clostridium botulinum type A neurotoxin complex, 0.5 milligrams of albumin (human), and 0.9 milligrams of sodium chloride in a sterile, vacuum-dried form without a preservative.

How was this found?

                         United States Food and Drug Aadmistration (FDA) approved BOTOX® over 10 years ago to treat certain problems with the eye muscle. Doctors observed that some wrinkles around the eyes looked better, too. The company that makes BOTOX® tested it. They showed the FDA that BOTOX® worked and was safe for treating some kinds of wrinkles. The lines that appear between your brows actually result from muscle movement and the passage of time. Two muscles are largely responsible for frown lines between the brows. When those muscles contract, they draw the brows together. As skin becomes less elastic over time, repeated frowning can result in those moderate to severe lines between your brows. BOTOX® works by blocking nerve impulses, which reduces the movement of those muscles. With less movement, the skin surface gradually smoothes out, and the moderate to severe frown lines between your brows begin to fade. A doctor skilled in the use of BOTOX® will inject little amounts of BOTOX® into the muscle. Only the `treated muscle can’t move. BOTOX® works for about four months. As the muscle returns to normal, you will see the wrinkle again.

 How is BOTOX® Tested for Potency?

                  The standard method for assessing the potency of BOTOX® is the LD50 Test. This test involves giving mice a single injection of the product into their abdominal cavity and seeing if they die within 3-4 days. The mice are first assigned to one of various dose groups. The aim of the test is to approximation the dose that kills 50% of the animals (hence the name “Lethal Dose 50%”). The LD value is designated as a unit (U) or a Mouse Unit (MU) of potency. The lower the LD50 value (or the U or MU), the higher the potency. Approximately 100 mice have conventionally been used to test each batch of botulinum toxins products, though Allergan has claimed that the company has significantly reduced this number.  Depending on the dose injected and the potency of the batch being assessed, the test animals experience differing levels of muscular paralysis and impaired vision. The end point of the LD50 Test is death, usually by suffocation after the respiratory muscle becomes paralyzed.

Background of hazardous bacterial toxin as a biological weapon

                                           The term “toxin” means the toxic material of plants, animals, microorganisms, viruses, fungi, or infectious substances, or a recombinant molecule, whatever its origin or method of production, including – (A) any poisonous substance or biological product that may be engineered as a result of biotechnology produced by a living organism; or (B) any poisonous isomer or biological product, homolog, or derivative of such a substance.

                       Over 60 years ago, botulinum toxin was first developed as a biological weapon; it can be aerosolised, or used to contaminate food, and the estimated lethal oral dose is 70 µg. Botulism is a rare paralytic disease caused by botulinum neurotoxin (BT), a protein produced by the soil anaerobic bacteria Clostridium botulinum. Eight types of Cl. botulinum have been identified (Types A, B, C1, C2, D, E, F, G) based on the immunological diversity in the toxins produced by them. The toxins are proteins with molecular weights of approximately 70,000 mgD. Botulinum toxin is the most poisonous substance known and the microorganism that manufactures it is relatively easy to isolate from nature and to culture. The only problem is making sure that oxygen is excluded from cultures. Despite its large size, the toxin molecule is stable enough to be dispersed as an aerosol. The toxin was considered for use as a major biological weapon by Britain and Japan during the 1930’s and there are suggestions that it was used by the Japanese to poison streams used as water sources by the Soviets. It has also been suggested that either the microorganism or the toxin were incorporated into grenades used in the assassination of the Reinhard Gehlen, Hitler’s likely successor and Reichsprotektor of Czechoslovakia.

                              A biological warfare attack with botulinum toxin delivered by aerosol will expected to cause symptoms similar in most respects to those observed with food-borne botulism. Commercially this organism attacked in canned food industry due to contamination. Proper sterilization will remove these organisms. In pure form, the toxin is a white crystalline substance, which is readily dissolvable in water, but decays rapidly in the open air. The lethal dose of toxin is very minute amount, for mice, its amount is 0.000,000,033 mg .The lethal dose of human being is probably 1-2 µg. It is a neurotoxin and acts gradually, taking several hours to kill.

                         Naturally-occurring cases of botulism arise from food spoiling in the absence of oxygen in canned foods. The commonest source historically has been spoiled meat, notably sausage (the species name derives from the Latin botulus meaning “sausage”) and poultry (especially duck), and improperly sterilized canned foods.  The neurotoxin produced from Clostridium tetani (tetanus toxin)another bacteria from some genus is encountered by humans as a result of wounds and remains a serious public health problem in developing countries around the world.

How long does the effect last?

        BOTOX® can help by reducing the over activity of the muscles very quickly. In fact, most people begin to see results within the first two weeks. Generally, BOTOX® provides release of spasticity in the injected muscle for over three months (12-14 weeks). Then patients begin to notice a gradual fading of its effects over several weeks. At this point the doctor can give another treatment. BOTOX® treatment may be required three or four times a year. It is not given more frequently than every three months.

Side effects may include:

                                 All prescriptions may have some side effects. With BOTOX®, side effects are usually transient and mild to moderate in nature. Side effects are usually temporary, and BOTOX® is generally well tolerated. Mild bruising or swelling may occur at the site of the injection. The treatment may also cause a brief headache. In rare cases, the injections may cause the eyebrow or upper eyelid to droop, but this condition is temporary and corrects itself without treatment within 2 to 3 weeks. The effects of      BOTOX® may be increased with the use of certain antibiotics or other drugs that interfere with neuromuscular transmission. Ensure your doctor is aware of any current medications you are taking. If you have any questions regarding the use of BOTOX® treatment, please consult your doctor. Some side effects includes

• Droopy eyelids, which can last for a few weeks

• Feeling like you have the flu

• Headache and upset stomach

• Risk of botulism (a life or death illness that makes it hard for a person to move the arms and legs or to breathe) is low with BOTOX® if used the right way

                      Most patients experience minimal pain or discomfort following injections. Swelling and puffiness at the site of the injection usually subside within a few days. Care must be taken to keep the head upright and avoid massaging the treated area, so that the BOTOX® affects only the desired areas.

Who Markets BOTOX®?

BOTOX® is marketed by Allergan, Incorporated, which describes itself as “a global specialty pharmaceutical company that develops and commercializes innovative products for the eye care, neuromodulator, skin care and other specialty markets.” Allergan made over $1.2 billion dollars in net BOTOX® sales in 2007. 50% of that ($606 million) came from BOTOX® COSMETIC. Total yearly sales of BOTOX® increased 406% between 2000 and 2007. Net sales in 2007 for all Allergan products reached $3.88 billion, of which BOTOX® and BOTOX® COSMETIC accounted for 31%.

In order to learn more specifically about the product BOTOX® please visit the official Allergan site, where you can also find listings of private offices offering a variety of services to treat many aesthetic issues, such as wrinkle reduction. Most wrinkles may be treated with BOTOX® injections.

References:

1. Wang YC, Burr DH, Korthals GJ, Sugiyama H. Acute toxicity of aminoglycoside antibiotics as an aid in detecting botulism. Appl Environ Microbiol 1984; 48:951-955.

2. Allergan (ELN 1145) BOTOX® COSMETIC (Botulinum Toxin Type A) Purified Neurotoxin Complex(Glabellar Lines) Manufactured by: Allergan Pharmaceuticals (Ireland) Ltd. A subsidiary of: Allergan, Inc. 2525 Dupont Dr. Irvine, California 92612.

3. Bazbek Davletov, Mark Bajohrs  and Thomas Binz, Beyond BOTOX: advantages and limitations of individual botulinum neurotoxins, TRENDS in Neurosciences Vol.28 No.8 August 2005.

4. Harimuraleedharan , Book: Bioterrorism (Releasing book  review),Tamilnadu book house,Chapter:Biological agents/clostridium.

5. Teguh Thajeb, Yi-Min Chen, Dao-Fu Dai, Daniel Daile Thajeb, Peterus Thajeb Botulism: A Frequently Forgotten Old Malady, Review Article, International Journal of Gerontology | September 2007 | Vol 1 | No 3.

6. Lewis, C. (2002) Botox Cosmetic: A look at looking good. FDA Consumer Magazine July–August, 1–4 .

Guidelines for BOTOX® Reconstitution

1.

Using the reconstitution needle, draw up the proper amount of saline (see dilution table) in the appropriately sized sterile syringe. A 21-gauge, 1®-inch needle is recommended for reconstitution. Reconstituted BOTOX® neurotoxin should be clear, colorless, and free of particulate matter.

2.

Insert the needle at a 45º angle and slowly inject the saline into the BOTOX® vial. Vacuum is present in the vial, which demonstrates that the sterility of the vial is intact. Do not use the vial if the vacuum does not pull the saline into the vial.

3.

Release the vacuum by disconnecting the syringe from the needle and allowing air to flow into the vial. Gently mix BOTOX® neurotoxin with the saline by moving vial side-to-side or rotating the vial.

4.

Draw the fluid into the injection syringe by placing the needle into the bottom corner of the vial for full extraction. Do not completely invert the vial.

5.

Disconnect the injection syringe from the vial and attach an appropriate needle for injection. A 25-, 27-, or 30-gauge needle may be used for superficial muscles, and a longer 22-gauge needle may be used for deeper musculature.

Note: Because the product and diluent do not contain a preservative, use within 4 hours once opened and reconstituted. During the 4 hours, BOTOX® solution should be stored in a refrigerator at 2ºC to 8ºC.