Regenerative biology deals with the “process of replacing, engineering or regenerating human cells, tissues or organs to restore or establish normal function. [Mason C and Dunnill P (2008). A brief definition of regenerative medicine. Regenerative Medicine. 3 (1): 15.]. Regenerative veterinary biology extends this concept to the animal world. In many cases, this emerging biotechnological discipline can be thought of as a branch of tissue engineering applied towards finding solutions for structural repair of tissue damage that best utilizes the body’s own processes.

There are many forms of regenerative technologies such as concentration, in vitro growth, banking, stimulation, or collection.

Platelet Rich Plasma (PRP)  PRP is a concentration technology that starts with a large quantity of whole blood drawn from an animal. The blood is centrifuged to concentrate platelets, the contents of which are sometimes extracted, and reintroduced into an animal at a site of injury. This autologous (self-derived) treatment is believed to help curb inflammation and rebuild damaged tissue due to the action of protease (protein-damaging) inhibitors and fibrin protein, amongst other important biomolecules present.

In vitro growth and transplantation  This process involves harvesting stem cells from an animal’s tissue (e.g. blood or fat tissue), expanding their numbers by allowing growth in tissue culture, and reintroducing them into the animal.

Tissue stimulation  Injured areas can be stimulated using mechanical devices such as Shockwave, Low Light, or Cold Lasers, to spur low-level inflammation and trigger the body’s own healing processes.

The newest biotechnology is the use of Amnion.

Amnion is the tissue that makes up the innermost membrane of the birth sac connected to the placenta that contains the developing animal, prior to birth.  The membrane and surrounding fluid serve to nourish, cushion, and protect the fetus by forming this barrier between the gestational environment and the outside world.  The tissues associated with the amnion are considered “immune privileged” since they do not evoke an immune response from the mother’s system that would otherwise reject the foreign body growing during pregnancy.  These tissues are also rich in structural building blocks important to nearly all tissues including a mix of biomolecules such as collagen, hyaluronic acid, carbohydrate-rich proteins (proteoglycans), and other assorted macromolecules.

Amnion is collected non-invasively during the birthing of healthy animals. Animal owners with pregnant pets consent to the collection during the normal birthing of newborns. All deliveries are veterinarian assisted with the health and safety of the animals as the paramount concern.  Do you have a pregnant mare or a hard-to-birth dog?  Please take a look at the AniCell Biotech Miracle Ranch Program.

Amniotic material has been used for centuries on humans with the earliest documented cases on skin burns and wounds dating back to the early 1900s. And ophthalmologists began using amnion for eye wounds in the 1940s. [de Ratth, A. (1940). Plastic repair of conjunctival defects with fetal membranes. Arch. Ophthalmol. 23, 522] The FDA recognizes human amnion as an HCT/P (human cells, tissues, and cellular and tissue-based products) when used as a wound covering, regulated under Section 361 of the Public Health Service Act. This designation was reaffirmed for ocular repair in 1999. [Silini AR, et al, (2015) The long path of human placenta and its derivatives in regenerative medicine. Frontiers in Bioengineering and Biotechnology 3: 162]

The amniotic membrane has several properties that make it suitable as a wound covering or biological bandage. These include:

1) Biocompatible  Low or non-immunogenicity of the tissue impedes rejection by the animal host [Hori J, et al. (2006) Immunological characteristics of amniotic epithelium. Cornea 25: S53-S58]

2) Durable  The complexity and unique arrangement of specialized structural proteins (collagen, laminin, elastin, proteoglycans, glycoproteins, glycosaminoglycans, etc.) provide substantial mechanical integrity and viscoelasticity, properties that are important for forming a barrier that protects and separates the wound from the outside environment. [Niknejad H, et al. (2008) Properties of the amniotic membrane for potential use in tissue engineering. Eur Cell Mater 15: 88-99]

3) Hydratable  The high concentration of carbohydrate side-groups in the extracellular matrix molecules that comprise the amniotic membrane allow for generous hydration of the biomaterial, which can be beneficial for decreasing wound dehydration and requiring fewer dressing changes [Loeffelbein DJ, et al. (2014) Evaluation of Human Amniotic Membrane as a Wound Dressing for Split-Thickness Skin-Graft Donor Sites. BioMed Res Int 2014: Article ID 572183]

4) Resorbable  Since the natural amniotic membrane biomaterial is resorbable, it does not have to be physically removed from the wound after application which could perturb the wound bed and generate greater trauma. [Branski LK, et al. (2008) Amnion in the treatment of pediatric partial-thickness facial burns. Burns 343: 393-399]

5) Adhesive  As a naturally adhesive substrate and biomaterial, the amnion is adherent to wounds and permissive to cell adhesion and ingrowth, acting as a scaffold when the animal’s body is building new tissue. [Koizumi N, et al. (2007) Comparison of intact and denuded amniotic membrane as a substrate for a cell-suspension culture of human limbal epithelial cells. Graefes Arch Clin Exp Ophthalmol 24: 123]

6) Semipermeable – Gas exchange through the membrane material provides a healthy environment for allowing the animal’s own healing mechanisms to progress. [Hopf HW and Rollins MD. (2006) Wounds: An overview of the role of oxygen. Antioxid Redox Signal 9: 1183-1192]

These products perform the same function in the recipient as they did in the donor. In the donor animal, the amniotic membrane provided multiple functions: 1) a barrier function, separating an external and internal environment; 2) a cover function, surrounding the enclosable area; and 3) a protection function, in a durable biocompatible material that is difficult to breach or penetrate. In the recipient, as a wound covering, AniCell’s amniotic membrane-based products provide the same functions. Whether applied to superficial wounds or ocular lesions, the products provide the same great benefits.

Most of the products can be effective with a single application and a one-time visit to the veterinarian. In eyes, the material is applied in the form of a membrane or drops and for superficial wounds, the material topically adheres to form a biological bandage. Recovery times can vary depending on the age and health of the animal.

If we draw a comparison between AniCell’s amniotic tissue-based products to those used in humans, there are few risks. Since 1910, there have been no documented serious adverse events in scientific literature.  Some patients experienced injection point selling or initial discomfort but all have subsided within 24-72 hours.

No. Due to its anti-reactive properties, amnion is not invasive at all. AniCell’s products can be banked by your veterinarian and used the same day as the diagnosis in the field during a farm call. These products are administered topically for superficial wounds during cleaning and debridement or as an implantable for tendon and ligament injuries.

Yes, when compared to the total cost of wound management, using amniotic material is extremely affordable for injuries due to its single-use application.  Expenditures on changes of dry wound dressings around the incorporated amniotic material can be reduced due to less exudate produced. [Fetterolf DE and Snyder RJ. (2012) Scientific and clinical support for the use of a dehydrated amniotic membrane in wound management. Wounds 24: 299-307.]

The use of amnion must be performed by a veterinarian. Simply ask your veterinarian about this type of regenerative technology and whether it could be a suitable alternative to the typical standard of care.

After use, the product, in some cases, is known to have a euphoric effect that lasts for 24-48 hours following treatment making the animal feel overly content.  Other cases have reported that their animal is lethargic and lacks an appetite for 24 hours.  In all cases of acute injury, we recommend 48 hours of stall rest for horses and kenneling for dogs.  All exercise should be controlled such as hand walking.  Due to the desire to maintain the inflammatory signaling, a vet-prescribed accelerated rehabilitation protocol should include an ever-increasing exercise regimen.  Follow-up visits can include concomitant therapies such as the shockwave and therapeutic ultrasound to aid in regeneration.   In less than 3 percent of the cases, some injection point swelling and moderate to severe soreness can occur for 24 to 48 hours following product use.  In these cases, the use of pain medication and animal movement lessens the stiffness and mobilizes the product.  Pain medications should be opioid-based and not steroidal or non-steroidal anti-inflammatory drugs due to their impact on inflammation and the mechanism of action for amnion-based products being that very inflammation.  In these cases, there is no impact to the product’s results.

One can expect one of the 4 profiles in the weeks following the use of the product and dependent on the injury:

• In 25% of the cases, the animal will experience almost immediate impact from the product returning to near soundness in 2-3 weeks.
• In 40% of the cases, the animal will steadily improve over the course of a 6-week time horizon.
• In another 25% of the cases, the animal will not see any substantial response for 4 weeks and then experience a hockey stick-like improvement over weeks 5 and 6.
• And in 10% of the cases, we simply do not see any substantial improvement warranting either additional applications or a separate course of treatment.

While we expect to see improvement over the 6-week time horizon, some cases have reported slow continual improvement over 6 months.  While these cases are abnormal, it seeks to prove that all animals respond differently to treatments.

 

Due to the propensity of the equine and canine species to be overly sensitive, in under 3% of AniCell’s cases, there have been reports of injection point swelling as well as some intense pain following the injection that lasts up to 72 hours.  Many times, these effects are due to implanting the amniotic material into connective tissue which creates extensive soreness. Pain has also been reported when the material is placed in synovial spaces.  In these cases where this occurs, do not use NSAIDs. For pain, use opioid-based pain medications such as Tramadol or Gabapentin.  We also suggest moving the animal around to better disperse the product.  All that having been said, the worst-case scenario is that the use does not yield the desired outcome.