Cleft palate reconstruction using collagen and nanofiber scaffold incorporating bone morphogenetic protein in rats

Absorbable collagen sponge (ACS) loaded with bone morphogenetic protein-2 (BMP-2) is approved for selected clinical applications; however, burst release limits its widespread use. Therefore, nanofiber (NF)-based scaffold with ACS backbone was developed to sustain release of loaded BMP-2 to improve the outcomes of bone grafting in a rodent model of cleft palate.

BMP-2 was loaded on ACS scaffold and then NF hydrogel with different densities (1-2%) was added to sustain the BMP-2 release. The release profiles of BMP-2 from constructs with different NF densities were evaluated in vitro to explore the optimum NF density that could recapitulate physiological bone healing process. Subsequently, scaffold with the appropriate NF density was implanted into a rodent model of cleft palate. Wistar rats, with surgically induced maxillary cleft defects, were then assigned to one of the following groups (n=6/group): no scaffold (control), ACS, ACS+BMP-2, NF+ACS, and NF+ACS+BMP-2. Micro-computed tomography (μCT) was utilized to evaluate percent bone filling (%BF) at defect site as well as changes in anteroposterior and transverse dimensions of the maxilla at weeks 0, 4, and 8. Histological assessment of bone healing was performed at week 8.

In vitro release experiments showed that scaffolds containing 2% NF exhibited a release profile conducive to the natural stages of bone healing and, hence, it was utilized for subsequent in vivo studies. Bone healing occurred at the defect margins leaving a central bone void in the control, ACS, and NF+ACS groups over the 8-week study period. BMP-2-treated groups demonstrated higher %BF as compared with other groups at week 8 (p<0.05). Whereas the NF+ACS+BMP-2 group showed bone bridging of the defect as early as 4 weeks, which was not evident in ACS+BMP-2 group. In all groups, bone grafts did not disrupt anteroposterior and transverse growth of maxilla. Based on histological evaluations together with μCT data, NF+ACS+BMP-2 treatment resulted in clinically significant and consistent bone healing throughout the implanted scaffold when compared with the ACS+BMP-2 group.

NF+ACS+BMP-2 constructs exhibited osteoinductive properties together with preparation simplicity, which makes it a novel approach for BMP-2 delivery for cleft palate reconstruction.