The molecular weight of hyaluronic acid influences metabolic activity and osteogenic differentiation of periodontal ligament cells

Abstract

Objective While HA is present naturally in periodontal tissues, its molecular weight can vary widely in vivo. The objective of this study was to directly compare the biological reactions of periodontal ligament cells to four distinct molecular weights of hyaluronic acid (HA). Materials and methods Immortalized human periodontal ligament cells (PDL-hTERT) were cultured for 21 days in culture medium alone (control) or enriched with osteogenic supplements (OS group). Other 4 experimental groups were cultured in OS medium with the addition of HA with different molecular weights (HMW, MMW, LMW, and ULMW). The cell morphology was examined daily. WST1 assays were performed to evaluate metabolic activity. Von Kossa staining and calcium deposition assay were used to analyze osteogenic differentiation and mineralization. Results Cell morphology remained unaltered in all groups. Cells stimulated with OS alone or with the addition of hyaluronan showed all the typical microscopic appearance of osteogenic differentiation. Metabolic activity increased in all groups over time. Hyaluronan stimulated greater metabolic activity than the control group, with LMW HA and MMW HA showing the most significant increase. All groups showed mineral deposits and calcium deposition after 21 days of stimulation. Conclusion Our results suggest that hyaluronan can promote metabolic activity and mineralization of PDL-hTERT cells, with LMW HA being the most effective. Clinical relevance These results shed light on how the various molecular weight fractions of HA promote tissue regeneration and repair, as well as help to identify an optimal molecular weight range for this application in periodontal tissues.

Evaluation of in vitro osteoblast and osteoclast differentiation from stem cell: a systematic review of morphological assays and staining techniques

“…A total of 14 articles related to the morphological approach, published between 2013 and 2023 were selected. The human cultured stem cells used for the in-vitro studies include both peripheral blood mononuclear cells (PBMSC) ( Kleinhans et al, 2015 ; Maria et al, 2018 ; Steller et al, 2020 ; Grünherz et al., 2020 ; Zainol Abidin et al., 2023 ) and mesenchymal stem cells (MSc) involved 5 studies ( Maria et al, 2017 ; Hashimoto et al, 2018 ; Liu et al, 2018 ; Ganguly et al, 2020 ; Nugraha et al, 2023 ) whereas periodontal ligament stem cells (PDLSC) ( Di Vito et al, 2020 ; Frasheri et al, 2023 ) and dental pulp stem cells (DPSC) ( Escobar et al, 2020 ; Escobar et al, 2023 ) involved two studies ( Table 3 ). The outcomes of the risk of bias assessment in Table 4 reflect the quality assessment analysis.…”

“…Seven studies focus solely on the staining of osteoblast differentiation ( Escobar et al, 2020 ; Escobar et al, 2023 ; Ganguly et al, 2020 ; Di Vito et al, 2020 ; Frasheri et al, 2023 ; Zainol Abidin et al., 2023 ; Nugraha et al, 2023 ), three studies only on morphological evaluation methods for osteoclast differentiation ( Kleinhans et al, 2015 ; Steller et al, 2020 ; Grünherz et al., 2020 ) and another four articles involved qualification analysis of both osteoblast and osteoclast differentiation ( Hashimoto et al, 2018 ; Maria et al, 2018 ; Liu et al, 2018 ) ( Table 3 ). All the staining techniques were proven to be successful in evaluating the presence of osteoblast and osteoclast differentiation in cells.…”

Reconstructive Approach in Residual Periodontal Pockets with Biofunctionalized Heterografts—A Retrospective Comparison of 12-Month Data from Three Centers

“…Via binding to its canonical receptors CD44 and RHAMM, it has been shown to increase proliferation, migration, and cell metabolism in periodontal ligament fibroblasts (PDLs), mesenchymal stromal cells (MSCs), and cementoblasts. In osteoblasts and cementoblasts, xHyA may also stimulate the expression of bone-specific genes, while it shifts macrophage polarization towards an anti-inflammatory M2 phenotype (Figure 5) [14,16,[21][22][23][24]. In a recent histomorphometric study in dogs, the combination of a resorbable matrix and xHyA was also shown to be superior, underlining the regenerative capacity of this biomaterial [25].…”

Insights and Advancements in Periodontal Tissue Engineering and Bone Regeneration

“…A recent study investigated the development of an in-situ tissue engineering scaffold (iTE-scaffold) using the coaxial electrospinning technique to sequentially deliver bFGF and BMP-2 [65][66][67][68]. The iTE-scaffold was found to effectively enhance the angiogenesis of periodontal ligament stem cells (PDLSCs) and induce macrophage polarization into the prohealing M2 phenotype, thereby modulating inflammation [7].…”

“…In this regard, the state of the art is that the microenvironment that initiates the generation of tissues supporting teeth is not maintained following tooth development, thereby posing challenges in the restoration of the lost periodontal ligament after reaching maturity [6]. As an advanced technology, the in situ tissue engineering approach, employing biomaterials for controlled delivery adaptable to recapitulating this environment, is considered a promising technology for the treatment of periodontal bone defects [7]. To achieve this objective, numerous biomaterials were established, significantly influencing oral tissue.…”