The Association Between Buccal Mucosa Thickness and Periimplant Bone Loss and Attachment Loss: A Cross-Sectional Study.

PURPOSE: The aim of this study was to assess if there is an association between buccal mucosa thickness and periimplant attachment loss after 1 year of function. MATERIALS AND METHODS: A total of 28 patients (14 periimplantitis implants and 14 healthy implants) were included. The buccal mucosal thickness was assessed using K-files at 3 mm apical to the soft tissue margin of the implant. Probing depth, recession (REC), clinical attachment level (CAL), bleeding on probing, and radiographic bone loss on mesial and distal sites of the implant were recorded. RESULTS: The data showed that there was a statistically significant difference in midfacial REC between thin and thick buccal mucosa groups. However, the CAL was not statistically significant different between both groups. In addition, there was no statistically significant difference in mesial and distal bone loss between implants with thin and thick mucosa. CONCLUSION: When the midfacial soft tissue thickness was thin, the midfacial REC was greater and the CAL also tended to be higher. There was no association between buccal mucosa thickness and periimplant bone loss on mesial and distal sites of the implant after 1 year of function.

Comparative analysis of dental implant treatment outcomes following mandibular reconstruction with double-barrel fibula bone grafting or vertical distraction osteogenesis fibula: a retrospective study.

PURPOSE: The purpose of this study was twofold: (i) to compare vertical bone height (VBH) after tumor resection through grafting with either a double-barrel fibula (DBF) technique or vertical distraction osteogenesis of the fibula (VDOF); (ii) to compare the performance of loaded dental implants following either DBF or VDOF with special focus on implant survival, implant success, and bone resorption. MATERIALS AND METHODS: This retrospective clinical study involved 19 patients who underwent implant placement following DBF (group A, n = 9) or VDOF (group B, n = 10) for mandibular reconstruction from March 2006 to May 2008. Clinical and radiographic assessments, including VBH, modified Plaque Index (mPI), modified Sulcus Bleeding Index (mSBI), and marginal bone level (MBL), were taken for both groups after delivery of the final prostheses and annually thereafter. RESULTS: Nine patients underwent DBF with 24 implants placed and 10 patients underwent VDOF with 27 implants placed for mandibular reconstruction after tumor resection. Overall, all DBF and VDOF procedures were successful for group A and group B. VBH for group A and group B were 20 and 17 mm. There was no statistically significant difference of mSBI scores between group A and group B in the 3-year follow-up (P = 0.40). In four cases with eight implants of group A and two cases with three implants of group B, granulomatous soft tissue grew. There was no statistically significant differences of MBL between group A and group B in the 3-year follow-up (p = 0.736). The cumulative survival and success rates of implants for group A were 100% and 87.5%, and for group B were 100% and 85.2% in 3-year follow-up, respectively. CONCLUSIONS: On the basis of the study of 19 patients who received a total of 51 implants, reconstruction of the mandible with DBF flap or VDOF flap, combined with dental implant therapy, was considered a predictable option. Compared with implants placed in VDOF bone, implants placed in DBF bone had a relative higher incidence of associated gingival inflammation. The DBF bone seems more resistant to peri-implant resorption processes than VDOF bone during functional loading.

Implant success remains high despite grafting voids in the maxillary sinus.

OBJECTIVES: Given that the nature and presence of voids present within grafted sinuses following maxillary sinus elevation procedures were not known, nor was the contribution of these factors to implant success, the purpose of this study was to investigate these parameters and their relationship to implant success. MATERIALS AND METHODS: This study evaluated data from 25 subjects who had a lateral window maxillary sinus augmentation procedure. Cone-beam computed tomography (CBCT) was performed at baseline and 4 months after surgery. CBCT images were used to evaluate grafted sites prior to implant placement. Using CBCT images, three examiners independently measured bone-grafted areas (BG), void areas (V), and percentage of void areas (V%) from six different sections within grafted sites. The six sections were defined as a cross-sectional (CS) midpoint, CS mesial point, CS distal point, horizontal section (HS) low point, HS midpoint, and HS high point. Implant success was also determined. RESULTS: The calculated V% (V/BG) for the CS midpoint, CS mesial point, CS distal point, HS low point, HS midpoint, and HS high point were 5.30 ± 6.67%, 5.79 ± 8.51%, 6.67 ± 7.12%, 2.07 ± 2.56%, 5.30 ± 6.62%, and 4.92 ± 5.17% respectively. Implant success after 6 months of follow-up approximated 100%. CONCLUSIONS: Although voids within grafts varied in terms of distribution and size, the V% within the HS low point were significantly smaller compared to those within the CS midpoint and CS distal point, which had the most intra-subject V%. Thus, more attention should be given to the distal aspect of the sinus when compacting graft materials in the lateral wall sinus augmentation procedure. Implant success was not influenced by the existence of voids as implant success remained high.

Periodontists and stem cell-based therapy for alveolar bone regeneration: A national survey.

BACKGROUND: Stem cell-based therapy for bone regeneration has received attention in medical settings but has not yet been used in clinical practice for treating alveolar bone defects. The objectives of this study were to explore whether periodontists had heard about this approach, and if so how, how interested they were to learn about it, which attitudes and behavioral intentions they had related to using stem cell-based grafting, and what they would like to know before using this approach. METHODS: Anonymous survey data were collected from 481 members of the American Academy of Periodontology (response rate: 19.41%). RESULTS: Responses showed 35.3% had heard about stem cell-based therapy, mostly from publications (9.6%) and meetings (8.3%); 76.1% wanted to learn about it through in-person continuing education (CE) courses, 68.6% in online CE courses, and 57.1% from manuals; 73% considered this approach promising; and 54.9% preferred it to traditional approaches. It was important to them that it would result in more bone volume (93%), better bone quality (90.4%), and accelerated healing (83.2%). Also, 60.1% considered it likely/very likely that they would adopt this approach, 54% that patients would prefer it, and 62.1% that it would benefit their practice. When asked what they would like to know about this approach, information about short- and long-term outcomes, cost, and logistical considerations were most frequently named. CONCLUSIONS: These findings provide the basis to develop educational interventions for periodontists about this novel approach and inform future research activities aimed to translate this approach to clinical practice.

Immunomodulatory differences between mesenchymal stem cells from different oral tissues.

Mesenchymal stem cells (MSCs) have recently been identified as having potentially therapeutic immunomodulatory properties. MSCs isolated from different oral tissues have similar morphology and immunophenotypes, however, direct comparisons of their gene expression and immunomodulatory properties have not been conducted. We isolated alveolar bone-derived MSCs (aBMSCs), dental pulp stem cells (DPSCs) and gingiva-derived MSCs (GMSCs) from the same patients and compared their immunophenotypes and transcriptomes. Additionally, we compared their production of soluble immunomodulatory cytokines as well as their immunoregulatory properties in coculture with THP-1 human monocytic cells. RNA sequencing revealed distinct gene expression in DPSCs while aBMSCs and GMSCs had less differentially expressed genes. DPSCs also had significantly less secretion of osteopontin compared to aBMSCs and GMSCs. Finally, DPSCs did not exhibit an immunosuppresive effect on THP-1 cells to the same degree as aBMSCs and GMSCs. These findings demonstrate that MSCs from different oral tissues have distinct transcriptomes and immunoregulatory properties.

The role of fibroblast growth factor-2 in modulating the differentiation of periodontal ligament and alveolar bone-derived stem cells.

OBJECTIVE: This study examined how range concentrations of Fibroblast Growth Factor-2 (FGF-2) influence the differentiation and activity of human-derived periodontal ligament (hPDLSCs) and alveolar bone-derived stem cells (haBMSCs). DESIGN: hPDLSCs and haBMSCs were cultured with varying concentrations of FGF-2 (0, 1, 2.5, 5, 10, 20 ng/mL) and monitored for osteogenic differentiation through alkaline phosphatase (ALP) activity and quantification of gene expression (qRT-PCR) for osteogenesis markers. Additionally, alizarin red staining and a hydroxyproline colorimetric assay evaluated and quantified osteogenic matrix mineralization and collagen deposition. Statistical analyses were performed using one-way ANOVA or two-way ANOVA for multiple comparisons between groups. RESULTS: At low FGF-2 concentrations, hPDLSCs differentiated toward an osteogenic lineage, whereas higher concentrations of FGF-2 inhibited osteogenesis and promoted fibroblastic differentiation. The effect of FGF-2 at the lowest concentration tested (1 ng/mL) led to significantly higher ALP activity than osteogenically induced positive controls at early time points and equivalent RUNX2 expression at early and later time points. FGF-2 supplementation of haBMSC cultures was sufficient, at all concentrations, to increase ALP activity at an earlier time point. Mineralization of haBMSC cultures increased significantly within 5-20 ng/mL FGF-2 concentrations under basal growth media conditions (α-minimal essential medium supplemented with 15 % fetal bovine serum and 1 % penicillin/streptomycin). CONCLUSIONS: FGF-2 has a dual capacity in promoting osteogenic and fibroblastic differentiation within hPDLSCs contingent upon the dosage and timing of administration, alongside supporting osteogenic differentiation in haBMSCs. These findings underscore the need for precision growth factors dosing when considering the design of biomaterials for periodontal regeneration.

Personalized bioceramic grafts for craniomaxillofacial bone regeneration.

The reconstruction of craniomaxillofacial bone defects remains clinically challenging. To date, autogenous grafts are considered the gold standard but present critical drawbacks. These shortcomings have driven recent research on craniomaxillofacial bone reconstruction to focus on synthetic grafts with distinct materials and fabrication techniques. Among the various fabrication methods, additive manufacturing (AM) has shown significant clinical potential. AM technologies build three-dimensional (3D) objects with personalized geometry customizable from a computer-aided design. These layer-by-layer 3D biomaterial structures can support bone formation by guiding cell migration/proliferation, osteogenesis, and angiogenesis. Additionally, these structures can be engineered to degrade concomitantly with the new bone tissue formation, making them ideal as synthetic grafts. This review delves into the key advances of bioceramic grafts/scaffolds obtained by 3D printing for personalized craniomaxillofacial bone reconstruction. In this regard, clinically relevant topics such as ceramic-based biomaterials, graft/scaffold characteristics (macro/micro-features), material extrusion-based 3D printing, and the step-by-step workflow to engineer personalized bioceramic grafts are discussed. Importantly, in vitro models are highlighted in conjunction with a thorough examination of the signaling pathways reported when investigating these bioceramics and their effect on cellular response/behavior. Lastly, we summarize the clinical potential and translation opportunities of personalized bioceramics for craniomaxillofacial bone regeneration.

Amino Acid-Based Poly(ester urea) Biodegradable Membrane for Guided Bone Regeneration.

Barrier membranes (BM) for guided bone regeneration (GBR) aim to support the osteogenic healing process of a defined bony defect by excluding epithelial (gingival) ingrowth and enabling osteoprogenitor and stem cells to proliferate and differentiate into bone tissue. Currently, the most widely used membranes for these approaches are collagen-derived, and there is a discrepancy in defining the optimal collagen membrane in terms of biocompatibility, strength, and degradation rates. Motivated by these clinical observations, we designed a collagen-free membrane based on l-valine-co-l-phenylalanine-poly(ester urea) (PEU) copolymer via electrospinning. Degradation and mechanical properties of these membranes were performed on as-spun and water-aged samples. Alveolar-bone-derived stem cells (AvBMSCs) were seeded on the PEU BM to assess their cell compatibility and osteogenic characteristics, including cell viability, attachment/spreading, proliferation, and mineralized tissue-associated gene expression. In vivo, PEU BMs were subcutaneously implanted in rats to evaluate their potential to cause inflammatory responses and facilitate angiogenesis. Finally, critical-size calvarial defects and a periodontal model were used to assess the regenerative capacity of the electrospun PEU BM compared to clinically available Cytoflex synthetic membranes. PEU BM demonstrated equal biocompatibility to Cytoflex with superior mechanical performance in strength and elasticity. Additionally, after 14 days, PEU BM exhibited a higher expression of BGLAP/osteocalcin and superior in vivo performance-less inflammation and increased CD31 and VWF expression over time. When placed in critical-sized defects in the calvaria of rats, the PEU BM led to robust bone formation with high expression of osteogenesis and angiogenesis markers. Moreover, our membrane enhanced alveolar bone and cementum regeneration in an established periodontal model after 8 weeks. We demonstrate that the PEU BM exhibits favorable clinical properties, including mechanical stability, cytocompatibility, and facilitated bone formation in vitro and in vivo. This highlights its suitability for GBR in periodontal and craniofacial bone defects.

Tissue-specific melt electrowritten polymeric scaffolds for coordinated regeneration of soft and hard periodontal tissues.

Periodontitis is a chronic inflammatory condition that often causes serious damage to tooth-supporting tissues. The limited successful outcomes of clinically available approaches underscore the need for therapeutics that cannot only provide structural guidance to cells but can also modulate the local immune response. Here, three-dimensional melt electrowritten (i.e., poly(ε-caprolactone)) scaffolds with tissue-specific attributes were engineered to guide differentiation of human-derived periodontal ligament stem cells (hPDLSCs) and mediate macrophage polarization. The investigated tissue-specific scaffold attributes comprised fiber morphology (aligned vs. random) and highly-ordered architectures with distinct strand spacings (small 250 µm and large 500 µm). Macrophages exhibited an elongated morphology in aligned and highly-ordered scaffolds, while maintaining their round-shape on randomly-oriented fibrous scaffolds. Expressions of periostin and IL-10 were more pronounced on the aligned and highly-ordered scaffolds. While hPDLSCs on the scaffolds with 500 µm strand spacing show higher expression of osteogenic marker (Runx2) over 21 days, cells on randomly-oriented fibrous scaffolds showed upregulation of M1 markers. In an orthotopic mandibular fenestration defect model, findings revealed that the tissue-specific scaffolds (i.e., aligned fibers for periodontal ligament and highly-ordered 500 µm strand spacing fluorinated calcium phosphate [F/CaP]-coated fibers for bone) could enhance the mimicking of regeneration of natural periodontal tissues.

Nanoscale ß-TCP-Laden GelMA/PCL Composite Membrane for Guided Bone Regeneration.

Major advances in the field of periodontal tissue engineering have favored the fabrication of biodegradable membranes with tunable physical and biological properties for guided bone regeneration (GBR). Herein, we engineered innovative nanoscale beta-tricalcium phosphate (ß-TCP)-laden gelatin methacryloyl/polycaprolactone (GelMA/PCL-TCP) photocrosslinkable composite fibrous membranes via electrospinning. Chemo-morphological findings showed that the composite microfibers had a uniform porous network and ß-TCP particles successfully integrated within the fibers. Compared with pure PCL and GelMA/PCL, GelMA/PCL-TCP membranes led to increased cell attachment, proliferation, mineralization, and osteogenic gene expression in alveolar bone-derived mesenchymal stem cells (aBMSCs). Moreover, our GelMA/PCL-TCP membrane was able to promote robust bone regeneration in rat calvarial critical-size defects, showing remarkable osteogenesis compared to PCL and GelMA/PCL groups. Altogether, the GelMA/PCL-TCP composite fibrous membrane promoted osteogenic differentiation of aBMSCs in vitro and pronounced bone formation in vivo. Our data confirmed that the electrospun GelMA/PCL-TCP composite has a strong potential as a promising membrane for guided bone regeneration.

Dual Digitally Guided Crown Lengthening in Esthetic Area Compromised by Disharmonic Implant Crown.

INTRODUCTION: A disharmonious smile results from excessive gingival exposure or gingival margin misalignment is a frequent finding in patients. The most common cause is altered passive eruption; however, in the case presented here, the esthetics of the smile is compromised also due to an inadequate implant placement and crown fabrication. CASE PRESENTATION: This case presented a combination of altered passive eruption and a buccally as well as deeply placed implant crown (#10) that led to disproportionate dimensions of an upper lateral incisor. Dual digitally guided crown lengthening surgical procedure for teeth #5 to #12 was performed aiming a better harmony of the smile. Treatment also included placement of tooth veneers and re-made of implant crown. A pleasant smile with appropriate colors and proportions was achieved. All esthetic and periodontal health parameters were maintained throughout the follow-up period (1 year). CONCLUSION: The use of dual digitally guided crown lengthening help to accomplish precision and stability of esthetic outcome, as it guides for both, bone and soft tissue removal. Particularly, in challenging cases with high esthetic demand and complicated factors present, such as the case presented here, the digital approach provides tools to attain excellent treatment outcome.

Three-dimensional printing of clinical scale and personalized calcium phosphate scaffolds for alveolar bone reconstruction.

OBJECTIVE: Alveolar bone defects can be highly variable in their morphology and, as the defect size increases, they become more challenging to treat with currently available therapeutics and biomaterials. This investigation sought to devise a protocol for fabricating customized clinical scale and patient-specific, bioceramic scaffolds for reconstruction of large alveolar bone defects. METHODS: Two types of calcium phosphate (CaP)-based bioceramic scaffolds (alginate/ß-TCP and hydroxyapatite/α-TCP, hereafter referred to as hybrid CaP and Osteoink™, respectively) were designed, 3D printed, and their biocompatibility with alveolar bone marrow stem cells and mechanical properties were determined. Following scaffold optimization, a workflow was developed to use cone beam computed tomographic (CBCT) imaging to design and 3D print, defect-specific bioceramic scaffolds for clinical-scale bone defects. RESULTS: Osteoink™ scaffolds had the highest compressive strength when compared to hybrid CaP with different infill orientation. In cell culture medium, hybrid CaP degradation resulted in decreased pH (6.3) and toxicity to stem cells; however, OsteoInk™ scaffolds maintained a stable pH (7.2) in culture and passed the ISO standard for cytotoxicity. Finally, a clinically feasible laboratory workflow was developed and evaluated using CBCT imaging to engineer customized and defect-specific CaP scaffolds using OsteoInk™. It was determined that printed scaffolds had a high degree of accuracy to fit the respective clinical defects for which they were designed (0.27 mm morphological deviation of printed scaffolds from digital design). SIGNIFICANCE: From patient to patient, large alveolar bone defects are difficult to treat due to high variability in their complex morphologies and architecture. Our findings shows that Osteoink™ is a biocompatible material for 3D printing of clinically acceptable, patient-specific scaffolds with precision-fit for use in alveolar bone reconstructive procedures. Collectively, emerging digital technologies including CBCT imaging, 3D surgical planning, and (bio)printing can be integrated to address this unmet clinical challenge.

Periodontal Tissue Bioengineering: Is the Future Now?

Periodontitis affects nearly half of the adult population in the United States and leads to periodontium destruction, tooth loss, and tooth mobility. Novel bioengineering has become an area of interest in dentistry, as various approaches aim to regenerate attachment apparatus around diseased teeth with the use of barriers, scaffolds, bone grafts, or biologics. This article emphasizes recent findings in the fields of stem cell/gene therapy, 3-dimensional printing, and innovative scaffold designs for future applications in clinical care.

Advanced 3D Models Cultured to Investigate Mesenchymal Stromal Cells of the Human Dental Follicle.

The human dental follicle (hDF) contains the developing tooth and is involved in regulating tooth maturation and eruption. To investigate the mesenchymal stromal cells of the dental follicle, 2 three-dimensional (3D) culture models were used, based on a dynamic bioreactor: the Rotary Cell Culture System (RCCS™) and the 3D culture of precursor cells isolated from follicular tissue (human dental follicle cells [hDFCs]). The hDFCs were obtained from impacted third molars of 20 patients. Two 3D culture models were tested. In the first model, intact hDF explants were cultured in 3D conditions, preserving the original tissue architecture; they were studied using histomorphological and molecular analyses. The second model involved the 3D culture of hDFCs, which were characterized to evaluate their multipotency in terms of differentiation capability. Of the biomarkers known to characterize hDFCs, hDF precursors were selected for our study. The immunophenotype and in situ immunocytochemistry were evaluated for markers CD44, CD90, CD146, CD105, CD31, CD34, and CD45 Ag. The results show that the conditions provided by the RCCS preserve the original organizational architecture of the cells. The 3D conditions of the model enhanced differentiation in response to adipogenic, osteogenic, and chondrogenic inductive growth media. The immunophenotype and the immunocytochemistry showed generally high expression of CD90, CD44, and CD105, while CD146 expression was more restricted to ∼30% of cells. No expression was observed for CD31, CD34, and CD45 Ags. Two 3D tissue- and cell-based ex vivo models of the hDF supported the long-term maintenance of hDF-specific cell phenotypes and their ability to recapitulate typical cellular differentiation states. As such, these ex vivo models could be used to study the physiopathology of human odontogenesis. In addition, in a therapeutic context, they could be used to examine the role of specific chemical signals (e.g., new therapeutic agents) in the processes of dental tissue repair and regeneration.

Stem cell therapy for reconstruction of alveolar cleft and trauma defects in adults: A randomized controlled, clinical trial.

BACKGROUND: Stem cell therapy with bone marrow-derived mesenchymal stem cells is a promising tissue engineering strategy to promote regeneration of craniofacial bone. PURPOSE: To determine whether cell therapy with ex vivo expanded stem cell populations would be safe and efficacious in the regeneration of large alveolar defects in patients with a history of cleft palate or craniofacial trauma. MATERIALS AND METHODS: Eighteen patients (10 patients with traumatic injury and 8 patients with cleft palate) presenting with missing teeth associated with horizontal alveolar bone deficiencies were included in this randomized controlled clinical trial. Patients were randomized to receive either conventional autogenous block grafts or stem cell therapy. After a healing period of 4 months the treated sites were re-entered and the bone width re-assessed prior to implant placement. Implant stability was evaluated through torque testing of the implant upon insertion and at 6 months postloading. RESULTS: The mean gain in bone width was 1.5 ± 1.5 mm in the stem cell therapy group and 3.3 ± 1.4 mm in the control group. Overall, bone gain was higher in trauma patients as compared to patients with cleft palate, for both the control and the stem cell therapy groups. Most postoperative complications were wound dehiscences and incision line openings. Implants were placed successfully in 5 out of 10 patients in the stem cell therapy group and in all 8 patients in the control group. One implant from the control/cleft palate group failed before loading, while the rest of the implants were loaded successfully and remained stable at 6 months. The patients who did not receive implants were re-treated with autogenous block bone graft. CONCLUSION: The ability of stem cells to treat large alveolar defects is safe, yet, their ability to completely reconstitute large alveolar defects is limited. This approach requires further optimization to meet the outcomes seen using current methods to treat large defects, particularly those resultant of cleft palate.

Dental Pulp Tissue Regeneration Using Dental Pulp Stem Cells Isolated and Expanded in Human Serum.

INTRODUCTION: Dental pulp-derived stem cells (DPSCs) have the potential to regenerate dentin and dental pulp tissue because of their differentiation capacity and angiogenic properties. However, for regenerative approaches to gain regulatory and clinical acceptance, protocols are needed to determine more feasible ways to cultivate DPSCs, namely, without the use of xenogeneic-derived components (animal sera) and exogenous growth factors. METHODS: In this study, human DPSCs were isolated from third molars and expanded in standard culture conditions containing fetal bovine serum (DPSCs-FBS) or conditions containing human serum (DPSCs-HS). After cell characterization and evaluation of their angiogenic secretome, DPSCs were seeded in tooth slice/scaffolds and implanted subcutaneously into immunodeficient mice. After 30 days, tooth slices were retrieved and evaluated for dental pulp tissue regeneration. Immunohistochemistry and confocal microscopy were used to quantify blood vessel formation and evaluate predentin and dentin formation. RESULTS: After culture, DPSCs-HS produced concentrations of angiogenic growth factors equivalent to DPSCs-FBS. Additionally, in DPSCs-HS, several angiogenic factors were produced in at least 1-fold higher concentrations than in DPSCs-FBS. In vivo, it was determined that DPSCs-HS produced a robust angiogenic response and regeneration of dentin equivalent to DPSCs-FBS. CONCLUSIONS: These findings show that DPSCs can be isolated and expanded to clinical scale numbers in media devoid of animal serum or exogenous growth factors and still maintain their pulp regenerative properties. The implications of these findings are significant for further development of clinical protocols using DPSCs in cell therapies.

Coating of VEGF-releasing scaffolds with bioactive glass for angiogenesis and bone regeneration.

Bioactive glasses are potentially useful as bone defect fillers, and vascular endothelial growth factor (VEGF) has demonstrated benefit in bone regeneration as well. We hypothesized that the specific combination of prolonged localized VEGF presentation from a matrix coated with a bioactive glass may enhance bone regeneration. To test this hypothesis, the capacity of VEGF-releasing polymeric scaffolds with a bioactive glass coating was examined in vitro and in vivo using a rat critical-sized defect model. In the presence of a bioactive glass coating, we did not detect pronounced differences in the differentiation of human mesenchymal stem cells in vitro. However, we observed significantly enhanced mitogenic stimulation of endothelial cells in the presence of the bioactive glass coating, with an additive effect with VEGF release. This trend was maintained in vivo, where coated VEGF-releasing scaffolds demonstrated significant improvements in blood vessel density at 2 weeks versus coated control scaffolds. At 12 weeks, bone mineral density was significantly increased in coated VEGF-releasing scaffolds versus coated controls, while only a slight increase in bone volume fraction was observed. The results of this study suggest that a bioactive glass coating on a polymeric substrate participates in bone healing through indirect processes which enhance angiogenesis and bone maturation and not directly on osteoprogenitor differentiation and bone formation. The mass of bioactive glass used in this study provides a comparable and potentially additive, response to localized VEGF delivery over early time points. These studies demonstrate a materials approach to achieve an angiogenic response formerly limited to the delivery of inductive growth factors.

Temporally regulated delivery of VEGF in vitro and in vivo.

The exposure duration and tissue distribution will likely dictate the success of vascular endothelial growth factor (VEGF) in therapeutic angiogenesis. We hypothesized that these variables can be regulated via the manner in which the VEGF is incorporated into polymer constructs (formed with a gas foaming technique) used for its delivery. VEGF was incorporated directly into poly(lactide-co-glycolide) (PLG) scaffolds or pre-encapsulated in PLG microspheres used to fabricate scaffolds. Protein release kinetics and tissue distribution were determined using iodinated VEGF. VEGF was positioned predominantly adjacent to scaffold pores when incorporated directly and was released rapidly (40-60% in 5 days). Pre-encapsulation led to the VEGF being more deeply embedded and resulted in a delayed release. Alterations in polymer composition, scaffold size, and matrix composition generated minor variations in release kinetics. In vivo, the released VEGF generated local protein concentrations above 10 ng/mL at distances up to 2 cm from the implant site for the 21 days of the experiment, with negligible release into the systemic circulation, and significantly enhanced local angiogenesis. These data indicate that VEGF can be administered in a sustained and localized fashion in vivo, and the timing of VEGF delivery can be altered with the mechanism of incorporation into polymer scaffolds used for its delivery.

Growth factor delivery for oral and periodontal tissue engineering.

The treatment of oral and periodontal diseases and associated anomalies accounts for a significant proportion of the healthcare burden, with the manifestations of these conditions being functionally and psychologically debilitating. Growth factors are critical to the development, maturation, maintenance and repair of craniofacial tissues, as they establish an extracellular environment that is conducive to cell and tissue growth. Tissue-engineering principles aim to exploit these properties in the development of biomimetic materials that can provide an appropriate microenvironment for tissue development. These materials have been constructed into devices that can be used as vehicles for delivery of cells, growth factors and DNA. In this review, different mechanisms of drug delivery are addressed in the context of novel approaches to reconstruct and engineer oral- and tooth-supporting structures, namely the periodontium and alveolar bone.

Combined angiogenic and osteogenic factor delivery enhances bone marrow stromal cell-driven bone regeneration.

UNLABELLED: Bone formation is a coordinated process involving various biological factors. We have developed a scaffold system capable of sustained and localized presentation of osteogenic (BMP-4) and angiogenic (VEGF) growth factors and human bone marrow stromal cells to promote bone formation at an ectopic site. Combined delivery of these factors significantly enhanced bone formation compared with other conditions. INTRODUCTION: Tissue regeneration entails complex interactions between multiple signals and materials platforms. Orchestrating the presentation of these signals may greatly enhance the regeneration of lost tissue mass. Bone formation, for example, is dependent on the signaling of BMPs, molecules initiating vascularization (e.g., vascular endothelial growth factor [VEGF]), and osteogenic precursor cells capable of responding to these cues and forming bone tissue. It was hypothesized that combined and concerted delivery of these factors from biodegradable scaffolds would lead to enhanced bone formation. MATERIALS AND METHODS: Poly(lactic-co-glycolic acid) scaffolds containing combinations of condensed plasmid DNA encoding for BMP-4, VEGF, and human bone marrow stromal cells (hBMSCs) were implanted into the subcutaneous tissue of SCID mice. Implants (n = 6) were retrieved at 3, 8, and 15 weeks after implantation. Bone and blood vessel formation was determined qualitatively and quantitatively by methods including histology, immmunostaining, and muCT. RESULTS: Scaffolds delivering VEGF resulted in a prominent increase in blood vessel formation relative to the conditions without VEGF. BMP-4 expression in scaffolds encapsulating condensed DNA was also confirmed at the 15-week time-point, showing the characteristic of long-term delivery in this system. Combined delivery of all three types of factors resulted in a significant increase in the quantity of regenerated bone compared with any factor alone or any two factors combined, as measured with DXA, X-ray, and histomorphometric analysis. Furthermore, bone formed with all three factors had elastic moduli significantly higher than any other condition. CONCLUSIONS: Concerted delivery of BMP-4, VEGF, and hBMSCs promoted greater bone formation relative to any single factor or combination of two factors. Materials systems that allows multifactorial presentation more closely mimic natural developmental processes, and these results may have important implications for bone regeneration therapeutics.