Activity of two hyaluronan preparations on primary human oral fibroblasts

Abstract

Background and Objective The potential benefit of using hyaluronan ( HA ) in reconstructive periodontal surgery is still a matter of debate. The aim of the present study was to evaluate the effects of two HA formulations on human oral fibroblasts involved in soft tissue wound healing/regeneration. Material and Methods Metabolic, proliferative and migratory abilities of primary human palatal and gingival fibroblasts were examined upon HA treatment. To uncover the mechanisms whereby HA influences cellular behavior, wound healing‐related gene expression and activation of signaling kinases were analyzed by qRT ‐ PCR and immunoblotting, respectively. Results The investigated HA formulations maintained the viability of oral fibroblasts and increased their proliferative and migratory abilities. They enhanced expression of genes encoding type III collagen and transforming growth factor‐β3, characteristic of scarless wound healing. The HA s upregulated the expression of genes encoding pro‐proliferative, pro‐migratory, and pro‐inflammatory factors, with only a moderate effect on the latter in gingival fibroblasts. In palatal but not gingival fibroblasts, an indirect effect of HA on the expression of matrix metalloproteinases 2 and 3 was detected, potentially exerted through induction of pro‐inflammatory cytokines. Finally, our data pointed on Akt, Erk1/2 and p38 as the signaling molecules whereby the HA s exert their effects on oral fibroblasts. Conclusion Both investigated HA formulations are biocompatible and enhance the proliferative, migratory and wound healing properties of cell types involved in soft tissue wound healing following regenerative periodontal surgery. Our data further suggest that in gingival tissues, the HA s are not likely to impair the healing process by prolonging inflammation or causing excessive MMP expression at the repair site.

Role of hyaluronan in regulating self-renewal and osteogenic differentiation of mesenchymal stromal cells and pre-osteoblasts

“…The investigated HA formulations were kindly provided by Regedent AG (Zurich, Switzerland) and previously described [31]. In brief, HA1 (hyaDENT) is a native non-crosslinked HA with MW of 2500 kDa, whereas HA2 (hyaDENT BG) is a formulation containing complexes of butanediol diglycidyl ether (BDDE)-crosslinked 1000 kDa-HA monomers and the above non-crosslinked form.…”

“…In brief, HA1 (hyaDENT) is a native non-crosslinked HA with MW of 2500 kDa, whereas HA2 (hyaDENT BG) is a formulation containing complexes of butanediol diglycidyl ether (BDDE)-crosslinked 1000 kDa-HA monomers and the above non-crosslinked form. Throughout the study, HA was used at a final concentration of 4 mg/ml in 0.3%FBS/DMEM as previously described [31]. The concentration was chosen based on (1) pilot experiments (data not shown) comparing 4 mg/ml-diluted HAs with the undiluted commercial preparations and (2) the assumption that each of the HA preparations will be naturally diluted by patients' blood and saliva during the periodontal surgical procedure.…”

“…Proliferation rates of HA-treated ST2 and MC3T3-E1 cells were determined using a 5-bromo-20-deoxyuridine (BrdU) chemiluminescent ELISA assay (Roche, Basel, Switzerland) as described [31,33]. In brief, after 24 h of starvation, cells were plated in triplicate at 2 × 10 3 cells/well in 3% FBS/ DMEM on Black 96-well microtiter plates (Thermo Fisher Scientific, Reinach, Switzerland) coated with HA at a final concentration of 4 mg/ml.…”

“…Before conducting such clinical studies, a better understanding is needed of the influence of HA on the tissues comprising the periodontium, i.e., cementum, alveolar bone, periodontal ligament, and gingival connective tissue. In a recent study, we have demonstrated diverse positive effects of two commercially available HA formulations on primary human palatal and gingival fibroblasts, two cell types involved in soft tissue healing/regeneration following periodontal reconstructive therapies that utilize palatal connective tissue or free gingival grafts [31]. The observed pro-proliferative, pro-migratory, and pro-wound healing properties of the two HAs speak in favor of their clinical potential.…”

Neues regeneratives Konzept für die Behandlung von Periimplantitis mit Verwendung von Hyaluronsäure als (Wund-)Heilungsbeschleuniger (Clean and Seal)

“…Bei Rezessionsdeckungen konnte mit der begleitenden HA-Anwendung (hyaDENT BG, REGEDENT GmbH) eine statistisch signifikant verbesserte Abdeckung der exponierten Zahnwurzeloberflächen erzielt werden. Mit Hyaluronsäure behandelte Patienten beanstandeten zudem statistisch signifikant weniger Unbehagen und Schwellungen [32,33].…”