Assessment of cellular materials generated by co-cultured 'inflamed' and healthy periodontal ligament stem cells from patient-matched groups.

Recently, stem cells derived from the'inflamed' periodontal ligament (PDL) tissue of periodontally diseased teeth (I-PDLSCs) have been increasingly suggested as a more readily accessible source of cells for regenerative therapies than those derived from healthy PDL tissue (H-PDLSCs). However, substantial evidence indicates that I-PDLSCs exhibit impaired functionalities compared with H-PDLSCs. In this study, patient-matched I-PDLSCs and H-PDLSCs were co-cultured at various ratios. Cellular materials derived from these cultures were investigated regarding their osteogenic potential in vitro and capacity to form new bone following in vivo transplantation. While patient-matched I-PDLSCs and H-PDLSCs could co-exist in co-culture systems, the proportion of I-PDLSCs tended to increase during in vitro incubation. Compared with H-PDLSC monoculture, the presence of I-PDLSCs in the co-cultures appeared to enhance the overall cell proliferation. Although not completely rescued, the osteogenic and regenerative potentials of the cellular materials generated by co-cultured I-PDLSCs and H-PDLSCs were significantly improved compared with those derived from I-PDLSC monocultures. Notably, cells in co-cultures containing either 50% I-PDLSCs plus 50% H-PDLSCs or 25% I-PDLSCs plus 75% H-PDLSCs expressed osteogenesis-related proteins and genes at levels similar to those expressed in H-PDLSC monocultures (P>0.05). Irrespective of the percentage of I-PDLSCs, robust cellular materials were obtained from co-cultures with 50% or more H-PDLSCs, which exhibited equivalent potential to form new bone in vivo compared with sheets generated by H-PDLSC monocultures. These data suggest that the co-culture of I-PDLSCs with patient-matched H-PDLSCs is a practical and effective method for increasing the overall osteogenic and regenerative potentials of resultant cellular materials.

Stem cells derived from "inflamed" and healthy periodontal ligament tissues and their sheet functionalities: a patient-matched comparison.

AIM: The aim of this study was to compare the properties of stem cells derived from "inflamed" and healthy periodontal ligament (PDL) tissues from patient-matched groups. MATERIAL AND METHODS: Patient-matched stem cells derived from root-attached "inflamed" and healthy PDL tissues from six donors, termed I-PDLSCs and H-PDLSCs, respectively, were investigated with regard to their stem cell properties, immunomodulatory effects and capacity to form robust cell sheets for therapeutic applications. RESULTS: We found that cells derived from both sources exhibited typical mesenchymal stem cell (MSC) characteristics. However, compared with H-PDLSCs, I-PDLSCs demonstrated an increased capacity to proliferate, a greater potential to migrate and a decreased capacity to differentiate into osteoblasts in vitro. When I-PDLSCs and H-PDLSCs were co-cultured with peripheral blood mononuclear cells, the MSCs derived from "inflamed" PDL tissues exhibited impaired immunomodulation. Although I-PDLSCs led to increased collagen type I, periostin and integrin ß1 content in the matrix, the cell sheets formed by I-PDLSCs were dysfunctional due to their impaired osteogenic/chondrogenic differentiation and tissue regeneration. CONCLUSIONS: These data provide additional evidence that I-PDLSCs are functionally compromised compared with H-PDLSCs. Nonetheless, their dominant abundance in the available tissues indicates that stem cells derived from damaged teeth extracted due to periodontitis warrant further exploration.

Cell Responses to Conditioned Media Produced by Patient-Matched Stem Cells Derived From Healthy and Inflamed Periodontal Ligament Tissues.

BACKGROUND: Periodontal ligament stem cells (PDLSCs) derived from clinically compromised teeth with periodontitis are considered a readily accessible cell source, but their impaired stem cell functionalities, as observed in various in vitro and in vivo models, necessitate further investigation of these inflamed cells before their translation into therapeutic applications. In this study, the effects of conditioned media (CM) produced by stem cells derived from human healthy periodontal ligament tissues (H-PDLSCs) or inflamed periodontal ligament tissues (I-PDLSCs), referred to as H-CM and I-CM, respectively, on the biologic properties of H-PDLSCs and I-PDLSCs from the same donor are compared to explore the extent to which inflamed cells can be rescued by their extrinsic environment (i.e., by H-CM). METHODS: H-CM and I-CM were prepared from in vitro cell cultures, and the cellular responses of H-PDLSCs and I-PDLSCs to patient-matched H-CM and I-CM were investigated in terms of colony-forming ability, cell proliferation, and adipogenic/osteogenic differentiation. RESULTS: In H-CM and I-CM, H-PDLSCs and I-PDLSCs exhibited similar adipogenic potential. However, when incubated in I-CM, both cell types demonstrated an increased capacity to proliferate but a decreased capacity to differentiate into osteoblasts. Significantly, the impaired osteogenic differentiation of I-PDLSCs was partially rescued by incubation in H-CM under osteo-inducing conditions. CONCLUSION: The CM of patient-matched H-PDLSCs and I-PDLSCs differed, and the impaired osteogenic differentiation of inflamed stem cells had the potential to be rescued, at least partially, for therapeutic use via changing the cell culture microenvironment in vitro.

Stromal cell-derived factor-1-directed bone marrow mesenchymal stem cell migration in response to inflammatory and/or hypoxic stimuli.

Directing cell trafficking toward a target site of interest is critical for advancing stem cell therapy in clinical theranostic applications. In this study, we investigated the effects of inflammatory and/or hypoxic stimuli on the migration of bone marrow mesenchymal stem cells (BMMSCs) during in vitro culture and after in vivo implantation. Using tablet scratch experiments and observations from a transwell system, we found that both inflammatory and hypoxic stimuli significantly enhanced cell migration. However, the combination of inflammatory and hypoxic stimuli did not result in a synergistic effect. The presence of stromal cell-derived factor-1 (SDF-1) significantly enhanced cell migration irrespective of the incubation conditions, and these positive effects could be blocked by treatment with AMD3100. Based on a time course experiment, we found that preconditioning cells with either inflammatory or hypoxic stimuli for 24 h or with both stimuli for 12 h led to high levels of chemokine receptor type 4 (CXCR4) expression. In vivo studies further demonstrated that pretreatment of BMMSCs with inflammatory and/or hypoxic stimuli resulted in an increased number of systemically injected cells migrating toward skin injuries, and local SDF-1 administration significantly increased cell migration. These findings suggest that in vitro control of either inflammatory or hypoxic stimuli has significant potential to enhance SDF-1-directed BMMSC migration via the upregulation of CXCR4 expression. Although combining the stimuli did not necessarily lead to a synergistic effect, the potential to reduce the dose and time required for cell preconditioning indicates that combinations of various strategies warrant further exploration.