Impact of APRF+ in Combination with Autogenous Fibroblasts on Release Growth Factors, Collagen, and Proliferation and Migration of Gingival Fibroblasts: An In Vitro Study.

The present study aimed to compare the action of advanced platelet-rich fibrin (A-PRF+) alone with the action of A-PRF+ combined with autologous gingival fibroblasts. The components released from A-PRF+ conditioned with autogenous fibroblasts that were quantified in the study were fibroblast growth factor (FGF), vascular endothelial growth factor (VEGF), trans-forming growth factor-beta1 and 2 (TGFß1 and TGFß2), and soluble collagen. A-PRF+ combined with fibroblasts demonstrated significantly higher values of released VEGF at every time point and, after 7 days, significantly higher values of released TGFß2. A viability test after 72 h showed a significant increase in proliferation fibroblasts after exposition to the factors released from A-PRF+ combined with fibroblasts. Similarly, the degree of wound closure after 48 h was significantly higher for the factors released from A-RRF+ alone and the factors released from A-RRF+ combined with fibroblasts. These results imply that platelet-rich fibrin (PRF) enhanced with fibroblasts can be an alternative method of connective tissue transplantation.

In Vitro Gingival Wound Healing Activity of Extracts from Reynoutria japonica Houtt Rhizomes.

Rhizomes of Reynoutria japonica Houtt. are a traditional Chinese medicinal herb (Polygoni cuspidati rhizoma, hu zhang) used for treatment of numerous diseases including wound healing support. The aim of this study was to provide evidence for the value of this herbal drug's traditional use as a gingival healing treatment as well as to obtain the most active extract. In vitro studies were performed using primary human gingival fibroblasts (HGFs) with determination of viability (MTT assay), cell proliferation (the confocal laser scanning microscope (CLSM) was used to visualize histone 3 expression), cell migration (wound healing assay), and evaluation of the expression of collagen type III (immunocytochemical staining) after incubation with extracts from R. japonica rhizomes (25% or 40% ethanol or 60% acetone). In addition to these extracts, commercial dental rinse (containing chlorhexidine digluconate 0.2%) was tested as the gold standard of choice for gum healing in dental practice. The studied extracts were qualitatively and quantitatively characterized using the validated HPLC/DAD/ESI-HR-QTOF-MS method. Total phenols and tannins content were determined using the Folin-Ciocalteu assay. Low concentration of all extracts after 24 h incubation caused significant increase in HGF viability. This effect was most pronounced at a concentration of 50 µg/mL, which was selected for further experiments. All extracts (at 50 µg/mL) stimulated HGF to proliferate, migrate, and increase collagen III synthesis, but with different strength. The highest stimulated proliferation and migration activity was observed after incubation with 25% EtOH, which according to phytochemical analysis may be related to the highest content of resveratrol and an appropriate composition of procyanidins. The 25% EtOH extract from R. japonica rhizomes appears to be a promising gingival wound healing agent worthy of animal and clinical trials.

Assessment of Human Gingival Fibroblast Proliferation after Laser Stimulation In Vitro Using Different Laser Types and Wavelengths (1064, 980, 635, 450, and 405 nm)-Preliminary Report.

PURPOSE: to assess the effect of photobiomodulation (PBM) on human gingival fibroblast proliferation. METHODS: The study was conducted using the primary cell cultures of human fibroblasts collected from systemically healthy donors. Three different laser types, Nd:YAG (1064 nm), infrared diode laser (980 nm), and prototype led laser emitting 405, 450, and 635 nm were used to irradiate the fibroblasts. Due to the patented structure of that laser, it was possible to irradiate fibroblasts with a beam combining two or three wavelengths. The energy density was 3 J/cm2, 25 J/cm2, 64 J/cm2. The viability and proliferation of cells were determined using the (Thiazolyl Blue Tetrazolium Blue) (MTT) test conducted 24, 48, and 72 h after laser irradiation. RESULTS: The highest percentage of mitochondrial activity (MA = 122.1%) was observed in the group irradiated with the 635 nm laser, with an energy density of 64 J/cm2 after 48 h. The lowest percentage of MA (94.0%) was observed in the group simultaneously irradiated with three wavelengths (405 + 450 + 635 nm). The use of the 405 nm laser at 25 J/cm2 gave similar results to the 635 nm laser. CONCLUSIONS: The application of the 635 nm and 405 nm irradiation caused a statistically significant increase in the proliferation of gingival fibroblasts.

The effect of dental gel formulation on human primary fibroblasts - an in vitro study.

INTRODUCTION: In ordinary dental practice, the dentist often meets with patients suffering from ulcers, aphtha with edema, bleeding gums, bothersome burning, and dry mouth. These are prosthetic, orthodontic patients, after surgery, in various age ranges. Protefix® gel is a soothing and regenerating preparation aimed at patients with mucosal problems. The aim of our study was to evaluate the protective properties and the safety of Protefix® gel application after dental procedures in vitro. MATERIAL AND METHODS: Human gingival fibroblasts (HGFs) were isolated from normal gingival tissues, cultured to full monolayer and exposed to Protefix® gel in the concentration from 1 to 100%. The viability of cells was examined by MTT assays. Cell migration as a response of treated cells was assessed. The expression of collagen III was estimated by immunocytochemistry after 20 minutes or 24 hours incubation with Protefix® gel. RESULTS: The obtained results indicated that the verified gel significantly stimulated fibroblasts' proliferation, and mitochondrial activity determined by MTT assay increased almost two-fold for lower gel concentrations. The immunohistochemical detection of collagen III revealed an increased expression after incubation with 5% gel. CONCLUSIONS: The results proved that the gel is safe for cells derived from human gingiva and moreover has regenerative properties, which can be of great importance in the treatment of gingivitis after retraction and surgical procedures, or even ordinary daily injuries of oral cavity.

Chemical Composition of East Asian Invasive Knotweeds, their Cytotoxicity and Antimicrobial Efficacy Against Cariogenic Pathogens: An In-Vitro Study.

BACKGROUND Giant knotweeds originating from East Asia, such as Reynoutria japonica, and Reynoutria sachalinensis, and their hybrid such as Reynoutria x bohemica, are invasive plants in Europe and North America. However, R. japonica is also a traditional East Asian drug (Polygoni cuspidati rhizoma) used in Korean folk medicine to improve oral hygiene. The aim of this study was to evaluate the antibacterial activity of acetone extracts of Reynoutria species against dominant caries pathogen such as Streptococcus mutans and alternative pathogens, as well as characterize the phytochemical composition of extracts and examine their cytotoxicity. MATERIAL AND METHODS Ultrasonic extraction was used to obtain polyphenol-rich extracts. The extracts were characterized by HPLC-DAD-ESI-MS. To test bacterial viability, the minimal inhibitory concentration (MIC) and minimal bactericidal concentration (MBC) against S. mutans, S. salivarius, S. sanguinis, and S. pyogenes were determined. The cytotoxicity of the extracts to human fibroblasts derived from gingiva was evaluated using the MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay. RESULTS The R. japonica extract had the highest bacteriostatic and bactericidal activity against pathogens causing caries, mainly dominant caries pathogen S. mutans (mean MIC 1000 µg/mL and MBC 2000 µg/mL), which was most likely associated with a higher content of stilbene aglycons and anthraquinone aglycons in the extract. Moreover, the R. japonica extract demonstrated the lowest cytotoxic effect on human fibroblasts and exhibited cytotoxic activity only at the concentration causing the death of all S. mutans. CONCLUSIONS The results indicate that the R. japonica acetone extract can be considered as a natural, antimicrobial agent for caries control.