Enhanced Wound Healing Potential of Primary Human Oral Fibroblasts and Periodontal Ligament Cells Cultured on Four Different Porcine-Derived Collagen Matrices

Abstract

Xenogenic collagen-based matrices represent an alternative to subepithelial palatal connective tissue autografts in periodontal and peri-implant soft tissue reconstructions. In the present study, we aimed to investigate the migratory, adhesive, proliferative, and wound-healing potential of primary human oral fibroblasts (hOF) and periodontal ligament cells (hPDL) in response to four commercially available collagen matrices. Non-crosslinked collagen matrix (NCM), crosslinked collagen matrix (CCM), dried acellular dermal matrix (DADM), and hydrated acellular dermal matrix (HADM) were all able to significantly enhance the ability of hPDL and hOF cells to directionally migrate toward the matrices as well as to efficiently repopulate an artificially generated wound gap covered by the matrices. Compared to NCM and DADM, CCM and HADM triggered stronger migratory response. Cells grown on CCM and HADM demonstrated significantly higher proliferative rates compared to cells grown on cell culture plastic, NCM, or DADM. The pro-proliferative effect of the matrices was supported by expression analysis of proliferative markers regulating cell cycle progression. Upregulated expression of genes encoding the adhesive molecules fibronectin, vinculin, CD44 antigen, and the intracellular adhesive molecule-1 was detected in hPDL and hOF cells cultured on each of the four matrices. This may be considered as a prerequisite for good adhesive properties of the four scaffolds ensuring proper cell–matrix and cell–cell interactions. Upregulated expression of genes encoding TGF-β1 and EGF growth factors as well as MMPs in cells grown on each of the four matrices provided support for their pro-proliferative and pro-migratory abilities. The expression of genes encoding the angiogenic factors FGF-2 and VEGF-A was dramatically increased in cells grown on DADM and HADM only, suggesting a good basis for accelerated vascularization of the latter. Altogether, our results support favorable influence of the investigated collagen matrices on the recruitment, attachment, and growth of cell types implicated in oral soft tissue regeneration. Among the four matrices, HADM has consistently exhibited stronger positive effects on the oral cellular behavior. Our data provide solid basis for future investigations on the clinical application of the collagen-based matrices in surgical periodontal therapy.

Positive Effects of Three-Dimensional Collagen-Based Matrices on the Behavior of Osteoprogenitors

“…On contrary, both the mesenchymal stromal and pre-osteoblastic cells showed increased proliferative rates on the CCM matrix that were comparable to the cell growth rates on the non-crosslinked HADM and superior compared with the natural NCM and DADM matrices. The impact of the four collagen-based matrices on the migratory, adhesive, and proliferative properties of ST2 and MC3T3-E1 cells, with a better pronounced pro-proliferative effect of HADM and CCM, resembled the impact of the matrices on the functionality of primary human periodontal ligament cells and oral fibroblasts investigated in a recently published study (Lin et al, 2020). This suggests that the difference in the potency of the four matrices to induce changes in the cellular behavior is not cell-specific but rather dependents on their physicochemical characteristics.…”

“…Furthermore, the matrices have exhibited sustained growth factor release over 13 days with kinetics that will likely favor the long-term tissue regeneration following surgical reconstructive periodontal therapies. We have further demonstrated that the investigated collagen-based matrices successfully promote migration, adhesion, and proliferation of cell types involved in oral soft tissue regeneration, namely primary human oral fibroblasts and periodontal ligament cells ( Lin et al, 2020 ). The current study extends the in vitro investigations on the four 3D matrices in relation to their ability to trigger osteogenic differentiation.…”

A Novel Volume-Stable Collagen Matrix Induces Changes in the Behavior of Primary Human Oral Fibroblasts, Periodontal Ligament, and Endothelial Cells

“…Primary hPDLCs and hOFs were obtained from three donors each using the tissue explant technique as described previously [ 9 ]. The collection of donor tissue from systemically and periodontally healthy individuals below 35 years had received an approval by the Ethics Committee, Berne, Switzerland (ethical code ID 2018-00661 from 13.08.2018).…”

“…RNA, quantified spectrophotometrically and reverse transcribed by using the Transcriptor First Strand cDNA Synthesis Kit (Roche, Basel, Switzerland), was subjected to qPCR using the FastStart Universal SYBR Green Master ROX (Roche) and the primer sequences listed in Table 1 . The PCR was carried out in a QuantStudio 3 instrument (Applied Biosystems, Rotkreuz, Switzerland) as described [ 9 ]. The efficiency ∆∆Ct method was used to calculate gene expression levels normalized to GAPDH and calibrated to controls.…”

“…In agreement with the inherited ability of collagen to bind a number of proteins [ 7 ], we recently showed that the transforming growth factor-β1 (TGF-β1), fibroblast growth factor-2 (FGF-2), platelet-derived growth factor-BB (PDGF-BB), growth and differentiation factor-5 (GDF-5), or the bone morphogenetic protein-2 (BMP-2) can be adsorbed to commercially available porcine-derived collagen matrices with an efficiency close to 100%, and are then continuously released over time [ 8 ]. Furthermore, the four investigated collagen scaffolds have provided a favorable environment able to successfully promote the migration, adhesion, and proliferation of two oral cell types, namely primary human oral fibroblasts (hOF) and periodontal ligament cells (hPDLC) [ 9 ]. A number of clinical studies have demonstrated the successful application of acellular collagen matrices in periodontal surgical reconstructions, with low patient morbidity [ 10 , 11 , 12 , 13 , 14 , 15 ].…”