Safi IN, et al. J Periodontal Implant Sci. 2022 Jan;52:e5. https://doi.org/10.5051/jpis.2006080304

Per-implantitis is one of the implant treatment complications. Dentists have failed to restore damaged periodontium by using conventional therapies. Tissue engineering (stem cells, scaffold and growth factors) aims to reconstruct natural tissues. The paper aimed to isolate both periodontal ligament stem cells (PDLSCs) and bone marrow mesenchymal stem cells (BMMSCs) and use them in a co-culture method to create three-layered cell sheets for reconstructing natural periodontal ligament (PDL) tissue. BMMSCs were isolated from rabbit tibia and femur, and PDLSC culture was established from the lower right incisor. The cells were co-cultured to induce BMMSC differentiation into PDL cells. Cell morphology, stem cells and PDL-specific markers (CD90, CD34, and periostin) were also detected using immunofluorescent assay. Co-cultured cell monolayers were detached using temperature-responsive tissue culture dishes and collagen graft to create the three-layer construct. The 3D-engineered tissue was examined histologically and by field emission scanning electron microscopy (FESEM). BMMSCs co-cultured with PDLSCs successfully induced more PDL cells. The newly induced PDL cells exhibited periostin and CD90 expression. Fluorescence green intensity was measured for the co-cultured cells that were stained with periostin, the mean fluorescence green intensity (periostin expression) was significantly higher for the newly induced PDL cells after 1, 2, and 3 weeks when compared with control (BM-MSCs), at 21 days non-significant difference was measured when compared with control (PDLSCs).The results showed the successful formation of 3D multilayer PDL tissue. Histological cross-section showed cell sheets and the stable adhesion between them. FESEM examination was conducted for the cross-section, showing three-layered cell sheets with stable adhesion between cells. The results of this paper report that the three layered-cell sheets were successfully constructed by the novel use of collagen graft as a scaffold to be used in treatment of periodontitis and to envelop the dental implants to create biohybrid implant.

This work presents laser coating of grade 1 pure titanium (Ti) dental implant surface with sintered biological apatite beta-tricalcium phosphate (β-TCP), which has a chemical composition close to bone. Pulsed Nd:YAG laser of single pulse capability up to 70 J/10 ms and pulse peak power of 8 kW was used to implement the task. Laser pulse peak power, pulse duration, repetition rate and scanning speed were modulated to achieve the most homogenous, cohesive and highly adherent coat layer. Scanning electron microscopy (SEM), energy dispersive X-ray microscopy (EDX), optical microscopy and nanoindentation analyses were conducted to characterise and evaluate the microstructure, phases, modulus of elasticity of the coating layer and calcium-to-phosphate ratio and composition.Results showed that the laser power and scanning speed influenced coating adherence. The cross-sectional field-emission scanning electron microscopy images at low power and high speed showed poor adherence and improved as the laser power increased to 2 kW. Decreasing the scanning speed to 0.2 mm/s at the same power of 2 kW increased adherence. EDX results of the substrate demonstrated that the chemical composition of the coat layer did not change after processing. Moreover, the maps revealed proper distribution of Ca and P with some agglomeration on the surface. The sharp peaks on the X-ray diffraction patterns indicated that β-TCPs in the coat layer were mostly crystalline. The elastic modulus was low at the surface and increased gradually with depth to reach 19 GPa at 200 nm; this value was close to that of bone. The microhardness of the coated substrate increased by about 88%. The laser pulse energy of 8.3 J, pulse peak power of 2 kW, pulse duration of 4.3 min, repetition rate of 10 Hz and scanning speed of 0.2 ms-1 yielded the best results. Both processing and coating have potential use for dental implant applications.

The strategies of tissue-engineering led to the development of living cell-based therapies to repair lost or damaged tissues, including periodontal ligament and to construct biohybrid implant. This work aimed to isolate human periodontal ligament stem cells (hPDLSCs) and implant them on fabricated polycaprolactone (PCL) for the regeneration of natural periodontal ligament (PDL) tissues. hPDLSCs were harvested from extracted human premolars, cultured, and expanded to obtain PDL cells. A PDL-specific marker (periostin) was detected using an immunofluorescent assay. Electrospinning was applied to fabricate PCL at three concentrations (13%, 16%, and 20% weight/volume) in two forms, which were examined through field emission scanning electron microscopy (FESEM). The isolated hPDLSCs were implanted on the fabricated PCL. After 21 days, FESEM was conducted to evaluate the implanted scaffolds, and an MTT assay was performed to characterize the biological response of the PCL scaffold at different cell exposure durations (24, 48, and 72 h). Periostin was expressed in the expanded PDL cells, and this result revealed that 20% weight/volume PCL scaffold with a pore size of more than 10 μm was the best. The growth rates of PDLSCs were high. Cytotoxicity test of fabricated PCL scaffold demonstrated no significant change in the cell viability when compared with the negative control and no deteriorating or inhibitory effect on growth after different durations. A cell sheet was successfully formed by using PCL as a scaffold to cover dental implants and promote PDL cell attachment, proliferation, and growth for biohybrid implant construction.