Combination therapies for the treatment of oral squamous cell carcinoma have been studied extensively and represent a synergistic approach with better outcomes than monotherapy. In this study, a novel combination therapy was investigated using gold nanoparticles (GNP) conjugated to programmed cell death protein ligand 1 (PD-L1) antibodies and nonthermal plasma (NTP). The present study describes the effectiveness of NTP using PD-L1 antibody conjugated to GNP in PD-L1 expressing SCC-25 cells, an oral squamous cell carcinoma line. Immunocytochemistry revealed higher levels of PD-L1 expression and an increase in the selective uptake of PD-L1 antibody + GNP on SCC-25 cells compared to HaCaT cells. In addition, cell viability analyses confirmed higher levels of cell death of SCC-25 cells after treatment with PD-L1 antibody, GNP, and NTP compared to HaCaT cells. Among the experimental groups, the highest cell death was observed upon treatment with PD-L1 antibody + GNP + NTP. Following the Western blot analysis and immunofluorescence staining, the expression of apoptosis-related proteins was found to increase after treatment with PD-L1 antibody + GNP + NTP among the other experimental groups. In conclusion, the treatment of SCC-25 cells with PD-L1 antibody + GNP + NTP significantly increased the number of dead cells compared to other experimental groups. The results of this in vitro study confirmed the therapeutic effects of PD-L1 antibody + GNP + NTP treatment on oral squamous cell carcinoma.

Whitlockite (WH; Ca 18 Mg 2 (HPO4) 2 (PO4) 12 ) is a calcium phosphate based ceramic that contains magnesium ions. As the second most abundant mineral in living bone, WH occupies 25−35 wt % of the inorganic portion of human bone. Compared to hydroxyapatite (HAp, Ca 10 (PO 4 ) 6 (OH) 2 ), WH possesses better mechanical properties, faster resorbability, and promotion behavior on the osteogenesis. In this article, we introduced a fabrication method of interconnected porous WH granules through vacuum filtration, followed by sintering treatment based on the thermal stability of WH synthesized using the tri-solvent system. This study presents a histological, radiological, and immunohistochemical evaluation of the bone healing potential of these WH granules in a 5 mm diameter calvarial bone defect in rats. The histological evaluation shows no inflammation or foreign body reaction in the WH group. The WH group displays newly formed bone at the same thickness as the original bone. On the contrary, bone formation is not observed in the nontreated (NT) group. Besides, immunohistochemistry (IHC) confirmed that WH granules promoted bone regeneration with the significantly higher expression of bone morphogenetic proteins-2 (BMP-2), alkaline phosphatase (ALP), osteopontin (OPN), and osteocalcin (OCN) compared to the NT group without the addition of exogenous cells or growth factors. These results suggest that WH has excellent potential for application in bone tissue regeneration.

The objective of this study was to evaluate the effect of non-thermal plasma (NTP) on the healing process of peripheral nerve crush injuries, which can occur during dental implant procedures. For this, a rat model of sciatic nerve crush injury (SNCI) was adopted. The rats were divided into three groups: non-nerve damage (non-ND), nerve damage (ND), and ND+NTP group. To evaluate the sciatic nerve (SN) function, the static sciatic index was calculated, and the muscle and SN tissues were subjected to a histologic analysis. The results showed that NTP effectively accelerated the healing process of SNCI in rats. In contrast to the ND group, which showed approximately 60% recovery in the SN function, the NTP-treated rats showed complete recovery. Histologically, the NTP treatments not only accelerated the muscle healing, but also reduced the edema-like phenotype of the damaged SN tissues. In the ND group, the SN tissues had an accumulation of CD68-positive macrophages, partially destroyed axonal fibers and myelinated Schwann cells. Conversely, in the ND+NTP group, the macrophage accumulation was reduced and an overall regeneration of the damaged axon fibers and the myelin sheath was accomplished. The results of this study indicate that NTP can be used for healing of injured peripheral nerves.

This experimental research aimed to investigate the effects of non-thermal plasma on nerve regeneration after transected nerve damage using the sciatic nerve in Wistar albino (A) rats. The experiments were performed on 27 Wistar A rats. The rats underwent surgery for right sciatic nerve exposure and were divided into three groups (each group, n = 9) according to sciatic nerve transected injury (SNTI) and non-thermal plasma application: a non-nerve damage (non-ND) group, a only nerve damage without non-thermal plasma application (ND) group, and a nerve damage with non-thermal plasma application (ND + NTP) group. Subsequent to SNTI and immediate suture, non-thermal plasma was administered three times per week for eight weeks. Evaluation for functional recovery was performed using the static sciatic index measured over the full treatment period of eight weeks. The sciatic nerve specimens were obtained after euthanasia and third day from the last non-thermal plasma application. The sciatic nerve tissues were subjected to histological analysis. Behavior analysis presented that the ND + NTP group showed improved static sciatic index compared with the nerve damage group. Histopathological findings demonstrated that the ND + NTP group had more dense Schwann cells and well-established continuity of nerve fibers, greater than the nerve damage group. Immunohistochemistry showed that the ND + NTP group had increased levels of markers for microtubule-associated protein 2 (MAP2), tau, S100 calcium-binding protein B, and neurofilament-200 and regulated the overexpression of CD68 and MAP2. These results indicated that non-thermal plasma enhanced the motor function and restored the neuronal structure by accelerating myelination and axonal regeneration. Additionally, non-thermal plasma was confirmed to have a positive effect on the recovery of SNTI in rats.

Background Whitlockite (WH; Ca18Mg2(HPO4)2(PO4)12) as the second most abundant ceramic of human bone after hydroxyapatite (HA; Ca10(PO4)6(OH)2), possessing high osteogenic activity, has been known that it can stimulate osteogenic differentiation as well as suppress osteoclastic activity, particularly, in the preparatory stage of bone regeneration. Methods In this work, we construct a unique ceramic structure by layering WH on the surface of HA granules (HAGs)s via dip-coating and sintering method. Results The cell proliferation of the WH-coated HAGs (WHHAGs) group in the cell counting kit (CCK-8) was about 1.15-fold higher than that of the NT group at 72 h after incubation. The western blot and qPCR results demonstrated that WHHAGs treatment readily stimulated the transcription of osteogenic genes by regulating the downstream signaling pathway of BMP and WNT receptors.This study presents a histological and radiological evaluation of the bone healing potential of the WHHAGs in a 7 mm diameter calvarial bone defect in rats, compared with/without the BMP and non-treated (NT) groups at the 8 weeks after surgery. The bone volume fraction (bone volume/tissue volume) of the WHHAGs group was about 7.32- and 3.56- fold higher than that of the NT group with/without the BMP, respectively. Besides, histological evaluation confirmed that the WHHAGs with BMP promoted bone regeneration. Conclusion These results suggest that the WHHAGs present the remarkable potential for application in dental and orthopedic bone regeneration.