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Bioengineering
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Tissue Engineering for Regenerative Dentistry
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Tissue Engineering for Regenerative Dentistry

A special issue of Bioengineering (ISSN 2306-5354). This special issue belongs to the section "Regenerative Engineering".

Deadline for manuscript submissions: 30 April 2025 | Viewed by 9559

Special Issue Editors

Prof. Dr. Shahram M. Ghanaati

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Guest Editor
Clinic for Maxillofacial and Plastic Surgery, Johann Wolfgang Goethe University, FORM-Lab (Frankfurt Oral Regenerative Medicine), 60596 Frankfurt Am Main, Germany
Interests: cell culture; molecular biology; cell biology; biomaterials; tissue engineering; biocompatibility; molecular pathology; scaffolds
Special Issues, Collections and Topics in MDPI journals
Dr. Carlos Fernando Mourão

E-Mail Website
Guest Editor
Department of Periodontology, Dental Research Administration, Tufts University School of Dental Medicine, Boston, MA 02111, USA
Interests: membranes/barries; blood-derived growth factors; guide bone regeneration; soft tissue regeneration; biomaterials; regenerative medicine; drug delivery systems
Special Issues, Collections and Topics in MDPI journals

Special Issue Information

Dear Colleagues,

The field of dentistry is constantly progressing and there is great promise in using autologous blood concentrates, stem cells, and biomaterials to advance tissue regeneration.

Autologous blood concentrates have the ability to improve wound healing and tissue regrowth, particularly in oral and maxillofacial surgery. This is due to their composition of growth factors and cytokines. Stem cells have potent regenerative capabilities and can mobilize the body's own reparative mechanisms for tissue restoration.

Biomaterials, such as bioactive ceramics, polymers, and composites, can fortify these biological resources and mimic the natural extracellular matrix. They promote cell attachment, growth, and differentiation, thereby laying the foundation for successful integration into the human body.

The purpose of this Special Issue is to showcase the potential of this powerful combination for tissue engineering in dentistry. The goal is to highlight innovative research developments in bioengineering and tissue regeneration. Ultimately, dentistry can shift from traditional treatments to a more regenerative model, which will enhance the future of dental care and improve the quality of life for patients.

Prof. Dr. Shahram Ghanaati
Prof. Dr. Carlos Fernando Mourão
Guest Editors

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Published Papers (7 papers)

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12 pages, 3234 KiB  
Article
PRF Lysates Modulate Chemokine Expression in Oral Squamous Carcinoma and Healthy Epithelial Cells
by Zohreh Afradi, Layla Panahipour, Salman Abbas Zadeh and Reinhard Gruber
Bioengineering 2024, 11(8), 746; https://doi.org/10.3390/bioengineering11080746 - 24 Jul 2024
Viewed by 958
Abstract
Platelet-rich fibrin (PRF), originally used to support soft tissue healing, is also considered a therapeutic option for treating oral lichen planus and leukoplakia. The progression from the two premalignant lesions to the aggressive malignant oral squamous cell carcinoma involves an inflammatory process linked [...] Read more.
Platelet-rich fibrin (PRF), originally used to support soft tissue healing, is also considered a therapeutic option for treating oral lichen planus and leukoplakia. The progression from the two premalignant lesions to the aggressive malignant oral squamous cell carcinoma involves an inflammatory process linked to chemokine expression. Thus, there is a rationale for studying how PRF modulates the expression of chemokines in oral squamous carcinoma cells. To this aim, we expose the oral squamous carcinoma cell line HSC2 to IL1β and TNFα either alone or in the presence of lysates obtained from solid PRF membranes. We report here that in HSC2 cells, PRF lysates significantly reduce the forced transcription of chemokines, e.g., CXCL1, CXCL2, CXCL8, CXCL10, and CCL5. Moreover, PRF lysates attenuate the nuclear translocation of p65 in HSC2 oral epithelial cells when exposed to IL1β and TNFα. PRF lysates further reduce chemokine expression provoked by poly:IC HMW. Even though less pronounced, PRF lysates reduce IL1β- and TNFα-induced chemokine expression in TR146 cells. In primary oral epithelial cells, however, PRF lysates increase the basal expression of CXCL1, CXCL2 and CXCL8. Thus, PRF can exert a biphasic effect on chemokine expression in oral squamous cell carcinoma cell lines and primary oral epithelial cells. These findings suggest that PRF may reduce inflammation in a malignant environment while provoking an immunological response in healthy oral epithelium. Full article
(This article belongs to the Special Issue Tissue Engineering for Regenerative Dentistry)
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13 pages, 1583 KiB  
Article
In Vitro Bioassay for Damage-Associated Molecular Patterns Arising from Injured Oral Cells
by Layla Panahipour, Chiara Micucci, Benedetta Gelmetti and Reinhard Gruber
Bioengineering 2024, 11(7), 687; https://doi.org/10.3390/bioengineering11070687 - 5 Jul 2024
Cited by 1 | Viewed by 858
Abstract
Gingival fibroblasts are a significant source of paracrine signals required to maintain periodontal homeostasis and to mediate pathological events linked to periodontitis and oral squamous cell carcinomas. Among the potential paracrine signals are stanniocalcin-1 (STC1), involved in oxidative stress and cellular survival; amphiregulin [...] Read more.
Gingival fibroblasts are a significant source of paracrine signals required to maintain periodontal homeostasis and to mediate pathological events linked to periodontitis and oral squamous cell carcinomas. Among the potential paracrine signals are stanniocalcin-1 (STC1), involved in oxidative stress and cellular survival; amphiregulin (AREG), a growth factor that mediates the cross-talk between immune cells and epithelial cells; chromosome 11 open reading frame 96 (C11orf96) with an unclear biologic function; and the inflammation-associated prostaglandin E synthase (PTGES). Gingival fibroblasts increasingly express these genes in response to bone allografts containing remnants of injured cells. Thus, the gene expression might be caused by the local release of damage-associated molecular patterns arising from injured cells. The aim of this study is consequently to use the established gene panel as a bioassay to measure the damage-associated activity of oral cell lysates. To this aim, we have exposed gingival fibroblasts to lysates prepared from the squamous carcinoma cell lines TR146 and HSC2, oral epithelial cells, and gingival fibroblasts. We report here that all lysates significantly increased the transcription of the entire gene panel, supported for STC1 at the protein level. Blocking TGF-β receptor 1 kinase with SB431542 only partially reduced the forced expression of STC1, AREG, and C11orf96. SB431542 even increased the PTGES expression. Together, these findings suggest that the damage signals originating from oral cells can change the paracrine activity of gingival fibroblasts. Moreover, the expression panel of genes can serve as a bioassay for testing the biocompatibility of materials for oral application. Full article
(This article belongs to the Special Issue Tissue Engineering for Regenerative Dentistry)
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11 pages, 1527 KiB  
Article
Biomimetic Remineralization of Artificial Caries Lesions with a Calcium Coacervate, Its Components and Self-Assembling Peptide P11-4 In Vitro
by Basel Kharbot, Haitham Askar, Dominik Gruber and Sebastian Paris
Bioengineering 2024, 11(5), 465; https://doi.org/10.3390/bioengineering11050465 - 8 May 2024
Cited by 1 | Viewed by 1400
Abstract
The application of calcium coacervates (CCs) may hold promise for dental hard tissue remineralization. The aim of this study was to evaluate the effect of the infiltration of artificial enamel lesions with a CC and its single components including polyacrylic acid (PAA) compared [...] Read more.
The application of calcium coacervates (CCs) may hold promise for dental hard tissue remineralization. The aim of this study was to evaluate the effect of the infiltration of artificial enamel lesions with a CC and its single components including polyacrylic acid (PAA) compared to that of the self-assembling peptide P11-4 in a pH-cycling (pHC) model. Enamel specimens were prepared from bovine incisors, partly varnished, and stored in demineralizing solution (DS; pH 4.95; 17 d) to create two enamel lesions per sample. The specimens were randomly allocated to six groups (n = 15). While one lesion per specimen served as the no-treatment control (NTC), another lesion (treatment, T) was etched (H3PO4, 5 s), air-dried and subsequently infiltrated for 10 min with either a CC (10 mg/mL PAA, 50 mM CaCl2 (Ca) and 1 M K2HPO4 (PO4)) (groups CC and CC + DS) or its components PAA, Ca or PO4. As a commercial control, the self-assembling peptide P11-4 (CurodontTM Repair, Credentis, Switzerland) was tested. The specimens were cut perpendicularly to the lesions, with half serving as the baseline (BL) while the other half was exposed to either a demineralization solution for 20 d (pH 4.95; group CC + DS) or pHC for 28 d (pH 4.95, 3 h; pH 7, 21 h; all five of the other groups). The difference in integrated mineral loss between the lesions at BL and after the DS or pHC, respectively, was analyzed using transversal microradiography (ΔΔZ = ΔZpHC − ΔZbaseline). Compared to the NTC, the mineral gain in the T group was significantly higher in the CC + DS, CC and PAA (p < 0.05, Wilcoxon). In all of the other groups, no significant differences between treated and untreated lesions were detected (p > 0.05). Infiltration with the CC and PAA resulted in a consistent mineral gain throughout the lesion body. The CC as well as its component PAA alone promoted the remineralization of artificial caries lesions in the tested pHC model. Infiltration with PAA further resulted in mineral gain in deeper areas of the lesion body. Full article
(This article belongs to the Special Issue Tissue Engineering for Regenerative Dentistry)
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11 pages, 7515 KiB  
Article
Human versus Rat PRF on Collagen Membranes: A Pilot Study of Mineralization in Rat Calvaria Defect Model
by Karol Ali Apaza Alccayhuaman, Patrick Heimel, Stefan Tangl, Stefan Lettner, Carina Kampleitner, Layla Panahipour, Ulrike Kuchler and Reinhard Gruber
Bioengineering 2024, 11(5), 414; https://doi.org/10.3390/bioengineering11050414 - 23 Apr 2024
Viewed by 1386
Abstract
Platelet-rich fibrin, the coagulated plasma fraction of blood, is commonly used to support natural healing in clinical applications. The rat calvaria defect is a standardized model to study bone regeneration. It remains, however, unclear if the rat calvaria defect is appropriate to investigate [...] Read more.
Platelet-rich fibrin, the coagulated plasma fraction of blood, is commonly used to support natural healing in clinical applications. The rat calvaria defect is a standardized model to study bone regeneration. It remains, however, unclear if the rat calvaria defect is appropriate to investigate the impact of human PRF (Platelet-Rich Fibrin) on bone regeneration. To this end, we soaked Bio-Gide® collagen membranes in human or rat liquid concentrated PRF before placing them onto 5 mm calvarial defects in Sprague Dawley rats. Three weeks later, histology and micro-computed tomography (μCT) were performed. We observed that the collagen membranes soaked with rat PRF show the characteristic features of new bone and areas of mineralized collagen matrix, indicated by a median mineralized volume of 1.5 mm3 (range: 0.9; 5.3 mm3). Histology revealed new bone growing underneath the membrane and hybrid bone where collagen fibers are embedded in the new bone. Moreover, areas of passive mineralization were observed. The collagen membranes soaked with human PRF, however, were devoid of histological features of new bone formation in the center of the defect; only occasionally, new bone formed at the defect margins. Human PRF (h-PRF) caused a median bone volume of 0.9 mm3 (range: 0.3–3.3 mm3), which was significantly lower than what was observed with rat PRF (r-PRF), with a BV median of 1.2 mm3 (range: 0.3–5.9 mm3). Our findings indicate that the rat calvaria defect model is suitable for assessing the effects of rat PRF on bone formation, but caution is warranted when extrapolating conclusions regarding the efficacy of human PRF. Full article
(This article belongs to the Special Issue Tissue Engineering for Regenerative Dentistry)
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18 pages, 7997 KiB  
Article
Three Milliliters of Peripheral Blood Is Sufficient for Preparing Liquid Platelet-Rich Fibrin (PRF): An In Vitro Study
by Sarah Al-Maawi, Eva Dohle, Robert Sader and Shahram Ghanaati
Bioengineering 2024, 11(3), 253; https://doi.org/10.3390/bioengineering11030253 - 4 Mar 2024
Viewed by 1252
Abstract
Platelet-rich fibrin (PRF) has assumed an important role in supporting tissue regeneration in different fields. To date, the standard protocol for liquid PRF requires at least 10 mL of peripheral blood. The present study aimed to analyze the composition, growth factor release, and [...] Read more.
Platelet-rich fibrin (PRF) has assumed an important role in supporting tissue regeneration in different fields. To date, the standard protocol for liquid PRF requires at least 10 mL of peripheral blood. The present study aimed to analyze the composition, growth factor release, and effects on the cell proliferation of PRF samples produced using 3 mL vs. 10 mL of peripheral blood in vitro. Peripheral venous blood from six healthy donors was used to prepare liquid PRF using either 3 mL or 10 mL tubes. Three different centrifugation protocols were used according to the low-speed centrifugation concept. The cellular distribution was evaluated using immunohistology and automated cell count. ELISA was used to determine the release of different growth factors (EGF, TGF-β1, and PDGF) and interleukin 8 at different time points. Primary human osteoblasts (pOBs) were cultivated for 7 days using PRF-conditioned media acquired from either 3 mL or 10 mL of peripheral blood. The results showed that 3 mL of peripheral blood is sufficient to produce a liquid PRF concentrate similar to that acquired when using 10 mL blood. The concentrations of platelets and leukocytes were comparable regardless of the initial blood volume (3 mL vs. 10 mL). Similarly, the release of growth factors (EGF, TGF-β1, and PDGF) and interleukin 8 was often comparable in both groups over 7 days. The cultivation of pOBs using PRF-conditioned media showed a similar proliferation rate regardless of the initial blood volume. This proliferation rate was also similar to that of pOBs treated with 20% FBS-conditioned media. These findings validated the use of 3 mL of peripheral blood to generate liquid PRF matrices according to the low-speed centrifugation concept, which may open new application fields for research purposes such as in vivo experiments and clinical applications such as pediatric surgery. Full article
(This article belongs to the Special Issue Tissue Engineering for Regenerative Dentistry)
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12 pages, 2209 KiB  
Article
Platelet-Rich Fibrin-Conditioned Medium as an Alternative to Fetal Bovine Serum Promotes Osteogenesis of Human Dental Pulp Stem Cells
by Ayano Hatori, Daiki Yamakawa, Sarah Al-Maawi, Eva Dohle, Jin Chikira, Yasuyuki Fujii, Megumu Miki, Robert Sader, Daichi Chikazu, Shahram Ghanaati and Yoko Kawase-Koga
Bioengineering 2023, 10(10), 1196; https://doi.org/10.3390/bioengineering10101196 - 14 Oct 2023
Cited by 1 | Viewed by 1747
Abstract
Human dental pulp stem cells (DPSCs) exhibit multilineage differentiation capabilities and superior clonogenic and proliferative properties. However, the use of animal-derived components such as FBS raises concerns regarding the clinical application of stem-cell-based therapies. Platelet-rich fibrin (PRF) derived from human blood is rich [...] Read more.
Human dental pulp stem cells (DPSCs) exhibit multilineage differentiation capabilities and superior clonogenic and proliferative properties. However, the use of animal-derived components such as FBS raises concerns regarding the clinical application of stem-cell-based therapies. Platelet-rich fibrin (PRF) derived from human blood is rich in fibrin, platelets, and growth factors and acts as a bioactive scaffold for grafting with biomaterials. In this study, we assessed the efficacy of PRF-conditioned medium (CM) in promoting DPSCs proliferation and osteogenic differentiation compared with the standard culture medium supplemented with FBS. A comparison of DPSCs cultured in FBS and PRF-CM revealed no differences in characteristics or morphology. However, cells cultured with PRF-CM exhibited inferior proliferation rates and cell numbers during passage in comparison with those cultured with FBS. In contrast, DPSCs cultured in PRF-CM showed significantly higher levels of calcification, and RT-PCR confirmed that the gene expression levels of markers associated with osteoblast differentiation were significantly increased. The PRF-CM approach offers a convenient, straightforward, and advantageous method for culturing DPSCs, without relying on animal-derived components. In summary, this study introduces a novel application of PRF-CM for enhancing the osteogenesis of DPSCs, which provides an alternative to FBS culture medium and addresses concerns associated with the use of animal-derived components in clinical settings. Full article
(This article belongs to the Special Issue Tissue Engineering for Regenerative Dentistry)
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16 pages, 2530 KiB  
Systematic Review
Changes of the Alveolar Bone Ridge Using Bone Mineral Grafts and Collagen Membranes after Tooth Extraction: A Systematic Review and Meta-Analysis
by Nansi López-Valverde, Bruno Macedo de Sousa and José Antonio Blanco Rueda
Bioengineering 2024, 11(6), 565; https://doi.org/10.3390/bioengineering11060565 - 3 Jun 2024
Cited by 1 | Viewed by 1151
Abstract
Background: Alveolar preservation techniques for esthetic or functional purposes, or both, are a frequently used alternative for the treatment of post-extraction sockets, the aim of which is the regeneration of the lesion and the preservation of the alveolar bone crest. Methods: Studies published [...] Read more.
Background: Alveolar preservation techniques for esthetic or functional purposes, or both, are a frequently used alternative for the treatment of post-extraction sockets, the aim of which is the regeneration of the lesion and the preservation of the alveolar bone crest. Methods: Studies published in PubMed (Medline), Web of Science, Embase, and Cochrane Library databases up to January 2024 were consulted. Inclusion criteria were established as intervention studies, according to the PICOs strategy: adult subjects undergoing dental extractions (participants), with alveoli treated with bone mineral grafts and collagen membranes (intervention), compared to spontaneous healing (comparison), and observing the response to treatment in clinical and radiological measures of the alveolar bone crest (outcomes). Results: We obtained 561 results and selected 12 studies. Risk of bias was assessed using the Cochrane Risk of Bias Tool, and methodological quality was assessed using the Joanna Briggs Institute. Due to the high heterogeneity of the studies (I2 > 75%), a random-effects meta-analysis was used. Despite the trend, no statistical significance (p > 0.05) was found in the experimental groups. Conclusions: The use of bone mineral grafts in combination with resorbable collagen barriers provides greater preservation of the alveolar ridge, although more clinical studies are needed. Full article
(This article belongs to the Special Issue Tissue Engineering for Regenerative Dentistry)
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