Discovery and characterization of two novel polyethylene terephthalate hydrolases: One from a bacterium identified in human feces and one from the Streptomyces genus.

Polyethylene terephthalate (PET) is widely used for various industrial applications. However, owing to its extremely slow breakdown rate, PET accumulates as plastic trash, which negatively affects the environment and human health. Here, we report two novel PET hydrolases: PpPETase from Pseudomonas paralcaligenes MRCP1333, identified in human feces, and ScPETase from Streptomyces calvus DSM 41452. These two enzymes can decompose various PET materials, including semicrystalline PET powders (Cry-PET) and low-crystallinity PET films (gf-PET). By structure-guided engineering, two variants, PpPETaseY239R/F244G/Y250G and ScPETaseA212C/T249C/N195H/N243K were obtained that decompose Cry-PET 3.1- and 1.9-fold faster than their wild-type enzymes, respectively. The co-expression of ScPETase and mono-(2-hydroxyethyl) terephthalate hydrolase from Ideonella sakaiensis (IsMHETase) resulted in 1.4-fold more degradation than the single enzyme system. This engineered strain degraded Cry-PET and gf-PET by more than 40% and 6%, respectively, after 30 d. The concentrations of terephthalic acid (TPA) in the Cry-PET and gf-PET degradation products were 37.7% and 25.6%, respectively. The discovery of these two novel PET hydrolases provides opportunities to create more powerful biocatalysts for PET biodegradation.

Toll-like receptors in health and disease.

Toll-like receptors (TLRs) are inflammatory triggers and belong to a family of pattern recognition receptors (PRRs) that are central to the regulation of host protective adaptive immune responses. Activation of TLRs in innate immune myeloid cells directs lymphocytes to produce the most appropriate effector responses to eliminate infection and maintain homeostasis of the body's internal environment. Inappropriate TLR stimulation can lead to the development of general autoimmune diseases as well as chronic and acute inflammation, and even cancer. Therefore, TLRs are expected to be targets for therapeutic treatment of inflammation-related diseases, autoimmune diseases, microbial infections, and human cancers. This review summarizes the recent discoveries in the molecular and structural biology of TLRs. The role of different TLR signaling pathways in inflammatory diseases, autoimmune diseases such as diabetes, cardiovascular diseases, respiratory diseases, digestive diseases, and even cancers (oral, gastric, breast, colorectal) is highlighted and summarizes new drugs and related clinical treatments in clinical trials, providing an overview of the potential and prospects of TLRs for the treatment of TLR-related diseases.

Preliminary study on reliability and validity of the Chinese version of the Asher-McDade aesthetic index.

OBJECTIVES: This study aims to test the reliability and validity of the Chinese version of the Asher-McDade aesthetic index and clarify its feasibility for the postoperative outcome evaluation of cleft lip in China. METHODS: The Chinese version of the Asher-McDade aesthetic index was established through translation, back translation, debugging, and pre-survey. Eighty postoperative photographs of patients with cleft lip admitted to the West China Hospital of Stomatology, Sichuan University were included. Ten healthcare professionals, including surgeons, nurses, and students, in the department of cleft lip and palate surgery finished the index to test its reliability and validity. RESULTS: The Cronbach's alpha coefficient and retest reliability of this index are 0.804 and 0.895, respectively. The item-level content validity index (I-CVI) and scale-level content validity index ave-rage (S-CVI/ave) of the index are 1.000 and 0.95, respectively. For this index, the Kaiser-Meyer-Olkin (KMO) test score is 0.706, the χ2 value of Bartlett's test for the consistency of the index is 962.260 (P<0.01), and the cumulative variance contribution rate is 63.095%. CONCLUSIONS: The Chinese version of the Asher-McDade aesthe-tic index has good reliability and validity and is applicable to the professional evaluation of the effect of postoperative photographs in Chinese patients with cleft lip.

Posttraumatic Growth and its Correlates Among Parents of Children With Cleft Lip and/or Palate.

This study examined the posttraumatic growth (PTG) about parents of children with cleft lip and/or palate (CL/P) and the correlates of PTG.A cross-sectional study.Parents (N = 388) of children with isolated CL/P (ages 3 months-18 years) who had at least one cleft surgery within an oral and maxillofacial surgery department of a university-affiliated tertiary hospital in a provincial capital in southwest China.Demographic information questionnaire, The Posttraumatic Growth Inventory, Simplified Coping Style Questionnaire, and Social Support Rating Scale were used for data collection.Parents' mean PTG score was in the moderate range (M = 65.7, SD = 13.73). PTG differed regarding the participants' sex, ethnicity, and educational background. Pearson's correlation analysis revealed that positive coping (r = 0.43, P < .01), negative coping (r = 0.13, P < .01), and social support (r = 0.26, P < .01) were positively correlated with PTG. Multiple regression model showed that 20.6% (P < .001) of the variance in PTG was explained by higher positive coping (ß = 0.35, P < .001), greater social support (ß = 0.13, P = .01), and for cleft lip compared to cleft palate (ß = -0.14, P ≤ .01), with no variance difference for cleft lip and palate.Parents of children with CL/P had moderate PTG. Potentially modifiable correlates of PTG suggest interventions to enhance parental positive coping and social support may increase PTG. Further studies are needed to confirm the PTG level and its affecting factors of parents of children with CL/P.

Current insights in the preclinical study of palatal wound healing and oronasal fistula after cleft palate repair.

Poor palatal wound healing after cleft palate repair could lead to unfavorable prognosis such as oronasal fistula (ONF), which might affect the patient's velopharyngeal function as well as their quality of life. Thus, restoring poor palatal wound healing for avoiding the occurrence of ONF should be considered the key to postoperative care after cleft palate repair. This review provided current insights in the preclinical study of poor palatal wound healing after cleft palate repair. This review comprehensively introduced the animal model establishment for palatal wound healing and related ONF, including the models by mice, rats, piglets, and dogs, and then demonstrated the aspects for investigating poor palatal wound healing and related treatments, including possible signaling pathways that could be involved in the formation of poor palatal wound healing, the related microbiota changes because of the deformity of palatal structure, and the studies for potential therapeutic strategies for palatal wound healing and ONF. The purpose of this review was to show the state of the art in preclinical studies about palatal wound healing after cleft palate repair and to show the promising aspects for better management of palatal wound healing.

Establishment of an Oronasal Fistula Mice Model.

This study presents a method utilizing heated ophthalmologic cautery to develop a viable model for investigating oronasal fistulas. C57BL/6 mice were used to establish the oronasal fistula (ONF) model. To create the ONF, the mice were anesthetized, immobilized, and their hard palates were exposed. During the surgical procedure, a 2.0 x 1.5 mm full-thickness mucosal injury was induced in the midline of the hard palate using ophthalmologic cautery. It was crucial to control the size of the ONF and minimize bleeding in order to ensure the success of the experiment. Verification of the ONF model's effectiveness was conducted on the 7th-day post-operation, encompassing both anatomical and functional assessments. The presence of the nasal septum within the oral cavity and the outflow of sterile water from the nostrils upon injection into the oral cavity confirmed the successful establishment of the ONF model. The model demonstrated a practical and successful oronasal fistula, characterized by a low mortality rate, significant weight changes, and minimal variation in ONF size. Future studies may consider adopting this methodology to elucidate the mechanisms of palate wound healing and explore novel treatments for oronasal fistulas.

ChatGPT for shaping the future of dentistry: the potential of multi-modal large language model.

The ChatGPT, a lite and conversational variant of Generative Pretrained Transformer 4 (GPT-4) developed by OpenAI, is one of the milestone Large Language Models (LLMs) with billions of parameters. LLMs have stirred up much interest among researchers and practitioners in their impressive skills in natural language processing tasks, which profoundly impact various fields. This paper mainly discusses the future applications of LLMs in dentistry. We introduce two primary LLM deployment methods in dentistry, including automated dental diagnosis and cross-modal dental diagnosis, and examine their potential applications. Especially, equipped with a cross-modal encoder, a single LLM can manage multi-source data and conduct advanced natural language reasoning to perform complex clinical operations. We also present cases to demonstrate the potential of a fully automatic Multi-Modal LLM AI system for dentistry clinical application. While LLMs offer significant potential benefits, the challenges, such as data privacy, data quality, and model bias, need further study. Overall, LLMs have the potential to revolutionize dental diagnosis and treatment, which indicates a promising avenue for clinical application and research in dentistry.

Development of postoperative velopharyngeal function in patients with cleft palate.

OBJECTIVE: Patients with a cleft palate often experience a velopharyngeal dysfunction known as velopharyngeal insufficiency (VPI). The purpose of this study was to examine the development of velopharyngeal function (VPF) following primary palatoplasty and the factors that are linked to it. METHODS: A retrospective study was conducted to examine the medical records of patients who had cleft palate, with or without cleft lip (CP ± L) and underwent palatoplasty at a Tertiary Affiliated Hospital between 2004 and 2017. Postoperative evaluation of VPF was conducted at two follow-up times (T1, T2) and was classified as either normal VPF, mild VPI, or moderate/severe VPI. The consistency of VPF evaluations between the two time points was then assessed, and patients were categorized into either the consistent or inconsistent group. The study collected and analyzed data on gender, cleft type, age at operation, follow-up duration, and speech records. RESULTS: The study included 188 patients with CP ± L. Out of these, 138 patients (73.4%) showed consistent VPF evaluations, while 50 patients (26.6%) showed inconsistent VPF evaluations. Among those with VPI at T1 (91 patients), 36 patients (39.6%) had normal VPF at T2. The rate of VPI decreased from 48.40% at T1 to 27.13% at T2, whereas the rate of normal VPF increased from 44.68% at T1 to 68.09% at T2. The consistent group had a significantly younger age at operation (2.90 ± 3.82 vs 3.68 ± 4.02), a longer duration of T1 (1.67 ± 0.97 vs 1.04 ± 0.59), and a lower comprehensive score of speech performance (1.86 ± 1.27 vs 2.60 ± 1.07) than the inconsistent group. CONCLUSIONS: It has been verified that there are changes in the development of VPF over time. Patients who underwent palatoplasty at a younger age were more likely to have confirmed VPF diagnosis at the first evaluation. The duration of follow-up was identified as a critical factor that affects the confirmation of VPF diagnosis.

Sommerlad-Furlow modified palatoplasty: A retrospective study.

The purpose of this study was to introduce the surgical process of Sommerlad-Furlow modified (S-F) palatoplasty and compare its surgical and functional outcomes with conventional Sommerlad (S) palatoplasty. Patients with non-syndromic cleft palate who had undergone either S-F palatoplasty or S palatoplasty were retrospectively reviewed. Data on the outcomes of velopharyngeal function and postsurgical palatal fistula incidence were collected for all patients. Data for preselected factors, including gender, age at palatoplasty, and cleft type, were also collected. Chi-square tests were conducted. 1254 patients were included. The postsurgical velopharyngeal competence (VPC) rate after S-F palatoplasty was significantly higher than after S palatoplasty (total, 70.5% vs 57.9%, p < 0.0001; age ≤ 1, 87.0% vs 69.2%, p < 0.0001; 1 < age ≤ 2, 78.3% vs 69.3%, p = 0.0479). With regard to different types of cleft palate, the postsurgical VPC rates after S-F palatoplasty were all significantly higher than for S palatoplasty in all patients younger than 2 years of age (complete cleft palate, 78.7% vs 62.4%, p = 0.0016; hard and soft palate cleft, 84.4% vs 74.8%, p = 0.0172; submucosal cleft and soft palate cleft, 96.6% vs 68.4%, p = 0.0114). The postoperative fistula rate after S-F palatoplasty was 4.3%. This modified palatoplasty technique provided adequate cleft palate closure, with satisfactory speech outcomes and low fistula rates, while older age at palatoplasty may affect the postsurgical outcomes. Within the limitations of the study it seems that the Sommerlad-Furlow modified technique is an option for cleft palate repair.

Evaluation of Postoperative Outcomes in Two Cleft Palate Repair Techniques without Relaxing Incisions.

BACKGROUND: A modified palatoplasty was established by incorporating the designs of both Sommerlad and Furlow techniques in addition to a novel incision on the medial pterygoid plate's surface, named the Sommerlad-Furlow modified technique. Thus, this study aimed to evaluate the clinical and functional outcomes of the Sommerlad-Furlow modified technique against an accepted standard, the Furlow technique. METHODS: A retrospective review was conducted for 212 consecutive nonsyndromic cleft palate patients who underwent Sommerlad-Furlow ( n = 106) and Furlow ( n = 106) repairs without relaxing incision on the hard palate between 2011 and 2016. The success of surgical procedures was estimated by the rate of postoperative fistula, speech outcomes, and velopharyngeal insufficiency (VPI)-related quality of life. The demographic and surgical data, including sex, age, cleft type, cleft width, and follow-up period were recorded. RESULTS: There was no statistically significant difference between the two treatment groups regarding demographic and surgical data, except the cleft width ( P < 0.001). The incidence of the fistula was 7.5% and 6.6% after the Sommerlad-Furlow and Furlow procedures, respectively. The two groups showed no significant differences in speech outcomes, and adequate velopharyngeal function was found in 84% and 82.1% in Sommerlad-Furlow and Furlow procedures, respectively. Besides, the rate of severe VPI was slightly lower in Sommerlad-Furlow (0.9%) than in Furlow (2.8%) procedures. Moreover, an adequate VPI-related quality of life was found in 80.4% of the Sommerlad-Furlow group and 78.6% of the Furlow group. CONCLUSION: The Sommerlad-Furlow technique has obtained acceptable postoperative outcomes and could be a choice for cleft palate repair, especially in wider clefts. CLINICAL QUESTION/LEVEL OF EVIDENCE: Therapeutic, III.

Depression in Chinese Patients with Cleft Lip and/or Palate: A Preliminary Study.

(1) Objectives: To investigate the difference in prevalence of depression between patients with CL/P (cleft lip and/or palate) and analyze the possible demographic factors that affect the prevalence of depression in Chinese patients with CL/P. (2) Methods: Patients with CL (cleft lip only), CP (cleft palate), and CLP (cleft lip and palate) were included in the study group. Non-CL/P individuals were included in the control group. The Patient Health Questionnaire (PHQ-9) was used to screen the depression of Chinese patients with CL/P. The different proportions of different depression groups between the CL/P group and the control groups were tested by the Fisher-Freeman-Halton test and Bonferroni correction. The scores between the study groups and the control group were analyzed by one-way ANOVA. In the study groups, demographic and clinical data of the patients, including diagnosis (CL, CP, CLP), gender, age, the only child or not, and region were collected to analyze whether they were the possible factors affecting depression through one-way independent-samples t-test. Pearson correlation analysis was used to analyze the correlation between monthly family income and depression. (3) Results: 111 and 80 valid questionnaires were collected from the study and control groups, respectively. The mean PHQ-9 score of the study group (5.459 ± 6.082) was relatively higher than the control group (4.362 ± 3.384), and the difference in proportions of depression groups was statistically significant between the CL/P group and the control group (p = 0.01), especially in the mild depression (p < 0.05) and moderately severe depression groups (p < 0.05). Statistically significant differences in PHQ-9 scores were observed between the individuals of different genders (p = 0.036) and ages (p = 0.007) in patients with CL/P, the individuals who were the only child or not in patients with CL (p = 0.007), and the individuals of different ages in patients with CP (p = 0.016). (4) Conclusions: The prevalence of depression in Chinese patients with CL/P was different compared with those without CL/P, while gender, age, the only child or not, and region played significant roles in affecting depression psychology.

Evaluation of late cleft palate repair by a modified technique without relaxing incisions.

OBJECTIVE: To evaluate the postoperative outcomes together with analyzing the associated influencing factors following a late cleft palate repair by the Sommerlad-Furlow modified technique (S-F). MATERIALS AND METHODS: In a retrospective cohort, 320 consecutive patients with cleft palate, who received S-F technique between 2011 and 2017, were reviewed. The patients were divided into three age groups, less than one year (143), one to two years (113), and greater than 2 years (64). The postoperative outcomes included wound healing (complete/fistula) and velopharyngeal function (VPF). RESULTS: The overall cleft width was 10 ± 3.07 mm. The overall rates of complete wound healing and proper velopharyngeal function were 96.6% and 81.56%, respectively. No significant difference was found between the age groups regarding wound healing, with an overall fistula rate of 3.4%. The VPF was significantly varied among the age groups (P<0.001). In context, the rates of velopharyngeal insufficiency (VPI) were 9.8%, 14.2%, and 45.4% among patients repaired at ˂1, 1-2, and >2 years old, respectively. The cleft type was the most potential factor associated with fistula. The age at repair was identified as the most implicating factor for VPI. CONCLUSIONS: The S-F technique had achieved low fistula rate and satisfactory speech outcome, especially in early repair group and even in the wide cleft palate. The older age at repair and severe cleft type had a major impact on the postoperative outcomes.

The pilot study of a blended learning module using porcine tongue for surgical simulation of double-opposing Z-plasty.

PURPOSE: Simulation course serves as an efficient supplement to the experiential learning inside the operation room in surgical residency programs. Furlow double-opposing Z-plasty (DOZ) remains an important and challenging surgical technique in cleft palate repair, but the simulation is currently limited. This study aims to develop a blended learning module with low-cost and easy-to-promote DOZ simulation. METHODS: Comparison among the common rodent experimental animals and the readily available porcine tongue was made to select the ideal animal for surgical simulation, with which a DOZ learning module was developed. Oral surgery residents were enrolled to attend the blended learning module. An after-session questionnaire and a quiz were distributed to the participants to collect their feedback of the course perceptions and to assess their learning outcomes. RESULTS: The porcine tongue had more advantages in simulating the human soft palate over the common experimental animals. The surgical procedure of DOZ could be well designed and simulated on the porcine tongue. The vast majority of the participant liked the learning module regarded it as beneficial for the comprehension if DOZ principles and procedures. With the help of the porcine tongue simulator, the blended learning module helped the participants grasp the designing principles of DOZ procedure and promote their associative learning. CONCLUSIONS: The blended learning module incorporating the porcine tongue simulator can help oral surgery residents master the principles and procedures of DOZ palatoplasty.

The Necessity of Nostril Retention Application After Secondary Unilateral Cleft Rhinoplasty.

BACKGROUND: This study aims to investigate the necessity of nostril retention after secondary unilateral cleft rhinoplasty. METHODS: Seventy four patients who underwent secondary unilateral cleft rhinoplasty were categorized into control group, nasal retainer group, and nasal clip group. A nasal retainer or nasal clip was applied 7 days after secondary correction and retained in the nostrils of the patients for 6 to 12 months. Measurements from pre- and post-operative photos and patient-reported outcomes were used in this study. RESULTS: After 6 to 12 months, although relapse still occurred in three groups, the nasal retainer group showed a better effect in the ratio of nostril height, whereas the nasal clip group showed a significant improvement in the ratio of nasal base. And both the two groups also showed significant improvement in the ratio of nasal columella. Patients from the control group generally had lower Derriford Appearance Scale (DAS) scores than the nasal retainer group and nasal clip group. Wearing a nasal retainer may negatively influence self-concept, facial self-consciousness of appearance and physical state, and significant differences in mean scores were also found in 9 items. CONCLUSIONS: The nostril retention after secondary unilateral cleft rhinoplasty could help preserve nasal morphology and reduce postoperative relapse but worsen the patients' quality of life. The necessity of applying the nostril retention should consider the forms of nasal deformity and the potential influence on the quality of life. LEVEL OF EVIDENCE: 3 Laryngoscope, 133:1618-1623, 2023.

Application of a Novel Nasal Clip for Nostril Retention After Primary Unilateral Cleft Rhinoplasty.

OBJECTIVE: This study aimed to investigate the outcomes of using a novel nasal clip designed for nostril retention after primary unilateral cleft rhinoplasty. DESIGN: This is a retrospective study. SETTING: Department of Cleft Lip and Palate in a stomatological hospital. PATIENTS: A retrospective study was conducted on 57 patients who had a unilateral complete cleft lip without cleft palate and underwent primary surgical repair. INTERVENTIONS: The patients were categorized into 3 groups: the group in which the conventional nasal retainer was used, that in which the nasal clip was used, and the control group in which no postoperative nasal retainer was used. The nasal retainer or our nasal clip was applied 7 days after primary surgical repair and kept in the nostrils of children from 6 to 12 months. MAIN OUTCOME MEASURES: Noses underwent photogrammetry preoperatively, postoperatively, and at follow-up. The nose was also subjectively scored at the follow-up. RESULTS: The nasal retainer and the nasal clip significantly minimized relapse as determined by nasal tip deviation. The nasal clip sustained nasal symmetry, as determined by the postoperative nostril width ratio, better than did the conventional retainer. After 6 to 12 months, the nasal clip achieved better columellar morphology and a more symmetric nasal base than did the conventional nasal retainer. Statistical differences in subjective scores between the control and nostril retention groups were noted. CONCLUSION: The application of the novel nasal clip after primary unilateral cleft rhinoplasty preserves nasal morphology and reduces postoperative relapse without the need for adhesive tape.

Rapid Additive Manufacturing of a Superlight Obturator for Large Oronasal Fistula in Pediatric Patient.

This study developed a novel digital workflow to fabricate a 3D printed hollow obturator for the prosthetic reconstruction of palatal fistula. It will provide cleft surgeons and therapists a choice for treating children with large palatal fistula before the appropriate age for surgical reconstruction. Laryngoscope, 133:1507-1512, 2023.

The Sommerlad-Furlow Modified Palatoplasty Technique: Postoperative Complications and Implicating Factors.

OBJECTIVE: To explore the incidence of postoperative complications and investigate the impact of preselected factors on functional and quality of life outcomes following the Sommerlad-Furlow modified palatoplasty technique. STUDY DESIGN: Retrospective cohort. METHODS: A total of 429 patients with cleft palate, who received Sommerlad-Furlow modified technique between 2011-2017 were enrolled. The postoperative complications including oronasal fistula (ONF), velopharyngeal insufficiency (VPI), and inadequate quality of life (QOL) were collected. Data of preselected factors including gender, age at palatoplasty, cleft type, cleft width, palatal width, pharyngeal cavity depth, and operation duration were also collected. RESULTS: Among 429 patients, 40.1% were males whereas 59.9% were females. The mean age at palatoplasty was 1.23 ± 0.69 (0.42-4) years, and the average cleft width was 10.15 ± 2.95 (4-27) mm. The cleft types had recorded rates of about 6.8%, 69.5%, 17.7%, and 6.1% of Veau I, II, III, and IV, respectively. The overall incidence rates of ONF, VPI, and inadequate QOL were 2.3%, 19.4%, and 31.3%, respectively. In both the univariate and multivariate analyses, the cleft type was significantly implicated in ONF formation (p = 0.023 and 0.032, respectively) whereas the velopharyngeal function was impacted by the palatoplasty age (p Ë‚ 0.001). The receiver operating characteristic curve indicated that age of palatoplasty ≥1.3 years (area under the curve = 0.611, p = 0.002) was the cutoff value for predicting the incidence of VPI. CONCLUSIONS: The Sommerlad-Furlow modified technique appears to have appropriate postoperative outcomes, even in the wide cleft palate. The older age at palatoplasty has a major impact on the overall postoperative outcomes. LEVEL OF EVIDENCE: 4 Laryngoscope, 133:822-829, 2023.

Comparative Analysis of Nucleic Acid-Binding Polymers as Potential Anti-Inflammatory Nanocarriers.

Conventionally, nanocarriers are used to regulate the controlled release of therapeutic payloads. Increasingly, they can also be designed to have an intrinsic therapeutic effect. For example, a positively charged nanocarrier can bind damage-associated molecular patterns, inhibiting toll-like receptor (TLR) pathway activation and thus modulating inflammation. These nucleic acid-binding nanomaterials (NABNs), which scavenge pro-inflammatory stimuli, exist in diverse forms, ranging from soluble polymers to nanoparticles and 2D nanosheets. Unlike conventional drugs that primarily address inflammation symptoms, these NABPs target the upstream inflammation initiation pathway by removing the agonists responsible for inflammation. Many NABNs have demonstrated effectiveness in murine models of inflammatory diseases. However, these scavengers have not been systematically studied and compared within a single setting. Herein, we screen a subset of the most potent NABNs to define their relative efficiency in scavenging cell-free nucleic acids and inhibiting various TLR pathways. This study helps interpret existing in vivo results and provides insights into the future design of anti-inflammatory nanocarriers.

The Correlation between Periodontal Parameters and Cell-Free DNA in the Gingival Crevicular Fluid, Saliva, and Plasma in Chinese Patients: A Cross-Sectional Study.

Purpose: To investigate the correlation between periodontal parameters and cell-free DNA (cfDNA) concentrations in gingival crevicular fluid (GCF), saliva, and plasma. Methods: Full mouth periodontal parameters, including probing depth (PD), bleeding on probing (BOP), and plaque index (PI) were recorded from 25 healthy volunteers, 31 patients with untreated gingivitis, and 25 patients with untreated periodontitis. GCF, saliva, and plasma samples were collected from all subjects. Extraction and quantification assays were undertaken to determine cfDNA concentrations of each sample. Results: GCF and salivary cfDNA levels were increased with aggravation of periodontal inflammation (GCF p < 0.0001; saliva p < 0.001). Plasma cfDNA concentrations in patients with periodontitis were significantly higher than those in healthy volunteers and patients with gingivitis. GCF and salivary cfDNA were positively correlated with mean PD, max PD, BOP, and mean PI (p < 0.0001), whereas plasma cfDNA was not correlated with BOP (p = 0.099). Conclusion: GCF, saliva, and plasma concentrations of cfDNA were significantly elevated in patients with periodontal disease. There were also positive correlations between cfDNA levels in GCF and saliva and periodontal parameters.