Abstract
ObjectivesHematoma quality (especially the fibrin matrix) plays an important role in the bone healing process. Here, we investigated the effect of interleukin-1 beta (IL-1β) on fibrin clot formation from platelet-poor plasma (PPP).MethodsFive-milliliter of rat whole-blood samples were collected from the hepatic portal vein. All blood samples were firstly standardized via a thrombelastograph (TEG), blood cell count, and the measurement of fibrinogen concentration. PPP was prepared by collecting the top two-fifths of the plasma after centrifugation under 400 × g for 10 min at 20°C. The effects of IL-1β cytokines on artificial fibrin clot formation from PPP solutions were determined by scanning electronic microscopy (SEM), confocal microscopy (CM), turbidity, and clot lysis assays.ResultsThe lag time for protofibril formation was markedly shortened in the IL-1β treatment groups (243.8 ± 76.85 in the 50 pg/mL of IL-1β and 97.5 ± 19.36 in the 500 pg/mL of IL-1β) compared to the control group without IL-1β (543.8 ± 205.8). Maximal turbidity was observed in the control group. IL-1β (500 pg/mL) treatment significantly decreased fiber diameters resulting in smaller pore sizes and increased density of the fibrin clot structure formed from PPP (P < 0.05). The clot lysis assay revealed that 500 pg/mL IL-1β induced a lower susceptibility to dissolution due to the formation of thinner and denser fibers.ConclusionIL-1β can significantly influence PPP fibrin clot structure, which may affect the early bone healing process.
Highlights
IL-1β (500 pg/mL) treatment significantly decreased fiber diameters resulting in smaller pore sizes and increased density of the fibrin clot structure formed from platelet-poor plasma (PPP) (P < 0.05)
The clot lysis assay revealed that 500 pg/mL IL-1β induced a lower susceptibility to dissolution due to the formation of thinner and denser fibers
Fracture hematoma that is formed immediately after injury is suggested to play an important role in fracture union, because the removal of a blood-clot during operative stabilization can impair the initial phase of healing [1]
Summary
Fracture hematoma (blood-clot) that is formed immediately after injury is suggested to play an important role in fracture union, because the removal of a blood-clot during operative stabilization can impair the initial phase of healing [1]. Numerous studies have focused on bone biology and fracture healing, to our knowledge, little reported literature is available on early bone healing, such as studies characterizing the fracture hematoma fibrin network and the factors that impact fibrin clot quality. The importance of hematoma at a fracture site is increasingly being recognized for its supportive role in providing a transient fibrin matrix to allow cell infiltration, proliferation, and differentiation, as well as serving as a short-term ‘reservoir’ for growth factors released from activated platelets and adjacent tissues [5]. It has been revealed that hematomas composed of loosely-woven fibrin structure with thicker fibers can better expedite the egress of mesenchymal stem cells (MSCs) and endothelial cells into injured sites, diffusion of oxygen and nutrients, and removal of metabolic waste [15, 16]
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