Soft gingiva is often compromised in gingival health; however, the underlying biological mechanisms remain unknown. Extracellular matrix (ECM) stiffness is involved in the progression of various fibroblast-related inflammatory disorders via cellular mechanotransduction. Gingival stiffness might regulate cellular mechanotransduction-mediated proinflammatory responses in gingival fibroblasts. This in vitro study aims to investigate the effects of substrate stiffness on proinflammatory responses in human gingival fibroblasts (hGFs). The hGFs isolated from two healthy donors cultured on type I collagen-coated polydimethylsiloxane substrates with different stiffnesses, representing soft (5 kPa) or hard (25 kPa) gingiva. Expression levels of proinflammatory mediators, prostaglandin E2 or interleukin-1β, in hGFs were significantly higher with the soft substrate than with the hard substrate, even without and with lipopolysaccharide (LPS) to induce inflammation. Expression levels of gingival ECM and collagen cross-linking agents in hGFs were downregulated more with the soft substrate than with the hard substrate through 14 days of culture. The soft substrate suppressed the expression of mechanotransduction-related transcriptional factors and activated the expression of inflammation-related factors, whereas the hard substrate demonstrated the opposite effects. Soft substrate induced proinflammatory responses and inhibition of ECM synthesis in hGFs by inactivating cellular mechanotransduction. This supports the importance of ECM stiffness in gingival health.

Oxidative stress hinders tissue regeneration in cell therapy by inducing apoptosis and dysfunction in transplanted cells. N-acetyl-L-cysteine (NAC) reinforces cellular antioxidant capabilities by increasing a major cellular endogenous antioxidant molecule, glutathione, and promotes osteogenic differentiation. This study investigates the effects of pretreatment of osteoblast-like cells with NAC on oxidative stress-induced apoptosis and dysfunction and bone regeneration in local transplants. Rat femur bone marrow-derived osteoblast-like cells preincubated for 3 h with and without 5 mM NAC were cultured in a NAC-free osteogenic differentiation medium with continuous exposure to 50 μM hydrogen peroxide to induce oxidative stress. NAC preincubation prevented disruption of intracellular redox balance and alleviated apoptosis and negative impact on osteogenic differentiation, even under oxidative stress. Autologous osteoblast-like cells with and without NAC pretreatment in a collagen sponge vehicle were implanted in critical-size defects in rat femurs. In the third week, NAC-pretreated cells yielded complete defect closure with significantly matured lamellar bone tissue in contrast with poor bone healing by cells without pretreatment. Cell-tracking analysis demonstrated direct bone deposition by transplanted cells pretreated with NAC. Pretreatment of osteoblast-like cells with NAC enhances bone regeneration in local transplantation by preventing oxidative stress-induced apoptosis and dysfunction at the transplanted site.

Purpose: The hazards of aerosols generated during dental treatments are poorly understood. This study aimed to establish visualization methods, discover conditions for droplets/aerosols generated in simulating dental treatments and identify the conditions for effective suction methods.Methods: The spreading area was evaluated via image analysis of the droplets/aerosols generated by a dental air turbine on a mannequin using a light emitting diode (LED) light source and high-speed camera. The effects of different bur types and treatment sites, reduction effect of intra-oral suction (IOS) and extra-oral suction (EOS) devices, and effect of EOS installation conditions were evaluated.Results: Regarding the bur types, a bud-shaped bur on the air turbine generated the most droplets/aerosols compared with round-shaped, round end-tapered, or needle-tapered burs. Regarding the treatment site, the area of droplets/aerosols produced by an air turbine from the palatal plane of the anterior maxillary teeth was significantly higher. The generated droplet/aerosol area was reduced by 92.1% by using IOS alone and 97.8% by combining IOS and EOS. EOS most effectively aspirated droplets/aerosols when placed close (10 cm) to the mouth in the vertical direction (0°).Conclusions: The droplets/aerosols generated by an air turbine could be visualized using an LED light and a high-speed camera in simulating dental treatments. The bur shape and position of the dental air turbine considerably influenced droplet/aerosol diffusion. The combined use of IOS and EOS at a proper position (close and perpendicular to the mouth) facilitated effective diffusion prevention to protect the dental-care environment.