Release kinetics and mitogenic capacity of collagen barrier membranes supplemented with secretome of activated platelets - the in vitro response of fibroblasts of the periodontal ligament and the gingiva

Eva-Maria Mozgan,Klara Janjić,Michael Edelmayer,Andreas Moritz,Manuela Pensch,Michael B Fischer,Hermann Agis

doi:10.1186/s12903-017-0357-6

Abstract

BackgroundPlatelet preparations can stimulate the healing process and have mitogenic properties. We hypothesized that collagen barrier membranes (CBM), clinically used in guided bone regeneration and guided tissue regeneration, can serve as carriers for platelet secretome.MethodsSecretome was generated from washed platelets and unwashed platelets (washed/unwashed PSEC) and lyophilized onto CBM. Overall appearance of CBM was evaluated by scanning electron microscopy. The impact of PSEC on cell attachment was measured based on fluorescence microscopy with DiI-labeled cells. To assess the release kinetics, supernatants of CBM were collected and medium was replaced at hour 1–48. The mitogenic effect was evaluated with periodontal fibroblasts. Furthermore, the release of total protein, platelet-derived growth factor (PDGF)-BB, and transforming growth factor (TGF) β1 was measured.ResultsCBM overall appearance and cell attachment was not modulated by PSEC. Supernatants taken after one hour induced a mitogenic response in fibroblasts and showed the highest levels of total protein, TGFβ1 and PDGF-BB. These effects decreased rapidly in subsequent supernatants. While supernatants of CBM loaded with unwashed PSEC induced a stronger mitogenic response than supernatants of CBM loaded with washed PSEC this difference between the PSEC preparations was not observed when cells were seeded on 48–hours-washed CBM.ConclusionsCBM release platelet-derived factors in continuously declining release kinetics.

Highlights

Platelet preparations can stimulate the healing process and have mitogenic properties
To reveal if supplementation of collagen barrier membranes (CBM) with unwashed and washed Platelet secretome (PSEC) modulates cellular attachment kinetics to the membranes, DiI-labeled Gingival fibroblasts (GF) and Periodontal ligament fibroblasts (PDLF) were seeded onto the smooth side of the CBM which is the side facing the soft tissue when applied clinically (Fig. 1d, e)
There was no significant difference between the numbers of fibroblasts attached to unwashed PSECloaded, washed PSEC-loaded, and the unloaded CBM after 1 h, 3 h, and 24 h of incubation

Summary

Introduction

Platelet preparations can stimulate the healing process and have mitogenic properties. In guided tissue regeneration collagen barrier membranes (CBM) are used as scaffolds to provide structural support for repair cells and prevent the ingrowth of soft tissue into periodontal and bone defects [1,2,3]. The success of this approach depends on the regenerative. Wound healing and bone regeneration are initiated by the formation of a blood clot In these blood clots, activated platelets release a myriad of growth factors that stimulate migration and proliferation of repair cells [10,11,12]. The finding of mitogenic effects of activated platelets induced the development of regenerative strategies based on platelet concentrates [6,7,8,9, 22]

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