Adipose-Derived Mesenchymal Stromal Cells Persist in Tissue-Engineered Vocal Fold Replacement in Rabbits

Abstract

Objectives Cell therapies using mesenchymal stromal cells (MSCs) have been proposed as a promising new tool for the treatment of vocal fold scarring. However, the mechanisms by which MSC promote healing as well as their duration of survival within the host vocal fold have yet to be defined. The aim of this work was to assess the persistence of embedded MSCs within a tissue-engineered vocal fold mucosal replacement in a rabbit model of vocal fold injury. Methods Male rabbit adipose-derived mesenchymal stromal cells (ASC) were embedded within a three-dimensional fibrin gel, forming the cell-based outer vocal fold replacement (COVR). Four female rabbits underwent unilateral resection of vocal fold epithelium and lamina propria and reconstruction with COVR implantation. Polymerase chain reaction and fluorescent in situ hybridization for the sex-determining region of the Y chromosome (SRY-II) in the sex-mismatched donor-recipient pairs sought persistent cells after 4 weeks. Results A subset of implanted male cells was detected in the implant site at 4 weeks. Many SRY-II negative cells were also detected at the implant site, presumably representing native female cells that migrated to the area. No SRY-II signal was detected in contralateral control vocal folds. Conclusions The emergent tissue after implantation of a tissue-engineered outer vocal fold replacement is derived both from initially embedded adipose-derived stromal cells and infiltrating native cells. Our results suggest this tissue-engineering approach can provide a well-integrated tissue graft with prolonged cell activity for repair of severe vocal fold scars.

A Tissue Engineered Construct for Laryngeal Regeneration: A Proof‐of‐Concept Device Design Study

“…The donor cells were identified by positive staining of the sex-determining region of the Y chromosome (SRY-II), using fluorescent in situ hybridization. 38 These cells were located in both the lamina propria and epithelial layer. Interestingly, many SRY-II negative cells were also detected, suggesting that native female cells had migrated to the area of emergent tissue.…”

Stem cell therapy for vocal fold regeneration after scarring: a review of experimental approaches

“…In the majority of studies, the VF defect is created with a cold microinstrument (forceps or scissors); in some cases, a surgical coagulator or a laser is used. The injury of the VF mucosa is inflicted in the region of the anterior and middle third of the VF and includes the superficial layers of the thyroarytenoid muscle; however, in a number of studies, e.g., in the works by Shiba et al (2016) and Goel et al (2018), the mucosa is removed along the whole VF length [ 29 , 31 , 47 , 48 ].…”

“…The implant administration is performed via an injection; as a rule, the endolaryngeal access is used, rarely—an exterior access [ 41 , 49 ]. When using hydrogel-based systems or tissue engineered constructs, they are implanted in a subepithelial pocket of the VF via the endolaryngeal route, or fixed in the region of the resected VF after a median thyrotomy [ 24 , 33 , 48 , 50 ]. In the prevailing number of studies, the product is administered to the acute primary wound of the VF immediately post-injury.…”

“…In the case of human cells, the method of fluorescent in situ hybridization (FISH) is frequently applied which uses labeled human DNA [ 22 , 23 , 26 ]. This technique along with PCR may detect male cells through the Y-chromosome DNA after their injection into the female VF, this concept was demonstrated in the study by Goel et al [ 48 ]. Such a cross-sex experimental design with the use of allogeneic cells allows one to avoid the restrictions of the traditional cell visualization methods related to the reduction of the dye concentration due to cell division and unstable expression of fluorochrome proteins.…”

Pathophysiology of Fibrosis in the Vocal Fold: Current Research, Future Treatment Strategies, and Obstacles to Restoring Vocal Fold Pliability

“…The ASCs constructs did not provoke increases in collagen expression, suggesting that grafts would be stable in the long term. Goel et al used a rabbit VF-injury model to assess the persistence of embedded ASCs within a tissue-engineered VF mucosal replacement [98]. The tissue produced after implantation of a tissue-engineered outer VF replacement was derived from both the embedded ASCs and infiltrating native cells.…”