Purpose: To evaluate the biochemical composition of bone nodules deposited by gingival mesenchymal stem cells (GMSCs) over titanium machined surfaces in vitro.
Materials and methods: GMSCs were isolated from healthy gingival tissues of patients undergoing crown-lengthening surgical procedures. GMSCs were characterized following the International Society for Cellular Therapy guidelines. After incubation of the GMSCs with titanium discs, osteogenic differentiation was induced for 28 days. Osteogenic lineage was confirmed by means of Alizarin Red S staining. Bone nodule morphology and deposition by GMSCs were characterized by scanning electron microscopy (SEM). An elemental analysis of the bone nodules was done using energy-dispersive x-ray spectroscopy (EDS). The biochemical composition of these nodules was further characterized via Raman spectroscopy, with native alveolar bone used as a control.
Results: GMSCs adhered and proliferated on the titanium discs and exhibited a spindle-shaped fibroblast-like morphology under standard culture conditions. Their phenotype was confirmed by the expression of CD105, CD90, CD73, and CD146, observed using flow cytometry. Deposits of calcium bone nodules were evident in the cultures after staining with Alizarin Red S, but were absent in the controls. Calcium and phosphate, the major components of hydroxyapatite, were present in the bone nodules, as shown by means of the EDS analysis. The results obtained from Raman spectra of these nodules showed the phosphate ions (ν[PO₄³⁻], ~960 cm¹), amide III (δ[NH], ~1,245 cm⁻¹), CH₂ scissors (~1,451 cm⁻¹), amide I (ν[C = O], ~1,667 cm⁻¹), and ν(CH) (2,800-3,100 cm⁻¹) bands were similar to those observed in native bone.
Conclusion: GMSCs can deposit a bone-like mineral highly similar to native bone (HA) over titanium surfaces. Ongoing studies are aimed at determining whether GMSCs can deposit a similar bone matrix/tissue over removed failed dental implants. If HA can be placed over removed failed dental implants, it may be possible to re-osseointegrate dental implants that are failing as a result of peri-implantitis in vivo.