Transforming Growth Factor-β3/Recombinant Human-like Collagen/Chitosan Freeze-Dried Sponge Primed With Human Periodontal Ligament Stem Cells Promotes Bone Regeneration in Calvarial Defect Rats

Shiyi Huang; Fenglin Yu; Yating Cheng; Yangfan Li; Yini Chen; Jianzhong Tang; Yu Bei; Qingxia Tang; Yueping Zhao; Yadong Huang; Qi Xiang

doi:10.3389/fphar.2021.678322

Transforming Growth Factor-β3/Recombinant Human-like Collagen/Chitosan Freeze-Dried Sponge Primed With Human Periodontal Ligament Stem Cells Promotes Bone Regeneration in Calvarial Defect Rats

Front Pharmacol. 2021 Apr 23:12:678322. doi: 10.3389/fphar.2021.678322. eCollection 2021.

Authors

Shiyi Huang¹, Fenglin Yu¹, Yating Cheng¹, Yangfan Li¹, Yini Chen¹, Jianzhong Tang², Yu Bei², Qingxia Tang³, Yueping Zhao³, Yadong Huang^{1

2}, Qi Xiang^{1

2}

Affiliations

¹ Institute of Biomedicine and Guangdong Provincial Key Laboratory of Bioengineering Medicine, Jinan University, Guangzhou, China.
² Biopharmaceutical R and D Center of Jinan University, Guangzhou, China.
³ Department of Stomatology, Jinan University Medical College, Guangzhou, China.

Abstract

Patients with a skull defect are at risk of developing cerebrospinal fluid leakage and ascending bacterial meningitis at >10% per year. However, treatment with stem cells has brought great hope to large-area cranial defects. Having found that transforming growth factor (TGF)-β3 can promote the osteogenic differentiation of human periodontal ligament stem cells (hPDLSCs), we designed a hybrid TGF-β3/recombinant human-like collagen recombinant human collagen/chitosan (CS) freeze-dried sponge (TRFS) loading hPDLSCs (TRFS-h) to repair skull defects in rats. CFS with 2% CS was selected based on the swelling degree, water absorption, and moisture retention. The CS freeze-dried sponge (CFS) formed a porous three-dimensional structure, as observed by scanning electron microscopy. In addition, cytotoxicity experiments and calcein-AM/PI staining showed that TRFS had a good cellular compatibility and could be degraded completely at 90 days in the implantation site. Furthermore, bone healing was evaluated using micro-computed tomography in rat skull defect models. The bone volume and bone volume fraction were higher in TRFS loaded with hPDLSCs (TRFS-h) group than in the controls (p < 0.01, vs. CFS or TRFS alone). The immunohistochemical results indicated that the expression of Runx2, BMP-2, and collagen-1 (COL Ⅰ) in cells surrounding bone defects in the experimental group was higher than those in the other groups (p < 0.01, vs. CFS or TRFS alone). Taken together, hPDLSCs could proliferate and undergo osteogenic differentiation in TRFS (p < 0.05), and TRFS-h accelerated bone repair in calvarial defect rats. Our research revealed that hPDLSCs could function as seeded cells for skull injury, and their osteogenic differentiation could be accelerated by TGF-β3. This represents an effective therapeutic strategy for restoring traumatic defects of the skull.

Keywords: freeze-dried sponge; periodontal ligament stem cells; skull bone defect repair; stem cell therapy; transforming growth factor 3.