Modulation of vocal fold scar fibroblasts by adipose-derived stem/stromal cells.
OBJECTIVE
To explore whether adipose-derived stem/stromal cells (ASCs) have therapeutic potential for treating scarred superficial lamina propria through the effects of secreted hepatocyte growth factor (HGF) on scar fibroblasts.


METHODS
In vitro study using coculture system.


METHODS
Scar fibroblasts (SFs) were isolated from ferret vocal folds electrocauterized 2 weeks previously. ASCs were isolated from ferret lipoaspirated subcutaneous abdominal fat. For coculture experiments, the two cell types were combined in Transwell plates for 6 days, followed by 1 or 3 days of monoculture after removing the upper chamber. Assays were then performed on cells and media from the bottom chamber. We measured: 1) the production of hyaluronic acid (HA), collagen and HGF via enzyme-linked immunosorbent assays, 2) the expression of alpha-smooth muscle actin (alpha-SMA), 3) cell proliferation, and 4) apoptosis of SFs (2, 3, and 4 via flow cytometry). Other experiments examined the effects of HGF on SFs and the effects of HGF neutralization in the coculture system.


RESULTS
Coculture led to significant decreases in SF collagen production (P < .05), cell proliferation (P < .05), and alpha-SMA expression (P < .05), whereas HA production increased (P < .05). Coculture also increased HGF secretion from ASCs (P < .05). Neutralization of HGF abolished the inhibitory effects of ASCs on SF collagen synthesis (P < .05).


CONCLUSIONS
ASCs influence SFs to adopt a less fibrotic profile. It appears that HGF is at least one of the soluble factors responsible for this effect. Implanted ASCs could potentially ameliorate vocal fold scar by acting as long-term, intrinsic sources of HGF.
Kumai
Yoshihiko
Y
Department of Surgery, Harvard Medical School, Center for Laryngeal Surgery and Voice Rehabilitation, Massachusetts General Hospital, Boston, Massachusetts 02114, USA.
Kobler
James B
JB
Park
Hyoungshin
H
Galindo
Marilyn
M
Herrera
Victoria L M
VL
Zeitels
Steven M
SM
D000199:Actins / Q000096:biosynthesis; D000273:Adipose Tissue / Q000166:cytology*; D000818:Animals; D017209:Apoptosis; D002921:Cicatrix / Q000473:pathology*; D018920:Coculture Techniques; D003094:Collagen / Q000096:biosynthesis; D005289:Ferrets; D005347:Fibroblasts / Q000378:metabolism* / Q000473:pathology / Q000502:physiology; D017228:Hepatocyte Growth Factor / Q000096:biosynthesis* / Q000494:pharmacology; D006820:Hyaluronic Acid / Q000096:biosynthesis; D007150:Immunohistochemistry; D008297:Male; D013234:Stem Cells / Q000502:physiology*; D017154:Stromal Cells / Q000502:physiology*; D014827:Vocal Cords / Q000473:pathology*
C541116:ACTA2 protein, human; D000199:Actins; D017228:Hepatocyte Growth Factor; D006820:Hyaluronic Acid; D003094:Collagen