Recellularization of Decellularized Allograft Scaffolds in Ovine Great Vessel Reconstructions

doi:10.1016/j.athoracsur.2004.09.033

The Annals of Thoracic Surgery

Volume 79, Issue 3, March 2005, Pages 888-896

https://doi.org/10.1016/j.athoracsur.2004.09.033 Get rights and content

Background

Decellularized allograft tissues have been identified as a potential extracellular matrix (ECM) scaffold on which to base recellularized tissue-engineered vascular and valvular substitutes. Decreased antigenicity and the capacity to recellularize suggest that such constructs may have favorable durability. Detergent/enzyme decellularization methods remove cells and cellular debris while leaving intact structural protein “scaffolds.” Allograft pulmonary artery tissues decellularized with an anionic detergent/enzyme methodology were tested in a long-term implantation model that used arterial wall repairs in the great vessels of juvenile sheep.

Methods

Twelve test sheep were implanted (n = 4) for each of three different scaffold protocols that compared traditional dimethylsulfoxide cryopreservation, cryopreservation followed by decellularization, and decellularization of fresh tissue. Four additional sheep served as controls (n = 2 sham, n = 2 fresh tissue). Patches were fashioned and implanted into pulmonary artery and aortic defects. Panel reactive antibodies (PRA) were measured over time (10 to 20 weeks). Explant histopathology determined recellularization morphology as well as calcium, collagen, and elastin distribution within explanted tissue.

Results

Unlike traditionally cryopreserved tissues, the decellularized tissues contained no residual cells or cellular debris before implantation, which correlated with measurable reductions in PRA. Decellularized explants demonstrated time-dependent migration of recipient cells through matrix, typically staining positive for α-smooth muscle actin with no calcification.

Conclusions

The properties demonstrated seem consistent with characteristics necessary for implantable tissue-engineered scaffolds. The decellularization method described appears to create a biologically suitable ECM scaffold for in vivo migration of phenotypically appropriate cells while avoiding antigenicity and calcification.

Section snippets

Tissue Procurement and Dissection

Sheep tissues were obtained from Animal Technologies (Tyler, TX). Hearts were transported on wet ice in Roswell Park Memorial Institute (RPMI) 1640 medium supplemented with polymyxin B. Warm ischemic time was less than 3 hours, and cold ischemic time didn't exceed 24 hours. Twenty-four pulmonary conduits were dissected from the heart and truncated immediately distal to the leaflets. They were then placed in RPMI 1640 supplemented with polymyxin B, cefoxitin, lincomycin, and vancomycin at 4°C

Results

Pulmonary allograft patches were prepared for implant by three different methodologies: (1) classically cryopreserved (control) tissue, (2) cryopreserved, thawed, then decellularized tissue and (3) fresh, decellularized tissue. Tissues in groups 2 and 3 were cryoconserved for storage before implant. At explant surgery, all patches appeared to be nicely incorporated into the great vessel repairs without aneurysms or infection. Animals were sacrificed at the 10-week and 20-week designated

Comment

Acellular tissues have been identified as a possible tissue-engineered solution for creating valvular and vascular tissue replacements. Infiltration of autologous, biologically active, phenotypically appropriate cells into an acellular collagen matrix could potentially provide reparative and functional advantages over current clinical nonviable prosthetics. If this recellularization process can occur in vivo, then perhaps preimplant bioreactor-based (cell seeding) recellularization methods may

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Original articleCardiovascularRecellularization of Decellularized Allograft Scaffolds in Ovine Great Vessel Reconstructions

Background

Methods

Results

Conclusions

Section snippets

Tissue Procurement and Dissection

Results

Comment

J Thorac Cardiovasc Surg

J Surg Res

Ann Thorac Surg

J Thorac Cardiovasc Surg

J Thorac Cardiovasc Surg

J Thorac Cardiovasc Surg

J Thorac Cardiovasc Surg

J Thorac Cardiovasc Surg

J Thorac Cardiovasc Surg

J Thorac Cardiovasc Surg

Surgery

Original article
Cardiovascular
Recellularization of Decellularized Allograft Scaffolds in Ovine Great Vessel Reconstructions