In situ tissue engineering for tracheal reconstruction using a luminar remodeling type of artificial trachea.

BACKGROUND: After successful trials of tracheal reconstruction using mesh-type prostheses in canine models, the technique has been applied clinically to human patients since 2002. To enhance tissue regeneration, we have applied a new tissue engineering approach to this mesh-type prosthesis. METHODS: The prosthesis consists of a polypropylene mesh tube reinforced with a polypropylene spiral and atelocollagen layer. The cervical tracheas of 18 beagle dogs were replaced with the prosthesis. The collagen layer was soaked with peripheral blood in 6 of the dogs, with bone marrow aspirate in another 6, and with autologous multipotential bone marrow-derived cells (mesenchymal stem cells) in another 6. The dogs were humanely killed at 1 to 12 months after the operation. RESULTS: All 18 dogs survived the postoperative period. Bronchoscopically, 3 of 4 dogs in the peripheral blood group showed stenosis, whereas no stenosis was evident in all 8 of the dogs in the bone marrow and mesenchymal stem cell groups 6 months after the operation. Faster epithelialization and fewer complications, such as mesh exposure and luminal stenosis, were observed in these two groups than in the peripheral blood group. Histologically, the cells from autologous bone marrow were found to proliferate into the tracheal tissue during the first month. Cilial movement in these two groups was faster than that in the peripheral blood group and recovered to 80% to 90% of the normal level. CONCLUSIONS: Bone marrow aspirate and mesenchymal stem cells enhance the regeneration of the tracheal mucosa on this prosthesis. This in situ tissue engineering approach may facilitate tracheal reconstruction in the clinical setting.

Histologic and electrophysiological study of nerve regeneration using a polyglycolic acid-collagen nerve conduit filled with collagen sponge in canine model.

OBJECTIVES: To determine the rate of achieving electrophysiologically proved functional recovery by autonomic nerve regeneration, with the aid of an artificial nerve conduit. METHODS: A polyglycolic acid (PGA) collagen nerve conduit filled with collagen sponge was interposed in a 10-mm-long gap of the right hypogastric nerve (HGN) in 16 dogs. Histologic evaluation of nerve regeneration and electrophysiological analysis at 2 weeks and 2, 3, 4, 5, 6, 7, and 8 months (n = 2, each) after surgery was performed, measuring the responses for the spermatic ducts (SD), bladder neck (BN), and prostate contraction, by stimulating the right lumbar splanchnic nerves (LSNs) from L2 to L4, after transection of the left HGN to eliminate substitutive pathways. RESULTS: Two months after implantation, the regenerated neurofilaments were successfully extended through the graft from the proximal-to-distal direction. In 2 control dogs, electrostimulation of the right LSNs induced elevation of the intraluminal pressure of the SD, elevation of the BN pressure, and prostate contraction. No responses were observed in all dogs up to 6 months of follow-up after implantation. In 1 dog with a 7-month follow-up, electrostimulation elicited elevation of BN pressure alone. In both dogs with an 8-month follow-up, electrostimulation induced similar responses to control in all SD, BN, and prostate; however, after excision of the area of the interposed right HGN, no response was observed. CONCLUSIONS: These results proved that regeneration of a 10-mm gap of the HGN, using a novel PGA-collagen nerve conduit could be achieved within 8 months.

Regeneration of central nervous tissue using a collagen scaffold and adipose-derived stromal cells.

Adipose-derived stromal cells (ASCs) include stem cells, which have the potential to differentiate into a variety of cell lineages. The regeneration of central nerves was examined using ASCs and a collagen scaffold. A cerebral cortex defect (3 x 4 x 3 mm(3)) was created in the left frontal lobe of 16 male rats. In one group (n = 8), collagen (3 x 4 x 3 mm(3)) seeded with DiI-labeled ASCs was implanted in the defect. In order to seed the ASCs, a combination of the rotary cell culture system and pressing the collagen scaffold gently several times with a glass rod was applied. In the control group (n = 8), collagen was implanted without ASCs. The rats were sacrificed at 1 month after the scaffold implantation. Histologically, 0.2% of the implanted ASCs were positive for anti-human/rat microtubule-associated protein 2 (MAP2) antibody and microvessels were present at a density of 4.6 +/- 1.2/mm(2) within the collagen scaffold-implanted area in each coronal section. In the control group, no MAP2-positive cells were detected and the microvessel density was 0.6 +/- 0.4/mm(2). These data suggest that ASCs seeded into a collagen scaffold may have the potential to promote regeneration of nervous tissue after cerebral cortex injury.

Experimental repair of phrenic nerve using a polyglycolic acid and collagen tube.

OBJECTIVE: The feasibility of a nerve guide tube for regeneration of the phrenic nerve with the aim of restoring diaphragmatic function was evaluated in a canine model. METHODS: The nerve tube, made of woven polyglycolic acid mesh, had a diameter of 3 mm and was filled with collagen sponge. This polyglycolic acid-collagen tube was implanted into a 10-mm gap created by transection of the right phrenic nerve in 9 beagle dogs. The tubes were implanted without a tissue covering in 5 of the 9 dogs (group I), and the tubes were covered with a pedicled pericardial fat pad in 4 dogs (group II). Chest x-ray films, muscle action potentials, and histologic samples were examined 4 to 12 months after implantation. RESULTS: All of the dogs survived without any complications. x-ray film examination showed that the right diaphragm was paralyzed and elevated in all dogs until 3 months after implantation. At 4 months, movement of the diaphragm in the implanted side was observed during spontaneous breathing in 1 dog of group I and in 3 dogs of group II. In the dogs showing diaphragm movement, muscle action potentials were evoked in the diaphragm muscle, indicating restoration of nerve function. Regeneration of the phrenic nerve structure was also examined on the reconstructed site using electron microscopy. CONCLUSION: The polyglycolic acid-collagen tube induced functional recovery of the injured phrenic nerve and was aided by coverage with a pedicled pericardial fat pad.

Experimental study on the regeneration of peripheral nerve gaps through a polyglycolic acid-collagen (PGA-collagen) tube.

We have developed a bioabsorbable polyglycolic acid (PGA) tube filled with collagen sponge (PGA-collagen tube) as a nerve connective guide, and compared its effectiveness with that of autograft in terms of nerve regeneration across a gap. The PGA-collagen tube was implanted into 24 beagle dogs across a 15-mm gap in the left peroneal nerve. The right peroneal nerve was reconstructed with the autograft harvested from the left side, as a control. After the surgery, the connective tissue extended from both cut ends in the PGA-collagen tube and connected again at the center. Pathologically, the collagen sponge in the tube provided adequate scaffolding for nerve tissue extension, and the nerve tissue reconnected within 3 weeks. Electrophysiologically, muscle-evoked potentials (MEPs) and compound nerve action potentials (CNAPs) were detected 18 days after the surgery. For up to 6 months postsurgery, CNAPs and somatosensory-evoked potentials (SEPs) on the PGA-collagen side had a shorter latency and larger peak voltage than those on the autograft side. The myelinated axons on the PGA side were larger in diameter than those on the autograft side. It is suggested that the PGA-collagen tube has the potential to be an effective alternative to conventional autografting for the repair of some peripheral nerve defects.