RT Journal Article
SR Electronic
T1 Phenotypic Differences in Adult and Fetal Dermal Fibroblast Responses to Mechanical Tension
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 2020.09.30.319749
DO 10.1101/2020.09.30.319749
A1 Umang M. Parikh
A1 Walker D. Short
A1 Natalie Templeman
A1 Alexander Blum
A1 Daniel Colchado
A1 Harrison Strang
A1 Aditya Kaul
A1 Hui Li
A1 Xinyi Wang
A1 Monica M. Fahrenholtz
A1 Swathi Balaji
YR 2020
UL http://biorxiv.org/content/early/2020/10/01/2020.09.30.319749.abstract
AB The regenerative wound healing phenotype observed in the fetal wounds is characterized by a unique hyaluronan rich wound microenvironment. Fetal fibroblasts produce a pericellular matrix which is abundant in high molecular weight HA. In vivo, fetal skin has negligible resting tension and heals regeneratively when small wounds are made. However, critically large fetal wounds heal with increased scar. We hypothesize that higher mechanical tension will differentially alter fibroblast-mediated HA metabolism in adult and fetal fibroblasts, leading to a profibrotic fetal fibroblast phenotype. C57BL/6J fetal (FFB, E14.5) and adult (AFB, 8 wk) dermal fibroblasts were cultured on silicone membranes +/-10% static strain (1, 3, 6, and 12h). Monolayers were analyzed via PCR for HA-synthesis (HAS1-3), HA-remodeling (HYAL1-2, KIAA1199), and HA-receptor (CD44) genes, normalized to static FFB. Total HA was analyzed by gel electrophoresis at 12h. Data is reported as mean+/-SD (n=3), with p-values calculated by two-way ANOVA (post-hoc Tukey’s test). FFB pericellular matrix was reduced after tension conditions, and HA profile shifted from HMW-HA to LMW-HA. Under static conditions, AFB had increased expression of HAS 1 and 2, as well as increased expression of KIA1199, HYAL1 and 2 when compared to FFB. Overall, tension resulted in an increase in HAS1, HAS3, and HYAL1 expression in FFB, and decreased HAS2 but increased HYAL1 for AFB under tension. Staining for cytoskeletal components demonstrates that tension results in organization of F-actin and aSMA in FFB to resemble AFB. Expression of aSMA is increased in AFB with tension, while CD26 expression is increased in FFB with tension. These insights into the intrinsic differences between regenerative FFB and fibrotic AFB may yield targets to attenuate fibrosis.Competing Interest StatementThe authors have declared no competing interest.LIST OF ABBREVIATIONSHAHyaluronanEN1Engrailed-1ECMExtracellular MatrixMWMolecular WeightLMW-HALow Molecular Weight HyaluronanHMW-HAHigh Molecular Weight HyaluronanHASHyaluronan SynthaseHYALHyaluronidaseFFBFetal FibroblastsAFBAdult Fibroblasts