A Gingiva-Derived Mesenchymal Stem Cell-Laden Porcine Small Intestinal Submucosa Extracellular Matrix Construct Promotes Myomucosal Regeneration of the Tongue

Abstract

In the oral cavity, the tongue is the anatomic subsite most commonly involved by invasive squamous cell carcinoma. Current treatment protocols often require significant tissue resection to achieve adequate negative margins and optimal local tumor control. Reconstruction of the tongue while preserving and/or restoring its critical vocal, chewing, and swallowing functions remains one of the major challenges in head and neck oncologic surgery. We investigated the in vitro feasibility of fabricating a novel combinatorial construct using porcine small intestinal submucosa extracellular matrix (SIS-ECM) and human gingiva-derived mesenchymal stem cells (GMSCs) as a GMSC/SIS-ECM tissue graft for the tongue reconstruction. We developed a rat model of critical-sized myomucosal defect of the tongue that allowed the testing of therapeutic effects of an acellular SIS-ECM construct versus a GMSC/SIS-ECM construct on repair and regeneration of the tongue defect. We showed that the GMSC/SIS-ECM construct engrafted at the host recipient site, promoted soft tissue healing, and regenerated the muscular layer, compared to the SIS-ECM alone or nontreated defect controls. Furthermore, our results revealed that transplantation of the GMSC/SIS-ECM construct significantly increased the expression of several myogenic transcriptional factors and simultaneously suppressed the expression of type I collagen at the wounded area of the tongue. These compelling findings suggest that, unlike the tongue contracture and fibrosis of the nontreated defect group, transplantation of the combinatorial GMSC/SIS-ECM constructs accelerates wound healing and muscle regeneration and maintains the overall tongue shape, possibly by both enhancing the function of endogenous skeletal progenitor cells and suppressing fibrosis. Together, our findings indicate that GMSC/SIS-ECM potentially served as a myomucosal graft for tongue reconstruction postsurgery of head and neck cancer.

SIS-ECM Laden with GMSC-Derived Exosomes Promote Taste Bud Regeneration

“…The healing of the tongue defects was individually observed at different time points. Similar to our previous report (Xu et al 2017), all rats were able to survive after the surgery without obvious body weight loss during the experimental process (data not shown), and the transplanted SIS-ECM scaffold was completely resorbed by day 7 after transplantation (Fig. 2A).…”

“…Strips (6 × 6 mm) of decellularized porcine SIS-ECM 2.0 (Cook Biotech, Inc.) was prepared as described previously (Xu et al 2017). For detailed procedures of animal surgery and transplantation of SIS-ECM with GMSCs or exosomes, see the Appendix.…”

“…We have successfully established a critical-sized tongue defect model in rats and have demonstrated that the transplantation of small intestinal submucosa–extracellular matrix (SIS-ECM) constructs seeded with gingival MSCs (GMSCs) could promote muscle regeneration and reduce fibrosis during tongue reconstruction (Xu et al 2017). Considering the multilayered structure of ablative tongue tissue as well as its specialized taste sensory function, we herein explored whether GMSCs and their derivative exosomes laden with SIS-ECM matrix could facilitate taste bud regeneration during tongue reconstruction.…”

A preclinical study—systemic evaluation of safety on mesenchymal stem cells derived from human gingiva tissue

“…In vitro, single colony-derived GMSCs have been shown to differentiate into adipocytes and osteoblasts [18]. Using porcine small intestinal submucosa extracellular matrix (SIS-ECM) and human GMSCs as a GMSC/SIS-ECM tissue graft for the tongue reconstruction and the constructs accelerates wound healing and muscle regeneration and maintains the overall tongue shape [20]. In addition, based upon their known immunomodulatory properties, we have previously reported that adoptive transfer of GMSCs in a mouse model of rheumatoid arthritis not only significantly reduced the severity of the arthritis and downregulated the production of Th1 and Th17 inflammatory cytokines (i.e., interferon-γ and interleukin-17A) but also enhanced Treg cell differentiation [3].…”

Oral Rehabilitation of Patients Sustaining Orofacial Injuries: The UPenn Initiative

“…Systemic administration of GMSCs facilitates the regeneration of epithelial layers in colitis, skin, and oral mucosal healing models (Zhang et al 2009; Zhang et al 2010). Our recent studies showed that local implantation of porcine small intestine submucosa extracellular matrix (SIS-ECM) laden with GMSCs or GMSC-derived exosomes promotes regeneration of tongue muscle, epithelial layer, and taste buds in a critical-sized rat tongue defect model, possibly via activating the endogenous skeletal muscle, epithelial, and taste bud progenitor cells (Xu et al 2017; Zhang et al 2019). Several lines of evidence have implicated the potential use of GMSCs in repair/regeneration of gingival and periodontal defects (Fawzy El-Sayed et al 2012; Li et al 2018).…”