Dong Xue, Landford Wilmina N, Hart James, Risolino Maurizio, Kaymakcalan Omer, Jin Julia, Toyoda Yoshiko, Ferretti Elisabetta, Selleri Licia, Spector Jason A
New York and Ithaca, N.Y.; and San Francisco, Calif.
From the Laboratory for Bioregenerative Medicine and Surgery, Division of Plastic Surgery, and the Department of Cell and Developmental Biology, Weill Cornell Medicine; the Nancy E. and Peter C. Meinig School of Bioengineering, Cornell University; and the Program in Craniofacial Biology, Institute of Human Genetics, Departments of Orofacial Sciences and Anatomy, University of California San Francisco.
Plast Reconstr Surg. 2017 Jul;140(1):75-85. doi: 10.1097/PRS.0000000000003417.
Cleft lip with or without cleft palate is present in approximately one in 500 to 700 live births, representing the most common congenital craniofacial anomaly. Previously, the authors developed a unique murine model with compound Pbx deficiency that exhibits fully penetrant cleft lip with or without cleft palate. To investigate the possibility of tissue repair at an early gestational stage, the authors designed a minimally invasive surgical approach suitable for intrauterine repair using Wnt9b-soaked collagen microspheres to restore craniofacial developmental programs for cleft correction.
Collagen microspheres with diameters ranging from 20 to 50 μm were fabricated to serve as a delivery vehicle for Wnt9b. At gestational day 11.5, wild-type and Pbx-deficient murine embryos were isolated. Microspheres soaked in murine purified Wnt9b protein were microsurgically implanted at the midface lambdoidal junction. Embryos were cultured in a 37°C modified whole-embryo culture system.
Targeted release of Wnt9b resulted in augmented Wnt expression at the lambdoidal junction. Microsurgical implantation of Wnt9b-soaked microspheres resulted in cleft correction in 27.1 percent of the Pbx-deficient embryos. The difference in the ratio of the areas of clefting between implanted and nonimplanted embryos was significant (p < 0.05).
Ex utero correction of cleft lip with or without cleft palate in the authors' murine model by means of microsurgical intervention and targeted delivery of Wnt proteins is an innovative and promising strategy. Although further refinement and optimization of this technique will be required to improve efficacy, the authors believe that this approach will open new avenues toward unconventional prenatal interventions for patients with cleft lip with or without cleft palate, and provide future approaches for prenatal repair of other congenital head and neck disorders.
唇裂伴或不伴腭裂在每500至700例活产婴儿中约有1例出现，是最常见的先天性颅面畸形。此前，作者开发了一种独特的复合Pbx缺陷小鼠模型，该模型表现出完全显性的唇裂伴或不伴腭裂。为了研究妊娠早期组织修复的可能性，作者设计了一种微创外科手术方法，适用于子宫内修复，使用浸泡Wnt9b的胶原微球来恢复颅面发育程序以矫正腭裂。
制备直径为20至50μm的胶原微球作为Wnt9b的递送载体。在妊娠第11.5天，分离野生型和Pbx缺陷型小鼠胚胎。将浸泡在小鼠纯化Wnt9b蛋白中的微球显微手术植入中面部人字缝交界处。胚胎在37°C改良全胚胎培养系统中培养。
Wnt9b的靶向释放导致人字缝交界处Wnt表达增加。显微手术植入浸泡Wnt9b的微球使27.1%的Pbx缺陷型胚胎腭裂得到矫正。植入和未植入胚胎的腭裂面积比差异有统计学意义（p<0.05）。
通过显微手术干预和Wnt蛋白的靶向递送对作者的小鼠模型中的唇裂伴或不伴腭裂进行子宫外矫正，是一种创新且有前景的策略。尽管需要进一步完善和优化该技术以提高疗效，但作者认为这种方法将为唇裂伴或不伴腭裂患者的非常规产前干预开辟新途径，并为其他先天性头颈部疾病的产前修复提供未来方法。
