Fetal tissue engineering: Diaphragmatic replacement

doi:10.1053/jpsu.2001.20034

Journal of Pediatric Surgery

Volume 36, Issue 1, January 2001, Pages 146-151

Presented at the 31st Annual Meeting of the American Pediatric Surgical Association, Orlando, Florida, May 25-29, 2000.

https://doi.org/10.1053/jpsu.2001.20034 Get rights and content

Abstract

Background/Purpose: Prosthetic repair of congenital diaphragmatic hernia has been associated with high complication rates. This study was aimed at applying fetal tissue engineering to diaphragmatic replacement. Methods: Fetal lambs underwent harvest of skeletal muscle specimens. Once expanded in vitro, fetal myoblasts were suspended in a collagen hydrogel submitted to controlled radial tension. The construct was then placed in a bioreactor. After birth, all animals underwent creation of 2 diaphragmatic defects. One defect was repaired with the autologous-engineered construct placed in between 2 acellular supporting membranes and the other with an identical construct but without any cells. Each animal was its own control (graft, n = 10). Animals were killed at different time-points postimplantation for histologic examination. Statistical analysis was by analysis of variance (ANOVA). Results: Fetal myoblasts expanded up to twice as fast as neonatal cells. Hydrogel-based radial tension enhanced construct architecture by eliciting cell organization within the scaffold. No eventration was present in 4 of 5 engineered constructs but in 0 of 5 acellular grafts (P <.05). At harvest, engineered constructs were thick and histologically resembled normal skeletal muscle, whereas acellular grafts were thin, floppy, and showed low cell density with increased fibrosis. Conclusions: Unlike acellular grafts, engineered cellular diaphragmatic constructs are anatomically and histologically similar to normal muscle. Fetal tissue engineering may be a viable alternative for diaphragmatic replacement. J Pediatr Surg 36:146-151. Copyright © 2001 by W.B. Saunders Company.

Section snippets

Materials and methods

The Harvard Medical School animal management program is sanctioned by the American Association for the Accreditation of Laboratory Animal Care (AAALAC, file # 000009) and meets National Institutes of Health standards as set forth in the Guide for the Care and Use of Laboratory Animals (National Research Council Publication, Revised 1996).

Results

Fetal and neonatal survival rates were 100%. There were no perioperative complications with either harvesting method (open or videofetoscopic), or involving construct implantations. Fetal myoblasts expanded at faster rates than have been reported from neonatal cells (up to 2-fold).¹³ Desmin immunocytochemistry before construct assembly showed that approximately 80% of isolated cells were myogenic in identity. The remaining cells had a fibroblastic appearance. Continuous radial tension enhanced

Discussion

At the current stage of fetal intervention, the application of the concept of autologous fetal tissue engineering for treatment of birth defects could only be justified in life-threatening congenital anomalies. Preterm labor (mostly), as well as other general complications and risks of fetal surgery would render other applications of this concept ethically unacceptable, at least for now. In CDH, the diaphragmatic defect per se is not life threatening and, hence, would not be eligible for repair

Acknowledgements

The authors are indebted to Jeffrey Pettit for his excellence in laboratory assistance.

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^☆: This research was supported by grants from the United States Surgical Corporation and the Plastic Surgery Educational Foundation. Dario O. Fauza was supported by grants from the Harvard Medical School's Center for Minimally Invasive Surgery and the United States Surgical Corporation. Jennifer J. Marler was supported by a Faculty Research Grant from the American College of Surgeons and a Scholars-in-Medicine Award from Harvard Medical School. Both authors contributed equally to the study.

^☆☆: Address reprint requests to Joseph P. Vacanti, MD, Massachusetts General Hospital, 55 Fruit St, Warren 1157, Boston, MA 02114.

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Fetal tissue engineering: Diaphragmatic replacement☆,☆☆

Abstract

Section snippets

Materials and methods

Results

Discussion

Acknowledgements

J Pediatr

J Pediatr Surg

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J Pediatr Surg

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Delayed surgical repair and ECMO improves survival in congenital diaphragmatic hernia

Ann Surg

Congenital diaphragmatic hernia. Stabilization and repair on ECMO

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Thorax

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Semin Pediatr Surg