In Vitro Cell Proliferation and Migration Properties of Oral Mucosal Fibroblasts: A Comparative Study on the Effects of Cord Blood- and Peripheral Blood-Platelet Lysate

Abstract

Cord blood-platelet lysate (CB-PL), containing growth factors such as a platelet-derived growth factor, has a similar efficacy to peripheral blood-platelet lysate (PB-PL) in initiating cell growth and differentiation, which makes it a unique alternative to be implemented into oral ulceration healing. This research study aimed to compare the effectiveness of CB-PL and PB-PL in promoting oral wound closure in vitro. Alamar blue assay was used to determine the optimal concentration of CB-PL and PB-PL in enhancing the proliferation of human oral mucosal fibroblasts (HOMF). The percentage of wound closure was measured using the wound-healing assay for CB-PL and PB-PL at the optimal concentration of 1.25% and 0.3125%, respectively. The gene expressions of cell phenotypic makers (Col. I, Col. III, elastin and fibronectin) were determined via qRT-PCR. The concentrations of PDGF-BB were quantified using ELISA. We found that CB-PL was as effective as PB-PL in promoting wound-healing and both PL were more effective compared to the control (CTRL) group in accelerating the cell migration in the wound-healing assay. The gene expressions of Col. III and fibronectin were significantly higher in PB-PL compared to CB-PL. The PDGF-BB concentration of PB-PL was the highest and it decreased after the wound closed on day 3. Therefore, we concluded that PL from both sources can be a beneficial treatment for wound-healing, but PB-PL showed the most promising wound-healing properties in this study.

Effects of Platelet Lysate Gels Derived from Different Blood Sources on Oral Mucosal Wound Healing: An In Vitro Study

“…PDGF-BB concentrations of CB-PL and PB-PL before gel polymerization were 35,250.71 ± 395.80 pg/mL and 53,015 ± 741.65 pg/mL, respectively, as published in our previous paper [ 37 ]. In this study, the protein level of PDFG-BB released from gels of CB-PL (905.48 ± 69.65 pg/mL) and PB-PL (1303.10 ± 343.96 pg/mL) was higher than the baseline of PDGF-BB in the control group (626.90 ± 8.01 pg/mL) on day 3 ( Figure 6 ).…”

“…The plasma layer was collected into a new centrifuge tube for the second centrifugation at 600× g for 5 min at room temperature. Following this, ⅔ of the upper part of the centrifuged plasma (platelet-low plasma) was removed and ⅓ of the centrifuged plasma (PRP) was retained [ 37 ]. A total of 1 mL of CB-PRP was sent to the Pathology unit, UKMMC to determine the platelet count using the SYSMEX Hematological Analyzer (SYSMEX, Tokyo, Japan).…”

“…Cell characterization of HOMFs (passage 2) was carried out through cell morphology identification and gene expression of the fibroblast marker (vimentin). The data of cell characterization were published in our previous paper [ 37 ]. All assays were performed using HOMFs at passage 4.…”

“…The total platelet counts of PRP obtained from cord blood (CB-PRP) and peripheral blood (PB-PRP) were 378 ± 59 9 /L and 486 ± 50 9 /L as published in our previous paper [ 37 ]. Both types of PRP were standardized to the platelet count of 300 × 10 9 /L.…”