Tutoplast^® Process

Donor selection – Prior to beginning the Tutoplast^® process, all grafts go through a comprehensive donor-screening regimen to assure graft safety and eliminate the potential of using high-risk donors. The screening includes a medical/social history review, a detailed interview with next of kin and an extensive donor physical evaluation.

Serological testing – Comprehensive serological testing is performed by third-party CLIA (Clinical Laboratory Improvement Amendment) certified laboratories, using FDA approved test methodology. Tutogen’s exclusionary criteria are substantive and surpass FDA regulations and AATB standards.  Following a thorough quality assurance data review and acceptance by a licensed physician, donor grafts are released for Tutoplast^® processing. The serological screening includes testing for transmissible diseases as required by FDA & AATB standards.

This blood sample screening is a significant step in reducing potential disease transmission by eliminating any donor that may have had a prior medical condition or been involved in high-risk behavior. Tutogen’s medical director, a Board certified physician, oversees the implementation of our screening guidelines by tissue recovery agencies and releases the grafts for processing.

Donor grafts are additionally tested for microbial growth prior to the commencement of the preservation and sterilization process. This step determines the level of bacterial loading in the graft. Any grafts that exhibit unacceptably high levels of specific contaminants or highly pathogenic microbes is eliminated from processing. 

Grafts that pass this rigorous screening and testing process already present a very remote risk to the patient. The Tutoplast^® process further reduces the contamination risk of transplanted grafts until it is virtually eliminated, by subjecting it to additional processing steps that destroy, remove or inactivate disease causing agents.

The Tutoplast^® process is comprised of numerous phases as shown in the following example:

1. Lipids are removed in an ultrasonic acetone bath. Removal of lipids is important, as they may interfere with the healing process and stimulate bacterial growth. This step also inactivates enveloped viruses such as HIV and HCV, as well as reducing Prion activity by 2 log.¹
   
2. Bacteria are destroyed utilizing a series of alternating hyperosmotic saline and deionized water baths. This process ruptures the cell membranes, killing bacteria, washes out cellular debris, removes antigens (usually found in the membranes) and exposes any intracellular viruses that may be present, which can then be addressed in the subsequent step.
3. Soluble proteins are eliminated, and non-enveloped viruses and bacterial spores are destroyed using an oxidative treatment with hydrogen peroxide (H₂O₂). This treatment has been confirmed to inactivate viruses, including enveloped and non-enveloped, DNA and RNA viruses.²
   
4. A final acetone wash assures that any residual Prions are removed and enveloped viruses are inactivated. The acetone wash, followed by vacuum extraction, dehydrates the tissue, allowing it to be stored at room temperature.

Sterilization – After Tutoplast processing, the grafts are cut to shape and size and placed in double barrier packaging, then terminally sterilized using low dose gamma irradiation. This step eliminates any ancillary microbial contamination that may result from post-Tutoplast process handling and packaging and yields a Sterility Assurance Level (SAL) of 10^-6. This final step, alone, reduces the chance of viable microorganisms on Tutogen products to one (1) in one (1) million.³

References

1. Mould, et al. Scrapie in Mice. (1965). Vet Sci.
2. Viral Inactivation Study During the Tutoplast Allografts Manufacturing Process. (1994). Institut Pasteur Texcel. Paris, France. On file at Tutogen Medical, Inc.
3. 10^-6 SAL radiation dose validation study on file at Tutogen Medical, Inc.