Cryopreserved human umbilical cord patch for in-utero spina bifida repair.
OBJECTIVE
To identify a patch system to repair surgically created spina bifida in a sheep model for its efficacy in healing the skin defect, protecting the underlying spinal cord and reducing the Chiari II malformation.


METHODS
Spina bifida was created surgically in 16 fetuses from eight timed-pregnant sheep at gestational age of 75 days. Two fetuses did not survive the procedure. Repeat hysterotomy was performed at 95 days' gestation to cover the defect with either biocellulose film with underwater adhesive (BCF-adhesive) (n = 7) or human umbilical cord with suture (HUC-suture) (n = 7). Three fetuses without formation of the defect served as reference controls. The skin healing was examined by direct visualization after a planned Cesarean section at term, followed by histological analysis using hematoxylin and eosin and Masson's trichrome stains. Mid-sagittal sections of the fetal cranium and upper cervical spine were analyzed by a pediatric neuroradiologist who was blinded to the type of patch received.


RESULTS
Three fetuses that received the BCF-adhesive and six fetuses that received the HUC-suture survived to term for final analysis. As a result of dislodgment of the BCF-adhesive, all spina bifida defects repaired using BCF-adhesive were not healed and showed exposed spinal cord with leakage of cerebrospinal fluid. In contrast, all spinal defects repaired by HUC-suture were healed with complete regrowth of epidermal, dermal and subdermal tissue components, with no exposed spinal cord. The maximal skin wound width was 21 ± 3.6 mm in the BCF-adhesive group but 3 ± 0.8 mm in the HUC-suture group (P < 0.001). The spinal cord area (P = 0.001) and the number of anterior horn cells (P = 0.03) was preserved to a greater degree in the HUC-suture group than in the BCF-adhesive group, whilst psammoma bodies, signifying neuronal degeneration, were only observed in the BCF-adhesive group. Anatomic changes, indicative of Chiari II malformation, were seen in all three fetuses of the BCF-adhesive group but in none of the HUC-suture group (P < 0.01).


CONCLUSIONS
Cryopreserved umbilical cord graft is a promising regenerative patch for intrauterine repair of spina bifida.
Department of Obstetrics, Gynecology and Reproductive Medicine, Division of Maternal-Fetal Medicine, UT Health School of Medicine, Houston, TX, USA.|Fetal Center, Houston, TX, USA.
R
R
Papanna
Department of Obstetrics, Gynecology and Reproductive Medicine, Division of Maternal-Fetal Medicine, UT Health School of Medicine, Houston, TX, USA.|Fetal Center, Houston, TX, USA.
K J
KJ
Moise
Department of Obstetrics, Gynecology and Reproductive Medicine, Division of Maternal-Fetal Medicine, UT Health School of Medicine, Houston, TX, USA.|Department of Pathology and Laboratory Services, UT Health School of Medicine, Houston, TX, USA.
L K
LK
Mann
Department of Pediatrics, Division of Pediatric Neurosurgery, UT Health School of Medicine, Houston, TX, USA.
S
S
Fletcher
Department of Pediatric Surgery, UT Health School of Medicine, Houston, TX, USA.
R
R
Schniederjan
Department of Pediatric Surgery, UT Health School of Medicine, Houston, TX, USA.
M B
MB
Bhattacharjee
Department of Bioengineering, University of Utah, Salt Lake City, UT, USA.
R J
RJ
Stewart
Department of Bioengineering, University of Utah, Salt Lake City, UT, USA.
S
S
Kaur
Department of Radiology, Harvard Medical School, Boston Children's Hospital, Boston, MA, USA.
S P
SP
Prabhu
Ocular Surface Center and TissueTech, Inc., Miami, FL, USA.
S C G
SC
Tseng
D000818:Animals; D001139:Arnold-Chiari Malformation / Q000196:embryology / Q000209:etiology / Q000601:surgery; D002482:Cellulose; D015925:Cryopreservation; D005260:Female; D046128:Fetal Therapies / Q000379:methods*; D005333:Fetus; D005865:Gestational Age; D006801:Humans; D023421:Models, Animal; D011247:Pregnancy; D012756:Sheep; D013116:Spinal Cord; D016135:Spinal Dysraphism / Q000150:complications / Q000196:embryology / Q000601:surgery*; D014014:Tissue Adhesives / Q000627:therapeutic use*; D014470:Umbilical Cord / Q000637:transplantation*
D014014:Tissue Adhesives; D002482:Cellulose
neural tube defect; regenerative healing; repair; umbilical cord