Purpose: The aim of the study is to histologically evaluate new bone formation in extraction sockets augmented with autologous demineralized dentin bone (ADDB). Materials and Methods: Teeth were extracted from 19 patients and prepared as ADDB and then grafted in 27 extraction socket sites. The graft was covered with membrane made of concentrated growth factors (membrane) and the wounds were closed without periosteal releasing incision (open-membrane technique). Bone biopsy was performed at each implant site after 4 months of healing. Results: Wound healing was uneventful. Histologically, bone reformation was observed in all the augmented sockets, and ADDB showed favorable integration with newly formed bone. Conclusion: Immediate graft after extraction using ADDB is recommended for socket preservation.
Background: The use of Vitamin D3, as an alternative drug, combined with common therapeutic strategies to treat nonmelanoma skin cancers, has recently attracted attention. However, in vitro data on Vitamin D3action on different tumor cell lines compared to healthy cells are lacking. Aims and Objectives: In this context, the present study aimed to investigate the potential role of Vitamin D3's ability as an antitumor treatment. Materials and Methods: Cell growth, cell viability, and apoptosis as well as cell cycle distribution were comparatively assessed in a squamous cell carcinoma (SCC) cell line, a basal cell carcinoma (BCC) cell line, and healthy primary normal human epidermal keratinocytes (NHEK) in response to various Vitamin D3concentrations. Results: Tumor and healthy cells clearly responded differently to Vitamin D3application with regard to metabolic activity and apoptosis. The application of Vitamin D3reduced the metabolic activity of the BCC and SCC cancer cell lines (and not NHEK) and induced cell cycle arrest. Furthermore, Vitamin D3-mediated increased apoptosis was observed in tumor cells but not in healthy primary keratinocytes. Conclusions: Our findings indicate an antiproliferative and proapoptotic Vitamin D3-dependent effect on skin cancer cells in vitro, highlighting Vitamin D3as a potential and beneficial alternative drug for further studies with respect to possible clinical strategies to treat nonmelanoma skin cancers.
In the last decade, an increasing awareness was directed to the role of Vitamin D in nonskeletal and preventive roles for chronic diseases in different fields. Vitamin D deficiency was reported in many countries worldwide and is considered as a pandemic. However, no consensus exists about whether and how supplementation of Vitamin D may be beneficial as a preventive or adjuvant therapy. Thereby, this review aimed to deliver an overview about the administrated doses of Vitamin D in randomized controlled clinical studies, in order to evaluate the currently available clinical evidence. In addition, focus was placed on the recent advances on Vitamin D nonskeletal actions. The results sometimes showed a great discrepancy between the recommended Vitamin D dose by different guideline authorities, which are from 400 to 4000 IU/day, and the used doses in recent randomized controlled clinical studies, which were up to 100,000 IU/day. Different studies showed the positive effect of Vitamin D in supporting the immune system and preventing different chronic and infectious diseases. These findings reflect the need to rethink existing reference ranges and intake recommendations. Based on the analyzed range of clinically applied doses, we recommend a Vitamin D supplementation based on three different ranges, which include <40 ng/ml, >40 <80 ng/ml, and >80 ng/ml with oral Vitamin D intake of 10,000 IU/day, 5000 IU/day, and 1000 IU/day, respectively. A 25-hydroxyvitamin D blood serum monitoring is furthermore recommenced every 3 months to re-adjust the Vitamin D dose based on the above-mentioned concept. Ongoing clinical studies will have to further prove this concept for different patient groups.
After a dental extraction procedure, the extracted teeth are discarded disregarding their properties as an excellent graft material. The use of extracted teeth as a graft material was first performed by Dr. Marshall Urist in 1967, where he discovered and verified that the decalcified dentin matrix can induce bone formation, but his decalcification method took at least 5 days to accomplish before being able to use it as a graft material. Currently, an ultrasonic technology with temperature and vacuum control, named VacuaSonic® System reduces the decalcification time dramatically (≤80 min) converting the tooth into graft material at chairside immediately after the dental extraction with the result being an autogenous tooth graft material (ATG). The aim of this article is to introduce two clinical cases: one case of alveolar ridge augmentation, and the other, socket preservation using ATG mixed with liquid-phase concentrated growth factors (LP-CGF) prepared at chairside on the same day of the dental extraction procedure. LP-CGF is obtained from the same patient, which was collected in blood collection tubes and processed by a special centrifuge device. The result obtained by mixing ATG with LP-CGF is a graft matrix named “Gummy Tooth Graft.”
The present note aims to alert the importance of oral healthcare in patients under mechanical ventilation by COVID-19. Patients hospitalized for long periods and needing the support of mechanical ventilators are susceptible to secondary infections. Thereby, many patients may have already died from infections, secondary to pneumonia caused by COVID-19.
Odontogenic myxoma (OM) is a rare, benign neoplasm and is often located in the maxillofacial region. Clinical, radiological, and histopathological features should be considered when making a diagnosis. Several of these characteristics overlap with those of other benign and some malignant tumors. Soft-tissue localization that is classified as a peripheral myxoma is rarely seen than central localization. Peripheral myxoma is slowly growing and less aggressive compared to central myxoma and has a low recurrence rate. The probability that small peripheral OMs (POMs) are interpreted as edematous irritation fibromas may contribute to the small number of POMs recorded in literature. The treatment plan should consider the age and sex of the patient and the site and size of the lesion. In this report, the application of platelet-rich fibrin in the management of an unusual case of POM located on the gingiva in the posterior maxillary gingival region is presented.
The aim of this case report was to describe an alternative treatment for the healing and maintenance of peri-implant mucosa in esthetical region. Managing the peri-implant soft tissue is one of the most challenging tasks in anterior implant esthetics, especially when replacing a failing tooth. However, not all patients underwent extraction, and immediate implant placement can receive loading, conducting to uncertain predictability. This is a case report that describes an alternative treatment for the healing and maintenance of the peri-implant mucosa in an esthetically significant region, through the use of a customized abutment to obtain an emergency profile when there is insufficient torque for immediate implant loading. The present technique was able to ensure not only healthy peri-implant soft and hard tissues, but also esthetic maintenance of the anterior region for immediate implant placement. This clinical case demonstrates a different manipulation technique from the peri-implant soft tissues in immediate implants of the esthetic region favoring the peri-implant health and clinical results, promoting treatment success, and patient satisfaction.
Background: The importance of the amniotic fluid (AF) to the fetus is clear. However, very few studies have been published to examine the potential uses of this fluid in various areas such as tissue regeneration. AF contains epidermal growth factor, transforming growth factor-alpha, transforming growth factor beta-1, insulin-like growth factor-I, erythropoietin and granulocyte colony-stimulating factor, as well as hyaluronic acid and hyaluronic acid-stimulating factor. Previous studies suggest that AF can increase fibroblast proliferation and chemotaxis, and decrease apoptosis as well as promote wound healing. Furthermore, evidence showed that human AF inhibits hyaluronidase, elastase, and cathepsin. The current study examined the effects of human AF on periodontal ligament fibroblasts (PDLF) in terms of cell toxicity, cell proliferation, and cytokine/growth factor expression. Materials and Methods: Cytotoxicity of AF on PDLF was determined using lactate dehydrogenase assays. PDLF proliferation was determined using water-soluble tetrazolium-1 assays. Cytokine/growth factor expression was determined on AF-treated PDLF, AF alone, and PDLF alone utilizing protein arrays. Results: Human AF at 10% and below did not affect cell growth and was not toxic. AF-treated PDLF cells showed a decrease in cytokine/growth factor levels compared to the sum of cytokine/growth factor levels in AF only and cells only for 39 of the 80 proteins examined (48.8%). Of the 39 examined cytokines, 20 inflammatory cytokines, 11 cell cycle cytokines, 1 anti-inflammatory cytokine, and 7 other cytokines were decreased. Conclusion: Human AF at the examined concentrations was not toxic to PDLF cells and did not influence their proliferation. In addition, AF (10%) caused a decrease in the total protein levels of cytokines/growth factors expressed in 39 of the 80 proteins examined (48.8%). Of the 39 examined cytokines, 20 inflammatory cytokines, 11 cell cycle cytokines, 1 anti-inflammatory cytokine, and 7 other cytokines were decreased.
Investigations of endogenous blood-derived growth factors have increased in the past two decades. The initial protocols for blood concentrates, such as platelet-rich-plasma, utilized anticoagulants followed by bovine thrombin to catalyze fibrin polymerization. Subsequently, platelet-rich-fibrin (PRF) protocol was developed to eliminate anticoagulants and thrombin. The PRF production was described in a vertical rotor centrifuge with an angle of ~33°. Many commercial enterprises have attempted to replicate this protocol with a multitude of different centrifuges. These attempts have utilized revolutions per minute (RPM) to develop their protocol to generate PRF. However, RPM is a variable parameter, as it depends on the design and radius of the centrifuge. The separation of blood components is highly dependent on the relative centrifugal force (RCF) generated, which is based on the applied centrifugal force, expressed as multiples of earth's gravitation (g) field. RCF is a function of the acceleration due to the gravity of the earth, g = 9.81 m/s2 RCF, not RPM, is the key factor for the sedimentation of the cells and proteins within blood concentrates. RCF is determined using the maximum centrifuge radius for calculation. The initial PRF was generated by applying an RCF of 700 ×g. Accordingly, so far, a large number of studies existing about PRF are not comparable, as they were technically prepared with different RCFs. In addition, due to the nonstandardized measurement methods, in some published studies, incorrect RCF values are published as the authors did not know how to calculate the RCF correctly. This is the case for a widespread blood concentrate called leukocyte-PRF, which is commercially available. Recently, we introduced the low-speed centrifugation concept (LSCC) for the production for solid and liquid PRF matrices. This concept is based on the above-mentioned initial PRF protocols, which represents a relatively high RCF. It shows that a systematic reduction of 700 ×g to 44 ×g can significantly increase the cells and growth factors within the same blood concentrates. The LSCC concept was established initially for a fixed rotor centrifuge with a radius of ~110 mm. The present narrative review highlights the necessity of standardization in the generation of blood concentrates, which utilizes RCF, rather than commercial protocols.
Introduction: Human dentin matrix could be successfully used for bone grafting procedures. It was well accepted that dentin grafts can induce osteoblast proliferation. An innovative preparation method, using the dedicated automated device Tooth Transformer®, which can transform autologous teeth in suitable grafting material, has been recently introduced. The aim of the present article is to analyze the histologic outcomes in 11 consecutive human cases, in which autologous tooth graft materials, starting from the whole tooth of the patient, were used for bone regeneration. Results: The bone defects were completely filled by newly formed tissue after 4 months of healing. Histologic analysis revealed no inflammatory or infective reactions against the tooth graft. Tooth granules were surrounded by newly formed bone. Some tooth granules were incorporated in the bony trabeculae, and they appeared partially resorbed. This fact testified that tooth grafts underwent remodeling processes just like the native bone. Discussion: Results from the present histologic case series analysis revealed that tooth graft appeared well integrated in the regenerative tissue without any inflammatory or infective reaction. The tooth of the patient may be used as an autologous regenerative material, avoiding any foreign graft material.
Aim: The aim of this pilot study was to assess the performance of autologous tooth matrix, used as a graft material for maxillary sinus augmentation, after at least 1-year of follow-up. Settings and Design: The patients included in this prospective case series study were treated in four clinical centers using standardized clinical procedures. Materials and Methods: Patients with atrophic posterior maxilla in need of sinus augmentation before rehabilitation with implant-supported prostheses, and with compromised teeth to be extracted, were included. The extracted tooth was cleaned and processed by a recently introduced automated device, which allows fragmentation and partial demineralization of the tooth matrix, and used as a graft material for sinus augmentation. A covering membrane was used to protect the graft. Implants were placed after 6 months of healing. Five bone biopsies of the grafted sites were taken at the time of implant surgery, for histological analysis. Implants were followed for at least 1 year after placement. Cone-beam computed tomography and/or standardized periapical radiographs were used to assess the ridge height before and after grafting, up to 1-year postimplantation. Statistical Analysis: Descriptive statistics were used to synthesize the results, using mean values and standard deviations. Results: Twenty-three patients (9 males, mean age at surgery 57.1 ± 9.4 years) were treated and 40 implants were placed in grafted sites. Residual ridge height was 5.22 ± 2.04 mm and increased to 14.72 ± 2.83 mm after grafting. One implant failed during healing phase. Cumulative implant survival rate was 97.5% after 19.1 ± 8.0 months of follow-up (range 12.3–44.1 months). After 6 months of healing, the graft height appeared stable. No signs of sinus infection were present. The histologic analysis revealed neither inflammatory nor infective reaction against tooth graft. Granules appeared surrounded by newly formed bone and partially resorbed, indicating ongoing remodeling. Conclusion: Autologous human tooth matrix can be successfully used as graft material in sinus augmentation procedure.
The use of platelet-rich fibrin has gained importance as an autologous scaffold for tissue regeneration. One of its features is, when placing implants in fresh extraction sockets, to function as a selective scaffold and also as a barrier in order to correct marginal bone defects and/or to avoid collapse of surrounding soft tissue. However, its handling and placement require specific manipulation skills, to avoid unintentional slipping and dislodging during the suture procedure. In order to avoid these problems and to allow easier and faster use of the PRF matrix, a simple device for providing a self-stabilized fibrin plug has been developed. This plug surrounds, as an elastic ring, the transmucosal extension of the prosthetic components for conical internal connection implants, thus enhancing the effectiveness of the fibrin matrix expected performance.
Context: Periodontitis is an infectious disease characterized by an inflammatory response and bone resorption, which can cause tooth loss. The bone destruction may be caused by deregulation of the receptor activator of nuclear factor κB/receptor activator of NF-κB ligand/osteoprotegerin (RANK/RANKL/OPG) system, stimulating osteoclastogenesis. After the treatment of the active disease, a formal program of periodontal maintenance is implemented according to each patient needs. Aims: This study aimed to evaluate the RANKL/OPG relation in patients under periodontal maintenance. Setting and Design: Thirty individuals were selected and divided into the test group, comprised of patients who received periodontal treatment and were under periodontal maintenance showing active sites (n = 15), and nonactive sites of the disease (n = 15), and the control group, comprised of healthy periodontium (n = 15). Materials and Methods: The GCF was collected by absorption into paper cones. The measurement of RANKL and OP was performed by xMAP technology and a Luminex 200 flow luminometer system. Statistical Analysis: Data were compared through Kruskal–Wallis test, at 5% significance. Results: There was no intrapatient difference on the RANKL/OPG ratio, but it was significantly lower in the active sites of patients treated of periodontitis, under a maintenance program, compared with the control (P < 0.05), even tough all ratios were under the range reported for healthy individuals. Conclusions: Patients in periodontal maintenance have a significantly lower RANKL/OPG ratio than healthy indivduals, demonstrating its importance for patients' periodontal health.
Context: MC3T3-E1 cells are preosteoblasts from mouse calvaria widely used on in vitro studies of biocompatibility under the influence of different biomaterials for bone tissue therapy, through test that includes the determination of biomarkers such as alkaline phosphatase (ALP). However, many studies use cell densities ranging from 1 × 103 to 2 × 105 cells per well even though these cells exhibit decreased proliferation when worked at high cell initial cell densities. Aims: To determine the initial parameters for the determination of ALP activity employing commercial kits and MC3T3-E1 cells for cellular differentiation studies of bone substitute biomaterials. Materials and Methods: Twenty-four well plates were seeded at concentrations of 1 × 102 to 1 × 104 at passage 8 and cultivated from 1 to 32 days. Cell density at each time was estimated through a crystal violet dye exclusion assay, cell morphology was evaluated through light microscopy, and colorimetric ALP assays were performed each day. Results: The initial cell densities of 1 × 102 and 2.5 × 102 cells per well were determined as ideal for still maintaining desired cellular characteristics for long time trials while higher initial densities demonstrated certain saturation and cell death on the 24th day. The activity of secreted ALP can be adequately measured in vitro up to 32 days after the seeding of MC3T3 cells, through commercial kits employing initial cell densities of 100–250 cells per well. Conclusion: The present protocol may provide an interesting tool for the determination of differentiation for the preclinical evauation of biomaterials.
Platelet-rich plasma (PRP) and platelet-rich fibrin (PRF) regenerative therapy lacks evidence to make clinical decisions. This weakness cannot be addressed only by standardization of preparation protocols and devices. In this article, we have emphasized on the necessity for quality testing to obtain strong evidence supporting PRP/PRF therapy.
A larger number of senior patients want to eliminate the use of conventional overdenture such as Conventional Complete Denture or Removable Partial Denture and replace it by an Implant-supported fixed prosthesis or at least implant-supported overdenture with the purpose of restoring and improving the masticatory function, aesthetics besides helping them gain confidence to talk and smile. Most of those patients have lost their teeth and been posteriorly edentulous for extended periods of time, causing severe bone resorption and Pneumatized Maxillary Sinus that makes it impossible for conventional dental implant placement. This report reviews two successful cases of rehabilitation of the atrophic posterior maxilla with previous bilateral ridge augmentation in Pneumatized Maxillary Sinuses using solely Concentrated Growth Factors (CGF) without bone graft augmentation with simultaneous implant placement. Where CGF is obtained from the patient's which was collected in blood collection tubes and processed by a special centrifuge device.
Peri-implant mucosae inflammation during osseointegration period can promote host response imbalance and bone resorption by bacteria infiltration. Aim: To evaluate the association between EGF and EGFR gene expressions in the peri-implant tissue with mucosae inflammation during osseointegration period. Material and Methods: Forty-nine participants, with 59 endosseos implants, were recruited for this study. All participants included were rehabilitated with implants in two stages surgical protocol, presenting favorable bone quality and quantity. Osseointegration was evaluated one month after exposure surgery, wich was performed three months (mandible) and 6 months (maxillary) after implant placement. The criteria to consider the proper osseointegration were: Implant immobility; absence of peri-implant radiolucency; and no clinical signs of inflammation. Based on clinical and radiographic characteristics of peri-implant sites, participants were characterized as (i) having healing without complications (with proper osseointegration without mobility of the implant, and without clinical signs of mucosal inflammation) (control group) or (ii) failure peri-implant healing (with inadequate osseointegration characterized by signs of mucosae inflammation and/or implant mobility) (test group). Gingival biopsies were collected from 49 participants after osseointegration period, during the exposure procedure. Total RNA from gingival samples was isolated using the Trizol® reagent. The reaction of reverse transcription of PCR was performed for the synthesis of complementary DNA from 300ng RNA using Improm-II Reverse Transcription System™. Specific primers for EGF (NM_001963.4) and EGFR (NM_005228.3) were based on the BLAST data. The Livak method (2-ΔΔCT) was used to determine the relative quantification of the expression of EGF and EGFR. The values were normalized by relative expression of β-actin. Results: There was no difference between control and test groups to race, sex, age, alcohol consumption, general medical conditions, current medications, edentulism and periodontal phenotype. All RNA samples revealed proportions A260 nm/A280 nm more than 1.9. EGF and EGFR showed significant lower expression in gingival tissues removed from regions with failure healing (test group). EGF showed mRNA expression with an average of 44.53 ± 79.16 and 01.02 ± 1:33 in the control groups and test, respectively (P = 0.008). Similarly EGFR expression was significantly higher in the control group (102.03 ± 329.57) compared to the test group (7.85 ± 4.16) (P = 0:04). Conclusion: Low levels of EGF and EGFR are associated with inadequate healing of mucosal peri-implant during the osseointegration period.
A common problem encountered in every dental practice, root recession presents as an esthetic issue as well as creating exposed root surfaces that may illicit sensitivity and have greater caries potential than covered roots. Frequently, patients avoid treatment due to fear related to the traditional flap surgery approach. Technology, via biologics from the patient being treated has shifted surgical techniques to become minimally invasive, allowing lower morbidity and post-operative discomfort. The Gum Drop Technique (GDT) a papilla preservation technique, with a less invasive surgical approach utilizing PRF as the graft material with improved healing compared to packaged materials will be discussed as an alternative to traditional flap surgical approaches for root recession treatment.
